method for solubilizing and separating carboxylic acids with a solubilizing compound

专利摘要:
use of a compound to solubilize carboxylic acids in an aqueous or organic medium, aqueous microemulsion or aqueous nanoemulsion, device and method for solubilizing and separating carboxylic acids with a solubilizing compound. the present invention is directed to solubilizing compounds, devices and a method for solubilizing and removing carboxylic acids and especially fatty acids from oils, fats, aqueous emulsion, aqueous medium and organic solutions. devices using the inventive method must be used to separate carboxylic acids from oils, fats, aqueous emulsion, lipophilic medium or organic solutions, respectively by preparing an aqueous micro or manoemulsion of carboxylic acids especially fatty acids and the solubilizing compound containing at least one fatty group and the solubilizing compound containing at least one amidine and / or gianidine group. solubilizing effects of solubilizing compounds combined with the inventive use of carboxylic acid separation methods can be used to treat people in need of fatty acid removal or to analyze blood carboxylic acids or to process other solutions in the food, chemical, biofuel or other industrial processing.
公开号:BR112012032341B1
申请号:R112012032341
申请日:2011-06-22
公开日:2020-01-28
发明作者:Dietz Ulrich
申请人:Dietz Ulrich；
IPC主号:

专利说明:

METHOD FOR SOLUBILIZING AND SEPARATING CARBOXYLIC ACIDS WITH A SOLUBILIZING COMPOUND
A. Fundamentals of the Invention
The present invention is directed to solubilizing compounds, a device and a method for solubilizing and removing carboxylic acids and especially fatty acids from oils, fats, aqueous emulsions, aqueous or organic solutions. Devices using the inventive method must be used to separate carboxylic acids from oils, fats, aqueous emulsions, lipophilic medium, aqueous medium or organic solutions, respectively, thus changing their reaction conditions. An application is a device for removing fatty acids from the blood of subjects in need of them. It should be used additionally for analysis, respectively objectives of diagnosis of concentrations of fatty acid in body fluids of subjects, food or pharmaceutical preparations. In addition, this technique should be applied for the removal of carboxylic acid residues in industrial solutions, for example as they grow in the food and oil industry.
In general, fatty acids are highly lipophilic molecules that are sparingly soluble in aqueous solutions. Therefore, only small concentrations of fatty acids can be solubilized in aqueous solutions while all fatty acid molecules exceeding this concentration are present in the form of micelles, form an emulsion by phase separation, or are absorbed into the walls of the container and / or other lipophilic or amphilic molecules such as proteins in the solution. Above the critical micelle concentration (CMC) of esterified and non-esterified carboxylic acids, the concentration of free fatty acids in an aqueous medium remains unchanged.
Fatty acids tend to form emulsions in an aqueous medium. In the presence of proteins or fatty acids, cell structures can be absorbed by them or absorb into them. Solubilization of such immobilized fatty acids depends mainly on the concentration of critical micelle (CMC) of the fatty acid in the surrounding aqueous medium. Emulators and detergents are able to raise the CMC of hydrophobic substances and thus help to detach immobilized lipophilic molecules. These emulators and detergents can convert into mini, micro or nano emulsions. Here, the contact area of fatty acids
Petition 870190048064, of 05/22/2019, p. 8/13
2/118 solubilized with the aqueous phase is high. This allows for better separability and extractability of the solubilized fatty acids. Consequently, reactivity with other molecules is also increased.
Emulsions of fatty acids esterified and not esterified with an aqueous medium can be completely separated only by means of an organic solvent. Without the help of a membrane this can be achieved only by transferring the fatty acids in an organic phase mixing with an organic solvent. Extraction is also possible by absorbing an acceptor. In the presence of adsorbent molecules such as proteins, the separation of fatty acids into an emulsion or suspension by phase separation or extraction is often incomplete. In addition, the capacity of this technique is limited and usually not suitable for online (continuous) processing. When filtering such emulsions the aqueous fraction can be filtered almost entirely, also hydrophilic molecules, in particular large proteins, are retained and separated along with the organic phase. Molecular separation can be achieved with chromatographic methods. These methods, however, are time-consuming and limited in their capabilities.
Another method for separating carboxylic acids from the aqueous or organic medium is distillation. However, this procedure has a high energy demand and can generate isomerization of carboxylic acids or denatured organic components within the medium. An additional method is saponification. Added salts are often difficult to remove from the organic as well as from the aqueous solution during further processing.
Consequently, there is a need for continuous and selective extraction of 25 fatty acids from emulsions of aqueous or organic solutions. The purpose of the present invention is to provide a simple, rapid and biocompatible separation of fatty acids from aqueous emulsions or organic media.
It has been found that this objective can be achieved by adding a solubilizing compound to the aqueous emulsion or aqueous medium such as blood, lipophilic medium or organic medium containing carboxylic acids or mixtures of carboxylic acids with other organophilic molecules. An inventive solubilizing compound having the characteristics as defined herein is capable of solubilizing carboxylic acids and converting the emulgated carboxylic acids into micro or nano emulsions that allow a separation by means of separation methods such as dialysis, filtration and electrophoresis.
3/118
Thus, the issue is resolved by subsequent technical teaching of the independent claims of the present invention. Additional advantageous embodiments of the invention result from the dependent claims, the description and the examples.
Fatty acids
In general, fatty acids have a carboxyl head group and a long aliphatic chain. Depending on the presence of the double bonds they are differentiated into saturated and unsaturated fatty acids. There are different definitions in the literature on fatty acids. One definition states that carboxylic acids with 4 carbon atoms or more are considered to be fatty acids. Naturally occurring fatty acids, however, have at least 8 carbon atoms. In these carbon atoms, at least one nitro group can replace hydrogen atom (s) and convert them into nitro fatty acids. Also nitro fatty acids can carry additional substituents, as listed above.
Examples of linear saturated fatty acids are octanoic acids (caprylic acid), decanoic acid (caprinic acid), dodecanoic acid (lauric acid), tetradecanoic acid (myristic acid), hexadecaoic acid (palmitic acid), heptadecanoic acid (marginic acid), octadecanoic (stearic acid), axesanoic acid (arachidic acid), docosanoic acid (behenic acid) and tetracosanoic acid (lignicic acid).
According to the invention a preferred subgroup of saturated fatty acids to be separated are myristic acid, palmitic acid and stearic acid.
Examples of monoolefinic fatty acids are cis-9-tetradecenoic acid (myristoleic acid), cis-9-hexadecenoic acid (palmitoleic acid), cis-6-hexadecenoic acid (salpenic acid), cis-6-octadecenoic acid (petroselinic acid), cis acid -9-octadecenoic acid (oleic acid), cis-11-octadecenoic acid (vaccenic acid), 12-hydroxy-9-cis-octadecenoic acid (ricinoleic acid), cis-9eicosenoic acid (gadoleinic acid), cis-11-eicoseneic acid (gondóico acid), cis-13-docosenoic acid (erucic acid), cis-15-tetracosenoic acid (nervous acid), t9-octadecenoic acid (elaidic acid), t11-octadecenoic acid (tvacenic acid) and t3-hexadecenoic acid. According to the invention, a preferred subgroup of the unsaturated fatty acids to be separated are the transisomeric t9-octadecenoic acid, t11-octadecenoic acid and t3hexadecenoic acid.
4/118
Examples of polyolefinic fatty acids are 9,12-octadecadienoic acid (linoleic acid), 6,9,12-octadecatrienoic acid (γ-linoleic acid), 8,11,14eicosatrienoic acid (di-homo-Y-linoleic acid), 5,8,11,14-eicosatrienoic acid (arachidonic acid), 7,10,13,16-docosatetraenoic acid, 4,7,10,13,16docosapentaenoic acid, 9,12,15-octadecatrienoic acid (α-linolenic acid) , 6,9,12,15-octadecatetraenic acid (stearidonic acid), 8,11,14,17 eicosatetraenoic acid, 5,8,11,14,17-eicosapentaenoic acid (EPA), 7,10,13,16 acid, 19-docosapentaenoic acid (DPA), 4,7,10,13,16,19-docosahexaenoic acid (DHA), 5,8,11-eicosatrienoic acid (mead acid), 9c 11t 13t eleostearinoic acid, 8t 10t 12c calenic acid, 9c 11t 13c catpathic acid, 4, 7, 9, 11, 13, 16, 19 docosaheptadecanoic acid (stelaheptaenoic acid), taxolic acid, pinolenic acid and sciadonic acid.
According to the invention, a preferred subgroup of unsaturated fatty acids to be separated are the transisomers of linoleic acid, γlinoleic acid, EPA and DPA.
Examples of acetylenic fatty acids are 6-octadecinoic acid (taric acid), t11-octadecen-9-inoic acid (santálbico or ximênico acid), 9octadecinoico acid (stearolic acid), 6-octadecen-9-innoic acid (6,9octadecenoic acid ), t10-heptadecen-8-inoic acid (pyrulic acid), 9octadecen-12-inoic acid (crepeic acid), t7, t11-octadecadiene-9-inoic acid (heisteric acid), t8, t10-octadecadiene-12- inoic acid, 5,8,11,14eicosatetrainoic acid (ETYA).
It should be noted that, according to the invention, also the bases, respectively salts of fatty acids mentioned above, must be subsumed in the general terms of fatty acids or free fatty acids.
Examples for organic and inorganic bases suitable for salt formation are bases derived from metal ions, for example aluminum, alkali metal ions, such as potassium sodium, alkaline earth metal ions such as calcium or magnesium, or a amine salt ion or alkaline or alkaline earth hydroxides, carbonates or - bicarbonates. Examples include aqueous sodium hydroxide, lithium hydroxide, potassium carbonate, ammonia and sodium bicarbonate, ammonia salts, primary, secondary and tertiary amines, such as, for example, lower alkylamines such as methylamine, t-butylamine, procaine , ethanolamine, arylalkylamines such as dibenzylamine and N, Ndibenzylethylenediamine, lower alkylpiperidines such as N-ethylpiperidine, cycloalkylamines such as cyclohexylamine or dicyclohexylamine, morpholine,
5/118 glucamine, N-methyl-e N, N-dimethylglucamine, 1-adamantylamine, benzathine, or salts derived from amino acids such as lysine, ornithine or originally neutral amides or amino acid or other acids.
The following carboxylic acids are preferred examples of fatty acids:
octanoic acid (caprylic acid), decanoic acid (capric acid), dodecanoic acid (lauric acid), tetradecanoic acid (myristic acid), hexadecanoic acid (palmitic acid), heptadecanoic acid (marginic acid), octadecanoic acid (stearic acid), eicosanoic (arachidic acid), docosanoic acid (behenic acid), tetracosanoic acid (lignoceric acid), cis-9-tetradecenoic acid (myristoleic acid), cis-9-hexadecenoic acid (palmitoleic acid), cis-6-octadecenoic acid (petroselinic acid) , cis-9octadecenoic acid (oleic acid), cis-11-octadecenoic acid (vaccenic acid), cis-9-eicosenoic acid (gadoleic acid), cis-11-eicosenoic acid (gondoic acid), cis-13-docosenoic acid ( erucic acid), cis-15-tetracosenoic acid (nervous acid), t9-octadecenoic acid (elaidic acid), t11-octadecenoic acid (t-vaccenic acid), t3-hexadecenoic acid, 9,12-octadecadienoic acid (linoleic acid) , 6,9,12-octadecatrienoic acid (γ-linoleic acid), 8,11,14 eicosatrienoic acid (di-homo-y-linolenic acid), 5,8,11,14-eicosatetraenoic acid (arachidonic acid), 7,10 , 13,16-docosatetraenoic acid, 4,7,10,13,16docosapentaenoic acid, 9,12,15-octadecatrienoic acid (α-linoleic acid), 6,9,12,15-octadecatetraenoic acid (stearidonic acid), 8 , 11,14,17 eicosatetraenoic acid, 5,8,11,14,17-eicosapentaenoic acid (EPA), 7,10,13,16,19-docosapentaenoic acid (DPA), 4,7,10,13,16 acid, 19docosahexaenoic acid (DHA), 5,8,11-eicosatrienoic acid (mead acid), 9c 11t 13t eleostearic acid, 8t 10t 12c calenic acid, 9c 11t 13c catolic acid, 4, 7, 9, 11, 13, 16, 19 docosaheptadecanoico (stelaheptaenoico acid), taxoleic acid, pinolenic acid, sciadonic acid, 6-octadecinoic acid (tartaric acid), t11-octadecen-9-inoic acid (santálbico or ximenínico acid), 9octadecinoic acid (stearolic acid), 6 -9-inoic (á 6,9octadeceninoic acid), t10-heptadecen-8-inoic acid (pyrulic acid), 9octadecen-12-inoic acid (crepeninic acid), t7, t11-octadecadiene-9-inoic acid (hysteric acid), t8, t10- acid octadecadiene-12-inoic acid, 5,8,11,14eicosatetrainoic acid (ETYA), eleosearic acid, calendic acid, catalpic acid, stelaheptaenoic acid, taxoleic acid, retinoic acid, isopalmitic acid, pristanic acid, phytic acid, 11,12-methyleneoctadecan acid , acid 9.10
6/118 methylenehexadecanoic, coronary acid, (R, S) -lipoic acid, (S) -lipoic acid, (R) -lipoic acid, 6,8-bis- (methylsulfanyl) -octanoic acid, 4.6bis (methylsulfanyl) ) -hexanoic acid, 2,4-bis- (methylsulfanyl) -butanoic acid, 1,2ditioliol carboxylic acid, (R, S) -6,8-octanoic acid, (R) -6,8-octanoic acid, acid (S) -6,8-octanoic dithian, cerebronic acid, hydroxinervonic acid, ricinoleic acid, lesquerolic acid, brassylic acid and tapsic acid.
Qraxic acids in the blood
In mammals, fatty acids serve as physiologically important energy substrates and play a key role in energy metabolism. In addition, they are important substrates for the synthesis of membrane phospholipids and biologically active agents, such as eicosanoids and leukotrienes. The mammalian body relies heavily on fatty acids as suppliers of stored chemical energy, building blocks of cell membranes and signal transducers. The main source of fatty acids is dietary lipid, digested in the gastro-intestinal tract by the catalytic action of hydrolytic pancreatic enzymes. Part of the fatty acids is produced by the liver with carbohydrates as a substrate. A large percentage of fatty acids, however, are stored in the fat cells (adipocytes) that make up adipose tissue in the form of triacylglycerol.
The concentration of esterified and non-esterified fatty acids in the blood depends on several factors, such as food intake or the release of adipose tissue. Fatty acids can be attached or attached to other molecules, such as triglycerides or phospholipids, or for a smaller percentage of fatty acids, non-binding occurs. In any case, fatty acids are insoluble in water and must be linked to a water-soluble component for transport in the body. Fatty acids are transported in the body through the lymphatic and vascular system. Basically, two forms of transport are at hand: Fatty acids can be transported as triacylglycerols, which is the main component of circulating lipoproteins, such as chylomicrons and lipoproteins of much lower density, or as non-esterified fatty acids that are bound to proteins plasma concentrations, in particular plasma albumin. Free fatty acids that are completely turned off have very low solubility and occur only in very low concentrations.
7/118
The composition, distribution and concentration of fatty acids in human blood can vary widely, and be composed of the sum of different plasma fractions: cholesterol ester, phospholipids, and triacylglycerols as well as fatty acids linked to albumin. The saturated fatty acids in human blood are mainly made up of myristic acid (14: 0), palmitic acid (16: 0) and stearic acid (18: 0). The main type of monounsaturated fatty acids belongs to the group of oleic acid (18: 1) and palmitoleic acid (16: 1). Polyunsaturated omega-3 fatty acids include linolenic acid (18: 3), eicosapentaenoic acid (20: 5), docosapentaenoic acid (22: 5) and docosahexaenoic acid (22: 6). Polyunsaturated omega-6 fatty acids are mainly linoleic acid (18.2), eicosadienoic acid (20: 2), dihomogamalinolenic acid (20: 3), arachidonic acid (20: 4), adrenic acid (22: 4) and docosapentaenoic acid ( 22: 5). The concentration of other fatty acids is usually very low in whole blood, but it can vary depending on genetics, nutrition and lifestyle.
Concentrations of fatty acids in the blood are increased in obese patients and contribute to type 2 diabetes, hepatic steatosis and several cardiovascular disorders such as atherosclerosis. The pathogenic role of fatty acids in the development of atherosclerosis and associated diseases, such as cerebral, myocardial, renal, erectile dysfunction has been elucidated. Not intended to be exhaustive, some aspects should be described below. An increase in fatty acids was considered responsible for the increase in the formation of reactive oxygen radicals causing endothelial dysfunction, which can be attenuated by an antioxidant (Pleiner et al, FFA-induced endothelial dysfunction can be corrected by vitamin C. J Clin Endocrinol Metab 2002, 87, 2913-7). This effect is enhanced by trans fatty acids, which are suspected of having additional harmful effects (Lopez García et al, Consumption of trans-fatty acids is related to plasma biomarkers of inflammation and endothelial dysfunction. J Nutr 2005, 135, 562-566; Mozaffarian et al, Health effects of trans-fatty acids: experimental and observational evidence. Eur J Clin Nutr 2009, 63 Suppl 2, S5-21). They are accused of increasing blood pressure and considered a pathogenic factor in hypertension (Zheng et al, Plasma fatty acid composition and 6-year incidence of hypertension in middle-aged adults: the Atherosclerosis Risk in Communities (ARIC) Study. Am J Epidemiol 1999, 150, 492-500). Trans fatty acids have been considered to increase the risk of myocardial infarction and sudden cardiac death (Ascherio et al, Trans-fatty acids intake and
8/118 risk of myocardial infarction. Circulation 1994, 89, 94-101; Baylin et al, High 18: 2 trans-fatty acids in adipose tissue are associated with increased risk of nonfatal acute myocardial infarction in Costa Rican adults. J Nutr 2003, 133, 1186-1191). Together with a chronic rise in blood fatty acid concentrations, they are responsible for insulin resistance and the development of diabetes mellitus (Krachler et al, Fatty acid profile of the erythrocyte membrane preceding development of Type 2 diabetes mellitus. Nutr Metab Cardiovasc Dis 2008, 18 , 503-510; Lionetti et al, From chronic overnutrition to insulin resistance: the role of fat-storing capacity and inflammation.Nutr Metab Cardiovasc Dis 2009, 19, 146-152; Yu et al, Mechanism by which fatty acids inhibit insulin activation of insulin receptor substrate-1 (IRS-1) -associated phosphatidylinositol 3-kinase activity in muscle. J Biol Chem 2002, 277, 50230-50236). In total, a high fatty acid delivery as a result of chronic over-nutrition is now believed to be the most important pathomechanism in the development of the most common diseases in industrialized countries (Bays, Sick fat, metabolic disease, and atherosclerosis. Am J Med 2009, 122, S26-37). Medical treatment for effective overweight reduction is lacking (Aronne et al, When prevention fails: obesity treatment strategies. Am J Med 2009, 122, S24-32). However, obese people who are successful in reducing body weight and thus a significant reduction in fatty acid-induced disorders can be found (Lien et al, The STEDMAN project: biophysical, biochemical and metabolic effects of a behavioral weight loss intervention during weight loss, maintenance, and regain.Omics 2009, 13, 21-35; Schenk et al, Improved insulin sensitivity after weight loss and exercise training is mediated by a reduction in plasma fatty acid mobilization, not enhanced oxidative capacity. 2009, 587, 4949-4961). Therefore, a medical device to effectively reduce the total amount of fatty acids and preferably those with increased pathogenicity is desirable.
Surgical extraction of subcutaneous adipose tissue was considered ineffective in reducing concentrations of circulating fatty acid or its qualitative content. The removal of the lipoprotein fraction with high cholesterol concentrations by direct blood adsorption can be achieved by adsorption or filtration of these particles. Those online blood purification procedures are called LDL apheresis. Although designed to lower LDL cholesterol, they also adsorb triglycerides. At the
9/118 However, the amount of triglycerides extracted is not sufficient for an effective reduction in the body fatty acid content.
The fatty acid content of the blood is lower in the fixation state at rest. However, the significant increase is seen during lipolysis (see below). Due to insolubility in an aqueous medium, the transport of non-esterified fatty acids is carried out by proteins and cellular structures (Spector et al, Utilization of long-chain free fatty acids by human platelets. J Clin Invest 1970, 49, 1489-1496). The main transport protein in the blood is albumin. The presence of at least 10 specific binding sites for fatty acids has been documented. However, the binding capacity can increase dramatically by the formation of micellar structures with fatty acids in a condition of excess fatty acids or other lipids (Schubiger et al, Mixed micelles: a new problem-free solution for omega- ¹²³ l-heptadecanoic acid in comparison, Nuklearmedizin 1984, 23, 27-28).
With a molarity of albumin of about 600 pmol / l a binding capacity of at least 0.006 mol / l for fatty acids would exist which is equivalent to about 0.0035 kg / l (Berk and Stump, Mechanisms of cellular uptake of long chain free fatty acids. Mol Cell Biochem 1999,192,17-31).
In addition, fatty acids are transported in esterified form as mono-, di- or triacyl glycerols. The serum fixation concentration varies considerably. However, normal values are set to be below 150 mg / dl (1.7 mmol / l). Postprandial exercise or during concentration can increase several times and even exceed 1000 mg / dl (11.3 mmol / l).
There are only replacement reports that investigate differences in lipid content at various locations within the circulation. In these investigations, significantly higher values for fatty acids and triglycerides were considered to be present in the central venous system (vena cava), in comparison with other measurement sites (Wiese et al, Lipid composition of the vascular system during infancy, childhood, and young adulthood , J. Lipid Res. 1967, 8, 312-320; Zauner et al, Pulmonary arterial-venous differences in lipids and lipid metabolites. Respiration 1985, 47, 214-219). There are no reports on changes in central venous lipid content during exercise and induced lipolysis.
It has now been found that, during exercise, the lipid content increases markedly in the abdominal central veins that exhibit a difference
10/118 increase in lipid content between the plant, compared to a peripheral evaluation site as described below.
Thus, the reduction of the fatty acid content in the blood, using the methods and devices and the solubilizing compounds disclosed here is useful for treating the aforementioned diseases associated with a high level of fatty acids in the blood or in the body.
Thus, the present invention relates to the treatment and prophylaxis of disorders induced by fatty acids, such as type 2 diabetes, hepatic steatosis, cardiovascular diseases, such as arterial hypertension, myocardial infarction, stroke, sudden cardiac death, atherosclerosis, diseases associated with atherosclerosis, such as cerebral, myocardial, renal and erectile dysfunction, as well as for weight reduction and cholesterol reduction and also for the prevention of insulin resistance and the prevention of the development of diabetes mellitus, using the compounds solubilizers disclosed here in order to remove fatty acids from the blood.
Lipolysis
Plasma fatty acids are an important energy substrate. The availability of fatty acids is determined predominantly by their mobilization from the triacylglycerol reserves of adipose tissue by the lipolysis process. In man, lipolysis of adipose tissue is regulated by a number of hormonal, paracrine and / or autocrine signals. The main hormonal signs can be represented by catecholamines, insulin, growth hormone, natriuretic peptides, thyroxine and some adipocytokines (Stich and Berlan, Physiological regulation of NEFA availability: lipolysis pathway. Proc Nutr Soc 2004, 63, 369-374). The absolute levels and relative importance and contribution of these signals vary in different physiological situations, with diet and exercise being the main physiological variables that affect hormonal signaling. A family of enzymes called lipases with distinct functions is responsible for the breakdown of triglycerides stored inside fat cells for energy storage. Carbohydrates and fatty acids are the main energy fuels for muscle contraction. During exercise training lipolysis releases body weight of 7.1 +/- 1.2 micromol x min (-1) x kg (-1), which could result in a release of 4200pmol fatty acids per hour in a person with a weight of 100 kg, which is equal to 0.15 kg of fatty acids (Coggan et al, Fat metabolism during high intensity exercise in endurance-trained and untrained men. Metabolism 2000, 49,
11/118
122-128). However, stimulation of lipolysis by pharmacological intervention and / or local physical measures can further increase lipolytic capacity. Lipolysis was increased up to 3 times by systemic application of natural receptor agonists or drugs (Riis et al, Elevated regional lipolysis in hyperthyroidism. J Clin Endocrinol Metab 2002, 87, 4747-4753; Barbe et al, In situ assessment of the role of the beta 1-, beta 2- and beta 3-adrenoceptors in the control of lipolysis and nutritive blood flow in human subcutaneous adipose tissue (Br J Pharmacol 1996, 117, 907-913). Adrenergic receptor agonists exhibiting stimulus lipolysis: adrenaline, norepinephrine, isoprenaline, ephedrine, isoproteriol, salbutamol, theophylline, phenoterol, orciprenaline, a.o.
Lipolytic effects have also been described from physical changes in adipose tissue. The researchers found that ultrasound had a liquefying effect on adipose tissue leading to a reduced adipose tissue content when performed during hunger (Faga et al, Ultrasound-assisted lipolysis of the omentum in dwarf pigs. Aesthetic Plast Surg 2002, 26 , 193-196; Miwa et al, Effect of ultrasound application on fat mobilization (Pathophysiology 2002, 9,13).
Although for all of the measures mentioned above an increase in lipolysis has been documented, the measurable effect on non-esterified fatty acid concentrations was small. In a pilot investigation it was found that, after stimulation of lipolysis, the fatty acid content increased dramatically when measured using the intent method for fatty acid solubilization. In addition, it was found that the fatty acid content was much higher within the abdominal venous system than in the peripheral circulation. This finding is surprising, since it was not seen in animal studies when measuring different blood collection sites simultaneously.
Thus, stimulation of lipolysis when purifying blood from fatty acids by the procedure of intention and using an abdominal central vein as an access site is a preferred embodiment of the present invention.
The extraction fraction could be increased if the content of esterified and non-esterified fatty acids transported in the blood could be increased during the procedure.
This issue could be resolved by increasing lipolysis using the method of the invention.
Solvation behavior and adherence of fatty acids in aqueous medium
11/12
The solubility of carboxylic acids in water is minimal when ο the carbon chain length exceeds 4 carbon atoms and in the absence of hydroxy groups (-OH), carboxyl groups (-COOH) or other polar or charged hydrophilic groups and / or by introduction alkyl substituents or other 5 lipophilic groups.
Solubility can be increased by detergents that penetrate fatty acid micelles, thereby reducing their stability and reducing their size and increasing the number of free fatty acid molecules in the aqueous medium. Both, free fatty acids and micelles, tend to bind to lipotropic structures. Among them are carbon, metals, ceramics, synthetic and natural polymers. In addition, organic structures carry lipophilic regions, some of which are designed to specifically bind fatty acids, which form membranes or lipid transport proteins. Steric binding sites are coated primarily with hydrophobic amino acids.
In the blood, lipids are electrostatically bound to specialized transport proteins. Fatty acids are mainly transported by albumin. The binding of fatty acids in the albumin molecule is also based on electrostatic forces that are located in hydrophobic bags. The binding energy of the pockets varies, however, the pKa for all of them is substantially higher than the CMC of fatty acids. Thus, the fatty acids remain in the surrounding environment, even after the complete removal of the free fatty acids. The extraction of fatty acids from albumin was considered almost complete when organic solvents were used for its release because of the better dissolution in organic solvents. However, these solvents alter the structure of the protein making them unsuitable for further processing or use in a living organism. To use albumin for medical or other purposes, it is necessary to reduce its fatty acid content, without changing the structure and functionality of albumin. This issue can be solved by activated carbon particles that have a higher binding affinity to fatty acids than albumin. However, this process requires additional steps for the purification of albumin. Therefore, until now, there is no procedure that allows the quick release and solubilization of the entire fatty acid content of an albumin molecule in an aqueous medium, which does not alter the ultrastructure and function of the albumin molecules.
11/13
Carboxylic acids are also transported within phospholipid vesicles. Electrostatic interactions between the hydrocarbon chains of carboxylic acids and those phospholipids retain carboxylic acids from diffusion to a surrounding aqueous medium.
Mutatis mutandis, this also applies to other organic solutions, biomass or organic waste water. In organic solutions intended for further improvement, purification or use where it is desirable not to use an organic solvent, an alternative biocompatible procedure is desirable. So far, that procedure is missing.
This objective can be achieved by using at least one solubilizing compound as described herein comprising at least one amidino and / or at least a fraction of guanidino and especially solubilizing compounds of the general formula (I), (II) and (III) and more especially arginine and its derivatives.
The carboxylic acids that must be removed are normally contained in an aqueous medium or aqueous solution such as blood or blood plasma or in an aqueous emulsion, such as milk or in an organic medium, such as fuel, gas, biodiesel, gasoline, oil and others, or in oils, such as vegetable oils such as linseed oil, walnut oil, linseed oil, evening primrose oil, sunflower oil, sunflower seed oil, soybean oil, canola oil, olive oil, olive oil virgin, palm oil, palm kernel oil, peanut oil, cottonseed oil, coconut oil, corn oil, grape seed oil, hazelnut oil, rice bran oil, safflower oil, sesame oil, as well as oils of animal origin, such as fish oil or contained in fats, such as butter, oil or margarine.
In case the carboxylic acid is contained in water, an aqueous medium, an aqueous emulsion or an aqueous suspension, the at least one solubilizing compound can be directly added to the aqueous medium, emulsion or suspension, or the at least one solubilizing compound can be dissolved in water and this aqueous solution can be added to the aqueous medium, emulsion or suspension containing the carboxylic acids. After this addition, the formation of a nanoemulsion and / or microemulsion is observed.
In the case that the carboxylic acids are contained in an organic medium or a lipophilic organic medium, the solubilizing compound is dissolved in water and the solution of the water solubilizing compound is added to the organic medium. The two-phase mixture is obtained and the carboxylic acids are transferred to the
14/118 aqueous phase. It is assumed that a complex or aggregated carboxylic acid of a molecule with a molecule solubilizing compound or a dimer or trimer thereof is formed which makes the carboxylic acid soluble in water. Thus, it is preferred to stir or stir the two-phase mixture of the organic and aqueous layer in order to obtain an intensive mixing of the two layers. The carboxylic acids contained in the aqueous phase can be removed by phase separation. If desired, the extraction method can be repeated.
In case the carboxylic acids are contained in an oil or fat, the solubilizing compound is dissolved in water and the solution of the water solubilizing compound is added to the oil or fat. If desired, an organic solvent could be added to the oil or fat in order to reduce the viscosity of the oil or fat to make the oil or fat better agitated. The mixture of oil or fat and the aqueous solution of the solubilizing compound is stirred. The carboxylic acid is transferred to the aqueous phase and the aqueous phase can be removed by decantation or phase separation. The extraction process can be repeated several times if desired.
Thus, the invention also relates to an aqueous microemulsion and / or an aqueous nanoemulsion containing at least one solubilizing compound and at least one carboxylic acid in a micromemulgated or nanoemulgated form.
If the solubilizing compound is used in excess of 1.2 to 2.8, preferably 1.5 to 2.5 and more preferably in excess of 1.7-2.3 mol equivalents, it is possible to remove more than 90% of the acids carboxylic acids in an extraction step. If the extraction steps are repeated twice, up to 99% of the carboxylic acids can be removed.
Carboxylic acids that can be removed are especially carboxylic acids, with more than 5 carbon atoms, more preferably with more than 7 carbon atoms and especially preferred with more than 9 carbon atoms. Preferably, the carboxylic acids are the fatty acids as described herein and also other lipophilic compounds containing a carboxyl group or carboxylic acid group, such as drugs or toxins can be removed by this method. A carboxylic acid that is expressly excluded from the present invention is naproxen. Furthermore, it is not the intention of the present invention to provide methods and compounds or devices for the solubilization of pharmaceutical products, in order to prepare galenic formulations. Especially preferred is the removal and solubilization of naphthenic acid from oil,
11/158 oil, gas and fuel. In addition, preferred carboxylic acids are carboxylic acids that contain double and / or triple bonds, such as unsaturated and polyunsaturated fatty acids. Even more preferred are carboxylic acids and in particular those physiological carboxylic acids that occur in humans. For industrial purposes, unsaturated fatty acids are preferably removed and solubilized from the source material, such as oils and fats although for medical purposes, saturated fatty acids are preferably removed from the patient's blood. In addition, these carboxylic acids are preferred, which occur in oils and fats of the aforementioned origin, especially from animals, such as fish, corn, olives, corn, crops, rice, soy and the like. In the case of carboxylic acids that are removed from the organic medium, such as fats, waxes, oil, fuel, petroleum and others are contained in an esterified form (that is, they are bound in esters), a saponification step can be performed before removal and inventive solubilization be carried out. Such saponification is preferably carried out in a mixture of water solvent and at least one second water-miscible solvent. Even more preferred carboxylic acids are perfluorinated carboxylic acids such as perfluoropropionic acid, perfluorooctanoic acid (PFOA), perfluorodecanoic acid, perfluorododecanoic acid, perfluorohexadecanoic acid as well as perfluorinated carboxylic acids and propyrinic acid.
The present invention also relates to the solubilization, respectively the removal of aromatic carboxylic acids that belong to the above mentioned target groups, such as benzoic acid, 4-aminobenzoic acid, anthranilic acid, benzyl acid, cinnamic acid, salicylic acid, phenylacetic acid, 4-methoxy-phenylacetic acid, gallic acid, phthalic acid, terephthalic acid, abietic acid, bicinconinic acid, quinic acid, chorisic acid, clavulanic acid, fusaric acid, fusidic acid, uric acid, hypuric acid, ibotenic acid, indole-3 acid -acetic, mandelic acid, stiphonic acid, ussic acid, abscisic acid, tropic acid, benzoquinonatetracarboxylic acid, bosonic acid, caffeic acid, carminic acid, chenodeoxycholic acid, cumharic acid, chromoglycic acid, cinnary, meclophenamic acid, 2,4-dichlorophenoxy acid , domoic acid, pipemidic acid, ferulic acid, acid o 5-hydroxyiferulic, isophthalic acid, mefenamic acid, meta-chloroperoxybenzoic acid, peroxybenzoic acid, protocatechuic acid, nalidixic acid, synapic acid, sucronic acid.
11/16
Especially preferred is the removal and solubilization of carboxylic acids from the blood, which lead to various diseases caused and / or associated by a high and / or unhealthy level of such carboxylic acids and especially fatty acids.
Carboxylic acids are preferably lipophilic and preferably have a partition coefficient between n-octanol and water (also known as log Kow or octanol-water partition coefficient) of> 2.0, preferably of> 3.0 and more preferably> 4.0. (For example: log Kow of acetic acid is -0.17, butyric acid is 0.79, octanoic acid is 3.05 and decanoic acid is 4.09).
It is also preferred if the carboxylic acids, which must be removed, have a pKs value> 4.85, preferably> 4.87. (For example: acetic acid has pKs of 4.76, butyric acid of 4.82, pentanoic acid of 4.84 and octanoic acid of 4.89).
Thus, the present invention provides a method for the separation of carboxylic acids that are not at all or not well soluble in water, which can be solubilized in water, by means of solubilizing compounds disclosed herein, preferably in the form of nano or microemulsions. . Once transferred to the aqueous phase, fatty acids can be removed by different techniques disclosed here.
Thus, the present invention relates to the use of a solubilizing compound for solubilizing carboxylic acids in an aqueous or organic medium, wherein said solubilizing compound contains at least one amidino group and / or at least one guanidino group and in which the compound has a partition coefficient between n-octanol and Kow water <6.30.
The term solubilizing carboxylic acids in an aqueous or organic medium should be understood as follows: the carboxylic acids that are solubilized are contained in an organic medium, such as oils or fuel or in an aqueous medium such as blood or milk and are solubilized through the use of a solubilizing compound in the aqueous phase.
Thus, it can also be stated that the present invention is directed to the use of a solubilizing compound to solubilize carboxylic acids from an aqueous or organic medium in the aqueous phase, wherein said solubilizing compound contains at least one amidino group and / or at least one guanidino group and the compound has a partition coefficient between noctanol and Kow water <6.30.
11/178
In addition, the present invention relates to the use of a solubilizing compound to solubilize lipophilic carboxylic acids in an aqueous medium, wherein said solubilizing compound contains at least one amidino group and / or at least one guanidino group and in which the compound has a partition coefficient between n-octanol and Kow water <6.30.
In the case where carboxylic acids are contained in the aqueous phase, such as blood, only very few amounts of free carboxylic acids are present in the blood, since these carboxylic acids and especially fatty acids are poorly soluble in water. Most of the carboxylic acids that must be removed from the blood are bound to other compounds, such as albumin, and are no longer free carboxylic acids. However, there is a balance between the very small amount of free carboxylic acids in the blood and the otherwise bound or deposited carboxylic acids that are considered to be no longer free. If, by means of the inventive method, the free carboxylic acids are complexed by the solubilizing compound, these free carboxylic acids are removed from the balance and albumin-bound carboxylic acids are released into the blood which can then be removed again by the inventive method so that finally almost all of the carboxylic acids contained in the blood in a free or bound form can be removed. Especially dialysis is suitable for such a continuous process of removing carboxylic acids and especially fatty acids from the blood.
The solubilizing compounds disclosed herein comprise at least one amidino group or at least one guanidino group or at least one amidino group and at least one guanidino group. If the amidino group is not substituted it can be represented by the following formula H ₂ NC (NH) -. But it is also possible that all three hydrogen atoms are replaced by substituents R, R 'and R as represented by the following general formula (R) (R') NC (NR) -. It is preferred that two of the three hydrogen atoms are replaced by a substituent as represented by the following formula: (R ') NH-C (NR) - or (R) (R') NC (NH) -. Thus, amidino groups with at least one hydrogen are preferred. If the guanidino group is not substituted it can be represented by the following formula H ₂ NC (NH) -NH-. But it is also possible that all four hydrogen atoms are replaced by substituents R, R ', R and R', as represented by the following formula (R) (R ') NC (NR) -N (R) -. It is preferable that three of the four hydrogen atoms are replaced by a substituent
18/118 as represented by the following formula: (R ') NH-C (NR) -N (R) - or (R) (R') NC (NH) -N (R) - or (R) (R ') NC (NR) -NH-. Thus, guanidino groups with at least one hydrogen and preferably with two hydrogens are preferred.
The solubilizing compound comprises or contains at least one amidino group and / or at least one guanidino group, while guanidino groups are preferred. In addition, the solubilizing compound preferably comprises or contains not more than 15 carbon atoms, more preferably not more than 14, more preferably not more than 13, more preferably not more than 12, more preferably not more than 11, more preferably not more than 10, more preferably not more than 9, and more preferably not more than 8 carbon atoms and more preferably the solubilizing compound is an arginine derivative. In the case of polymeric or oligomeric solubilizing compounds it is preferred that no more than 10 carbon atoms per fraction of amidine or guanidine fraction and more preferably no more than 8 carbon atoms are present.
In addition, the solubilizing compound is hydrophilic and may preferably contain one or more of the following substituents:
-NH ₂ , -OH, -PO3H2, -PO3H, -PO ₃ ² ', -OPO3H2, -OPO3H, -OPO3 ² ', COOH, -COO ', -CO-NH2, -NH ₃ ⁺ , -NH-CO -NH2i -N (CH3) 3 ⁺ , -N (C2H ₅ ) ₃ ⁺ , N (C3H7) 3 ⁺ , -NH (CH3) ₂ ⁺ , -NH (C2H5) 2 ⁺ , -NH (C3H ₇ ) ₂ ⁺ , -nhch3, -nhc2h5, -nhc3h7, -nh2ch3 ⁺ , -nh2c ₂ h ₅ ⁺ , -nh2c3h7 ⁺ , -so3h, -so ₃ ', -so ₂ nh ₂ , -co-cooh, -oCO-NH ₂ , -C (NH) -NH ₂ , -NH-C (NH) -NH ₂ , -nh-cs-nh ₂ , -nh-cooh.
Also preferred are solubilizing compounds that are derived from arginine or that are dipeptides or tripeptides and polypeptides containing the amino acid arginine or an arginine derivative.
It is also possible that the amidino group or a guanidino group is part of a heterocyclic ring system as in imidazole, histidine, clothianidin or 4- (4,5-dihydro-1H-imidazol-2-ylamino) -butyric acid.
The solubilizing compounds are hydrophilic and have a partition coefficient between n-octanol and water (also known as Kow or octanol-water partition coefficient) of Kow <6.30 (log Kow <0.80), preferably K _O w <1.80 (log Kow <0.26), more preferably Kow <0.63 (log Kow <-0.20) and more preferably Kow <0.40 (log Kow <-0.40).
Preferred solubilizing compounds are:
11/198
L-2-Amino-3-guanidinopropionic acid, L-arginine, L-NIL, H-homoarg-OH, histidine, Νω-nitro-L-arginine, Ν-ω-hydroxy-L-norarginine, D-arginine methyl ester , nomega-monomethyl-L-arginine, NG.NG-dimethylarginine, D - (+) - octopine, argininosuccinic acid, free base of L-canavanine, creatine, guanidinoacetic acid, 3-guanidinopropionic acid, 4- (4,5 -dihydro-1 H-imidazol-2-ylamino) butyric acid, (S) - (-) - 2-guanidinoglutaric acid, 6-guanidinohexanoic acid, guanidino, sulfaguanidine, agmatinsulfate, 4-guanidinobenzoic acid, 1,3-di-o-o- tolylguanidine, clothianidin, L-ornithine, N-guanylurea, cimetidine, 1- (o-tolyl) biguanide, chlorhexidine, L-dimethylbiguanide, proguanil, polyhexanide, poly-L-arginine (70,000150,000 mw), diminazene, melanin, 4 - (4,6-diamino-2,2-dimethyl-2H- [1,3,5] triazine-
1-yl, imidazole, methylimidazole, Tyr-Arg (chiotorphin), Arg-GIn, Gly-Ang, Arg-Phe, ArgGlu, Lys-Arg acetate, His-Arg, Arg-Gly-Asp (RGD), Arg- Phe-Ala, Thr-Lys-ProArg (tuftsin), Gly-Gly-Tir-Arg, Gly-His, argatroban, L-NMMA (L-NG-monomethylarginine), L-NAME (L-nitro-arginine-methylester) , L-hydroxy-arginine-citrate, dimethylarginine (ADMA), D-homoarginine, noraginine, L-canavanin acid (2 amino-4- (giianidinooxy)-butyric), 4-guanidino-phenylalanine, 3-guanidinophenylalanine, Oa-hypurilic acid -L-arginine, H-Arg-AMC (L-arginine-7-starch-4methylcoumarin), L-TAME (P-tosyl-L-arginine-methylester), diphenylacetyl-D-Arg-4hydroxybenylamide, agmatine (argamine, 1 -amino-4-guanidinobutansulfate), Larginine-ethylester, L-arginine-methylester, guanidine, guanidinacetate, guanidincarbonate, guanidinitrate, guanidintiocyanate, guanyl urea, guanyl urea phosphate, 2-guinyl dihydrate, guanidinyl dihydrate, 2-guanidinea dihydrate -guanidinobenzimidazole, S - ((2-guanidino-4-thiazolyl) methyl l) -isothio urea, guanidinobutyldehyde, 4-guanidinobenzoic acid, leonurin (4-guanidino-n-butylsyringate), ambazon ([4- (2- (diaminomethyliden) -hydrazinyl) phenyl] iminothio urea), amiloride (3,5-diamino- N-carbamimidoyl-6-chlorpyrazin-2-carbamide), aminoguanidine, amitrol (3-amino-1,2,4-triazole), nitroguanidine, argininosucinate, barettin «2S, 5Z) -cycle - [(6-brom-8 -en-tryptophan) -arginine]), lysine, chlorhexidine (1, Thexamethylenbis [5- (4-chlorophenyl) -biguanide]), cimetidine, (2-cyano-1-methyl-3- [2- (5methyl-imidazole -4-ylmethylsulfanyl) -ethyl] -guanidine, clonidin (2 - [(2,6-dichlorophenyl) imino] imidazolidine), clothianidin ((E) -1 - (2-chloro-1,3-thiazol-5-ylmethyl ) -3-methyl-2-nitroguanidine), 2,4-diaminopyridine, Ν, Ν'-di-o-tolylguanidine, guanetidine, creatine, creatinine, kiotorphine (L-tyrosyl-L-arginine), lugdunam, acid (N- ( 4-cyanophenyl) -N (2,3-methylendioxybenzyl) guanidinacetic, metformin (1,1-dimethylbiguanide), octopine (N ^a - (1-carboxyethyl) -arginine), polyhexanide (polyhexamethylenbiguanide (PHMB )), proguanil (1- (4-chlorophenyl) -5-isopropylbi-guanide), sulfaguanidine (420/118 amino-N- (diaminomethylene) benzensulfon-amide), tetrazene (4-amidino-1 (nitrosaminoamidino) -l - tetrazene), L-arginine-4-methoxy-p-naphthylamide, L-arginine-βnaftilamide, L-arginine-hydroxamate, L-arginine-p-nitroanilide, Na-benzoyl-DLarginine, Νω-nitro-L-arginine, robenidine , (1,3-bis [(4-chlorobenzyliden) amino] guanidine, 1- (2,2-diethoxyethyl) -guanidine, 1- (P-tolyl) -guanidine nitrate.
CL

N
melamine clonidine amitrol
NH NH polyhexanide
NH NH
1,1-dimethylbiguanide
The invention can be used effectively over a wide range of concentration ratios of the solubilizing compound and the fatty acids to be solubilized. Often, the fatty acid content of a solution is not exactly known. Therefore, the ratio of the solubilizing compound to be added must be estimated. Inventive solubilization of carboxylic acids and especially fatty acids can be achieved when the molar ratio of the solubilizing compound to fatty acids (free and limited) is in the range of 1: 1000 to 1000: 1. A range of 1: 100 to 100: 1 is preferred. A range from 1:10 to 10: 1 is most preferred. Even more preferable a range of 1: 2 to 2: 1. A ratio of 1: 1 to 2: 1 is most preferred. It is preferred that the solubilizing compound is used in a molar excess of 3% or 5% or 7% or 8% or 10% or 12% or 15% or 20% or 25% or 30% or 35% or 40% or 45% or 50% or 55% or 60% or 70% or 80% or 90% or 100% or 120% or 140% or 160% or 180% or 200%. In addition, a molar ratio of fatty acid to solubilizing compound of 1: 1 to 1: 200 is preferred. More preferred is a molar ratio of fatty acid to the solubilizing compound in the range of 1: 1 to 1: 100, more preferably from 1: 1a to 1:50, even more preferable from 1: 1 to 1:30, even more preferred from 1: 1 to 1:25, even more preferred from 1: 1 to 1:20, even more preferred from 1: 1 to 1:15, even more preferred from 1: 1 to 1:10, even more preferred from 1: 1 to 1: 9, and
21/118 even more preferred from 1: 1 to 1: 8, even more preferred from 1: 1 to 1: 7, even more preferred from 1: 1 to 1: 6, even more preferred from 1: 1 to 1: 5 , even more preferred from 1: 1 to 1: 4, even more preferred from 1: 1 to 1: 3, even more preferred from 1: 1 to 1: 2, even more preferred from 1: 1 to 1: 1.8 , even more preferred from 1: 1 to 1: 1.6, even more preferred from 1: 1 to 1: 1.5, even more preferred from 1: 1 to 1: 1.4, also preferred from 1: 1 to 1: 1.3, also preferred from 1: 1 to 1: 1.2, also preferred from 1: 1 to 1: 1.1, also preferred from 1: 1 to 1: 1.05, also preferred from 1: 1.2 to T.2.8, also preferred from 1: 1.4 to 1: 2.6, also preferred from 1: 1.6-1: 2.4, also preferred from 1: 1.8 to 1 : 2.2, most preferred from 1: 1.9 to 1: 2.1 and most preferred is a molar ratio of fatty acid to the solubilizing compound in the range of 1.0: 2.0. These molar ratios are preferably for solubilizing compounds with an amidino group or a guanidino group. If the solubilizing compound contains two amidino groups or two guanidino groups or an amidino group and a guanidino group, only half the amount of the solubilizing compound is preferably used. Thus, in this case, a molar ratio of fatty acid to the solubilizing compound of 1: 0.5 to 1:25, preferably 1: 0.6 to 1: 1.4, also preferably 1: 0.7 to 1 : 1.3, also preferred from 1: 0.8 to 1: 1.2, also preferred from 1: 0.9 to 1: 1.1, more preferably from 1: 0.95 to 1: 1.05 and most preferred is a molar ratio of fatty acid to the solubilizing compound in the 1.0: 1.0 range.
The solubilization is carried out preferably at a pH value> 7.0 and, more preferably within a pH range of 7.0 to 9.0. However, depending on the medium from which the carboxylic acids are to be separated, pH values up to 14 can be used, while a pH range between 7.0 and 8.0 is preferably used if the carboxylic acids are to be separated. removed from the blood. However, if complete solubilization is not achieved, the more solubilizing compound can be added or the pH value can be increased or the aqueous phase can be separated and the extraction process is repeated or a combination of these three possibilities is used.
Some of the solubilizing compounds of the present invention can be represented by the following general formula (I) and formula (II):
NR NR
Λ λ Λα
RR’tV X R’Hn X
22/118 formula (I) formula (II) in which
R ', R, R' and R independently represent each other -H, -OH, -CH = CH ₂ , -CH ₂ -CH = CH ₂ , -C (CH ₃ ) = CH ₂ , -CH = CH- CH ₃ , -C ₂ H ₄ -CH = CH ₂ , -CH ₃ , C ₂ H ₅ , -C ₃ H ₇ , -CH (CH ₃ ) ₂ , -C ₄ H ₉ , -CH ₂ -CH (CH ₃ ) ₂ , -CH (CH ₃ ) -C ₂ H ₅ , -C (CH ₃ ) ₃ , C ₅ Hn, -CH (CH ₃ ) -C ₃ H ₇ , -CH ₂ -CH (CH ₃ ) - C ₂ H ₅ , -CH (CH ₃ ) -CH (CH ₃ ) ₂ , -C (CH ₃ ) ₂ C ₂ H ₅ , -CH ₂ -C (CH ₃ ) _3i -CH (C ₂ H ₅ ) ₂ , -C ₂ H ₄ -CH (CH ₃ ) _2i -C ₆ H- | _3i -C ₇ H ₁₅ , cycle-C ₃ H ₅ , cycle-C ₄ H7, CYCL0-C5H9, cycle-CôHn, —PO ₃ H ₂ , —PO ₃ H ', - PO ₃ ² , -NO ₂ , - C = CH, -C = C-CH ₃ , -CH ₂ -CsCH, -C ₂ H ₄ -C = CH, -CH2-C = C-CH ₃ or R 'and R together form the -CH ₂ residue -CH ₂ -, -CO-CH ₂ -, -CH ₂ CO-, -CH = CH-, -CO-CH = CH-, -CH = CH-CO-, -CO-CH ₂ -CH2-, - CH ₂ -CH ₂ CO-, -CH ₂ -CO-CH ₂ - or -CH ₂ -CH ₂ -CH ₂ X represents -NH-, -NR, -O-, -S- or -CH ₂ - or a substituted carbon atom; and
L represents a hydrophilic substituent selected from the group comprising or consisting of
-nh ₂ , -oh, -po ₃ h ₂ , -po ₃ h-, -po ₃ ² -, -opo3h2, -opo3h-, -opo3 ² ·, -COOH, COO ', -CO-NH2, -NH ₃ ⁺ , -NH-CO-NH2, -N (CH3) 3 ⁺ , -N (C2H ₅ ) 3 ⁺ , -N (C3H7) 3 ⁺ , NH (CH3) ₂ ⁺ , -NH (C2H5) 2 ⁺ , -NH (C3H ₇ ) 2 ⁺ , -NHCH3, -NHC2H5, -nhc3h7, -nh2ch3 ⁺ , -NH2C2H ₅ ⁺ , -NH2C3H7 ⁺ , -SO3H, -SO ₃ -, -SO2NH2, -CO-COOH, -O- CO-NH2, C (NH) -NH2, -NH-C (NH) -NH2, -NH-CS-NH2, -NH-COOH, or
or
L represents a linear or branched and saturated or unsaturated C1 to Ce carbon chain with at least one substituent selected from the group comprising or consisting of
-nh ₂ , -oh, -po ₃ h ₂ , -po ₃ h ', -po ₃ ² ', -opo3h2, -opo3h, -opo3 ^2_ , -COOH, -COO ', -CO-NH2, -NH3 ⁺ , -NH-CO-NH2, -N (CH3) 3 ⁺ , -N (C2H ₅ ) ₃ ⁺ , -N (C3H7) 3 ⁺ , -NH (CH3) ₂ ⁺ , -NH (C2H5) 2 ⁺ , - NH (C3H ₇ ) ₂ ⁺ , -nhch ₃ , -nhc ₂ h ₅ , -nhc ₃ h ₇ , nh ₂ ch ₃ ⁺ ,
-NH ₂ C ₂ H ₅ ⁺ , -NH2C3H7 ⁺ , -SO3H, -SO ₃ ', -SO2NH2, -CO-COOH, -O-CO-NH2, -C (NH) -NH ₂ , -NH-C ( NHJ-NH ₂ , -NH-CS-NH2, -NH-COOH, or
11/23
or
L represents a benzene ring and preferably a pair of benzene ring substituted with at least one substituent selected from the group comprising or consisting of
-NH ₂ , -OH, -PO3H2, -PO ₃ H ', -PO ₃ ² ', -OPO3H2, -OPO3H, -OPO3 ² ', -COOH, -COO', -CO-NH2, -NH ₃ ⁺ , -NH-CO-NH2i -N (CH3) 3 ⁺ , -N (C2H ₅ ) ₃ ⁺ , -N (C3H7) 3 ⁺ , -NH (CH3) ₂ ⁺ , -NH (C2H5) 2 ⁺ , -NH ( C3H ₇ ) ₂ ⁺ , -nhch ₃ , -nhc ₂ h ₅ , -nhc ₃ h ₇ , -nh ₂ ch ₃ ⁺ ,
-NH ₂ C ₂ H ₅ ⁺ , -NH2C3H7 ⁺ , -SO3H, -SO ₃ ', -SO ₂ NH _2i -CO-COOH, -O-CO-NH ₂ , -C (NH) -NH ₂ , -NH -C (NH) -NH ₂ , -NH-CS-NH ₂ , -NH-COOH, or
However, these compounds are not preferred and can be excluded from the present application, where X represents -O- or -S- and L represents -NH ₂ , -OH, -OPO ₃ H ₂ , -OPO ₃ H ', -OPO ₃ ² ', -NH ₃ ⁺ , -NH-CO-NH2, -N (CH3) 3 ⁺ , -N (C2H ₅ ) ₃ ⁺ , -N (C3H7) 3 ⁺ , -NH (CH3) ₂ ⁺ , -NH (C2H5) 2 ⁺ , -NH (C3H ₇ ) ₂ ⁺ , -NHCH ₃ , -NHC ₂ H ₅ , -nhc ₃ h ₇ , -nh ₂ ch ₃ ⁺ , -nh ₂ c ₂ h ₅ ⁺ , -nh2c3h7 ⁺ , -so3h, -so ₃ ", -so ₂ nh ₂ , -cocooh, -o-co-nh ₂ , -NH-C (NH) -NH ₂ , -NH-CS-NH ₂ , -NH -COOH,
Also excluded are compounds where X represents -ΝΗ- or -NR- and L represents -OPO ₃ H ₂₎ -ΟΡΟ ₃ Η ^_ , -OPO3 ² ', -NH-CO-NH2, -CO-COOH, -O -CONH2, -NH-C (NH) -NH ₂ , -NH-CS-NH ₂ or -NH-COOH.
Residue L can be further substituted by substituents as defined as R ¹ to R ¹³ . Residue L preferably consists of 1 to 10 carbon atoms, more preferably 1 to 6 carbon atoms and more preferably 2 to 4 carbon atoms. Carbon atoms of any substituents such as -COOH present in residue L are included in the above-mentioned number of carbon atoms. Thus, residue L contains a straight or branched carbon atom chain, or a phenyl ring that can be substituted with one or more saturated or unsaturated and linear or branched alkyl substituents and / or substituents defined as R ¹ to R ¹³ .
11/24
It is preferred that the L carbon chain is in the range of Ci to C ₇ , more preferably in the range of Ci to C ₆ and most preferably in the range of Ci to C5.
The compounds of general formula (I) or (II), which can be used to solubilize fatty acids in aqueous medium or in water are represented by the following formula (I) or (II):
NR NR
Λ Λ Λ
RR-rr x R'Htr χ formula (I) formula (II) where
RR, R 'and R independently represent each other -H, -OH, CH = CH ₂ , -CH ₂ -CH = CH ₂ , -C (CH ₃ ) = CH _2i -CH = CH-CH _3i -C ₂ H ₄ -CH = CH ₂ , -CH _3i C ₂ H ₅ , -C ₃ H _7i -CH (CH ₃ ) ₂ , -C4H9, -CH ₂ -CH (CH ₃ ) ₂ , -CH (CH ₃ JC ₂ H ₅ , -C (CH ₃ ) ₃ , C5H11, -CH (CH ₃ ) -C ₃ H ₇₁ -CH ₂ -CH (CH ₃ ) -C ₂ H ₅ , -CH (CH ₃ ) -CH (CH ₃ ) ₂ , -C (CH ₃ ) ₂ C ₂ Hs, -CH ₂ -C (CH ₃ ) _3i -CH (C ₂ H ₅ ) _2i -C ₂ H ₄ -CH (CH ₃ ) _2i -CeH-is, -C7H15, cycle-C ₃ H5, cycle-C ₄ H ₇ , CYCL0-C5H9, cycle-CeHn, —PO ₃ H ₂ , —PO ₃ H, —PO ₃ ² , —NO ₂ , -C = CH, CsC -CH ₃ , -CH ₂ -CeCH, -C ₂ H ₄ -C = CH, -CH ₂ -CsC-CH ₃ , or R 'and R together form the residue -CH ₂ -CH ₂ -, -CH = CH- or -CH ₂ CH ₂ -CH ₂ X represents -NH-, -NR-, -O-, -S- or -CH ₂ - or a substituted carbon atom; and
L represents -CR ¹ R ² R ³ , -CR ⁴ R ⁵ -CR ¹ R ² R ³ , -CR ⁶ R ⁷ -CR ⁴ R ⁵ -CR ¹ R ² R ³ , -CR ⁸ R ⁹ CR ⁶ R ⁷ -CR ⁴ R ⁵ -CR ¹ R ² R ³ , -CR ^1o R ¹¹ -CR ⁸ R ⁹ -CR ⁶ R ⁷ -CR ⁴ R ⁵ -CR ¹ R ² R ³ , -CR ¹² R ¹³ CR ¹⁰ R ^{1 1} _cr ⁸ R ⁹ -CR ⁶ R ⁷ -CR ⁴ R ⁵ -CR ¹ R ² R ³ ;
R *. R ^# , R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² . R ¹³ independently represent the following substituents: -nh2, -oh, -po3h2) -po3h; -po3 ² ·, -opo3h ₂ , -ορο ₃ η; -pot ₃ ² ; -cooh, -COO ', —CO — NH2, —NH3 ⁺ , -NH — CO — NH2i —N (CH ₃ ) ₃ ⁺ , —N (C2H5) 3 ⁺ , —N (C3H ₇ ) ₃ ⁺ , -NH (CH ₃ ) ₂ ⁺ , -NH (C2H5) 2 ⁺ , -NH (C3H ₇ ) ₂ ⁺ , -nhch ₃ , -nhc ₂ h ₅ , -NHC ₃ H _7i -nh ₂ ch ₃ ⁺ , -nh ₂ c ₂ h ₅ ⁺ , -nh2c3h7 ⁺ , -so3h, -SO ₃ -, -SO ₂ NH _2i -co-cooh, -oCO-NH _2i -C (NH) -NH ₂ , -NH-C (NH) -NH ₂ , -nh-cs-nh ₂ , -nh-cooh, -h,
11/25
OCH _3i -OC ₂ H ₅₎ -OC ₃ H ₇ , -O-cyclo-C ₃ H ₅ , -OCH (CH ₃ ) ₂ , -P (O) (OCH ₃ ) ₂ , Si (CH ₃ ) ₂ ( C (CH ₃ ) ₃ ), -OC (CH ₃ ) ₃ , -OC4H9, -OPh, -OCH ₂ -Ph, -OCPh ₃ , -SH, -SCH ₃ , SC ₂ H ₅ , -SC ₃ H ₇ , -S-cycle-C ₃ H ₅ , -SCH (CH ₃ ) ₂ , -SC (CH ₃ ) ₃ , -NO ₂ , -F, -Cl, -Br, -I, P (O) (OC ₂ H ₅ ) ₂ , -P (O) (OCH (CH ₃ ) ₂ ) ₂ , -C (OH) [P (O) (OH) ₂ ] ₂ , -Si (C ₂ H ₅ ) ₃ , -Si (CH ₃ ) ₃ , n ₃ , -cn, -ocn, -nco, -scn, -ncs, -cho, -coch ₃ , -coc ₂ h ₅ , -coc ₃ h ₇ , CO-cycle-C ₃ H ₅ , -COCH (CH ₃ ) ₂ , -COC (CH ₃ ) ₃ , -COCN, -COOCH _3i -COOC ₂ H ₅ , COOC ₃ H ₇ , -COO-cyclo-C ₃ H ₅ , -COOCH (CH ₃ ) ₂ , -COOC (CH ₃ ) ₃ , -OOC-CH ₃ , OOC-C ₂ H ₅ , -OOC-C ₃ H ₇ , -OOC-cyclo-C ₃ H ₅ , -OOC-CH (CH ₃ ) ₂ , -OOC-C (CH ₃ ) ₃ , CONHCH ₃ , -CONHC ₂ H _5i -CONHC ₃ H ₇ , -CONH-cycle-C ₃ H ₅ , -CONH [CH (CH ₃ ) ₂ ], CONH [C (CH ₃ ) ₃ ], -CON (CH ₃ ) ₂ , -CON (C ₂ H ₅ ) ₂ , -CON (C ₃ H ₇ ) _2i -CON (C ₃ H ₅ cycle) ₂ , CON [CH (CH ₃ ) ₂ ] ₂ , -CON [C (CH ₃ ) ₃ ] _2i -nhcoch ₃ , -nhcoc ₂ h ₅ , -nhcoc ₃ h ₇ , NHCO-cyclo-C ₃ H ₅ , -NHCO-CH (CH ₃ ) ₂ , -NHCO-C (CH ₃ ) _3i -NHCO-OCH _3j -NHCOOC ₂ H ₅ , -NHCO-OC ₃ H ₇ , -NHCO-O-cyclo-C ₃ H ₅ , -NHCO-OCH (CH ₃ ) ₂ , -NHCOOC (CH ₃ ) ₃ , -NH-cyclo-C ₃ H ₅ , -NHCH (CH ₃ ) ₂ , -NHC (CH ₃ ) ₃ , -N (CH ₃ ) ₂ , -N (C ₂ H ₅ ) ₂ , N (C ₃ H ₇ ) ₂ , - N (C-cycle ₃ H5) ₂ , - N [CH (CH ₃ ) ₂ ] ₂ , —N [C (CH ₃ ) ₃ ] _2i - SOCH _3i —SOC ₂ Hs, - SOC ₃ H ₇ , -SO-cycle-C ₃ H5, -SOCH (CH ₃ ) ₂ , -SOC (CH ₃ ) ₃ , -SO ₂ CH ₃ , -SO ₂ C ₂ Hs , SO ₂ C ₃ H ₇ , - SO ₂ -cyclo-C ₃ H5, —SO ₂ CH (CH ₃ ) ₂ , —SO ₂ C (CH ₃ ) _3j —SO ₃ CH _3i —SO ₃ C ₂ H5, - SO ₃ C ₃ H ₇ , -SO ₃ -cyclo-C ₃ H ₅ , -SO ₃ CH (CH ₃ ) ₂ , -SO ₃ C (CH ₃ ) ₃ , -SO ₂ NH ₂ , -OCF _3i OC2F5, - O-COOCH3, -O-COOC ₂ H ₅ , -O-COOC3H7, -O-COO-cyclo-C ₃ H ₅ , -OCOOCH (CH ₃ ) ₂ , -O-COOC (CH ₃ ) ₃ , -nh- co-nhch ₃ , -nh-co-nhc ₂ h ₅ , -nhCS-N (C ₃ H ₇ ) _2i -NH-CO-NHC3H7, -NH-CO-N (C ₃ H ₇ ) _2i -NH-CO -NH [CH (CH ₃ ) ₂ ], NH-CO-NH [C (CH ₃ ) ₃ ], -NH-CO-N (CH ₃ ) _2i -NH-CO-N (C ₂ H ₅ ) _2i - NH-CO-NH-cycloC ₃ H ₅ , -NH-CO-N (cyclo-C ₃ H ₅ ) ₂ , -NH-CO-N [CH (CH ₃ ) ₂ ] ₂ , -NH-CS-N (C ₂ H ₅ ) _2i -NHCO-N [C (CH ₃ ) ₃ ] _2i -NH-CS-NH2, -NH-CS-NHCH3, -NH-CS-N ( CH ₃ ) ₂ , -NH-CSNHC ₂ H _5i -NH-CS-NHC3H7, -NH-CS-NH-CÍCI0-C3H5, -NH-CS-NH [CH (CH ₃ ) ₂ ], NH-CS-NH [C (CH ₃ ) 3], -NH-CS-N (cyclo-C ₃ H ₅ ) ₂ , -NH-CS-N [CH (CH ₃ ) ₂ ] 2, -NH-CSN [C (CH ₃ ) ₃ ] ₂ , -NH-C (= NH) -NH ₂ , -NH-C (= NH) -NHCH ₃ , -NH-C (= NH) -NHC ₂ H ₅ , NH-C (= NH) -NHC3H7, -O-CO-NH-CICIO-C3H5, -NH-C (= NH) -NH-cyclo-C3H5, NH-C (= NH) -NH [CH (CH ₃ ) ₂ ], -0- CO-NH [CH (CH ₃ ) ₂ ], -NH-C (= NH) -NH [C (CH ₃ ) ₃ ], NH-C (= NH) -N (CH ₃ ) ₂ , -NH-C (= NH) -N (C ₂ H ₅ ) ₂ , -NH-C (= NH) -N (C3H ₇ ) ₂ , -nhC (= NH) -N (C3H- ₅ cycle) ₂ , -O-CO -NHC3H7, -NH-C (= NH) -N [CH (CH3) ₂ ] 2, -NHC (= NH) -N [C (CH3) 3] 2, -0-C0-NHCH3, -O-CO -NHC ₂ H ₅ , -O-CO-NH [C (CH ₃ ) ₃ ], O-CO-N (CH ₃ ) ₂ , -O-CO-N (C ₂ H ₅ ) ₂ , -O-CO -N (C ₃ H ₇ ) _2i -O-CO-N (cycle-C ₃ H ₅ ) ₂ , O — CO — N [CH (CH3) ₂ ] ₂ , - O — CO — N [C (CH ₃ ) ₃ ] ₂ , —O — CO — OCH3, - O — CO — OC ₂ H5, —O— CO-OC3H7, -O-CO-O-CICIO-C3H5, -O-CO-OCH (CH ₃ ) ₂ , -O-CO-OC (CH ₃ ) 3, CH ₂ F, -CHF ₂ , -CF3, -C H ₂ CI, -CH ₂ Br, -CH ₂ I, -CH ₂ -CH ₂ F, -CH ₂ -CHF ₂ , -CH ₂ -CF ₃ ,
11/26
-CH ₂ -CH ₂ CI, -CH ₂ -CH ₂ Br, -CH ₂ -CH ₂ I, cyclo-C ₃ H ₅ , cyclo-C ₄ H ₇ , cyclo-CsHg, cycloCeHn, CyCI0-C7H13, cyclo- CsHiõ, -Ph, -CH ₂ -Ph, -CPh ₃ , -CH3, -C ₂ H5, -C3H7, CH (CH ₃ ) _2j -C4H9, -CH ₂ -CH (CH ₃ ) ₂ , -CH (CH ₃ ) -C ₂ H5, -C (CH ₃ ) _3> -C ₅ Hh, -CH (CH ₃ ) C ₃ H ₇₎ -CH ₂ -CH (CH ₃ ) -C ₂ H _5i -CH (CH ₃ ) -CH (CH3) 2-C (CH3) 2-C ₂ H5-CH ₂ -C (CH3) 3, -CH (C ₂ H ₅ ) ₂ , -C ₂ H ₄ -CH (CH ₃ ) ₂ , -CeHis, -C7H15, -CeHn, -C ₃ H6-CH (CH3) ₂ , - Ο ₂ Η ₄ - CH (CH ₃ ) -C ₂ H _5i -CH (CH ₃ ) -C4H _9i -CH ₂ -CH (CH ₃ ) -C3H7, -CH (CH ₃ ) -CH ₂ CH (CH ₃ ) _2i -CH (CH ₃ ) -CH (CH3) -C ₂ H5, -CH ₂ -CH (CH ₃ ) -CH ( CH ₃ ) _{2 (} -CH ₂ -C (CH ₃ ) 2C2H5, -C (CH ₃ ) ₂ -C ₃ H ₇ , -C (CH ₃ ) 2-CH (CH ₃ ) 2, -C ₂ H ₄ - C (CH ₃ ) _3i -CH (CH ₃ ) -C (CH3) ₃₎ -CH = CH ₂ , -CH ₂ -CH = CH ₂ , -C (CH ₃ ) = CH ₂ , -ch = ch-ch _3i -c ₂ h ₄ -ch = ch _2> -ch ₂ CH = CH-CH _3t -CH = CH-C ₂ H ₅ , -CH ₂ -C (CH ₃ ) = CH ₂₁ -CH (CH ₃ ) - CH = CH, CH = C (CH ₃ ) _2i -C (CH ₃ ) = CH-CH3, -CH = CH-CH = CH ₂ , -C ₃ H6-CH = CH ₂₎ -c ₂ h ₄ CH = CH-CH ₃₁ -CH ₂ -CH = CH-C ₂ H ₅ , -CH = CH-C ₃ H ₇ , -ch ₂ -ch = ch-ch = ch _2i CH = CH-CH = CH-CH ₃ , -CH = CH-CH ₂ -CH = CH _2> -C (CH ₃ ) = CH-CH = CH ₂ , CH = C (CH ₃ ) -CH = CH _2i -CH = CH-C (CH ₃ ) = CH ₂ , -C ₂ H ₄ -C (CH ₃ ) = CH ₂ , -ch ₂ CH (CH ₃ ) -CH = CH ₂ , -CH (CH ₃ ) -CH ₂ -CH = CH _2> -CH ₂ -CH = C (CH ₃ ) ₂ , -CH ₂ C (CH ₃ ) = CH-CH3, -CH (CH ₃ ) - CH = CH-CH _3i -CH = CH-CH (CH ₃ ) ₂ , -CH = C (CH ₃ ) C ₂ H ₅₎ -C (CH ₃ ) = CH-C ₂ H _5i -C (CH ₃ ) = C (CH ₃ ) _2i -C (CH ₃ ) 2-CH = CH _2i -CH (CH ₃ ) C (CH ₃ ) = CH _2i -C (CH ₃ ) = CH-CH = CH _2i -CH = C (CH ₃ ) -CH = CH ₂ , -ch = chC (CH ₃ ) = CH _2i -C ₄ H ₈ -CH = CH ₂ , -C ₃ H6-CH = CH-CH ₃ , -c ₂ h ₄ - ch = ch-c ₂ h ₅ , -ch ₂ CH = CH-C ₃ H _7> -CH = CH-C ₄ H ₉ , -C ₃ H6-C (CH ₃ ) = CH ₂ , -C ₂ H ₄ -CH (CH ₃ ) -CH = CH _2i CH ₂ -CH (CH ₃ ) -CH ₂ -CH = CH _2i -C ₂ H ₄ -CH = C (CH ₃ ) ₂ , -CH (CH ₃ ) -C ₂ H4-CH = CH _2i C ₂ H ₄ -C (CH ₃ ) = CH-CH ₃ , -CH ₂ -CH (CH ₃ ) -CH = CH-CH3, -CH (CH ₃ ) -CH2-CH = CHCH _3i -CH ₂ -CH = CH-CH (CH ₃ ) _2i -CH ₂ -CH = C (CH ₃ ) -C ₂ H5, -CH ₂ -C (CH ₃ ) = CHC ₂ h _{5 (} -CH ( CH ₃ ) -CH = CH-C ₂ H5, -ΟΗ = ΟΗ-ΟΗ2-ΟΗ (ΟΗ3) 2, -CH = CH-CH (CH ₃ ) C ₂ h _{5 (} -CH = C (CH ₃ ) -C ₃ H7, -C (CH ₃ ) = CH-C ₃ H ₇ , - CH2-CH (CH ₃ ) -C (CH ₃ ) = CH ₂ , C [C (CH ₃ ) ₃ ] = CH ₂ , -CH (CH ₃ ) -CH ₂ -C (CH ₃ ) = CH2, -CH (CH ₃ ) -CH (CH ₃ ) -CH = CH ₂₁ CH = CH-C ₂ H ₄ -CH = CH ₂ , -CH ₂ -C (CH ₃ ) ₂ -CH = CH ₂ , -C (CH ₃ ) ₂ -CH ₂ -CH = CH _2> ΟΗ2-Ο (ΟΗ ₃ ) = Ο (ΟΗ ₃ ) _2ι -CH (CH ₃ ) -CH = C (CH ₃ ) ₂ , -C (CH ₃ ) ₂ -CH = CH-CH _3> CH = CH-CH2-CH = CH-CH _3i -CH (CH ₃ ) -C (CH ₃ ) = CH-CH ₃ , -CH = C (CH ₃ ) CH (CH ₃ ) ₂ , -C (CH ₃ ) = CH-CH (CH ₃ ) _2i -C (CH ₃ ) = C (CH ₃ ) -C ₂ H _5> -CH = CH-C (CH ₃ ) _3i C (CH ₃ ) ₂ -C (CH ₃ ) = CH ₂ , -CH (C ₂ H ₅ ) -C (CH ₃ ) = CH ₂ , -C (CH ₃ ) (C ₂ H ₅ ) -CH = CH ₂ CH (CH ₃ ) -C (C ₂ H5) = CH ₂ , -CH2-C (C ₃ H ₇ ) = CH ₂ , -CH2-C (C ₂ H ₅ ) = CH-CH ₃ CH (C ₂ H ₅ ) -CH = CH-CH ₃ , -C (C ₄ H ₉ ) = CH ₂ , -C (C ₃ H ₇ ) = CH-CH _3i -C (C ₂ H ₅ ) = CH-C ₂ H ₅ , -C ( C ₂ H ₅ ) = C (CH ₃ ) ₂ , -C [CH (CH ₃ ) (C ₂ H ₅ )] = CH _2> -C [CH ₂ -CH (CH ₃ ) ₂ ] = CH ₂ , - c ₂ h ₄ ch = ch-ch = ch ₂ , -CH ₂ -CH = CH-CH ₂ -CH = CH ₂ , -C ₃ H ₆ -CEC-CH ₃ , -ch ₂ CH = CH-CH = CH -CH ₃ , -CH = CH-CH = CH-C ₂ H ₅ , -CH2-CH = CH-C (CH ₃ ) = CH _2j 27/118
CH2-CH = C (CH ₃ ) -CH = CH ₂₎ -CH ₂ -C (CH ₃ ) = CH-CH = CH _2i -CHÍCHsJ-CHz-CsCH, -CH (CH ₃ ) -CH = CH-CH = CH ₂ , -CH = CH-CH ₂ -C (CH ₃ ) = CH ₂ , -CH (CH ₃ ) -C = C-CH _3i -CH = CH-CH (CH ₃ ) -CH = CH ₂ , -CH = C (CH ₃ ) -CH ₂ -CH = CH ₂ , -C ₂ H ₄ -CH (CH ₃ ) C = CH, -C (CH ₃ ) = CH-CH ₂ -CH = CH ₂ , - CH = CH-CH = C (CH ₃ ) ₂ , -CH ₂ -CH (CH ₃ ) CH ₂ -CeCH, -CH = CH-C (CH ₃ ) = CH-CH _3i -CH = C (CH ₃ ) -CH = CH-CH ₃ , -ch ₂ CH (CH ₃ ) -C = CH, -C (CH ₃ ) = CH-CH = CH-CH ₃₎ -CH = C (CH ₃ ) -C (CH ₃ ) = CH _2i C (CH ₃ ) = CH-C (CH ₃ ) = CH ₂ , -C (CH ₃ ) = C (CH ₃ ) -CH = CH ₂ , -ch = ch-ch = chCH = CH _2i -CECH, -CeC-CH ₃ , -CH ₂ -CeCH, -C ₂ H ₄ -CeCH, -CH ₂ -CeC-CH ₃ , CeC-C ₂ H ₅ , -C ₃ He-CECH, -C ₂ H ₄ -CEC-CH ₃ , -CH ₂ -CeC-C ₂ H _5i -CeC-C ₃ H ₇ , CH (CH ₃ ) —C = CH, - C ₄ Hg — C = CH, —C ₂ H ₄ - C = C — C ₂ Hs, —CH ₂ —C = C — C ₃ H _7i —C = C— C ₄ H ₉ , -CEC-C (CH ₃ ) ₃ , -CH (CH ₃ ) -C ₂ H4 -CECH, -CH ₂ -CH (CH ₃ ) -CeC-CH ₃₁ CH (CH ₃ ) —CH ^ C = C — CH _3j —CH (CH ₃ ) —C = C — C ₂ Hs, - CH ^ C = C — CH (CH ₃ ) ₂ , —C = C— CH (CH ₃ ) -C ₂ H5, -CeC-CH ₂ -CH (CH ₃ ) _2> -CH (C ₂ H ₅ ) -CEC- CH ₃ , -C (CH ₃ ) ₂ -CeCCH ₃ , -CH (C ₂ H5) -CH ₂ -C = CH, -CH ₂ -CH (C ₂ H5) -C = CH, -C (CH ₃ ) ₂ -CH ₂ -CeCH, CH2-C (CH ₃ ) ₂ -CECH, -CH (CH ₃ ) -CH (CH ₃ ) -CeCH _i -CH (C ₃ H ₇ ) -CECH, C (CH ₃ ) ( C ₂ H ₅ ) -CECH, -CH ₂ -CH (CeCH) ₂ , -C = CC = CH, -CH ₂ -CeC-CeCH, CeC-CeC-CH ₃ , -CH (CeCH) ₂ , -C ₂ H ₄ -CeC-CeCH, -CH ₂ -CeC-CH ₂ -CeCH, C = C — C ₂ H ₄ —C = CH, —CH ₂ —C = C — C = C — CH _3i —C = C— CH ^ C = C — CH _3i - C = CC = C— C ₂ H ₅₁ -C (CECH) ₂ -CH ₃ , -CeC-CH (CH ₃ ) -CeCH, -CHiCHabCEC-CECH, CH (CECH) - CH ₂ -CECH, -CH (CECH) -CEC-CH _3> -CH = CH-Ph, -NH-CO-CH2COOH, -NH-CO-C ₂ H ₄ -COOH, -NH-CO-CH2-NH ₂ , -nh-co-c ₂ h ₄ -nh ₂ , -nhCHiCOOHhCHr-COOH, -NH-CH ^ COOH, -NH-C ₂ H ₄ -COOH, -NH-CH (COOH) C ₂ H ₄ -COOH , -NH-CH (CH ₃ ) -COOH;
wherein preferably at least one of the substituents R *, R ^# , R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ . R ¹² . R ¹³ is selected from the following substituents:
-NH ₂ , -OH, -PO ₃ H _2i -po ₃ h ', -po ₃ ² ·, -OPO3H2 (-OPO3H, -OPO3 ² ', -COOH, -COO, -CO-NH2i -NH ₃ ⁺ , -NH-CO-NH2, -N (CH3) 3 ⁺ , -N (C2H ₅ ) ₃ ⁺ , -N (C3H7) 3 ⁺ , -NH (CH3) ₂ ⁺ , -NH (C2H5) 2 ⁺ , -NH (C3H ₇ ) ₂ ⁺ , -nhch3 (-NHC2H5i -NHC3H7i nh2ch3 ⁺ , -nh2c ₂ h ₅ ⁺ , -nh2c3h7 ⁺ , -so3h, -so ₃ ', -so ₂ nh ₂ , -co-cooh, -oC0- NH2, -C (NH) -NH _2> -NH-C (NH) -NH ₂ , -nh-cs-nh ₂₎ -nh-cooh
Also preferred are the compounds of the general formula (III) as shown below:
11/28
NH where residues X and L have the meanings as disclosed herein.
Preferably, the compounds of general formula (I), (II) and (III) have a partition coefficient between n-octanol and water (also known as K _O w or octanol-water partition coefficient) of K _O w < 6.30 (log Kow <0.80), preferably Kow <1.80 (log Kow <0.26), more preferably Kow <0.63 (log Kow <-0.20) and more preferably Kow <0.40 (log Kq _W <-0.40).
In addition, the compounds of general formula (I), (II) and (III) have the same number of preferred carbon atoms as described above, the same preferred pH range for the solubilization reaction, the same preferred acid molar ratio solubilizing compound carboxylic and the same preferred reaction conditions, as described above for solubilizing compounds in general.
The partition coefficient is the ratio of the concentrations of the non-ionized compound between the two solutions. To measure the partition coefficient of ionizable solutes, the pH of the aqueous phase is adjusted in such a way that the predominant form of the compound is non-ionized. The logarithm of the ratio between the concentrations of non-ionized solute in the solvents is called log P:
, P, j ^ i)
The distribution coefficient is the ratio between the sum of the concentrations of all forms of compound (ionized plus non-ionized), in each of the two phases. For distribution coefficient measurements, the pH of the aqueous phase is buffered to a specific value such that the pH is not significantly disturbed by the introduction of the compound. The logarithm of the ratio between the sum of the concentrations of the various forms of the solute in one solvent, the sum of the concentrations of its forms in the other solvent is called log D:
j _n _ j ([sohráe] o _etonl Q _f A ^{9 9} k
In addition, log D is pH dependent, so the pH must be specified at which log D was measured. Of particular interest is the log D at pH = 7.4
29/118 (the physiological pH of blood serum). For non-ionizable compounds, log P = log D at any pH.
Arginine
Arginine (2-amino-5-guanidinopentanoic acid) is an α-amino acid. The arginine amino acid side chain consists of a linear 3-carbon aliphatic chain, the distal end of which is covered by a complex guanidine group. According to the invention, L-arginine, D-arginine, as well as racemates thereof, can be used.

arginine L-arginine
With a pKa of 12.48, the guanidine group is positively charged in neutral, acidic and even more basic environments, and thus transmits basic chemical properties to arginine. Because of the conjugation between the double bond and the solitary nitrogen pairs, the positive charge is relocated, allowing the formation of multiple ligações bonds. Arginine can be protonated by carrying three additional charges, located in the side chain (pKa 12.48), in the amino group (pKa 8.99) and in the carboxyl group (pKa 1.82)
The L form is one of the 20 most common natural amino acids. In mammals, arginine is classified as a semi-essential or conditionally essential amino acid, depending on the individual's stage of development and health status. Babies are unable to meet their needs and thus arginine is nutritionally essential for babies.
Arginine is an amphiphilic molecule with a reactive carboxyl group.
The literature value for the log P _ow (see above) of arginine is - 4.20. For arginine, the distribution coefficient is approximately equal to the partition coefficient, because pH = 7 arginine is almost exclusively present in ionic form. Libby et al. (Mol. Pharmacol 1981, 20, 602-608) determined log D _ow = 4.08.
When investigating the solubilization capacity of fatty acids by aqueous amino acid systems, it was found that arginine completely dissolves oleic acid through the formation of micro- and nanoemulsions when greater than a molar ratio of 1: 1 (arginine: fatty acid) . Interestingly, no cosolvent is needed to solubilize carboxylic acids. Formation
Spontaneous 30/118 of a nanoemulsion was observed at room temperature. The pH value after self-assembly of a 1: 1 microemulsion is about 9.8. In nanoemulsions the particle size was about 2 nm in diameter and no aggregates larger than 25 nm were found. This self-assembly of nanoparticles is a central feature of a nanoemulsion. The nanoemulsion is completely transparent and stable for more than 6 months at temperatures between -20 and 100 ° C. Decreasing the pH by adding acid (HCI) reduces the solvation capacity, which can be overcome by adding arginine. However, the pH of the solution is critical to the arginine capacity of nanoemulsification which decreases below pH 8.
It was found that arginine exhibits its ability to solubilize in organic solutions as well. Oleic acid and albumin in various concentrations in an aqueous solution were investigated by adding arginine. The excess of fatty acids results in the dilution of the solution. By adding arginine, this effect can be completely reversed. Balance dialysis was performed with and without arginine. It could be demonstrated that the transfer of fatty acids through a 5000D cellulose membrane was up to 10 times greater when arginine was in the model solution. The same investigations performed with human plasma have shown comparable results (Example 1). The ability of arginine to release fatty acids linked to albumin was investigated by oleic acid labeled ³ H. Without arginine, about 40% of the radiolabelled fatty acid resides in the organic phase after extraction with n-hexane. The addition of fatty acids released from arginine. However, a higher molar concentration of arginine than that of fatty acid was necessary to achieve the maximum effect. The reduction of residual fatty acids up to 2% could be achieved. The efficiency was increased when the temperature was raised to 38 ° C, compared to the ambient temperature. Other hydrophilic amino acids (lysine, asparagine, asparagine acid, glutamine, glutamine, histidine) as well as the hydrophobic amino acids investigated by an identical process also provided a reduction in fatty acid residues. However, this was significantly less compared to arginine.
Arginine derivatives as solubilizing compounds
The term arginine derivatives refers to compounds having a carboxy group (-COOH) and an amidino group (H ₂ NC (NH) -), or substituted amidino group separated by at least one carbon atom or with a carboxy group ( COOH) and a guanidino group (H ₂ NC (NH) -NH-) or a guanidino group
31/118 substituted separated by at least one carbon atom. The above-mentioned compounds of general formula (I) are also derived from arginine.
Examples of arginine derivatives are, for example, amidinoacetic acid, amidinopropionic acid, amidinobutyric acid, guanidinopropionic acid, guanidinobutyric acid, oligoarginine, polyarginine, as well as
NH
NH CH ₃
H ₂ NN COOH
H
H ₂ NN COOH
H h ₂ n
NH ₂
N COOH
Guanidinoacetic acid
NH creatine
Glycocyanine
H ₃ CN
H
NH ₂
COOH
H ₅ C ₂
NH
N
NH ₂
COOH
NH
HN
COOH
NH ₂
H
N
NH ₂
COOH
NH
HN
COOH
NH ₂
NH ₂
COOH
NH
h ₃ c ^ ³ Ν N
nh ₂
COOH
NH
Ν N
NH ₂
COOH
NH
Ν
NH ₂
COOH
NH
Ν N
NH ₂
COOH
Η H
NH
H ₂ NN
H
H ₃ C
H
COOH
COOH
11/28
Interaction of carboxylic acids and the solubilizing compound
According to the invention, a preferred method for removing carboxylic acids from aqueous or organic solutions comprises the following key steps:
a) Add the solubilizing compound to a solution containing fatty acid;
b) Pass the solution along a surface that carries immobilized lipases or catalysts capable of releasing fatty acids from their esterified forms and which is implemented in a capillary micro- or nanofluidic system to obtain complete interaction with the esterified fatty acids;
c) Pass the solution along an interphase consisting of a separation membrane, a gel, or a hollow capillary assembly
11/33
d) Apply a degree across the phase separation interface, through a degree of concentration, an osmotic degree, a physical-chemical degree, a pH degree, a pneumatic degree, a temperature degree, an electrical degree or a combination of these;
e) Separate from the fatty acid fraction associated with the solubilizing compound;
f) Dissolve fatty acids in acceptor medium; and
g) optionally, remove the solubilizing compound from the solution.
Thus, the ability of a solubilizing compound to build micro- and nanoemulsions from carboxylic acids in aqueous or organic media is critical to the present invention. This fundamental principle can be used for a variety of medical, pharmaceutical, biochemical, industrial and environmental applications, which will be described below.
Formulation of solubilizing compounds
The solubilizing compound can be used as a pure solution, pH adjusted solution or complexed solution. It can be covalently or electrostatically linked to a peptide or protein, as well as a negatively charged organic or inorganic polymer or surface.
The solubilization effect can be improved by reducing the ionic strength of the solution, emulsion or oil to be treated, or analyzed, for example, through chelation, dialysis or electrodialysis, in order to reduce the concentration of the calcium. In addition, it may be useful to alter the interaction binding energy of an electrostatic carboxylic acid - carboxylic acid protein to be purified by altering the surface energy of proteins or by thiolisation of sulfide bonds, thereby altering the steric conformation of the protein. As stated in the examples, it can be demonstrated by a solubilizing compound such as arginine that there is a spontaneous and stoichiometric formation of an arginine adduct and ionized hydrophobic substances, in particular fatty acids, under suitable circumstances. Results in mini, micro or nanoemulsions.
According to the invention, these mini, micro or nanoemulsified carboxylic acids can be separated, resp. extracted from aqueous or organic medium by
1. an adsorption of the substance to be separated on surfaces such as aerolites, spheres, microspheres or ceolites, particularly if they exhibit surface characteristics that allow them to electrostatically or covalently bond with the substance to be separated;
11/34
2. complexation, that is, formation of salts
3. a diffusion of the substance to be separated in an accepting medium (in particular organogels);
4. a dialysis of the substance to be separated by means of a thermal, electrical or physical-chemical degree;
5. filtering of the substance to be separated by means of an electrical or physical-chemical thermal degree;
6. distillation techniques,
7. extraction of supercritical liquid,
8. nanofluidic separation techniques.
Hemodialysis
Hemodialysis is a procedure to purify the blood and is a method to treat patients suffering from kidney dysfunction. The dialysis principle is the diffusion of solutes through a semipermeable membrane (osmosis), through which the blood / plasma with water-soluble toxins, electrolytes, urea and other substances is on one side of the membrane (permeate) and a solution dialysis consisting of water and a number of important electrolytes in physiological concentrations, on the other side (dialysate). Small molecules such as water, electrolytes, urea and urates diffuse through small holes in the membrane throughout the degree of concentration, but not proteins and blood cells. In hemodialysis, blood is pumped from the patient, flows along the membrane in the dialyzer and the clean (retinal) blood is pumped back to the patient. The counter-current flow of blood and dialysate maximizes the degree of solute concentration between blood and dialysate.
The dialysis solution is prepared by mixing a concentrated solution of electrolytes and a buffer system with sterile deionized water (dialysate). The dialysate is heated, and released from the gas. There are basically two different types of mixing systems. In the volumetric mixing system, two set fixed pumps mix the concentrate and the water so that the amount of flow and the concentration of the dialysis solution do not change. Conductivity is a measure for the concentration of electrolytes in the solution. After mixing the conductivity of the solution is measured and the pumps are adjusted by hand, if necessary to change the amount of water or concentrate.
Dialysis machines generally provide the following functionality:
- aspiration of the patient's blood by means of roller pumps
- anticoagulation
11/35
- transport processes through the dialyzer (s)
- control of the flow speed of the dialysate and the filtrate
- control of pressure degrees between the dialysate and filtrate circulation
- conductivity control
- adjustment of tonic strength and pH
- removal of trapped air and particles
- recirculating blood quenching
- blood recirculation to the patient, using electrolyte roller pumps, urea, creatinine, phosphate, amino acids, active pharmacological substances and water are able to pass semipermeable membranes as used in hemodialysis.
B. Definitions
Dialyzer means a device that contains a phase separation interface allowing the diffusion and permeation of carboxylic acids in a solution from one side of the separation interface to the other in another solution through a physical and / or chemical degree.
Extractor means a device that contains materials that allow the physical and / or chemical interaction of carboxylic acids with an interface thus adsorbing and / or absorbing and / or complexing and / or separating them.
Capillary voids are continuous, linear, tubular open spaces within a material.
Porous refers to the characteristics of a material that has pores, openings or recesses, which allows the passage of defined molecules from one compartment to another compartment along a gradient. These pores, openings or recesses can be uniform or different in size, shape and distribution throughout the material, preferably a membrane.
The term blood, as mentioned herein, refers to blood, whole blood, blood plasma and serum.
Note that the term blood can also refer to blood components and blood substitutes here.
The term blood components as used herein are cellular and acellular components, comprising red and white blood cells, thrombocytes, proteins and peptides, as well as lipid fractions.
The term blood substitutes as used here, refers to blood substitutes, which may, at least partially, carry oxygen and
36/118 volume expanders used to increase the volume of blood that supports blood circulation, but cannot perform the physiological function of blood.
The term individual refers to any mammal, including humans. Human beings are preferred.
The term first entry, second entry, third entry, fourth entry, etc., has to be understood as the number of entries of the device, and not as the number of entries of a part of the device, such as a camera of the device. Thus, fourth entry of the second chamber does not mean that the second chamber has four entries, but that the second chamber has entry No. 4 of the device.
The term microemulsion, as used herein, refers to the characteristics of an emulsion of the solubilizing compound of the invention and a carboxylic acid comprising at least two of the following:
spontaneous self-assembly, optical translucency, turbidity <1.1 cm-1,> 80% of micelles being <200nm at 25 ° C, optical translucency stability in a temperature range between -40 and 99 ° C, optical translucency stability for at least 12 months, the surface tension <60dyn / s.
The term nanoemulsion as used herein refers to the characteristics of an emulsion of the solubilizing compound of the invention and a carboxylic acid that comprises at least two of the following:
Spontaneous self-assembly, optical transparency, turbidity <0.4 cm-1,> 80% of the micelles being <100 nm, at 25 ° C, the stability of optical transparency in a temperature range between -40 and 99 ° C, stability of optical transparency for at least 12 months, surface tension <50dyn / s
C. Methods for the separation and reaction of free carboxylic acids from aqueous or organic media
Separation of free carboxylic acids can be accomplished by physical or chemical methods, as is known in the art. This includes, but is not restricted to, one or a combination of the following methods: adsorption, complexation, filtration, dialysis, evaporation, segregation by gravity, electrophoresis, electrolytically, electroosmotically, electrokinetically, osmotically, thermally, by a degree of concentration or by a chemical reaction.
It is preferred that the separation is carried out by adsorption, filtering, dialysis, electrolysis, as well as complexation and segregation by gravity.
Methods for using the nano-emulsion effect
37/118
The nano-emulsion effect of the solubilizing compound in carboxylic acids allows its solvation and purification in aqueous media, allows its use as an electrolyte, allows chemical reactions in aqueous media, increases chemical reactivity, increases the dissolution capacity of carboxylic acids for hydrophobic and lipophilic substances. Nanoemulsification allows, respectively, to increase the dissolution and penetration of carboxylic acids in complexes, molecular, and organic or inorganic solids. In addition, it can be used to dissolve complexes of carboxylic acids with apolar or amphiphilic molecules in organic medium or emulsions.
Nanoemulsification
The term nanoemulsification refers to the formation of nanoemulsions. They can be used in a wide variety of applications. The optimal solubility of the two phases in an aqueous medium, the large surface area between the two phases, as well as the dimensions and geometric structures of these phases in the nanometer range make them a versatile vehicle for dissolving reagents, chemicals or pharmaceuticals. Solubilizing compounds, such as arginine or arginine derivatives have been shown to be a valuable additive in several communications reporting on nanoemulsions. However, the exclusive use of such solubilizing compounds for the preparation of a nanoemulsion has not been documented until now.
Methods for coupling and adsorption
Carboxylic acids that have been solubilized in an aqueous medium can be further processed, separating them by means of complexation or adsorption. In aqueous media, the materials for complexation must be proton donors, with an ability to build salts with carboxylic acids. A preferred modality is the use of calcium salts.
Adsorbents can be used in hydrophilic or hydrophobic media, can have hydrophilic or hydrophobic properties, be present in a solvated form or immobilized on a support material. Materials to be used are listed below (5. Acceptor-Zadabsorbent molecules / materials). In a preferred form of carbon, immobilized arginine or calcium is used. For the adsorption or complexation of fatty acids it may be necessary to protonize them in an aqueous organic medium or to deprotonize them in an organic solvent. In doing so, they can be easily adsorbed into an organic aqueous medium or solvent. A preferred modality is the use of n-hexane, triglycerides or cholesterol.
11/38
Methods for using fluid-fluid separation procedures
The solubilization effect of the invention can also be used to separate carboxylic acids from organic media, by means of a fluid-fluid extraction as known in the art. The principle of this method is the spontaneous or guided separation of the aqueous and organic phases. To separate or remove the solvated carboxylic acids in the organic phase, an aqueous solution containing the solubilizing compound is mixed with the organic phase. This mixing can be carried out using various physical means such as stirring, agitation, vibration, sonification, heating, bubbling, steam, as well as by means of laminar or turbulent fluid dynamics. Solubilized carboxylic acids are taken with the aqueous medium, thus separated and concentrated in aqueous solution. Phase separation can be accomplished by gravity, which can be forced by physical means, such as centrifugation or sonification. For the extraction of carboxylic acids so separated or aqueous or organic solution is removed by conventional methods. The dissolved carboxylic acids can be separated by acidifying the solution and extraction with an organic solvent. On the other hand, carboxylic acids dissolved in an organic solvent can be transferred or released to an aqueous medium by extraction with the solubilizing compound of the invention (s). This method can be used for preparation and analysis or applied to large-scale industrial facilities.
Electrokinetic and electrophoretic methods
A preferred embodiment is the electrophoretic separation of carboxylic acids. Carboxylic acids are weak electrolytes. Its ionic strength corresponds to its CMC in an aqueous system because of its low partition. Ionic and anionic detergents have been shown to increase partition by reducing CMC. However, greater ionic strength is achieved by ionic surfactants or the presence of counterions only. Theoretically, a counterion that makes it difficult to micellate would lead to both an optimal partition and an even better partition when the dissociation constant of the ion pair bond is high. Detailed analysis of the partition of carboxylic acids in nanoemulsions is missing. Superubilizing compounds have been found to exhibit these properties. In addition, its carboxylic acids are solubilized and allow an unexpectedly high electrophoretic mobility of carboxylic acids. It is likely that the lower adhesion between the carboxylic acid carbon chains during the interaction with such a molecule is responsible for the observed mobility. Thus, nanoemulgation of solubilizing compounds and acids
39/118 carboxylics allow electrophoresis in aqueous or organic media. In addition, these solubilizing compounds make carboxylic acids suitable for electrophoretic analysis and separation procedures as known in the art. A preferred embodiment is the use of gel electrophoresis. Hydro or organogels are suitable. Detection of electrophoretically separated carboxylic acids can be done using an inorganic chromophore, that is, chromate (254 nm), molybdate (230 nm), or aromatic acids with strong UV chromophores, that is, phthalic, trimellitic, pyromelitic, benzoic, pyridinedicarboxylic acid, cupric-acetate-pyridine and 4-aminobenzoate, by means of techniques known in the art.
Another preferred modality is the use of electro-osmosis.-dialysis, filtration. Carboxylic acids exhibit a tendency to form micelles in an aqueous medium, as previously mentioned. In organic solvents they move freely. However, since they are protonated in organic solvents electrokinetic movement is not possible. When present as a salt, in an aqueous medium, electrophoretic mobility is poor. In addition, although carboxylic acids are usually small molecules, due to the formation of micelles they pass through filter media only to a small extent. This finding is compounded by the fact that the available filter materials are either hydrophilic or hydrophobic. These properties, however, are not ideal for the partition of carboxylic acids. Therefore, electro-osmosis, dialysis, filtration is ineffective for this purpose.
The use of a solubilizing compound of the invention leads to micro or nanoemulgation of carboxylic acids and allows their separation through osmosis, dialysis, filtration, distillation or extraction of supercritical fluid using a degree of concentration, a thermal degree, an electrical degree, a physicochemical degree or a combination of these. In a preferred embodiment, an organophilic separation membrane is used. Said membranes must have a high proportion of organic molecules, when used for osmosis or dialysis, or the surface of the filter channels must have a high content of lipophilic molecules.
However, the separation means exhibiting such properties are lacking. The inventor found that the transport capacity and selectivity for carboxylic acids can be increased by selecting the molecules on the surface of the separation media while reducing the pore / channel dimensions.
11/40
The mechanism of the inventive procedure is believed to be the combination of the aforementioned effects: that is, the high partition of carboxylic acids and the dissociation rate of high arginine or other solubilizing compounds allow rapid movement in the electric field. Electrokinetic flow occurring within nanochannels can further increase the transport capacity by electrophoresis.
Nanofiltration methods
Carboxylic acids have a low molecular weight and small dimensions. Therefore, they are mainly suitable for nanofiltration. However, the low concentration of free carboxylic acids in aqueous media makes nanofiltration ineffective. In addition, the selective membranes of hydrophobic or lipophilic substances are inadequate or non-existent. The solubilization of fatty acids of the invention increases the fraction of free carboxylic acids and allows their nanofiltration. In recent years, nanofiltration membranes have become commercially available. However, their surfaces are hydrophilic making them unsuitable for carboxylic acids. Hydrophobization of channel surfaces only had little effect on their filtration rates. Since nanofiltration would offer decisive advantages in relation to the use of micro or ultrafiltration membranes, such as the specificity of the substrate or increased flow, efforts have been made to improve the partition of carboxylic acids in nanochannels. Therefore, the functionalization of the nanochannel surface was premeditated, in order to make them lipophilic. This objective can be achieved by several molecular classes: amino acids, polypeptides and carboxylic acids (see Chapter E, 4. Materials for surface functionalization.).
Therefore, a preferred modality is the use of a nanofiltration membrane that has a functionalized surface exhibiting low hydrophilic properties (contact angle for water> 100 ° to 25 ° C) and a high liffilic (contact angle for oleic acid <10 ° to 25 ° C). The average channel width should be in the range of 5 to 100 nm, more preferably between 10 and 50 nm. The channel length should be in the range of 0.1 to 10 pm, more preferably between 2 and 5 pm. Materials that can be used are listed below (Chapter E, 3.3 Separation membrane materials).
The preferred materials are aluminum oxide, titanium oxide, carbon, polycarbonate, polyethylene, silicates.
41/118
In a preferred embodiment, surface functionalization should be preceded by monolayer coating of the channel surface with a suitable polymer to react with the molecules used for functionalization called the bonding layer and molecules to be associated with this layer determining the surface properties, called the layer of functionalization.
1. Connection layer
This layer acts as an interface between the data support material and the functionalization layer. It may be necessary to introduce or activate molecular structures in the first support material. The bonding layer must completely cover the surface, which must be smooth after processing. There must be a dense count of reactive groups appropriate for the reaction with the molecules of the functionalization layer. However, a defined multilayer coating may be advisable in order to shape the channel structures. This can be achieved by polymers, as indicated below (Chapter E, 3.3 Separation membrane materials).
A preferred embodiment is the use of polymers, such as APTS, pentafluorophenyl acrylate (PFA), pentafluorophenyl methacrylate (PFMA), poly-Ntrimethyl-amino-ethyl methacrylate (PTMAEMA) and poly (2-dimethylamino) ethyl methacrylate (PDMA) .
2. Functionalization layer
This objective layer will achieve defined surface conditions.
The preferred surface condition resulting from the sum of the intermolecular forces of Coulomb, van der Waals, hydrogen bonds and hydrophobic interaction of the surface functionalization of the invention has a lipophilic effect allowing partition of the carboxylic acids. Interposition of charged groups, creating a surface charge, which can be positive or negative, is used in a preferred mode. An additional preferred modality is the institution of a hydrophobic or lipophilic surface force ranging from 2-20 nm, while having a neutral or positive surface charge. Suitable molecules can be carboxylic acids, amino acids, peptides, proteins, ternary or quaternary amides, aromatic hydrocarbons or cyclodextrins. (See Chapter E, 4. Materials for surface functionalization).
In a preferred embodiment of fatty acids, alanine, phenylalanine, arginine, lysine, amidine or guanidine are used as a central functional group within a molecular structure.
42/118
Thus, the present invention also relates to an ultra or nanofiltration separation panel for dialysis, filtration or nanofiltration of carboxylic acids that is functionalized with one or more of the substances of general formula (I) or (II) comprising at least one functionalization layer and, optionally, a bonding layer.
D. Applications of the inventive solubilization procedure
Blood fatty acid dialysis
Dialysis is a standard procedure in medicine, analysis and in the chemical or pharmaceutical industry. The procedure is used to reduce or eliminate substances from a feed solution, by a degree of passage conducted through an interface. The interface consists mainly of a porous membrane, which allows the passage of molecules up to a defined size. Available membranes vary in their hydrophilic and hydrophobic properties. However, the passage of free carboxylic acids is only possible in negligible amounts. One reason for this is that the amount of free fatty acids is small, even in the presence of high concentrations of detergents, since the micelles are still present, resulting in a particle size that does not allow it to pass through the membrane pores. . This problem can be overcome by forming a micro or nanoemulsion inventory. The method of the invention can be carried out by modifying a conventional blood dialysis, for example, a subject's blood system is coupled to the dialyzer via tubes. Alternatively, a certain amount of blood is taken as a sample, processed in a dialyzer modified according to the method of the invention, and then reinfused into the patient. In the latter case, the volume of the blood sample obtained is in the range of 10 ml to 0.5 I, preferably between 100 and 500 ml, more preferably between 300 and 500 ml.
It must be emphasized that all the methods described in the present invention can be carried out in vivo, that is, on the human and animal body and in-vitro, that is, not on the human and animal body.
In a preferred embodiment the method of the invention is used to remove free fatty acids from the blood of a subject in need. Therefore, there are medical and scientific applications of this method. According to the invention, a dialyzer, respectively extractor, is provided which applies the principle described above inventive to the subject's blood. For this purpose, the inventive device is coupled to the venous or arterial circulation of a
43/118 mammal, preferably a human, through cannulas, catheters and tubes. In some embodiments, a commercially available dialysis machine can be used here, for example, Plasmat futura® (BBraun Melsungen, Germany). The blood aspirated by this machine through an arterial or venous tube or supplied by a blood sample can be pumped into the dialysis unit (see schematic figures in Fig. 2: 213), using a roller pump ( 212), either after or without hemoseration. The dialysis unit consists of two subunits, which are interconnected by appropriate tubes (for example, Fresenius, Lifeline® Beta SN Set SRBL-R, Germany). The first subunit first consists of a dialyzer as described below (for a detailed description see section on dialysis / extraction procedures). The blood aspirated by the dialysis machine can be pre-treated by means of dialysis and or citrate anticoagulation (202) as is known in the art, or undergo a hemoseparation procedure or a sequence of these procedural steps. The prepared blood (plasma) is pumped by the dialysis machine into the entrance of the dialyzer (203), together with a constant infusion of the arginine solution or any other solution of a solubilizing compound, by means of an infusion pump (214) from a storage bag (215) containing a sterile solution of the solubilizing compound such as arginine. The final concentration of the solubilizing compound in the dialysate can be calculated from the current blood flow (plasma) and the concentration of this compound in the infusion solution. In the case of arginine, a final concentration of 100 to 300 mmol / l is targeted. When it is dissolved in the dialyser dispensing chamber (for detailed descriptions see the dialysis / extraction procedure section), the mixture of the blood-solubilizing compound (plasma) solution passes the hollow fiber membranes that allow micro or nanofluidic conditions. In a preferred embodiment, an arrangement of hollow chamber capillaries is used with a typical diameter of 200 pm and a length of 30 cm. The volume of blood (plasma) within the separation medium is 40-80ml, preferably 50-60ml. The material can be inorganic, organic or combinations of both. Materials that can be used are summarized in the interface material list (see chapter E, 3.4 hollow porous capillaries). The membranous interface has micro or nanopores being molecularly functionalized. Functionalization of the surface, pore and channel and properties are described below (see Chapter Ε, 4. Materials for surface functionalization). The duration of the passage of the feed solution should preferably be
44/118 between 20 to 60 s, more preferably between 30 and 40 s. The resulting duration of contact between the feed solution and the interface should allow for complete removal of solubilized fatty acids. The assembly of such filter cartridges is known in the art. The dialysis fluid is perfused through the dialysate inlet. It fills the space between the outside of the hollow fibers and the inside wall of the cartridge. The perfusion direction is opposite to that of the feed flow and is called the cross flow in the technique. The normal perfusion rate of the dialysis fluid is in the same range as that of the feed liquid. However, it may be necessary to vary the flow rate ratios, which can typically be between 1: 2.1: 1, 2: 1 or 3: 1, depending on the concentration of lipid in the feed solution. The dialysis liquid exits the cartridge through the outlet cone and is transferred to a secondary circulation unit, which is described below.
Blood (plasma) leaves the dialyzer through collection in the collection chamber and is transferred through the appropriate tubing to the second unit. The second unit consists of a standard dialyzer used for hemodialysis (201). The blood (plasma) is conducted through hollow chamber capillaries inside the dialyzer allowing balance of concentration of low molecular weight hydrophilic molecules and electrolytes against the dialysis solution (210), which is pumped through a roller pump (216) and penetrates the space between the outer surface of the fibers and the inner surface of the cartridge. The dialysate outlets of the standard dialyzer through an outlet are collected in a waste tank (211). The concentration, respectively, of the ionic strength of the dialysate and the pH is regulated by the dialysis machine. The purified plasma is fused with the blood cells, which have been separated (not shown in Fig. 2). The purified blood is pumped by means of a roller pump (212) back to the patient through the aforementioned catheter system.
Secondary circulation unit
As previously mentioned, the solution within the secondary circulation, called here the acceptor solution, must have a high capacity to bind to the fatty acids that have passed through the dialyzer interface (203) and connect them to an additional secondary extraction device (204), here called the fatty acid exchange module. The acceptor solution (see Chapter E, 7. Acceptor solutions) within the secondary circuit can be an aqueous solution with an organic or inorganic acceptor (see Chapter E, 5. Acceptor / adsorbent molecules / materials). The acceptor molecules can
45/118 be dissolved in free form or immobilized to ceolites or adsorbing surfaces (see Chapter E, 4. Materials for surface functionalization). Preferably, a solution of purified fatty acid binding proteins originating from human or synthetic sources should be used as an acceptor for fatty acids in the secondary circulation. The secondary circulation acceptor solution is driven by a pump (205), preferably a roller pump. The acceptor solution loaded with fatty acids leaves the dialyzer via the suction filtrate outlet of a roller pump (205) and is routed into the entrance of the fatty acid exchange module (204) through a suitable pipe and perfused through the common exchange chamber filled with the extension granulate. The acceptor solution is being purified while passing through the extraction granulate, then exits the exchange module through the outlet port and enters a pipe that is connected with the dialyzer at the entrance of the filter chamber.
The carboxylic acid exchange module (see fig. 3) consists of a cartridge that displays two inlets and two outlets located on the opposite sides of the cylindrical cartridge. Inlet (301) and outlet (303) of the secondary circulation are covered by a filter funnel (305) having a pore size below the lower range of the extraction granulate from the exchange circulation. The inlet (302) and outlet (304) for the tertiary circulation are large enough to allow an internal and external pressure flow from the granular extraction material. Within the exchange module, the secondary circulation exchange solution is in immediate contact with the tertiary circulation extraction granules. Perfusion of the exchange module with the secondary circulation solution and with the granule of the tertiary circulation is directed in the opposite direction between them. The cooled secondary circuit acceptor solution exits through the outlet (303) of the exchange module and is conducted to the dialyzer outlet connection via a pipe.
Tertiary circulation
The tertiary extraction circulation is driven by a pump (see fig. 2: 205) suitable for the use of granulated extraction material. Preferably, a double cylinder pump (207) can be used. The introduction of air into the circulation is reduced by simultaneously filling the system with small amounts of purified acceptor solution by means of a communication through a pipe that connects the pump with the outlet of the exchange module. A pump bypass port allows simultaneous filling of the system. The material of
46/118 extraction dissolved in the acceptor solution is pushed forward through the tubing which is connected to the fatty acid exchange module outlet. Residual air transported within this segment is removed by an air trap (208) mounted on top of an air exhaust port (217). The door is sealed with a filter plate that does not allow extraction material to pass through. After passing, the extraction material exchange module leaves the cartridge through the tertiary circulation outlet. This outlet is connected with a pipe that can be fixed in a vertical position located above the exchange module and the storage container for the acceptor solution (209). This piping is interconnected with the storage container, which is sealed with a filter plate that does not allow extraction material to pass through (218). However, the acceptor solution is allowed to flow back into this piping to the approximate hydrostatic level of the filling plane of the storage container. The extraction material is pushed forward through the tubing into a container (206). The container is part of a cleaning system for the purification of the extraction material. Purification can be operated by physical or chemical means. The purification process is followed by a final cleaning step using sterile water. The purified extraction material is then collected in a second container and connected to the tertiary circulation pump.
Alternatively, to purify the extraction material in a tertiary circulation, fresh extraction material can be fed from one reservoir and discharged into another reservoir after passing the exchange module using the same pipes as previously described.
According to the invention, the aforementioned procedure or parts of it can be combined with standard techniques in dialysis-type hemotherapy, hemoperfusion, hemofiltration, hemodiafiltration, plasma separation with centrifuge, plasma apheresis, cascade filtration and term filtration.
Here, the separation efficiency is increased by additional hydrolysis of esterified fatty acids, reinforcement of lipolysis and / or the use of a central venous blood aspiration site for blood purification. Thus, an especially preferred embodiment of the present invention is to remove carboxylic acids and especially fatty acids from the blood using the solubilizing compounds disclosed herein and preferably arginine and arginine derivatives. The most common method for blood purification is dialysis which
47/118 can also be used to remove fatty acids and also fatty acids limited to albumin from the blood.
Solubilization fluid formulations for human use
In a preferred way, the solubilization of fatty acids is achieved through the use of a solubilizing compound of the invention. It can be applied in its pure form, or in a pH-adjusted solution with HCI or other acids acceptable for use in humans. The preferred pH range is 7.5-10.0, most preferably between 8.0 and 9.0. It may be advantageous to use an additive such as a co-solvent or buffer, as listed below (see Chapter E, 8. Additives for preparations of arginine or its analogs). A preferred additive is ascorbic acid.
Clinical use
The method of the invention for the solvation and extraction of fatty acids from human blood can be applied by conventional techniques, as known to a person skilled in the art and can be part of a hemodialysis procedure performed on a patient in need of it , due to kidney or liver failure. The use of this procedure can be indicated in other indications as well. Medical indications include, but are not limited to, diagnoses or conditions, such as diabetes mellitus, metabolic syndrome, overweight, obesity, hypertension, hypertriglyceridemia, hypercholesterolemia, hyperuricemia, cellulitis, atherosclerosis, fatty liver, lipomatosis, ventricular extrasystoles, ventricular tachycardia , supraventricular fibrillation. A preferred modality is a venous access site by aspiration and blood recirculation. The aspiration site must be within the central venous system, more preferably, within the inferior vena cava. The purified blood can be returned to the patient, through the same access site having the orifice distally to the openings of the aspiration site. This is done in commercially available catheter systems such as BioCath (Bionic Medizintechnik, Friedrichsdorf, Germany). The access system should have a French size between 8 and 14, more preferably between 10 and 12. The most preferred site of venous access is the femoral vein.
The connections and filling of the tubes must be carried out as done in hemodialysis and known to those skilled in the art. Therapeutic anticoagulation is mandatory during the procedure. This can be done by co-infusing heparin or low molecular weight heparins using a dosage in order to achieve therapeutic blockage of the extrinsic pathway of the
48/118 blood clotting, measured by activated partial thromboplastin time or by the anti-factor Xa activity test, respectively, as is known to those skilled in the art. Alternatively, administration of citrate in order to complex calcium ions can be used for blood anticoagulation within the hemodialysis system. This procedure is provided by dialysis machines, such as Multifiltrate (Fresenius, Medical Care, Germany). Complexed calcium is dialyzed by an initial hemodialysis stage. During further processing, the blood cannot clot. Before returning the purified blood to the patient, a predefined dosage of an infusion containing 10 calcium ions is co-administered to the blood flow, restoring coagulation.
According to the inventive finding that the extraction fraction of the procedure can be increased by stimulating lipolysis in a person so treated, it is preferable to applying drugs that induce lipolysis as β1 -, β2 -, 33-adrenergic agonists, inhibitors of phosphodiesterase III, agonists a-1 and a-2 15 of adreno-receptor, donors of nitroxide or nitroxide, hormone sensitive lipase, leptin, natriuretic peptide, vasopressin, heparin and analogues, tyrosine, yohimbine.
In addition, the stimulation of inventive lipolysis refers to the regional improvement of lipolytic activity, through local subcutaneous infiltration of the referred lipolytic drugs, as well as anesthetics, liposomes, including phospholipids, vasodilators, including histamine. The additional increase in lipolysis can be achieved by stimulating the electric field or by applying ultrasound or pulse wave energy.
Solubilizing compounds, such as arginine, as well as 25 other small water-soluble molecules, readily pass through hydrophilic dialysis membranes. Despite being non-toxic, the removal of high non-physiological concentration of arginine from the blood / plasma through final dialysis using a standard dialyzer (high or low flow) is preferred. A final dialysis guarantees the restoration of a physiological electrolyte concentration, 30 osmolarity and pH.
It can be useful to increase the concentration of albumin in the blood, phospholipids or cyclodextrins, in order to increase the transport capacity of non-esterified fatty acids during the procedure.
During such a combined treatment, a control close to the hemodynamic parameters (blood pressure, heart rate, temperature, hemoglobin oxygenation), as well as metabolic parameters (blood glucose, pH,
49/118 sodium and potassium) is mandatory. The duration of a treatment episode depends on clinical parameters. Typically, a procedure takes between 3 and 12 hours, more preferably, the duration is between 4 and 6 hours. The amount of fatty acids extracted depends on the selectivity of the filtration membrane used for pathogenic fatty acids. The preferred amount of extracted fatty acids is between 100 and 2000 ml, more preferably between 500 and 1500 ml.
The purification procedure for clinical use can be performed as a method of dialysis, filtration, adsorption, precipitation or combinations thereof.
Large-scale extraction of free fatty acids in industry
Free carboxylic acids are often found in solutions or emulsions produced or used in large-scale installations. For example, fatty acids are present in crude vegetable oils, in the shell and wrapper of fruits, vegetables and corn; in biomass or wastewater; in crude mineral oil or occurring during processing, as well as in soils containing petroleum, in residual oil and grease elimination. In most cases, physical procedures are used to remove these carboxylic acids that require high energy consumption. In some cases, the separation procedure has undesirable consequences on the purified product. For example, for refining oils they are exposed to steam in order to distill volatile fatty acids. During such a procedure, exposure to heat can lead to the transisomerisation of esterified carboxylic acids. This is a potential hazard to the consumer. Therefore, it is desirable to avoid such procedures.
It was found that this task can be solved by the inventive procedure.
According to the invention, these aqueous or organic solutions to be purified arise from plants, microorganisms, fossil materials, natural or synthetic reaction mixtures.
A preferred embodiment is the purification of oils from free carboxylic acids by adding aqueous solutions of at least one solubilizing compound as described herein, such as the compounds of general formula (I) or arginine or arginine analogs and mixtures thereof compounds (Fig.4). The oil to be purified is poured from a storage tank (402) to a reaction tank (401). A predefined amount of a concentrated solution of the solubilizing compound from a storage tank (403) is added to the reaction tank. Preferably, the
50/118 solution is mixed using a mixing system (411). Then, the mixture is pumped (pump 412) to a collection tank (405), in which the aqueous phase and the oil phase separate spontaneously through gravity. The lower aqueous phase contains the carboxylic acids that dissipate in the nanoemulsion or microemulsion that is continuously removed through the outlet at the bottom of the tank. Alternatively, the mixture is transferred to a centrifuge or a membrane separator. This procedure can be repeated if a higher degree of purification is required. It has been shown to be advantageous to slightly heat the solutions while mixing them in order to reach the completion of the solubilization process. The mixing process is accelerated by the application of sonar waves. When the mixture is transferred through a membrane separator, it has been found to be advantageous to use primarily anion-selective membranes. The purified oil (triglyceride phase), usually does not contain any solubilizing compound, after removing the water. However, for highly purified oils, it may be useful to repeat the wash with water or to use cation adsorbents. The triglyceride phase purified from the upper phase of the collection tank is continuously removed through an outlet located at the top of the tank and transferred to a triglyceride storage tank (404). The aqueous solution is pumped from the bottom of the collection tank (405) to a second reaction tank (406). A predefined amount of acid is added from an acid storage tank (407). The solution in the reaction tank is mixed using a mixing system (411). Thereafter, the mixed solution is pumped (412) into a second separation tank (408). Fatty acids and solubilizing compounds solubilized in water are left to separate by gravity. However, other means of separation, as known in the art, can be used instead. The purified fatty acids that are concentrated in the upper part of the second separation tank are constantly transferred to a fatty acid storage tank (409). The aqueous solution of the solubilizing compound that is concentrated at the bottom of the second separation tank is continuously pumped into an electrodialysis unit (410). Electrodialysis can be applied in order to remove cations and / or anions by means of techniques and devices as is known in the art and by means of selective membrane ions (413). The purified solubilizing compound solution is pumped into the solubilizing compound storage tank (403). A solubilizing compound
51/118 p _re f _er e _nc j _a | _for this purpose it is a derivative of arginine and especially arginine.
Alternatively to separation by gravity, carboxylic acids solubilized within the aqueous phase can be separated by processes using electrophoresis, pneumatic filtration or nanofiltration, immobilization, aggregation, distillation or phase transfer, using an organic solvent. In a preferred embodiment, the adsorption of volatile carboxylic acids by carbon, complexation with calcium, phase transfer by means of organic solvents, electrodialysis and organo-nanofiltration is used. A preferred option is the use of stearases, using the inventive solubilization procedure in sewage processing or biodiesel production (see Chapter E, 2. Hydrolases). Their combined use increases the effectiveness and completeness of removing organic components, respectively oily components or chemical reactivity.
For carrying out the methods of the invention, carboxylic acids can be present in a solution and at least one solubilizing compound of the general formula (I) or (II) is added.
Alternatively, carboxylic acids are added to a macro-, micro- or nanoemulsion containing at least one solubilizing compound of the general formula (I) or (II), in order to use said emulsion to release, decomplex, remove, react, aggregate, complex, coagulate, flocculate, sediment or separate complexes containing carboxylic acids.
These methods of the invention can be used to initialize, increase, decrease or maintain a physiochemical or chemical reaction, allowing, increasing the absorption and transport of reaction products or components in processes of a biological or chemical reaction, removing, solubilizing, releasing, convecting, transporting substances through the absorption of vesicles, or allowing or improving the penetration of emulsified carboxylic acids through the medium or hydrophilic or amphiphilic solids.
Preferred industrial applications include
- the removal of fatty acids from solutions containing fatty acids resulting in oil or fuel processing. Particularly in the production and processing of mineral oils and fuels, respectively in biofuels, the method of the invention can be applied.
- the removal of fatty acids from solutions containing fatty acids resulting in industrial food processing. Particularly in
52/118 production of edible oils, processing of rice, corn and whey bran, vegetables, as well as dairy products and fish, low-fat and non-fat products, and organisms containing oil or producers, respectively, this method should be useful.
- the removal of fatty acids from aqueous solutions containing fatty acids resulting from the processing of sewage containing bioorganic compounds or in industrial sewage. For example, for the processing of sewage from bioreactors, this method can be applied.
- the removal of fatty acids from organic or aqueous solutions containing fatty acids resulting from the cleaning of industrial products, such as wool, cotton or other textiles; in the processing of sewage from industrial cleaning, such as tank installations, ships, car washes, slaughterhouses, a.o.
- the removal of fatty acids in chemical or pharmaceutical processing, such as the preparation of adhesives or paints.
- the removal of substances without carboxylic acid that aggregate or adhere to solubilized carboxylic acids, thereby co-solubilizing and being separated or removed together with the solubulizing substance and carboxylic acids of the invention as to be used in the purification of petroleum fats and oils or of organic or mineral origin, in order to remove complexing phospholipids, glycolipids sterols, pesticides being already solubilized or immobilized in organic or inorganic matter.
- the removal of adhesive substances, bound or complexed by the macro-, micro- or nanoemulsions of the solubilizing substances and carboxylic acids of the invention for the extraction of organic oil seeds, oily sands or rocks and oily deposits.
Application for the analysis of fatty acids in aqueous solutions
Qualitative and quantitative analysis of carboxylic acids is an uncomfortable task. Carboxylic acids with a carbon chain length greater than 6-10, depending on the presence of hydrophobic or hydrophilic substituents, cannot be measured in an aqueous medium, prohibiting their measurement by electrophoresis or conductivity. In addition, the analyzes can be hampered by the incomplete dissolution of carboxylic acids from organic compounds, even when organic solvents have been used. Standard analysis is performed with gas chromatography (GC). However, carboxylic acids must be methylated to be suitable for GC, making this
53/118 method time-consuming and susceptible to methodological flaws. These difficulties can be overcome by the solvation procedure of the invention.
According to the invention, this method can also be used for the qualitative and quantitative analysis of the fatty acid content in aqueous solution. It is also suitable for differentiating the relative content of esterified and non-esterified fatty acids.
A preferred modality of the inventive solvation process is its use for the analysis of carboxylic acids by electrophoresis, conductivity and spectrometry.
Preparation of analytical samples
Mixtures of oil and fatty acids, as well as mixtures with water such as oil-in-water (o / w) and water-in-oil (w / o) emulsions are transferred to a reaction chamber. The solution with a solubilizing compound of the invention is added. To determine esterified fatty acids, stearases can be added before, in conjunction with or after the addition of the solubilized compound in order to release them. The solutions should be incubated. It has been shown to be useful for moderately heating the sample volume, reducing ionic strength or reducing the pH before adding the solubilizing compound. Subsequent analyzes can be done
- with the solution in its current state,
- by precipitation of free carboxylic acids,
- by extraction with an organic solvent.
The use of analyzers resulting with standard analytical methods is described below.
Gel electrophoresis procedure
For the analysis of carboxylic acids by gel electrophoresis the aqueous analyte can be taken in its present form or as an electro-nanofiltration filtrate or dialysis being dissolved by the solubilizing compound as a micro- or nanoemulsion. A standard gel electrophoresis device can be used (for example, BIOTEC-FISCHER GmbH, PHERO-vert 1010-Ε) and a SDS-polyacrylamide gel. It may be useful to add practical solvents, such as ethanol to the analyte. In a preferred embodiment, an organogel is used (see 6. Organogels). Calibration and reading can be done as is known in the art.
Distillation
54/118
A solution containing non-esterified solubilized fatty acids in an aqueous solution of the solubilizing compound can be purified by distillation in one or two stages. This can be done by heating and vaporizing the solution under ambient air pressure or under vacuum conditions, in order to reduce the evaporation temperature of the fatty acids to be distilled. A preferred modality is the use of a thin film evaporator (Normag, Roatafil device).
Precipitation / complexation procedure
The precipitation or complexation of the solubilized carboxylic acids can be carried out as described above and known in the art. In particular, methods such as complexation with metal ions or cyclodextrins are preferred. The precipitate must be extracted and washed with water as is known in the art. The purified precipitate is then dissolved in a strong acid (HCI, acetic acid, carbonic acid) until complete dissolution and protonation of the carboxylic acids. The carboxylic acids are then extracted with an organic solvent (n-hexane, diethyl ether, chloroform, a.o.). The organic phase is carefully removed and processed for further analysis.
A preferred analytical method is liquid chromatography.
Solvent extraction procedure
The extraction of carboxylic acids from media not sensitive to acidification or exposure to organic solvents can be performed directly from an aqueous medium. The inventive use of the solubilizing compound has the decisive advantage that the extraction condition should not be so drastic as compared to single solvent extraction procedures. This is achieved by the carboxylic acids to be extracted being already dissolved in the aqueous phase by the inventive solvation process. Careful acidification in the presence of an organic solvent phase leads the protonated carboxylic acids to the solvent phase, without the need for rigorous mixing of solvent and medium. Then, solvent extraction is performed as known in the art. The solvent solution can be used for NIR- or IR, or distant IR spectrometry or liquid chromatography directly.
Electro-nanofiltration / diffusion procedure
An additional preferred method for analysis is filtration conducted electrophoretically or electrostatically by diffusion for the separation to be used with the inventive solubilization (Fig.5).
55/118
The organic or inorganic material, the solution / emulsion is prepared by the aforementioned solubilization. The pH value must be adjusted to values> 6.0, preferably to values between 8 and 11. In what follows, a defined sample volume is transferred to the donor chamber / reaction batch of the analysis device. This donor chamber (503) is arranged between a catholite-filled chamber (502) and a separation chamber (505). The donor chamber / reaction batch and the catholyte chamber are separated by a membrane (504). Preferably, this membrane is ion-selective. The separation chamber is filled with a chromatophore (for example, a gel, preferably an organogel). Alternatively, it can consist of a microfluidic system or a functionalized diffusion or nanofiltration membrane as described below (see Chapter E, 3. Membranes, and 4. Materials for surface functionalization) (510). On the other side of the membrane separator, an acceptor / container chamber (508) is arranged, which is filled with an arginine solution or a solution of any other solubilizing compound. This acceptor / container chamber is adjacent to an additional chamber / container (507) which serves to receive the anolyte. These chambers / containers are separated by a membrane (506). Alternatively, the separation panel is an organogel filled in a capillary. Preferably, the membrane is ion-selective. When voltage is applied between the cathode (501) and the anode (509), ionized carboxylic acid compounds present in the donor / recipient chamber, particularly fatty acids, such as anions, are conducted through the separation / membrane chamber. and thus transferred to the acceptor / container chamber. The solution in the acceptor / container chamber can be analyzed immediately. Preferably, the analysis is performed by conductivity, spectroscopy or mass detection methods. Alternatively, an additional agent (for example, an indicator, a derivative agent) is added and the analysis follows. Suitable anolytes and catholytes are the arginine solution, arginine derivative solutions, HCI, a.o. The addition, mixing and transfer of the agents is preferably carried out in a microfluidic system. This is particularly suitable for the development of a lab-on-the-chip analysis system.
The practical applications are medical - biochemical analyzes of fatty acid contents of body fluids taken as a sample from a subject. Such an analysis can serve as a diagnostic criterion. Medical diagnoses include, but are not restricted to atherosclerosis, hypertension, diabetes mellitus,
56/118 obesity, hyperlipoproteinemia, myocardial infarction, stroke, renal failure. Scientific applications include use in chemistry, biochemistry, pharmacy, pharmacology, materials science, biology, industrial food processing. This method of analysis can also be used in industrial applications as outlined in the previous paragraph on a large scale extraction of free fatty acids.
Dialysis / extraction devices and procedures
An object of the invention is an integrated dialyzer / extractor. For carrying out the solubilization and separation of carboxylic acids of the invention, in an aqueous or organic medium, with a solubilizing compound of the general formula (I) or (II), an integrated dialyzer / extractor must comprise the following essential key components, which are essential for most modalities, regardless of their application:
i) A first chamber for the reaction of the aqueous or organic medium containing carboxylic acid with the solubilizing compound of the general formula (I) or (II);
ii) A second chamber for receiving the solubilized carboxylic acids;
iii) A separation panel between said first chamber and said second chamber comprising a separation membrane or a hollow capillary assembly; and iv) Means for conducting said reactive solution from said first chamber to said second chamber through said separation panel by applying a concentration gradient, a thermal gradient, a gradient of physiochemical properties, a gradient pneumatic, an electric gradient or a combination thereof.
This device can be used for medical therapy, medical analysis, food analysis, food processing, oil processing, oil analysis, fuel processing, chemical processing and pharmacological or Pharmceutical treatment, analysis or science in the pharmaceutical or chemical industry, removal of carboxylic acids from sewage from private, commercial or industrial cleaning, removal of carboxylic acids from biological reactor processes, cleaning of oily solids, organogelatination or nanoemulsification of carboxylic acids.
For the use of such a device, a method with the following key steps is applied:
57/118
i) Provide the solution or emulsion or suspension containing the carboxylic acids;
ii) Addition of at least equimolar amounts of at least one solubilizing compound;
iii) Separation of the solubilized carboxylic acids from the solution or emulsion or suspension through phase separation, filtration, nanofiltration, dialysis, absorption, complexation, distillation and / or extraction.
More specifically, step iii) is preferably achieved by using one of the following separation methods or a combination thereof:
passing the carboxylic acids separately or together with at least one solubilizing compound through a separation membrane or a hollow capillary tube or assembly by applying a concentration gradient, a thermal gradient, a physiochemical properties gradient, a pneumatic gradient , an electric gradient or a combination thereof; or perform phase separation by combining two or more means by building phase separations; or passing the carboxylic acids together with the compound of at least one solubilizer through a phase separation interface that allows the passage of said carboxylic acids and said at least one solubilizing compound by applying a concentration gradient, a thermal gradient, a gradient of physiochemical properties, a pneumatic gradient, an electrical gradient, or a combination thereof, where the phase separation interface consists of a gel, an organogel or a solid material or a combination thereof; or filter the carboxylic acids using at least one solubilizing compound; or nanofilter the carboxylic acids using at least one solubilizing compound; or dialysis the carboxylic acids using at least one solubilizing compound; or adsorb carboxylic acids, using at least one solubilizing compound; or complexing the carboxylic acids using at least one solubilizing compound; or
58/118 distillate the carboxylic acids using at least one solubilizing compound; or separating the carboxylic acids using at least one solubilizing compound by extraction with supercritical fluid.
In the separation step, the gel and / or solid materials can be of organic or inorganic origin and they can be porous or non-porous.
According to the invention, the aforementioned device and method should be used in the following areas: medical therapy, medical analysis, food analysis, food processing, oil processing, oil analysis, fuel processing, modulation of chemical or physiochemical reactions , solubilization of poorly soluble molecules, chemical and pharmacological or pharmaceutical processing, analysis in the pharmaceutical or chemical industry or science, removal of sewage carboxylic acids from private, commercial or industrial cleaning, removal of carboxylic acids from processes of bioreactors or soils or plants, cleaning oily solid materials, organogel freezing or nanoemulsification of carboxylic acids. Said analytical methods can be quantitative or qualitative.
More specifically, preferential arrangements include the following parts:
a) A first chamber for reaction of the aqueous medium containing carboxylic acid, with the solubilizing compound, having a first entrance to said aqueous medium containing carboxylic acid;
b) A container for said solubilization compound, which has a second inlet to fill said container with said solubilization compound and to be connected to said first chamber through a third inlet;
c) A second chamber to receive the solution containing dialysed / filtered carboxylic acid;
d) A separation panel between said first chamber and said second chamber comprising a separation membrane or a hollow capillary assembly; and
e) Means for conducting said reactive solution from said first chamber to said second chamber through said separation panel by applying a concentration gradient, a thermal gradient, an electrical gradient, a physiochemical gradient or combinations thereof .
59/118
Optionally, you can also understand the following components:
f) Means for removing the associated carboxylic acid and solubilizing compound from said filtered solution by removing said filtered solution by convection of an acceptor solution being fed through a fourth inlet to said second chamber and left flow through a first outlet out of said second chamber; and
g) Means for removing the purified solution from said second chamber via a second outlet.
Such an integrated dializer / extractor is suitable for carrying out the solubilization and separation of carboxylic acids of the invention in an aqueous or organic medium in a wide range of medicinal and industrial applications. These key components build the centerpiece of devices designed for specific applications. Specific modalities for particular applications are described in more detail below.
It should be noted that, according to the invention, all entrances, exits and means of transport may have adjustment devices to control the respective flow or transfer rates. These adjustment devices will not be expressly mentioned for each modality. All adjustment devices known in the art will be suitable according to the invention.
The object of the invention is also a method that applies the key steps for the solubilization and separation of carboxylic acids in an aqueous or organic medium, using the above-mentioned integrated dializer / extractor. The core of the inventive methods is represented by the following steps:
a) The preparation of said solution through the reduction of ionic strength, by means of complexation, adsorption, separation or dialysis of bound and unbound cations;
b) Adjust the pH of the solution by adding an acid or a base;
c1) Adjust the molarity of the solubilizing compound to be in the range of 1:10 to 20:01 compared to the estimated concentration of the carboxylic acids to be solubilized; and
d) Adding said solubilization compound in a solid form or in a solution to said aqueous or organic solution containing carboxylic acid for the generation of a micro- or nanoemulsion.
60/118
Optionally, the method can also comprise any of the following steps:
a1) release of linked carboxylic acids by complexation or covalent bonding c2) If the solubilizing compound is administered in a solution, adjust the pH of that solution in order to optimize compatibility and reaction conditions with the carboxylic acids to be solubilized by means of acidification or alkalinization;
e) Addition of stearases, hydrolases or a complex builder;
f) Addition of water and / or a co-solvent to the solution; and / or
g) Optimization of reaction conditions, by heating and / or mixing the solution, thus generating an improved micro- or nanoemulsion.
In the above description of the devices and methods of the invention, as well as in the following modifications and modalities, not all features must be included, respectively not all steps must be performed, some of them are optional. In addition, in some modalities, some characteristics or stages are modified, respectively displaced by corresponding characteristics or stages. Therefore, the sequence of steps in the respective method must be read in alphabetical order first. Second, the numerical affix is decisive. For example, if a step c and a step c1 are present, step c1 will be performed after step c. In other words, step c1 will be interspersed between step c and step d. Similarly, if a step c1 and a step c2 are present in a method, this means that step c1 must be performed before step c2. In other words, step c1 is interspersed between steps b and c2. If in a modality, a step is modified, respectively replaced, in comparison with the previous modality, it may happen that, for example, in each modality, a different step g is listed. The sequence of these alternative steps must be read in the correct alphabetical order. Thus, if a modified step g is present in the list of steps, this means, of course, that a step g of another modality is not included in the present modality. If optional steps are included, the steps can be presented in a non-alphabetical order. This does not alter the line to understand the sequence of steps in an alphabetical order. The same applies to changes in the respective inventive devices.
61/118
In modalities in which at least two chambers are provided, the listing of the steps of the method of the invention can be complemented as follows:
g2) Conducting the reactive solution from a first chamber to a second chamber, through a separation panel using the nanofiltration technique by applying a concentration gradient, a chemical gradient, a pneumatic gradient, an electrical gradient or an combination thereof.
Optionally, the following steps can be understood in these modalities:
h) Removal of the carboxylic acid and solubilizing compound associates from the filtered solution through the convection of an acceptor solution being conducted through an inlet to said second chamber and allowed to flow through an outlet of said second chamber; and
i) removing the purified solution from said second chamber via an additional outlet.
Steps g2), h) and i) can be performed after step f), as described above. A very important application of the method of the invention is the purification of a patient's blood from volatile fatty acids.
Therefore, the respective modality of an inventive integrated extractor / dialyzer has the following modifications, respectively the additional characteristics (Fig. 6):
f) Means for conducting blood or plasma from said subject to said first chamber (610) of a dialyzer (603) through said first entry;
g) A pumping system and mixing system (602), which allows the solubilizing compound to be fed from said container (601) and mixing the solution;
h) Optionally, said first chamber contains support materials (604) in which the hydrolases are immobilized, in order to release the esterified fatty acids;
i) A first separation panel between said first chamber of a second dialyzer and the second chamber of a first dialyzer, comprising a separation membrane (605) or a hollow capillary assembly;
62/118
j) Means for conveying the solution containing carboxylic acid from said first chamber of the first dialyzer to a second chamber of the first dialyzer by applying a concentration gradient, a chemical gradient, a pneumatic gradient, an electrical gradient or a combination of the same;
k) Pumping means (606) of said filtered solution from said second chamber to a first chamber of a second dialyzer (607);
l) Means for the removal of carboxylic acid and solubulizing compound members that pass through said second separation panel of the second dialyzer (607) by means of a tertiary circulation;
m) An acceptor solution storage container;
n) Pumping means (612) of the carboxylic acid acceptor solution from said solution receiving storage container (609) to said second chamber of the second dialyzer;
o) the means for removing the carboxylic acid acceptor solution loaded into a waste container (608);
p) Means for returning the purified solution containing the solubilizing compound to leave said first chamber of the second dialyzer for entry to said second chamber of the first dialyzer; and
q) Means for returning blood fractions collected to leave the first chamber of the first dialyzer for the circulation of the subject (611).
In other preferred embodiments, standard blood dialysis precedes and / or succeeds the steps of the method of the invention. The advantage is to combine conventional blood dialysis as often performed on patients with kidney failure with special blood purification from volatile fatty acids in one procedure.
The respective method for applying a dialyzer / extractor such as for the purification of an ex vivo blood sample from volatile fatty acids includes the additional steps, respectively modifications:
g1) Release of fatty acids esterified in the blood of a subject by hydrolases immobilized on support materials inside the said first chamber, thereby generating a micro- or nanoemulsion;
h) Pumping the filtered solution from said second chamber to a first chamber of a second dialyzer;
63/118
i) Conducting the solution containing carboxylic acid from said first chamber of the second dialyzer to a second chamber of the second dialyzer through a second separation panel, applying a concentration gradient, a chemical gradient, a pneumatic gradient, an electrical gradient or a combination of them;
j) Removal of the associated carboxylic acid and solubulizing compound through the said second separation panel by means of a tertiary circulation;
k) Pumping the carboxylic acid acceptor solution from an acceptor solution storage vessel to said second chamber of the second dialyzer;
l) Removal of the loaded carboxylic acid acceptor solution into a waste container; and
m) Reconduction of the purified solution containing the solubilizing compound from said first chamber of the second dialyzer to the entrance of said second chamber of the first dialyzer.
A phase separation interface consists of porous membranes, gels with or without gaps or tubes with porous walls. The membrane configuration can be flat or round, processed in batches, rows or modules. The tubes can be single or have multiple channels. In a preferred embodiment, hollow chamber capillaries are used. They have diameters between 100 and 300gm, and a length between 200 and 400 mm. The number of hollow chamber capillaries ordered in parallel depends on the desired blood (plasma) flow rate. Typically, the number of capillaries within a dialyzer is between 10,000 and 40,000. The duration of blood (plasma) contact with the capillary wall should be between 2 and 50 seconds.
The interface material can be made up of inorganic or organic materials, or a combination of both. The materials are listed below (see Chapter E, 3. Membranes). A preferred modality is the use of a ceramic, polymeric, metallic or carbon support material. Even more preferred is aluminum oxide and polycarbonate. The material architecture can be symmetrical or asymmetric, as is known in the art. The intercepting channels / spaces / spaces can have a geometric or random configuration. The diameters of the channels can vary considerably, however, they must be in a range that allows for ultra-, micro- or nanofiltration. In
64/118 a preferred modality is the use of a nanofiltration membrane, as previously described (see chapter on nanofiltration methods).
In principle, the same membranes can be used for dialysis, filtration or osmosis. However, dialysis or osmosis membranes need to be more selective, respectively sealed. The use of a gel nested in said support structures is used in another preferred embodiment. Such gels can consist of hydrophilic or organophilic components or both. Gels showing self-assembly and display showing spans or nanostructured channel structures after formation, respectively, solvent extraction is used in a preferred modality.
An extractor used for biological materials, food, waste solution or industrial use may have different dimensions from the components mentioned above, however, the basic assembly is the same.
The concentration of carboxylic acids in solutions considered for analytical processing or purification can vary to a great extent. For optimal solubilization, a ratio of (solubilizing compound: carboxylic acid) 1: 1 to 4: 1, depending on the pH and ionic strength, must be adjusted. A smaller proportion will lead to incomplete solubilization, a larger proportion may interfere with further processing. However, the content of carboxylic acids may be completely unknown. This problem can be overcome by monitoring the turbidity of the aqueous solution, respectively emulsion. Emulsions are cloudy and miniemulsions exhibit turbidity when irradiated with UV light. Micro- and nanoemulsions are optically transparent. However, by means of nephelometric turbidity using unitary particles of multiple beams from 1 to 1000 nm can be detected. Therefore, solubilization progress can be monitored by measuring turbidity. For an individual application, it will be possible to calculate the amount of solubilizing compound that must be added if a defined transparency is achieved to achieve the desired ratio between the solubilizing compound and the carboxylic acids to be solubilized. In the event that the particles are not micelles of a carboxylic acid are present in the solution to be solubilized, it may be advantageous to filter those particles out or to centrifuge them out.
On the other hand, a less sophisticated use of the solubilization process can be used as well. Most solubilizing compounds, such as arginine, exhibit negligible toxicity. In addition, since they are highly soluble in non-physiological concentrations,
65/118 be removed by a dialysis step as known in the art. Therefore, an adjustment of the concentration of solution obtained during the mixing process can be chosen. This concentration should be in the range of 100 to 1000 mmol / l, the setting of an infusion pump providing the mixing segment with the solution of the solubilizing compound can be calculated from the blood flow rate (plasma) and the desired concentration .
A typical scheme for an integrated dialyzer / extractor is shown in Fig.
7. The module consists of a cylindrical cartridge (701). A reaction chamber (702) located on the inlet flow side is separated from the separation chamber (703) by the sealing plane A (704). The reaction chamber can house several systems for mixing fluids. The example shows corrugated lamellae (705). The reaction chamber has a separate flow inlet for the solubilizing compound (712).
A bundle of capillaries (706) comprising hollow membrane capillaries is incorporated at both ends of a sealing member in order to seal them out. Membrane capillaries are incorporated into a sealing compound of the sealing plane A and B (704, 707), opened with their ends towards the reaction chamber (702) and the collection chamber (708), respectively. The sealing planes are sealed so that the separation chamber (703) is separated. Both ends of the cylindrical housing are secured by a cover carrying an inlet / outlet with a connection plug (709). The housing has an additional entrance / exit (710, 711) intersecting the housing wall in the vicinity of both sealing planes that communicate with the separation chamber. The inlet / outlet tubes close on a connection plug (not shown). Housing and sealing material may consist of a polymer such as PU, PA, PE.
Another method for blood purification is hemofiltration. Therefore, the blood to be purified is pressurized by means of a bearing pumping system and a valve / flow limiter downstream of the extractor. Depending on the desired filtration fraction, a transmembrane pressure of up to 500 mm Hg can be adjusted, which is calculated using the formula
P inlet flow * P outflow / 2 ΠΟ Blood side - P inlet flow ⁺ P outflow / 2 on the filtrate side.
However, for industrial applications, higher pressures may be required.
66/118
Stearases can be immobilized, for example, on a composite membrane consisting of a polymeric binder, such as polysulfone, poly (tetrafluoroethylene and poly (vinylidene) fluoride, and metal oxides, such as TiO ₂ , SrO ₂ , HfO ₂ and ThO ₂ (WO 1990/15137) Alternatively, stearases can be covalently bonded to bi- or polyfunctional composites with a phosphate group between the composite and the aforementioned metal oxides (WO 1999/32549).
Esterified carboxylic acids cannot be solubilized directly by the procedure of the invention. In many cases, the hydrolysis of carboxylic acids can be indicated in order to make them suitable for solubilization. This can be achieved by hydrolases, more specifically by stearases, respectively lipases. There is a wide variety of this class of enzymes found in living organisms and plants. For use in blood or plasma, stearases that hydrolyze alkali residues from glycerin are of interest. It may be of interest to hydrolyse only carboxylic acids from mono-, di- or triglycerides using triglycerol hydrolases (EC 3.1.1.3) and to save phospholipids. However, in some situations, the removal of all classes of esterified carboxylic acids could be indicated, which can be carried out by the respective stearases (EC 3.1). In some indications, hydrolysis of certain fatty acids is desirable, for example, trans fatty acids, long chain saturated fatty acids. In general, hydrolysis of long-chain fatty acids (> 12 C atoms) is a preferred embodiment of the procedure of the invention when used for the purification of blood or plasma. Stearases must be immobilized to a support material, so that they cannot leave the reaction chamber. Suitable materials for transporting immobilized enzymes can be coverslips, meshes, membranes, tubes, spheres, ceolites or gels. The enzyme immobilization technique depends on the support material used and is not a subject of the invention. In a p _re f _er e _nC j _a l mode for use in an extractor for medical use, aluminum oxide or titanium oxide is used as a support material, configured as blocks containing tubular spaces with a width between 100 and 500 gm, more preferably between 200 and 400 μτη. Its surface is functionalized with an enzyme. Another preferred modality is the use of microspheres made of PMMA, PEEK, silicon, silicone or other materials. The preferred average diameter is between 100 and 500 gm, more preferably 200 to 400 gm. Its surface is functionalized with an enzyme. In an additional preferred mode,
67/118 carboxylic acids and / or triglycerides are released from phospholipid vesicles that transport those molecules into the blood. A preferred form of application for an extractor is to use materials containing enzymes as a separate unit that can be stored separately from the extractor. The advantage of this modular technique is that, in case it is necessary to store materials containing enzyme at a defined temperature, the space required for storage is reduced. In addition, in the case of loss of enzyme activity during the treatment procedure, this component can be renewed without the need to renew the other components.
Dialysis / extraction procedures - Variant II
In an even more preferred embodiment, the solubilizing compound is immobilized in the separation membrane or in the hollow capillaries. Thus, the dialyzer can be simplified and comprises the following parts:
a) means for conducting the subject's blood to a first cavity chamber;
b) a first cavity chamber
c) optionally, the immobilization of lipases used for the hydrolysis of fatty acids and the release of fatty acids adsorbed or bound to proteins, lipids or cell membranes;
d) a separation panel between the first cavity chamber and a second cavity chamber comprising a separation membrane or a hollow capillary assembly characterized by the fact that the solubilizing compound is immobilized in the separation membrane or inside the hollow capillaries ;
e) means for conducting the reactive solution from the first cavity chamber to the second cavity chamber through the application of a pneumatic gradient, an electric gradient or a combination thereof;
f) a second cavity chamber for receiving the filtrate / dialysate;
g) a tank to collect the purified filtrate / dialysate from the second cavity chamber with the fraction of residual blood from the first cavity chamber; and
h) means to return the collected blood fractions to the subject's circulation.
Applications for industrial use: two-chamber separators
68/118
According to the invention, a device is also provided for the removal of fatty acids from aqueous solutions resulting from the processing of crude oil, processing of industrial foods, in the processing of sewage containing bio-organic compounds or in any other industrial production or environmental technique. For these applications, preferably, a two-chamber system is provided, which comprises (Fig.8)
a) a first container (801) for receiving the aqueous solution containing fatty acids containing carboxylic acids being continuously poured into the container from a feed stream (803);
b) means for adding a solution of solubilizing compound to the first container (804) and mixing a solution with said aqueous solution containing fatty acids by means of an appropriate mixing system (805);
c) a separation membrane between the first container and a second container comprising a separation membrane (807) or a hollow tube or capillary assembly;
d) means for conducting the reactive solution from the first container to a second container through the application of a pneumatic gradient, an electrical gradient through an electric field between the cathode (806) and the anode (808), or a concentration gradient, a chemical gradient, or combinations thereof;
e) a second container (802);
f) means for removing the fatty acid solubilizing compound from the filtrate solution by removing the filtrate by convection of a suitable acceptor solution being fed through an inlet (809) and allowed to flow through an outlet (810 ); and
g) means for removing the purified solution from the first container through an outlet (811).
In another embodiment of this device for the removal of fatty acids from aqueous solutions resulting in the aforementioned industrial processes, the solubilizing compound is immobilized to the separation membrane or hollow tube or assembly of capillaries. Thus, the device can be simplified and comprises the following parts (Fig.9):
a) a first container (905) to receive the aqueous solution containing fatty acids (901) and an aqueous solution of solubilizing compound (902) after
69/118 mixing of both liquids using an appropriate mixing system (903, 904)
b) a separation membrane between the first container and a second container comprising a separation membrane or a hollow capillary assembly (906), optionally having the solubilizing compound immobilized in the separation membrane or inside the hollow tubes or the inner side a spiral module;
c) means for conducting the reactive solution from the first container to a second container by applying a concentration gradient, a chemical gradient, a pneumatic gradient, an electrical gradient or a combination thereof, and
d) a second container (908) receiving the reactive solution that is allowed to leave the chamber through the outlet (909);
e) a collection chamber (907) which is sealed to the first and the second container by a sealing plane (912, 913), being run through the hollow tubes or outer side of spiral modules, and
f) an inlet (911) and an outlet (910) flow from the collection chamber, allowing the perfusion of solutes through the collection chamber.
Another embodiment of the solubilization effect of the invention is the solubilization and separation of carboxylic acids from oils during pharmaceutical, chemical or industrial processing, by means of fluid-fluid separation. Carboxylic acids can be present in the form of an oil, an emulsion (O / W, W / O), or as a fluid / fluid system. Thus, this modality can additionally comprise the following parts:
h) Means for mixing the solution containing carboxylic acid and the solubilizing compound, by means of heating, sonification, turbulent or laminar flow conditions in said first chamber;
i) Means for transferring the mixed emulsion to said second chamber and separating the mixed emulsion by means of gravity or centrifugation;
j) A third chamber to receive the purified oil from the second outlet of the second chamber;
k) A fourth chamber to receive the members of carboxylic acid and the solubilizing compound from the first outlet of the second chamber;
l) A reservoir of water-soluble acid;
70/118
m) Means for suspending said water-soluble acid from said reservoir to said fourth chamber,
n) Means for mixing the solution in said fourth chamber;
o) A fifth chamber suitable for phase separation by gravity, to receive the mixed solution from said fourth chamber;
p) Means for conveying the mixed solution from said fourth chamber to said fifth chamber;
q) A sixth chamber for receiving purified carboxylic acids from said fifth chamber;
r) Means for conducting the purified carboxylic acids from said fifth chamber to said sixth chamber;
s) A seventh chamber to receive the solution containing the solubilizing compound and the water-soluble acid joining at the bottom of said fifth chamber;
t) Means for conducting the solution containing the solubilizing compound and the water-soluble acid to a seventh chamber;
u) Means for conveying the solution from the seventh chamber through an electrodialysis device or an ion exchanger to separate the solubilizing compound into a catholyte chamber and the added water-soluble acid to an anolyte chamber;
v) Means for conducting the solution from the catholyte chamber to said container for the solubilizing compound;
w) Means for conveying the solution from the anolyte chamber to the water-soluble acid reservoir;
x) Means for conducting the purified retinate solution after electrodialysis to a hydrophilic filter membrane; and
y) Means for reusing the aqueous filtrate, and optionally comprising
z) Means of suspension and the mixture of an organic solvent for the solution of the seventh chamber.
The corresponding method for applying a modified integrated dialyzer / extractor comprises the following respectively additional modifying steps:
f) Mix the solution containing carboxylic acid and the solubilizing compound by means of sonification, turbulent or laminar flow conditions in said first chamber;
71/118
g) Transfer the mixed emulsion to a second chamber;
h) Separate said mixed emulsion in said second chamber by means of gravity and centrifugation;
i) Conduct the purified oil through the second outlet of the second chamber to a third chamber;
j) Suspend a water-soluble acid from a reservoir to a fourth chamber;
k) Mix the solution in the fourth chamber;
l) Conduct the mixed solution from the said fourth chamber to a fifth chamber;
m) Perform phase separation by gravity in said fifth chamber with the solution received from the fourth chamber;
n) Conducting the purified carboxylic acids from said fifth chamber to a sixth chamber;
o) Conduct the solution containing the solubilizing compound and the water-soluble acid, joining the bottom of said quanta chamber to a seventh chamber;
p) Suspend and mix an organic solvent for the solution in the seventh chamber;
q) Conduct the solution from the seventh chamber through an electrodialysis device to separate the solubilizing compound into a catholyte chamber and the water-soluble acid added to an anolyte chamber;
r) Perform the electrodialysis in the solution from the seventh chamber;
s) Conducting the solution from the catholyte chamber to said container for the solubilized compound;
t) Conduct the solution from the anolyte chamber to the water-soluble acid reservoir;
u) Conduct the purified retinate solution after electrodialysis to a hydrophilic filter membrane; and
v) Store the aqueous filtrate for reuse.
It is also preferred that the device for the removal of fatty acids from aqueous solutions according to the two previous modalities additionally comprises means for the immobilization of stearases used for the release of fatty acids adsorbed or bound to other compounds present in aqueous solution or not watery.
72/118
Another modality of the solubilization effect of the invention is the solubilization and separation of carboxylic acids from organic solutions consisting of proteins, amino acids and other water-soluble molecules during pharmaceutical, chemical, biological or industrial processing, by means of fluid- fluid. Such a device can be simplified and comprise the following parts (Fig. 10):
a) a first container (1001) for receiving the organic matter / solution containing carboxylic acid (1009);
b) by suspending the solubilizing compound solution from a storage container (1010) for said solution and mixing the solution of the first container with the solubilizing compound solution using a mixing system (1011);
c) means for conveying the mixed solution to a second container (1002)
d) means for mixing the solution from the first container with a solution of CaCI ₂ or other complexing material from a storage container (1003), while suspending it to the second container by means of a pump (1005), which ensures a complete mixing of the two solutions, thus precipitating the carboxylic acids solubilized by the solubilizing compound;
e) means for filtering the purified organic solution upstream, in order to retain the precipitated particles (1006);
d) means for the continuous mechanical removal of the precipitate (1007);
e) means for transferring the precipitate to a third container and washing the transferred precipitate (1004);
f) means for acidifying the precipitate in the third vessel;
g) means for phase separation in the third container by means of organic solvents;
h) means for removing the upper phase of the third container containing the separated carboxylic acids;
i) means for transferring the purified organic solution from the second container (1002), to another container (1008); and
j) means for separating the solubilizing compound by electrodialysis, dialysis, using a cation exchange resin or cation chelation;
Optionally, the following steps can be merged, independently of each other:
73/118 d1) means for adding one or more complexation enhancers, adjusting the pH, and / or by adding organic solvents selected from methanol, chloroform and diethyl ether;
i1) means for performing one or more purification steps in order to remove the organic and / or inorganic matter still present in the purified aqueous organic medium.
E. Materials for using the inventive procedure
1. Phase separation and material separation interfaces
In general, all types of separation materials can be used for the separation of solubilized carboxylic acids according to the procedure of the invention. Since there is a wide field of applications, the phase separation interface must be adapted to the respective condition. In the case of a process in which the size of the solubilized carboxylic acids is smaller than the material, respectively of the compounds to be purified, classic filtration by size exclusion can be performed. The smaller the size difference between the molecules in the solution to be purified and the carboxylic acids, the more micro- and nanofluidic separation techniques must be employed. For its use, the surface properties of the interface determine the effectiveness of the separation. Since the carboxylic acids to be separated may vary according to the various applications, the surface properties required for high effectiveness may differ. In general, micro- or nanofluidic conditions exhibit the best conditions to be separated with a phase separation interface. Therefore, the interface materials can be composite materials consisting of a support material, a binder / filler material and functionalization material, being composed in various combinations. Support materials can be of organic or inorganic origin. Examples are listed in the Separation Membrane Materials section. Binding or filling materials can be organic or inorganic and selected from the list in the Separation Membrane Materials section. Preferred compounds to be functionalized on the interface surface are listed in the Functionalization Surface Materials section.
Surfaces, being in close contact with the standard solution to be purified, may have different needs for surface properties than the phase separation interface. This may be true for applications that are used for blood purification. In order to ensure
74/118 hemocompatibility, the surface in contact with blood or plasma must be covered with materials of known compatibility.
In addition, it may be advisable to immobilize the solubilizing compounds in materials from a reaction area or phase separation interface in order to avoid mixing the solubilizing compounds with the solution to be purified or to reduce the amount of the required solubilizing compound. The same is true for the use of hydrolases when their use is necessary, in combination with the inventive process.
2. Hydrolases
Hydrolases are an important group of enzymes (EC 3). They are able to cleave esters, ethers, peptides, glycosides, acid anhydrides and C-C bonds in a hydrolytic way. An important subgroup of hydrolases are stearases (EC 3.1). Stearases are enzymes that break down an ester bond to an alcohol and an organic acid (saponification). Among stearases, lipases (EC 3.1.1) build an important subgroup. Lipases are enzymes that catalyze the hydrolysis of ester bonds from water-insoluble lipid substrates, most of all triglycerides, into diglycerides, monoglycerides, fatty acids and glycerol. Therefore, lipases are a subclass of stearases. They play important physiological functions in the digestion of lipids in the diet, making the energy stored in these compounds available.
Industrial lipase applications involve lipases from fungi and bacteria that play important roles in human practices as old as yogurt and cheese fermentation. In more modern applications, lipases are used in baking, laundry detergents and even as biocatalysts in converting vegetable oil into fuel.
The immobilization of lipases offers the advantage of facilitating the recovery of the enzyme for reuse. In comparison with immobilization by means of methods such as adsorption or inclusion, covalent immobilization of lipophilic enzymes has the advantage that lipolytic activity cannot be removed by surfactants. It has been shown that lipases can be covalently immobilized on carbon nanotubes, so that they can be used as solid phase catalysts. Another application of lipase immobilization was shown cellulose-based organogels. Other examples of covalent immobilization of lipase include those in micron-sized magnetic spheres, in "sepabeads" and in polyphenylsulfone.
75/118
According to the invention, hydrolases can be used to release fatty acids from mono-, di- or triglycerides in the blood, body tissues, food or fuel and oil processing. In a preferred embodiment, stearases are used. Most preferred are lipases. Most preferred are triglcerol lipase (EC 3.1.1.3), phospholipase A ₂ (EC 3.1.1.4), cholinesterase (EC 3.1.1.8) and lipoprotein lipase (EC 3.1.1.34).
3. Membranes
a) Properties of membranes for use in dialysis
In principle, for membranes classified as micro-membranes, ultrananofilters can be used. The architecture can be symmetrical or asymmetrically porous or compact. They can consist of materials listed in the Support Materials or Polymers section, in the Separation Membrane Materials section. The membranes can be flat or have a hollow tube or fiber configuration. The surface of the transverse pores or channels and / or the interface with the supply current can be functionalized with the substances listed in the Materials for Surface Functionalization section.
In a preferred embodiment, the transmembrane openings consist of cylindrical or flattened channels or tubes with small variation in the diameter of the channel or tube (<20%) highly ordered, intersecting the membrane at right angles to the surface. A preferred embodiment is the use of membranes consisting of perpendicular nanotubes or filter membranes having layers of filter membrane having (double) layers of trapped lipids or structures similar to plasma membranes to seal the membrane surface.
Mass transport of carboxylic acids can be achieved by a concentration gradient, a chemical gradient, a pneumatic gradient, an electrical gradient or combinations thereof. Diffusion methods using a concentration gradient are most commonly used. The diffusion capacity can be increased by using acceptor means exhibiting a higher partition coefficient for the substance to be purified than in the donor solution. In principle, materials with a high affinity to accept organic anions are used in a preferred way. A preferred class are molecules that exhibit amino groups (primary, secondary, tertiary, quaternary), a phosphate or calcium group. The molecules, respectively, their structures, must present a minimum size larger than the lower range of openings (channels) of dialysis membrane or medium. If the size
76/118 is smaller, these molecules can be irreversibly immobilized in a matrix instead. In a preferred embodiment, the crosslinked polyester support functionality, ie, poly (trimethylammonium poly- (acrylamido-N-propyl chloride, poly [(3- (methacryloylamino) propyl] trimethylammonium chloride), is used. the use of macromolecules such as cyclodextrins and proteins, that is, albumin or fatty acid binding proteins. These proteins can be freely solubilized or immobilized on a matrix. The selection of matrix materials depends on the field of application. Materials can consist of solids , fibers, meshes, granules and ceolites. The use of microcounts and ceolites is preferred. Materials may consist of silicon, metals, ceramic materials or polymers. In preferred modalities, aluminum, titanium, silicone, polyacrylates, polylactates, polycarbonates, cellulose and its esters, cellulose acetate, polysulfone (PS), polyethersulfone (PES), polyamide (PA), polyvinylid fluoride ene (PVDF), polyacrylonitrile (PAN), polyetherimide (PEI) and / or polyethercetone (PEEK) are used.
3.1 Membranes with an immobilized solubilizing compound
According to the invention, a separation membrane is also provided, in which a solubilizing compound is immobilized on the membrane surface on the inflow side.
Influx side here means the side of the membrane from which the solution is conducted through the membrane. In dialyzers of the invention, this side refers to the first cavity chamber. In the two-chamber separators of the invention, this refers to the side of the first container.
The solubilizing compound can be immobilized directly to the membrane-forming polymer or can be attached via a binding molecule. Such a binding molecule can be an oligopeptide of 1 to 10 amino acids, or peptides of up to several hundred amino acids. These peptides are covalently linked to arginine and / or other solubilizing compounds. In the event that the solubilizing compound is arginine or a derivative thereof, the immobilized arginine should offer free access to its guanidine group to ensure that the interaction of the invention with a fatty acid can occur.
In a particularly preferred embodiment, arginine is immobilized within the pores of the membrane. Thus, it is ensured that the free fatty acids must pass close to an arginine when conducted through the membrane. This immobilization increases the effectiveness of the purification process.
77/118
Therefore, less arginine should be used. Alternatively, the physical parameters of the dialysis process can be adjusted accordingly. This can be particularly advantageous in blood dialysis, in order to conserve sensitive blood components.
According to the invention, membranes are also provided in which the solubilizing compound is immobilized on the surface of the membrane on the inflow side, as well as within the pores of the membrane.
According to the invention, a hollow capillary is also provided in which the solubilizing compound is immobilized inside the capillary. Depending on the polymer from which the capillary is formed, the solubilizing compound can be immobilized in a similar manner as within a pore of the membrane, as described above. The advantages of this modality have already been discussed in the previous paragraph.
Thus, the present invention also relates to a hollow capillary characterized by the fact that the solubilizing compound is immobilized within the capillary.
3.2 Membranes with immobilized hydrolases
In a particularly preferred embodiment, lipases are additionally immobilized on the inflow side of the separation membrane. Here arginine and / or other solubulizing compounds can be immobilized on the inflow side of the membrane as well, or within the pores of the membrane, or in a combination of both. The advantage of this modality is that additional means are not necessary for the lipases to be immobilized. In addition, the proximity between the lipase-based release of fatty acids to immobilized arginine (or solubilizing compound) increases the likelihood that a free fatty acid will interact with an arginine and / or other solubulizing compounds. As the distance increases, the likelihood is increased that the released fatty acid will re-absorb a hydrophobic structure before interacting with an arginine or a solubilizing compound.
3.3 Separation membrane materials
The following polymers proved to be suitable for use in separation membranes: polyolefins, polyethylene (HDPE, LDPE, LLPE), fluorinated ethylene, ethylene copolymers with butene-1, pentene-1, hexene-1, ethylene and propylene copolymers , EPR-rubber or EPT-rubber (third component with diene ao structure), dicyclopentadiene, ethylidene norbornene, methylene-domethylene hexahydronaphthalin, cis-cis-cyclooctadiene-1,5-hexadiene-1,4, hexyl- (1 - hexene
78/118 methylhexadiene), ethylene-vinyl acetate copolymer, ethylene methacrylic acid copolymer, ethylene-N-vinylcarbazole, methacrylamide-N copolymer, N'methylene-bis (methyl) acrylamide-alkylglycidyl ether, glycidyl (meth) acrylate, polymethacrylate, polyhydroxymethacrylate, styrene-glycidyl methacrylate copolymers, poly-methylpentene, poly (methyl-methacrylethyl-copolymer-methyl-methacryl-acrylate), poly-methyl-copolymer-acrylate, poly-acrylate, poly-acrylamide -mixtures of polyvinylpyrrolidone with crospovidone, ethylene-trifluorethylene, polypropylene, polybutene-1, poly-4- (methylpentene-1), polymethylpentane, polyisobutylene copolymer, isobutylene styrene copolymer, butyl rubber, polystyrene and styrene, polystyrene and styrene , sulfonated styrene, poly- (4-aminostyrene), styrene-acrylonitrile copolymer, styrene-butadiene-acrylonitrile copolymer, acryl copolymer itril-styrene-acrylester, styrene-butadiene copolymer, styrene-divinylbenzol copolymer, maleic acid styrene anhydride copolymer, polydienes in cis-trans configuration, in 1-2 and 3-4, butadiene, isoprene, purified natural rubber, styrene-butadiene copolymer (SBR), three-block polymers (SBS), NBR acrylonitrile-butadiene copolymer, poly- (2,3-dimethylbutadiene copolymer), a three-block copolymer of polybutadiene terminated with cycloaliphatic secondary amines, or - benzal-L-glutamate or polypeptides, or N-carbobenzoxy-L-lysine, poly (alkenamer) -polypentenamer, poly- (l-hexen-methyl-hexadiene), poly-phenylenes, poly- (p-xylylene), acetate polyvinyl, vinyl acetate-vinyl stearate copolymer, vinyl pivalate-vinyl acetate copolymer, vinyl chloride-vinyl acetate copolymer, polyvinyl alcohol, formal polyvinyl, polyvinyl butyral, polyvinyl ether, poly- (N-vinyl -carbazole), poly-N-vinyl-pyrrolidon a, poly- (4-vinyl pyridine), poly (2-vinyl-pyridinium oxide), poly- (2-methyl-5-vinyl-pyridine), butadiene copolymer (2-methyl-5-vinyl-pyridine) polytetrafluoroethylene, copolymer tetrafluoroetilenohexafluoropropileno copolymer, tetrafluoroethylene-perfluoropropilvinil ether, ethylene-tetrafluoroethylene copolymer, trifluoronitrosometano tetrafluoroethylene copolymer, tetrafluoroethylene perfluorometilvinil ether, tetrafluoroethylene copolymer, perfluoro (4-cianobutilvinil ether), poly (trifluoroclorometileno ), trifluorochlorethylene-ethylene copolymer, polyvinylidene fluoride, hexafluoroisobutylene-vinylidene fluoride, polyvinyl fluoride, polyvinyl chloride, impact resistant PVC by mixing ABS, MBS, NBR, dorado PE, FVAC or polyacrylate , Post-chlorinated PVC, copolymer of vinyl acetate-polyvinyl chloride, copolymer of chloride
79/118 vinyl-propylene, vinyl chloride-vinylidene chloride-vinyl chloride-polyvinyl chloride copolymer, vinylidene chloride-acrylonitrile copolymer, polyacrylic acid, acrylic acid-itaconic acid copolymer, acrylic acid-acid copolymer methacrylic, acrylic acid acrylnitrile ester copolymer, acrylic acid ester copolymer-ether-2-chloroethylenovinyl, poly- (1,1dihydroperfluoro-butylacrylate), poly- (3-perfluoromethoxy-1,1-dihydroperfluoropropyl acrylate), polyacrylate -sulfone, polyacrylamines, polyacrylamide, acrylamide-acrylic acid copolymer, acrylamide-maleic acid copolymer, acrylamide-hydroxymethylmethacrylate copolymer, acrylamide-methylamide-copolymer, acrylamide-methylacrylamide copolymer, methylamide-methylacrylamide copolymer methacrylic acid, acrylamideanilino-acrylamide copolymer, acrylamide- (N-acrylol-4-carboxyme) copolymer til-2,2dimethylthiazoline), polymethacrylamide, methacrylic acid-methacrylnitrile copolymer, methacrylic acid-3-fluoro-styrene copolymer, methacrylic acid copolymer 4-fluoro-styrene, methacrylic acid-3-fluoroanilide methacrylic acid copolymer, 3-fluoroanilide or fluoro-styrene or copolymers of methacrylic acid with 3,4-isothiocyanate-styrene, or N-vinyl-pyrrolidone with maleic acid anhydride, or polyvinyl alcohol and polyalyl alcohol, polyacrylonitrile, acrylonitrile-2-vinyl-pyridine, acrylonitrile copolymer, acrylonitrile, acrylate, acrylate and acrylate. copolymer of acrylonitrile-N-vinyl-pyrrolidone, hydroxy groups containing PAN, copolymer of acrylonitrile-vinyl-acetate, copolymer of acrylic acrylonitrile, polyalicylic compounds, polydiall-phthalates, polyitrialyl cyanurate, polyacyanacrylate, polydimethylmethyl and acrylate. methyl methacrylate-lauryl, copolymer of Pacetaminofeniletoximetacrilato-methylmethacrylate copolymer, glicoldimetilmetacrilato-methacrylate, poly-2-hydroxyethyl copolymer, methylmethacrylate-2hidroximetilmetacrilato, metacrilatoglicoldimetacrilato copolymer, poly-2hidroximetilmetacrilato, 2hidroximetilmetacrilato-methylmethacrylate copolymer, glicolmetacrilatoglicoldimetilmetacrilato copolymer, block copolymer and styrene-HEMA grafting, polyΝ, Ν-Ρ, Ρ-oxyphenylenemellitimide, polydiethylene glycol bisalylcarbonate, aliphatic polyethers, polyoxymethylenes, polyoxyethylenes, polyfluoral, polychloral, polyethylene oxides, polytetrahydrofuran, polypropylene oxide, polypropylene oxide, polypropylene oxide propylene-allylglycidyl ether, polyhepichlorohydrin, ethylene oxide-epichlorohydrin copolymer, ethylene-dichloromethyl poly-1,2-dioxide, poly-2,2-bis-oxacyclobutane-chloromethyl, epoxy resins, bis
80/118 phenol-A-diglycidyl ether, epoxidated-formaldehyde phenol, cresol-formaldehyde, resins, cross-links with carboxylic acid anhydrides, amines, such as diethyleneamine, isoforondiamines, 4,4-diaminodiphenyl-methane, aromatic polyethers, oxides of aromatics polyphenylene, polyphenols, phenoxy resins, aliphatic polyesters, polylactide, polyglycolide, ροϋ-β-propionic acid, ροϋ-β-D-hydroxybutyrate, polypivolactone, poly-e-caprolactone, polyethylene glycol adipate, polyethylene sebacate, polyethylene sebacate unsaturated maleic acid anhydride, phthalic acid anhydride, isophthalic acid, terephthalic acid or HET acid with ethylene glycol, 1,2propylene glycol, neopentyl glycol, oxetylated bisphenols or cyclododecanediol, unsaturated polyester resins or vinyl ester resins per copolymerization of unsaturated polyesters with styrene, methacrylate, vinyl monomers, vinyl acetate, methyl methacrylate, polycarbonate, polycarbon bisphenol A act and its derivatives and polyethers, polyesters, segmented bisphenol A polycarbonates and its aliphatic derivatives and polyethers, as well as aliphatic polyesters (see above), modified surface polyethylene terephthalate (PET) glycol grafted with acrylic acid or by partial hydrolysis of the PET surface, polyethylene terephthalate glycol, polyethylene glycol terephthalate-adipate-glycol, polyethylene terephthalate, segmented with polyether blocks and aliphatic polyester blocks and polytetrahydrofuran, poly-p-hydroxybenzoate blocks, acid copolymer hydroquinone-hydroxybenzoic, terephthalic-hydroxybenzoic acid copolymer, hydroxybenzoic acid-p, p-diphenyl, polyvinyl pyrrolidone copolymer, polyvinyl pyrrolidone-maleic anhydride copolymer, alkyd glycerol resins, trimethylpropane, pyridyl acid, pyridyl acetate, trimethylpropane, penta succinic, maleic acid, fumaric acid, adipinic acid and fatty acids from linseed oil, castor oil, soybean oil, coconut oil, polysulfides-aliphatics- (R-Sx-) = sulfur content, aromatic polysulfides, polythio-1,4-phenylene, aromatic polysulfide ether of phenol and thiophene, polyether sulfones, polysulfo-1,4-phenylene, poly-p-phenylenesulfone, polyimines, polyethyleneimine, branched polyethyleneimine, polyalkyleneamine, polyamide, polyhexamethylene adipamide, polyhexamethylene sebaceous amide, polyethylene diameters, dodecane, polyethylene diametre, dodecane, polyethylene dyane , versamides of vegetable oils with diamines and triamines, in polyamide of ω-aminocarboxylic acids with α, β, γ, δ-aminocarboxylic acids or lactams, terephthalic acid-m-aminobenzamide copolymer, polyamide hydrazide, for example, isophthalic acid and polyamide acid amide acid m-aminobenzohydrazide, polypiperazine, for example, fumaric acid and dimethylpiperazine, polybenzimidazois of terephthalic acid and benzene tetramino (substituted), or
81/118 diamino phenyl ethers and dichlorophenyl sulfone (substituted and cyclized), or mphenylene isophthalamide and terephthalamide, polyimides, for example, pyromelitic dianhydride, methoxy-m-phenylene-diamine, pyrrones, for example, pyromelitic dianhydride and benzene diamino, aromatic polyamides, poly-m-phenyleneisophthalamide, poly-p-benzamide, poly-p-phenylene terephthalamide, isophthalic acid m-amino-benzoic acid p-phenylene-diamine-copolymer, poly-4,4'- diphenylsulfone terephthalic acid terephthalic acid and hexamethylenetetramine, terephthalic acid and hexamethylenediamine trimethyl and 2,4,4-trimethyl hexamethylenediamine, from terephthalic acid, and diaminomethylene-norbonene and ε-caprolactam, from isomeric acid and from β-caprolactam to acid and isomeric acid. isophthalic and di-4- (cyclohexyl amino-3-methyl) -methane, from 1,12-diacid decane and 4,4'-dicyclohexylmethane diamine, aromatic polyamides with heterocycles, for example, acid dichloride the dicarboxylic, terephthalic acid and isophthalic acid, with heterocycles of diazinic oxazole, triazole, bitiazole and benzimidazole structures, 3- (p-aminophenyl) 7-amino-2,4- (1H, 3H) -chinazolinindione and isophthalic acid, polyamine acids, polymethyl-L-glutamate, poly-L-glutamic acid, etc., for example, copolipeptides from glutamic acid and leucine, phenylalanine and glutamic acid, glutamic acid and valine, glutamic acid and alanine, leucine and lysine, p -nitro-D, L-phenylalanine and leucine, etc., polyureas from diisocyanates with diamines and ureas, polyurethanes from aliphatics and aromatic diisocyanates and bifunctional and trifunctional hydroxyl polyesters containing (above) and aliphatic polyethers (above) and, optionally , modification with the bifunctional containing amino group, a hydroxyl group and a carboxyl group containing materials containing, for example, hexamethylene diisocyanate, diphenylmethane diisocyanate, toluene diisocyanate 2,4 and 2,6, tol idine diisocyanate, xylylene, glycerin, ethylene glycol, pentaerythrite, 3-dimethylamino-1,2-propanediol and aliphatic carbohydrates, and aromatic dicarboxylic acids and their derivatives, o, m, p-phenylenediamine, benzidine, methylene- bis-o-chloroaniline, p, p'-diamino-diphenylmethane, 1,2-diaminopropane, ethylene diamine, from urea resins and cyclic amino urea, melamine, thiourea, guanidine, urethane, cyanamide, acid amides and formaldehyde, as well as longer aldehydes and ketones, silicones, polydialkylsiloxane, diaryl siloxane and alkyl-aryl siloxanes such as dimethyl-, diethyl-, dipropyl-, diphenyl-, phenylmethyl siloxane, silicones containing functional groups, for example, allyl groups , γ-substituted fluorosilicones containing amino groups and vinyl groups, for example, aminopropyl triethoxysiloxane, 2-carboxy-propyl-methylsiloxane, the polymer block with dimethylsiloxane and polystyrene units or
82/118 polycarbonate blocks, styrene triblock copolymers, butyl acrylate, with α, ω-dihydroxy-polymethylsiloxane, 3,3,3-trifluoro propyl methylsiloxane, avocane (90% silicone and polycarbonate), copolymer blocks of silicone and polycarbonate, hydrophobic polymers with a hydrophilic polymer additive, for example polysulfone, polyvinyl pyrrolidone, cellulose and cellulose derivatives, for example, acetylcellulose, ethylcellulose perfluorobutyryl, perfluoroacetylcellulose, polyamide polyamide polymers, cellulose nitrate, cellulose nitrate, cellulose nitrate regenerated cellulose, cellulose regenerated from viscose cellulose derivatives and the like, agarose, polysaccharides such as carrageenan, dextran, mannan, fructosan, chitin, chitosan (diglycidyl ethyl glycol ether, chitosonEDGE), pectin, glycosamino glycans, starch, glycogen, alginic acids, and all deoxypolysaccharides and halogeno-deoxypolysaccharides and their derivatives, aminodeoxypoli saccharides or sulfhydryl-deoxypolysaccharides and their derivatives, murein, proteins, for example, albumin, gelatin, collagen l-XII, keratin, fibrin, fibrinogen, casein, plasmaproteins, milk proteins, crospovidone, structure proteins from animal tissues and plants, soy proteins, the proteins of the food industry.
The following polymers are preferred for the separation membranes:
silica, silicones, polyolefins, polytetrafluoroethylene, polyester urethane, polyether urethane, polyurethane, polyethylene terephthalates, polymethylpentane, polymethylpentene, polysaccharides, polypeptides, polyethylene, polyester, polystyrene, polysulfonates, polypropylene, polypropylene, polypropylene, polypropylene, polypropylene, polypropylene, polypropylene, polypropylene, polypropylene, polyethylene polyglycolic, polyiortoesters, polyaromatic polyamides, sepharose, carbohydrates, polycarbonate, copolymers of acrylates or methacrylates and polyamides, acrylic acid ester, methacrylic acid ester, acrylic acid amide, methacrylic acid amide, polyacrylic acrylate, glycol acrylate, copolymer ethylene glycol dimethacrylate and glycidyl acrylate or glycidyl methacrylate and / or allyl glycide ether, regenerated cellulose, acetylcellulose, hydrophobic polymers by adding hydrophilic polymers, for example, polyvinyl pyrrolidone polysulfone, derivatives and polymer copolymers above mentioned.
Poly (isohexyl cyanoacrylate) (PIHCA), poly (isobutyl cyanoacrylate) (PIBCA), poly (hexyl cyanoacrylate) (PHCA), poly (butyl cyanoacrylate) (PBCA), poly (2-dimethylamino) ethylmethacrylate ( PDMAEMA), polyimomethylaminoethylmethacrylate (PMMAEMA), poly-N-trimethylaminoethylmethacrylate (PTMAEMC), polyaminoethylmethacrylate (PAEMC), Polyaminoethylmethacrylamide (PAHMAC),
83/118
Polyaminohexyl-methacrylate (PAHMC), Polystyrene (PS), Polyvinylpyrrolidone (PVP), Polyvinylalcohol (PVA), poly (lactic-co-glycolic acid) (PLGA), Polyethylenimine (PEI).
Inorganic materials include, but are not restricted to, metals such as aluminum, iron, magnesium, copper, gold, zirconium, iridium, titanium, zinc, tin, as well as their oxides, silicon and their oxides, as well as silicon complexes such as silicon carbide (SiC), to be used alone or in combination with substances such as silicon nitride, aluminum nitride, molybdenum di-silicide and tungsten carbide, alternatively carbon and its oxides, as well as boron nitride ( BN), boron carbide (B4C).
3.4 Hollow porous tubes or capillaries
In general, all polymeric or ceramic materials as well as carbon tubes are suitable for separable membranes as listed above are also suitable for hollow capillaries.
Materials and dimensions vary according to the various applications.
For medical use
The length of the hollow fibers is between 30 - 500 mm, preferably between 50 and 300 mm. The outer diameter of such a hollow fiber has become 0.1 - 1.5 mm, the inner diameter is 0.01 - 1 mm and the wall thickness of the hollow capillary should be 5 - 200 ym, preferably 15-50 ym.
For industrial use
Hollow fibers or tubes can have a length between 150mm to 2000mm, preferably between 500mm and 1000mm. The outer diameter of such a hollow fiber or tube can be between 1.5 mm and 10 mm, the inner diameter between 1 mm and 4 mm and the thickness of the hollow capillary wall must be 200 ym to 500 ym, 300 um to 400ym preferential .
The walls of hollow capillaries or tubes may contain pores. The porosity of the internal and external surface of hollow capillaries or tubes made of gas-permeable membrane ranges from 10 to 90%. The average pore diameter is in the range of 0 - 5 ym and preferably 0-1.5 ym. Virtually all polymeric materials are suitable for building hollow capillaries or tubes. Especially preferred is polyacrylonitrile. Composite materials made from organogels and polymers are also suitable, as are ceramics, cellulose and combinations of these materials.
4. Materials for surface functionalization
The compounds being more appropriate depend largely on the carboxylic acids that must be separated. One or more compounds can be
84/118 used. In principle, the zeta-potential network of the interface must have a positive or neutral charge and the surface must have organophilic / lipophilic and hydrophobic properties. The compounds can be organic or inorganic, as well as their combinations. They include, but are not restricted to, aliphatic or cyclic hydrocarbons, as well as complex compounds such as cholesterol, cholic acid and its derivatives such as chenodeoxycholic acid and ursodeoxycholic acid, tetraether lipids and their conjugates.
The most preferred are molecules with a cationic charge such as cycloheptatrienyl cation, or with an electrophilic substituent such as iodine or bromine.
Additionally preferred are molecules that exhibit amino groups (primary, secondary, tertiary, quaternary) such as choline, ethanolamine, dimethylamine, triethylamine, betaine and the like.
Additionally preferred are aromatic carbon molecules with 2 or more nitrogen atoms like diazine like imidazole, inidazole, purine, pyrazole, pyrimidine, pyridazine, triazine, like atrazine, simazine, melamine, more specifically 2,4,6-triphenylpyrilio tetrafluoroborate (2 , 4,6-TPPT) and 1,3benzoditiolilio tetrafluoroborato (1,3-BDYT), bromobenzenodiazonio, nitronio, benzoditiolilio and triphenylpirilio tetrafluoroborato.
Additionally preferred compounds are arginine and its derivatives, such as:
5- (diaminomethylideneazanioyl) -2-oxopentanoate known as oxoarginine, (2S) -2-amino-5 - [(N'-methylcarbamimidoyl) amino] pentanoic acid, known as omega-methyl-arginine, 2-amino-5- ( diaminomethylideneamino) -N- (4nitrophenyl) pentanamide, known as arginine-4-nitroanilide; 2-benzamido-5 (diaminomethylideneamino) pentanoic acid, known as benzoyl-L-arginine; (2S) -2 - [[(2S) -2-amino-5- (diaminomethylideneamino) pentanoyl] amino] -5 (diaminomethylideneamino) pentanoic acid, known as arinylarginine, 2S) -2 [[(2S) -2-amino -3-phenylpropanoyl] amino] -5- (diaminomethylideneamino) pentanoic acid, known as phenylalanilarginine; (2S) -2-amino-4 (diaminomethylideneamino) butanoic acid, known as L-norarginine; [1 amino-4- (diaminomethylideneamino) -butyl] -hydroxy-oxophosphanium; (2S) -5- (diaminomethylideneamino) -2 - [(4-hydroxy-4-oxobutanoyl) -amino] pentanoic acid, known as succinyl-L-arginine; (2S) -2-amino-5 [[amino (dimethylamino) methylidene] amino] pentanoic acid, known as N, N-dimethyl-L-arginine; (2S) -2- (3-aminopropanoylamino) -5- (diaminomethylideneamino) pentanoic acid, known as beta-alanyl-L-arginine, 2-amino-5 - [[amino85 / 118 (phosphonoamino) methylidene] amino] pentanoic acid , known as phosphoarginine;
2 - [[(2R) -5- (diaminomethylidenoazanioyl) -1 -oxido-1-oxopentan-2yl] azanioyl] pentanedioate, known as nopaline; 5- (diaminomethylideneamino) 2 - [(1-hydroxy-1-oxopropan-2-yl) amino] pentanoic acid, known as octopine; (2S) -2-amino-5 - [[amino- (hydroxyamino) -methylidene] -amino] pentanoic acid, known as hydroxy arginine, (2S) -2- (2-carboxyethylamine) -5 (diaminomethylideneamino) pentanoic acid, known as L-N2- (2-carboxyethyl) arginine; [(4S) -4-azanioyl-5-hydroxy-5-oxopentyl] - (diaminomethylidene) azanium, known as arginedio; 4- (diaminomethylideneamino) butanamide, known as augmentin; and compounds containing arginine and arginine-related molecules, such as: arginyl-phenylalanine anilide, 2- (4-aminobutyl) guanidine, known as agmantine and its structural analogs, 2- (1-aminobutyl) guanidine, 2- (4aminobutyl) guanidine , 2- (4-aminobutyl) -1-bromoguanidine, 2- (4-aminobutyl) -1 chloroguanidine, 2- (1-amino-propyl) -guanidine, 2- (1-aminopropyl) -1- (diaminomethylidene) guanidine ; 2- (3-aminopropyl) -1 - (diaminomethylene) guanidine; 2- (3aminopropyl) -guanidine; 4-aminobutyl (diaminomethylene) azanium; diaminomethylene [3- (diaminomethylideneamino) propyl] azanium, 2- [3- (diaminomethylideneamino) propyl] guanidine; 4- (diaminomethylideneamino) butanamide, 2- (4-aminobutyl) -1 - (difluoromethyl) -guanidine; [1- (diaminomethylidene) piperidin-1-io-4-yl] methylazanium; [4 (aminomethyl) piperidine-1-io-1-ylidene] methanediamine; 3- (2-amino-ethyl) -2,5-dihydropyrrol-1-carboximidamide, 3- (2-aminoethylsulfanyl) -1 H-1,2,4-triazole-5-amine, 2 [3- (dimethylamino) propyl] guanidine; 3- (2-aminoethyl) -azetidine-1-carboximidamide; 2 (3-aminopropyl) -guanidine; 4- (aminomethyl) cyclohexane-1-carboxyidamide, 2- [2,2bis (sulfanyl) -ethyl] guanidine; 5- (aminomethyl) -thiophene-3-carboxyidamide; diaminomethylidene- [4- (diaminomethylideneazanioyl) butyl] azanium; [amino (diaminomethylideneamino) methylidene] -butylazanium, [amino (butylazanioylidene) methyl] (diamino-methylidene) azanium; butyl (diaminomethylene) azanium;
Additionally phenylalanine and its derivatives such as:
4-guanidinophenylalanine, N-guanyl-DL-phenylalanine, (2S) -2 - [[(2S) -2-amino-5 (diaminomethylideneamino) pentanoyl] amino] -3-phenylpropanoic acid, known as arginylphenylalanine; (2S) -2-amino-3- [4 [(diaminomethylideneamino) methyl] phenyl] propanoic acid; 2-amino-3-phenylpropane hydrazide; and polifemusin I or II.
Additionally guanidine and its derivatives, as
Urea; 2-methylguanidine; 2- [4- [4- (diaminomethylideneamino) phenyl] phenyl] guanidine, 3- (diaminomethylideneamino) -5 - [(diaminomethylidenoamino) methyl] acid
Benzoic 86/118; (2S) -2-amino-3- [4- (diaminomethylideneamino) phenyl] propanoic acid; 2- [2 (azocan-1-yl) ethyl] -guanidine known as sanotensin; N- (diamino-methylidene) -2 (2,6-dichloro-phenyl) acetamide known as guanfacin; 2 - [(3-iodo-phenyl) -methyl] guanidine; 2-methylguanidine; 2-butyl-1- (diaminomethylene) guanidine; 2 - [(E) - [(1 E) -1 (diaminomethylidenehydrazinylidene) propan-2-ylidene] amino] guanidine; 2- (3- (1 Himidazol-5-yl) propyl] -1 - [2 - [(5-methyl-1 H-imid azol-4-yl) methylsulfanyl] -ethyl] water nidine; 2 [ (3-iodanylphenyl) methyl] guanidine; 2- [iodine (phenyl) methyl] guanidine; 2-benzylguanidine; [(EJ-N'-iN'-benzylcarbamimidoilJcarbamimidoillazanio;
benzyl (diaminomethylene) azanium; 2 - [[4 [(diaminomethylideneamino) methyl] phenyl] methyl] guanidine; 4-phenyl-1,4-dihydro-1,3,5triazine-2,6-diamine, 2 - [[4 - [[[amino (diaminomethylideneamino) methylidene] amino] methyl] phenyl] methyl] -1 ( diaminomethylidene) guanidine; 2- (2H-tetrazol-5-yl) guanidine; 4- (5- (4carbamimidoifenoxy) pentoxy] benzenocarboximidamide; 2- [carbamimidoyl (methyl) amino] -acetic acid known as creatinine; 4- [2- (4-carbamimidoylphenyl) iminohydrazinyljbenzenecarboxyidamide; 1-cyano-2-methyl-3- [2 - [(5-methyl-1 Himidazol-4-yl) methylsulfanyl] -ethyl] guanidine hydrochloride; 2 - [(Z) - [(1Z) -1 (diaminomethylidenehydrazinylidene) propan-2-ylidene] amino] guanidine ; 1-N [amino (4-chloroanilino) methylidene] -1-N '- [N' - (4-chlorophenyl) carbamimidoyl] piperazine-
1,4-dicarboximidamide, methylglyoxal bis (guanylhydrazone) (known as mitoguazone);
and biguanidines like
3- (diaminomethylidene) -1,1-dimethylguanidine known as metformin, (1E) -2- [6 - [[amino - [(E) - [amino- (4-chloroanilino) methylidene] amino] methylidene] amino] hexyl ] -1- [amino- (4-chloroanilino) methylidene] guanidine, known as chlorhexidine; dimethyloctadecyl [3 (trimethoxysilyl) propyl] ammonium; octadecyl-guanidinium chloride; 1,10-bis (4-chlorophenyl) 1,3, 5,10,12,14-hexazadispir [5.2.5 ^A 9.2 ^A (6)] hexadeca-2,4,11,13-tetraene-
2,4,11,13-tetramine; 3,5-dimethyl-4-phenyldiazenylpyrazole-1-carboxyidamide hydrochloride; Ν, Ν'-dioctadecyl-guanidinium chloride; 2,2,8,8-tetraalkyl3,4,6,7,8,9-hexahydro-2H-pyrimido- [1,2-a] -pyrimidine; 3- (diaminomethylideneamino) propanoic acid; 2- [5- (diaminomethylidene-amino) pentyl] guanidine; 2- (4- (3aminopropylamino) butyl] guanidine: 2- (diamino-methylidenoamino) acetic acid; 3 (diaminomethylidenoamino) benzoic acid.
Additional amines like
87/118
Butanimidamide; decanimidamide hydrochloride; 4- (4- (4carbamimidoylphenyl) phenyl] benzenocarboximidamide; N, N-dimethyl-N'- (4phenylmethoxyphenyl) metanimidamide.
Additional amino acids, more preferably phenylalanine, isoleucine, leucine, valine, arginine, lysine, histidine, tryptophan, tyrosine, proline.
Additionally peptides consisting of one or more of these amino acids. The most preferred is the RDG-sequence peptide (Arg-AspGlic);
and proteins and macro molecules with known lipophilic properties, such as albumin, fatty acid binding protein, or with fatty acid binding properties such as apolipoproteins, lactoglobulins, casein. Additionally cyclodextrins or porphyrins and the like, and substances like chlorine and corpin.
Amines and polyamines, such as choline (2-hydroxyethyl (trimethyl) azanium); phosphocholine, betaine (2- (trimethylazanioyl) acetate); neostigmine; 2- [2,3-bis [2- (triethylazanioyl) ethoxy] phenoxy] ethyl-triethylazanium triiodide; [(2R) -2,4-dihydroxy-4-oxobutyl] -dimethyl- (trideuteriomethyl) azanium, known as carnitine, 3-hydroxy-4- (trimethyl-azanioyl) butanoate; 4-azanioylbutyl (3azanioylpropyl) azanium, known as spermidine; 3-azanioylpropyl- [4- (3azanioylpropylazanioyl) butyl] azanium, known as gerontine.
Additionally peptide-carboxylate conjugates, such as (2S) -2,5-bis (3-aminopropylamino) -N- [2 (dioctadecylamino) acetyl] pentanamide, known as transfectam; 6-amino-2 [[(2S) -2,5-bis (3-aminopropylamino) pentanoyl] amino] -N, N-dioctadecylhexanamide;
2-amino-6 - [[2- [3- [4- (3-aminopropylamino) butyl-amino] propylamino] acetyl] amino] N, N-dioctadecylhexanamide;
and peptides, such as (2S) -2 - [[2 - [[(2S) -2-amino-5 (diaminomethylideneamino) pentanoyl] amino] acetyl] amino] butanedioic, known as RGD-peptide; (2S) -2 - [[(2S) -2-amino-5- (diaminomethylideneamino) pentanoyl] amino] butanedioic acid, arinine-asparagine-dipeptides, and polypeptides, such as 2 - [[2 - [[2 - [[ 2-amino-5 (diaminomethylidenoamino) pentanoyl] amino] -5 (diaminomethylidenoamino) pentanoyl] amino] -5- (diaminomethylidenoamino) pentanoyl] amino] butanedioic acid, 2 - ((2- (2,6-diaminohexanoylamino) -5 ( diaminomethylidenoamino) pentanoyl] amino] butanedioic acid; 4-amino-2 - [[2
88/118 amino-5- (diaminomethylideneamino) pentanoyl] amino] -4-oxobutanoic acid; 2 - [[6 amino-2 - [[2-amino-5 (diaminomethylidenoamino) pentanoyl] amino] hexanoyl] amino] butanedioic acid, known as timotrinan; 2- [2 - [[2 - [[2-amino-5 (diaminomethylideneamino) pentanoyl] amino] -5- (diaminomethylideneamino) pentanoyl] amino] propanoylamino] butanedioic acid; 3 - [[2-amino-5 (diaminomethylideneamino) pentanoyl] amino] -4 - [(1-hydroxy-1-oxopropan-2-yl) amino] 4-oxobutanoic acid; (2R) -2 - [[(2S) -2-azanioyl-5 (diaminomethylidenoazanioyl) pentanoyl] amino] butanedioate.
Additional proteins such as: albumin, protamine, gelatin, natriuretic peptides.
5. Acceptable / adsorbent molecules / materials
The adsorption process is defined as the adhesion of molecules to a surface. Based on the nature of the connection between the molecule and the adsorption surface, phenomena can be divided into two categories: physiosorption and chemosorption. In physisorption, no chemical bonds are formed and the attraction between the adsorbent and the adsorbate is based only on intermolecular electrostatic forces, such as Van der Waals forces. In chemisorption, adsorbate adheres to the solid under the formation of chemical bonds with the surface.
If the associates of carboxylic acid and the solubilizing compound are adsorbed in a second cavity chamber in a second container, an adsorbent that is suitable to adsorb these associates must be provided.
The compounds and / or materials can be of natural or synthetic origin. Examples for groups of adsorbent materials that can be used in the present invention include, but are not limited to, zeolites, clays, activated carbon, activated alumina, natural and synthetic polymers, alkanes, proteins and silica gels. Absorbent materials can be used in different forms, such as spheres, membranes, fibers or coatings. The combination of different absorbent materials is also possible.
Microporous zeolites are aluminosilicate minerals with a porous structure that can accommodate a wide variety of cations. The selectively shaped properties of zeolites are also the basis for their use in molecular adsorption. They have the ability to preferentially adsorb certain molecules, while excluding others. Examples of zeolites include, but are not limited to, amicite, analcima, barrerite, bellbergite, bikitaite, boggsite, brewsterite,
89/118
Chabazite, clinoptilolita, cowlesite, dasiardita, edingtonite, epistilbite, erionita, faujasita, ferrierita, garronita, gismondina, gmelinita, gobbinsita, gonnardita, goosecreekita, harmotoma, herschelita, heulandita, laumontita, mite ,ita, mita natrolite, offretite, paranatrolite, paulingite, pentasil, perlialite, fillipsite, pollucite, scolecite, sodalite, sodium of the diadite, stelerite, stylbite, tetranatrolite, tomsonite, tschernichite, wairakita, wellsite, willhendersonite and yugawaralite.
Clays are a mineral substance made up of small crystals of silica and alumina. Clay minerals are divided into four main groups: the kaolinite group, the montmorillonite / smectite group, the illite group and the chlorite group. Examples of clays include, but are not limited to, kaolinite, diquite, nacrite, pyrophyllite, talc, vermiculite, sauconite, saponite, nontronite, montmorillonite, illite, amesite, baileichlor, bentonite, chamosite, kaemmererite, cookeite, daffodilite, corundite, daisy , gonierite, nimite, odinite, orthocamosite, penninite, pannantite, ripidolite, sudoita and touringite.
Activated carbon, also called activated charcoal or activated carbon, is a form of carbon that has been processed to become extremely porous and therefore has a very large surface area available for adsorption or chemical reactions. Based on its physical characteristics, it can be distinguished in powder, granular, extruded, impregnated and coated with carbon polymer.
Silica gel, an oxide of the silicon element is a highly porous, partially hydrated form of amorphous silica. Silica occurs naturally, but it can also be prepared synthetically. Crystalline silica is the anhydride of silicic acid and therefore, silica gel is a polymeric form of silicic acid. Also fumed silica such as Aerosil® or phyllosilicates, such as talc, montmorillonite hectorite and can be used, as well as their polymeric coatings.
Also useful are synthetic polymers that are composed of a large number of highly cross-linked microspheres. This macroreticular structure gives it a high surface area and uniform pore size. Examples of synthetic polymers include, but are not limited to, polyacrylate, polyamide, polyester, polycarbonate, polyamide, polystyrene, acrylonitrile-butadiene-styrene, polyacrylonitrile, polybutadiene, poly (butylene terephthalate), poly (ether-sulfone), poly (ether-sulfone), poly (ether-sulphone) ether ether ketone), polyethylene, poly (ethylene glycol), poly (ethylene terephthalate), polypropylene, poly (methyl methacrylate),
90/118 polyetheretherketone, polytetrafluoroethylene, acrylonitrile styrene copolymer resin, poly (trimethylene terephthalate), polyurethane, polyvinyl butyral, polyvinyl chloride, polyvinylidenodifluoride, poly (pyrrolidone vinyl).
Preferred are poly (methyl methacrylate) and polyetheretherketone.
The bonding capacity can be increased by functionalizing the surface of the materials mentioned above. This can be accomplished through molecules that exhibit organophilic properties. This property is shared with the molecules mentioned in the section materials for surface functionalization. They can be used in preferred embodiments.
A preferred class are molecules that exhibit amino (primary, secondary, tertiary, quaternary), calcium or magnesium groups.
6. Organogels
The most common definition of a gel is that of a macroscopic solid, a fluid that has no steady-state flow. In other words, a gel is a solid, gelatinous material that can have properties that range from soft and weak to rigid and resistant. The three-dimensional structural cross-linked network lends a structure to the gel.
Organogels are non-glassy, non-crystalline thermoreversible solid materials, constituted by an organic liquid phase that gelled in a three-dimensional network of cross-links. The properties of an organogel such as elasticity and firmness are determined by the solubility and dimensions of the particles. The liquid organic phase is gelled by gelling agents of low molecular weight, the so-called freezers. Organogels are often based on the self-assembly of organic molecules. Based on their characterization, organogels can be divided into solid matrix and fluid matrix organogels. Solid gels are strong systems, which form a rigid network persistent at a specific temperature, whereas fluid gels are formed by transient networks. If the gels are similar solids, characterized by rigid persistent networks, such as vulcanized rubber, or liquid type, characterized by transitory networks, such as non-vulcanized natural rubber, it can also be used to classify the gels.
Almost all organic solvents can be successfully gelled, as understood by the definition of aliphatic and aromatic hydrocarbons, such as heptane, octane, nonane, decane, benzene, toluene, ethylbenzene,
91/118 gasoline, kerosene, lubricating oils and similar oil fractions, and halogenated hydrocarbons such as methylene chloride, chloroform, carbon, tetrachloride, methylchloroform, perchlorethylene, propylene dichloride, methylene bromide, ethylene dibromide, butyl bromide , allyl chloride and propargyl bromide or chloride. Other types of organic liquids have been gelled, such as minerals and vegetable oils, lecithin, polysiloxanes, paraffins, nematic and liquid crystal crystalline materials, electrolytes, polymerizable liquids and others.
Organic fluids are gelled using low molecular weight organogelers or polymeric freezers. Organogeladores create a three-dimensional network by interweaving nanofibers or nanofibers formed by self-organization through non-covalent interactions, such as hydrogen, van der Waals bridges, π-stacking, and coordination. Due to the nature of the intermolecular interaction, gel formation is a thermo-reversible process.
A certain refrigerator can gel certain solvents. A requirement for the gelation process is a low solubility of the freezer, because a compound with low solubility and inclined to crystallize will most likely form a gel. The solvent is then trapped in the pores of the network and microcrystals of the refrigerator through capillary forces.
The molecules that can act as LMWG (low molecular weight freezer), include fatty acid derivatives, steroid derivatives, anthryl derivatives, refrigerators containing steroids and condensed aromatic rings, amino acid type organogelators, different types of refrigerators and systems two-component.
As LMWG, for example, hydrocarbons, haloalkanes, alcohols, acetone, alkanes, cycloalkanes, hexadecanes, cyclohexanes, toluene, pxylene, cyclohexanone, dichloroethane, DMSO, ethanol, propanol, mineral oil, kerosene, chlorobenzene, rapeseed oil are used. , ethyl acetate, dialkyl phthalates, DMF, DMA, dioxin, silicon oil, CHCI ₃ , CH ₂ CI ₂ , CCI ₄ , aromatic hydrocarbons, pyridine, acetonitrile, ketones, diethyl ether, halogenated alkanes, aliphatic hydrocarbons, aliphatic alcohols, amines aliphatics, methanol, oils, tetralin, dedecane long-chain aliphatic hydrocarbons, cycloalkanes, alkyl laureates, trialkylamines, methacrylates and additional metal soaps, fatty acids, steroids, perfluoroalkylalkanes, waxes, urea derivatives, bi-cyclic urea compounds, terephthalates, colamides, carbohydrates, surfactant twins, amino acid amides, camphoryl thiosemicarbazide, cyclopeptides, amino acid amides, protein such as albumin or acid-binding protein
92/118 fatty, anthranil, anthryl derivatives, phenazines, thiophenes, trimesamides, helicenes, lecithins, hydroxypropylmethylcellulose, nitrocellulose or gelatin, macrocyclic refrigerators, cyclohexylamides, cyclohexanols, cholesterol derivatives, amphiphilic balls, alkaline urates, nucleic acids, guanyl acids bromophenol blue, Congo red deoxycholates, cholates and others.
Organogels are prepared by supplying the refrigerator to the solvent that has to be gelled. This can happen by adding the refrigerator to the solvent, mixing the solution and, if necessary, heating it until a solution is obtained. When cool, gelation occurs (hot gelation). The freezer can also be added to a small amount of solvent and heated until dissolved, which is then added to the solvent, mixed and established (cold gelation).
Organogels are used and have the potential to be used in various applications, such as pharmacy, drug delivery, cosmetics, art conservation and research.
The ability of organogels to build self-organized structures offers great potential in nanotechnology. Scientists have created materials from organic gels that have nanoscale spans interconnected throughout mass formation. They allow the transfer of mass that is managed by capillary forces.
Organogels can also be used as an enzyme immobilization matrix. Lipase have been described as being encapsulated in lecithin and microemulsion-based organogels formulated with hydroxypropylmethyl cellulose or gelatin. The trapped lipase maintained its ability to catalyze esterification reactions.
It is preferable to use organogels exhibiting nanovans with lipophilic properties. An additionally preferred form of an organogel is the preparation of a xerogel.
7. Accepting Solutions
Accepting solutions can be aqueous, organic, or emulsions. Aqueous solutions must contain soluble or immobilized acceptor molecules, as listed in 4. Materials for surface functionalization or acceptor materials as listed in 5. Acceptable / adsorbent molecules / materials.
Organic solutions can comprise alkanes, alkenes, alkynes, carboxylic acids, esters, aldehydes, ketones, aromatic hydrocarbons, mono-, di- or triacylglycerols, silicone oils, organic solvents such as n
93/118 hexane, esters, tetrahydrofuran, ketones, lactones, acetone, acetonitrile, nitromethan, nitroarane, dimethylformamide, alcohol, methylsilanes, octamethylcyclotetrasiloxane, amides such as formamide, triethylamine.
8. Additives for preparations of solubilizing compounds
A number of solubilizing compounds such as arginine protonate in water, which results from its characteristic as a base. The resulting pH depends on the respective concentration. Therefore, this solubilizing compound can induce protein denaturation or cytolysis when used in high concentration. As these solubilizing compounds are mainly of amphiphilic molecules, they tend to aggregate through electrostatic interactions between the negatively charged carboxyl group and the positively charged α-amino group at pH values> 8, thus forming the polymers. Therefore, it may be advisable to protonize the solution of the invention with the solubilizing compound to a certain extent, in order to adjust the pH of the solution. Preferred protonating agents are short-chain, carboxylic acids, hydrochloric acid (HCI), hydrobromic acid (HBr), sulfuric acid, phosphoric acid, formic acid, acetic acid, propionic acid, butyric acid, pentanoic acid, citric acid, acid oxalic, malonic acid, salicylic acid, p-amino-salicylic acid, malic acid, fumaric acid, succinic acid, ascorbic acid, maleic acid, sulfonic acid, phosphonic acid, perchloric acid, nitric acid, gluconic acid, lactic acid, tartaric acid , hydroximalic acid, pyruvic acid, phenylacetic acid, benzoic acid, p-aminobenzoic acid, p-hydroxybenzoic acid, methanesulfonic acid, ethanesulfonic acid, nitrous acid, hydroxyethanesulfonic acid, ethylene sulfonic acid, p-toluene sulfonic acid, naphthylsulphonic acid , camphersulfonic acid, china acid, mandelic acid, o-methylmandel acid ico, hydrogen-benzenesulfonic acid, picric acid, adipic acid, Do-tolyl tartaric acid, tartronic acid, atolluic acid, (o, m, p) -tolenic acid, naphthyl aminosulfonic acid, and other mineral acids and, in particular, acetic acid , citric acid, lactic acid, acetylsalicylic acid and salicylic acid, benzoic acid, ascorbic acid, folic acid, or amino acids such as aspartic acid or glutamic acid.
A preferred pH for solubilizing fatty acids in the medium is between 7.4 and 10.
9. Micro or nanoemulsion
The present invention also relates to microemulsions and nanoemulsions in a hydrophilic solvent and especially in water of a carboxylic acid
94/118 including triacid diacids, as well as carboxylic tetra acids, and especially fatty acids, fatty diacids, fatty tracids, as well as tetra fatty acids and at least one compound solubilizer, wherein said solubilizing compound contains at least one amidino group and / or at least one guanidino group and in which the compound has a partition coefficient between n-octanol and Kow water <6.30. Thus, the microemulsion of the invention and the nanoemulsion of the invention contain at least one carboxylic acid and especially fatty acid and at least one solubilizing compound, as described herein.
The solubilizing compounds are disclosed in detail above and these solubilizing compounds form a microemulsion or a nanoemulsion with the carboxylic acid. The solubilizing compounds have the same number of the preferred carbon atom as described above, the same preferred structure, as described above, are preferably derived from arginine, as described here, have the same preferred pH range for the solubilizing reaction, the same proportion preferred molar carboxylic acid for solubilizing compound and the same preferred reaction conditions as described above for solubilizing compounds in general.
Main characteristics of micro and nanoemulsions as defined in section B.
Essential is the spontaneous molecular self-assembly of the solubilizing substance and the carboxylic acid that is constructed through electrostatic forces as mentioned above. They are composed of dimers that have a low tendency to aggregate to micelles. In nanoemulsions, all carboxylic acids can be linked to a solubilizing substance of the invention creating a monophasic solubilizing emulsion without micelles. Most of the vesicular structures measurable by dynamic light scattering (DLS) were <150nm for an emulsion with a 1: 1 molecular ratio, see also example 12. With a higher proportion of the solubilizing compound the measurable vesicles became smaller . At a 10: 1 ratio, 98% of the vesicles were <2 nm in diameter and no aggregates larger than 25 nm were found. This self-assembly of nanoparticles is a central feature of a nanoemulsion. The nanoemulsion is completely transparent and stable for more than 6 months at temperatures between 20 and 100 ° C. The decrease in pH by the addition of acid (HCI) reduces the solvation capacity, which can be overcome by adding arginine. However, the pH of the
95/118 solution is critical to the arginine's nanoemulsification ability that decreases below pH 8.
An additional central feature of the inventive use of micro or nanoemulsions is the solubilizing / releasing effect due to reduced surface tension. This allows the penetration and flooding of nanometric and capillary spans. The amphiphilic nature of 1: 1 aggregates allows adherence or adsorption to lipophilic or hydrophilic substances, respectively solid, thus altering the interfacial forces of the substances mentioned above, respectively solid and / or allowing partition of the molecules of said micro or nanoemulsions of the invention and / or molecules dissolved in said micro or nanoemulsions of the invention within or in the above mentioned substances, respectively solid, and allowing solvation, liquefaction, detachment or convection of the above mentioned substances, respectively solid, as shown in examples 5, 9 and 10.
A decisive advantage of arginine or a solubilizing compound, according to formula I or II, is that a co-solvent is not necessary to build micro or nanoemulsions. However, the addition of a co-solvent can increase the described solubilizing effect.
Microemulsions and nanoemulsions preferably have a pH value> 7.0 and, more preferably within a pH range of 7.0 to 9.0. However, depending on the medium from which the carboxylic acids are to be separated, the pH values of microemulsions and nanoemulsions up to pH 14 can be obtained, while a pH range between 7.0 and 8.0 is preferably used if the acids carboxylic acids must be removed from the blood. However, if complete solubilization is not achieved, which can be seen if the microemulsion or nanoemulsion is not clear and / or colorless, more solubilizing compound can be added or the pH value can be increased or both possibilities can be increased. be used until a clear and in most cases colorless microemulsion or nanoemulsion is obtained.
In addition, micro or nanoemulsion of carboxylic acids can be used to allow, respectively to increase or decrease, respectively to end their chemical reactions, as known in the art. Liquid non-ionic liquids or amphiphilic surfactants have been used to dissolve carboxylic acids for this purpose. However, the use of arginine or a solubilizing compound according to formula I or II for these changes in chemical reactivity has not been reported so far. It was found that the nano
96/118 emulgation increased the chemical reactions of both the alkyl and carboxyl groups, as shown in example 11. The low ionic strength and high stability of these emulsions are decisive advantages over ionic or non-ionic emulsifiers. In addition, they can be used as a reagent or product-specific emulsifier that can be easily removed from solubilized carboxylic acid as well as from an organic reaction solution (Example 9).
Figures
Fig. 1 is a flow chart of the solubilizing process of the invention and separation techniques.
Fig.2 is an integrated dialyzer / extractor for the purification of blood from volatile fatty acids.
Fig. 3 shows a carboxylic acid exchange module.
Fig.4 is an integrated dialyzer / extractor to solubilize carboxylic acid impurities to be used in the processing of industrial oil.
Fig.5 refers to an electrostatically or electrophoretically activated filtration or diffusion for the separation to be used with the solubilization of the invention.
Fig. 6 shows another modality of an integrated dialyzer / extractor for the purification of blood from volatile fatty acids.
Fig.7 shows a typical integrated dialyzer / extractor.
Fig.8 refers to a modality of an integrated dialyzer / extractor to be used for the processing of crude oil, industrial food processing, in the processing of residues containing bio-organic compounds or in any other industrial production or environmental technique.
Fig.9 shows the modality of a simplified integrated dialyzer / extractor for industrial applications.
Fig.10 refers to an additional modality of a simplified integrated extractor / dialyzer to be used for the solubilization and separation of carboxylic acids from organic solutions consisting of proteins, amino acids and other water-soluble molecules, during pharmaceutical, chemical processing , biological or industrial by means of fluid-fluid separation.
EXAMPLES
Example 1
Investigation of the viability for plasma fatty acid dialysis.
97/118
Non-esterified fatty acids, despite small molecules, are not dialyzable when suspended in an aqueous medium, because they form micelles because of their low CMC and, therefore, are too large to pass conventional micropores. The solubilization of the invention raises the CMC and allows the formation of a nanoemulsion showing a high partition of fatty acids. Thus, solubilized fatty acids are present as anions.
The use of the solubilization of the invention for refinement of plasma from volatile fatty acids was investigated in a model dialysis system. This system consisted of a dialyser cell connected to tubes, two reservoirs and two roller pumps. The dialysis cell consisted of two flat glass hemispheres with flattened margins. The margins of both hemispheres were pressed against a Teflon membrane support by a ferrule thereby sealing the cavities. The dialysis membrane support consisted of two Teflon sealing rings that had a groove and tongue design to take a 47 mm diameter membrane and seal the membrane upwards by compressing both rings together. Each of the glass hemispheres had a perforation at the opposite poles on which a glass funnel was mounted by being connected to the glass of the hemispheres with one end and with its base being directed towards the membrane. The funnel is operated as an inlet. An additional perforation of the hemispheres was connected to a glass tube operating as an outlet. The inlet and outlet were connected with PTFE tubes. The tubing connected to the inlet was intercepted by a silicone tube forming part of the roller pump. The ends of the inlet and outlet pipes were joined in a PTFE reservoir, which had a filling volume of 200 ml. The inlet tubing was interconnected with a Y adapter that had a pressure-type gasket through a pressure sensor wire that was advanced into the dialysis chamber and sealed to the outside. Investigations were performed in duplicate using one of the following membranes: Polycarbonate track-etch, 0.4pm (Satorius, Germany); polyarylethersulfone with a cut of 10,000 Dalton (Gambro, Germany); 40 kD PVDF (Rhône-Poulenc, France); PTFE, 0.05pm (Sartorius, Germany); aluminum oxide (Anodise), 0.02pm (Whatman, USA).
Plasma of blood of human origin was used for the experiments. Oleic acid (technical grade, Sigma, Germany) was added to achieve a 100 mmol solution. The prepared solution was left to dissolve while stirring at 37 ° C for 10 minutes. Then, 100 ml of deionized water or
98/118 arginine solution (0.5 mol / l) was added in a 1: 1 volume ratio. The donor site of the dialysis system was filled with 250 ml of that solution; care was taken to exclude air bubbles. The accepting site of the dialysis system was filled with 250 ml of a 10% albumin solution. The roller pumps transported a volume of 200 ml / min through both hemispheres. Care was taken to maintain an identical pressure within both dialysis chambers, the differences in hemisphere pressures were leveled off by an interconnected valve in the drain piping. Dialysis was performed for 30 minutes each. Sample volumes were removed after filling the system, every 10 minutes from the donor and acceptor. The aqueous samples were transferred in isooctane and dried through a stream of nitrogen. Methylation was performed by adding methanol containing 2% sulfuric acid and heating the sample for 15 minutes at 70 ° C. The samples were resolved in water and isooctane. The organic phase was separated after centrifugation and analyzed by GC.
The analysis showed that when dialysis was performed without arginine, there was no or only small amounts of oleic acid found in the acceptor solutions. In dialysis performed with arginine, there was a proportional increase in the oleic acid content of the accepting solution in relation to the oleic acid concentration in the donor solution and the duration of dialysis. This increase was smaller when hydrophilic membranes, such as cellulose or polyarylethersulfone were used, and higher when hydrophobic membranes were used as PVDF and PTFE membranes.
In an additional configuration the viability of electrodialysis was investigated. For this purpose, the dialysis cell described was modified by placing a platinum mesh in both funnels. They were connected to a high voltage generator (EasyPhor, Biozyme, Germany), by means of sealed wires that were placed through the Y connectors. Dialysis was carried out with the identical membranes as in the previous experiments and under the same conditions, but applying a DC variable with a fixed voltage of 40V. Sample preparation was carried out as described above. Investigations were repeated using stearic acid and linoleic acid. The results found for oleic acid could be confirmed.
Conclusion: Solubilization of fatty acids linked to plasma proteins, using an arginine solution and separation of fatty acids by means of an electrodialysis is feasible.
99/118
Example 2
Investigation of the use of arginine solvation for analysis of plasma fatty acid content
Non-esterified and esterified fatty acids have important functions in physiology. However, a disproportion of critical fatty acids is pathogenic in several diseases such as atherosclerosis, hypertension or diabetes mellitus. Thus, there is a need for its exact determination for the prevention, diagnosis and control of therapy. Routine clinical analysis is only established for the determination of triglycerides. Analytical methods available for the determination of volatile fatty acids in the blood are rare and expensive. The determination of triglycerides is done by default. However, the results can only estimate the actual esterified fatty acid content because the enzyme-released glycerin is determined. As the content of fatty acids esterified with glycerin can vary, the actual concentration is ambiguous. In addition, there is no universal method for characterizing fatty acids according to their classes.
A comparison of 10 blood samples between routine clinical methods for determining the fraction of triglycerides, the analytical procedure of the invention and a reference procedure were performed.
For all measurements, the blood of fasting people was absorbed into a serum Monovette. The samples were left to coagulate for 20 minutes and centrifuged at 3000U / min for 15min. The plasma was separated and homogenized before separating sample volumes in up to 3 Eppendorf tubes. Margarine acid was used as an internal standard in samples used for GC and nano-organogel electrophoresis.
Standard enzymatic procedure for the determination of triglycerides
The clinical procedure was performed by a standard enzymatic assay (Serum Triglyceride Determination Kit, Catalog Number TR0100, SigmaAldrich, USA). The samples were prepared as follows:
Free Glycerol Reagent (0.8 ml) was pipetted into each cuvette and 10 ml (0.01 ml) of water, Standard Glycerol were added. Then, they were mixed gently by inversion, and incubated for 5 minutes at 37 ° C. Initial absorbance (AI) of the blank, standard, and sample at 540 nm against water as a reference was recorded. Reconstituted Triglyceride Reagent (0.2 ml) was added to each cuvette, mixed, and the incubation continued at 37 ° C for an additional 5 minutes. Standard white final absorbance (FA) and sample at 540 nm
100/118 against water as a reference was recorded. The concentrations of glycerol, true triglycerides and total triglycerides in the sample were calculated.
The reaction steps are presented below:
Enzymatic Reactions for Triglyceride Assay
Lipoprotein Lipase
Triglycerides —---------------— ► Glycerol + Fatty acids
ΘΡΟ
G_1_P = O ₂ ----------------— ► DAP + H2O2
POD
H2O2 + 4-AAP + ESPA ----------- ► Quinoneimine tincture + H ₂ O
REAL CONCENTRATION OF SERIOUS TRtGLICERIDES REAL CONCENTRATION OF SERIC TRIGLYCERIDES (equivalent triolein concentration) =
FAa ^ tra - (IAamostka X F)) x Standard Concentration FApaDRÃO - (IAbrancO X F))
Where F = 0.81 / 1.01 = 0.80
Calculations:
Total Triglyceride Concentration in Serum or Plasma:
Total Serum Triglyceride Concentration (equivalent triolein concentration) =
Sample - (FAnRANcn) x Concentration of Standard FApaDRÃO - (FAeRANCO)
Glycerol in Serum or Plasma
Glycerol Concentration (equivalent triolein concentration) IAa ^ tra-IIAwhite) x IApaorão Standard Concentration - (IAbranco)
GC measurements of fatty acids
Samples for GC determination were extracted according to the modified Folch method (Folch, Lees & Sloan Stanley, 1957). In summary, 2 grams of a sample were weighed in a cup; 10 ml of methanol + 0.01% 10 BHT were added, stirred and pelleted for a few minutes. CHCI ₃ (20 ml) was added, the solution was stirred and pelleted for a few hours or overnight in a refrigerator under N2. The solution was filtered twice and 7 ml of KCI was added and filled to 250 ml with methanol. The smaller fraction was then extracted by filtration through Na ₂ SO ₄ into a clean, large centrifuge tube. The solvent is evaporated. Then, the lipid is transferred to a vial
101/118 GC by adding 0.5 ml of hexane + 0.01% BHT into the tube, well stirred in other dissolving lipids, then using Pasteur pipettes all samples were transferred to a flask of GC, GC was performed according to the following protocol: aqueous samples were transferred in isooctane and dried through a nitrogen stream. Methylation was carried out by adding methanol containing 2% sulfuric acid and heating the sample for 15 minutes at 70 ° C. The samples were resolved in water and isooctane. The organic phase was separated after centrifugation and analyzed by GC.
Analysis of fatty acids with the analytical procedure of the invention
The analytical device was built as described above. In summary, a donor chamber and an analytical chamber are separated by an opening for placing the separation medium. The opening edges have joints that allow complete sealing between the chambers and the removable phase separation interface. Chambers for the anolyte and catholyte are separated for the donor and acceptor chambers, respectively, located at the opposite locations to the phase separation interface. Anolyte and catholyte chambers are separated from the donor and acceptor chambers by an ion selective membrane. Platinum and electrolyte chambers were installed in the anolyte and catholyte chambers (Umicor, Germany), which were connected to a high voltage generator (EasyPhor, Biozyme, Germany).
The disposable phase separation interface consisted of a PTFE O-ring with a diameter of 2.5 cm and a depth of 3 mm, which was filled to the margins with a gel-mixing liquid. The organogel was prepared as described elsewhere (Suzuki et al ·. Two-component organogelators based on two L-Amino acids: Effect of Combination of L-Lysine with Various L-Amino Acids on Organogelation behavior, Langmuir, 2009, 25, 8579-8585). After gelation, the two sides of the organogel were covered with a polycarbonate Track-Etched membrane with pore diameters of 1.0 pm (Nucleopore, Whatman, USA), which were affixed to the margins of the O-ring. The phases were mounted firmly in the opening between the donor chambers and accepted and controlled for tightening.
Plasma (1 ml) and 1 ml of an arginine solution (200mmol / l) were vortexed for 2min., Incubated for 5min at 40 ° C and vortexed again for 2min. Three commercial lipases (lipases A1 and A2 were pre-gastric lipases and lipase A3 was a fungal lipase) dissolved in toluene were added and mixed gently for 15min at 37 ° C. The sample was
102/118 weighed before and after 100 pl of pipetting. The pipette volume was poured into the donor chamber of the analytical device. The acceptor chamber was filled with 100 μΙ of acetonitrile. Anolyte and catholyte chambers were filled with a 100 mmol / l arginine solution. DC at 40V potential was applied for 15 minutes. The volume of the acceptor chamber was pipetted and transferred to a probe. The acceptor chamber was washed with 100 μΙ of acetonitrile which was added to the probe volume. Then, the measurement of fatty acids was performed with an FT-NIR MPA spectrometer (Bucker Optic, Germany) From these measurements the fatty acid content of the entire sample was calculated.
Results: The comparison of the results of the three analysis methods shows that (1) the indirect method for the enzymatic determination of triglycerides underestimates the amount of fatty acid content in human blood when compared with both other methods, (2) The results of the process of the GC analytical invention and for the total amount of fatty acids are the same, and (3) the determination of fatty acids with different from the procedure of the analytical invention exhibits a high correlation with the results of GC.
Example 3
Investigation of the purification capacity of arginine and other solubilizing compounds for the refining of crude vegetable oils
Crude vegetable oils contain various amounts of non-esterified fatty acids. As these fatty acids reduce the stability of the oil, they are separated during refining processing to values below 0.5%. There are two methods used in oil processing: saponification and distillation. During these refining processes, changes in the esterified fatty acids can occur. Therefore, it was intended to investigate the applicability of the process of the present invention to solubilize and separate fatty acids not esterified with arginine and to analyze the effects of procedures related to the quality of esterified fatty acids.
Crude soy and rapeseed oils were investigated. For this purpose, 10 liters of crude oil were poured into a 50 liter tank. A 0.5 molar solution of arginine was prepared by adding 871 g of arginine to 10 liters of deionized water. The arginine was left to resolve by slow rotation and heating the solution at 40 ° C for 2 hours. Compliant investigations were
103/118 performed using L-NG-monomethyl-arginine, argininosuccinic acid, Lcanavanine, 2-guanidinoglutaric acid solutions. The solutions were added to the crude oil and the emulsions were stirred for 1 hour while heated to 40 ° C. Then, the emulsions were allowed to settle for 24 hours. Subsequently, the aqueous phases were drained through a hydrophilic sieve that cannot be passed through triglycerides and which was located in the conical bottom of the tank. The aqueous solutions were weighed and samples were taken from the organic and aqueous phases. Samples of the aqueous phases were acidified with sulfuric acid to a pH of approximately 3.
Samples of the organic phase were poured into a sample tube of a titration apparatus. To 5 ml of the organic phase, 20 ml of a mixture of ethanol / hexane (1: 1, v / v) with 3 drops of 1% phenolphthalein in ethanol were added and stirred until the solution was completely clear. Then, the samples were titrated with KOH in ethanol, until a pink tint appears, indicating the amount of non-esterified fatty acids. The fatty acid content was calculated according to the formula
CffA [mmol / l] ⁼ (Vkoh [I] * CKOH [lTIOl / l]) / Vamostra [l] * 1000
Vkoh / Ckoh: volume / concentration of KOH consumed in ethanol Vamostra: volume of sample applied Cffa: concentration of free fatty acids
Samples of edible oil from the same source as crude oil, after industrial purification were analyzed accordingly for their non-esterified fatty acid content.
In addition, samples from organic phases were hydrolyzed and analyzed by GC, as described above.
Results: analyzes showed that the content of non-esterified fatty acids in crude oil, which initially was 2.0 and 2.6%, can be reduced to 0.2 and 0.6%, using the technique of the invention, by means of fluid fluid extraction in combination with the solubilization of the invention. The values found for non-esterified fatty acids were not significantly lower than those after their industrial removal. Additionally, the comparison between triglyceride fatty acids analyzed by GC differed in their content between the extraction methods. In comparison with the aforementioned fatty acids from the organic phase of the fluid-fluid extraction of the oil purified by saponification, they exhibited a lower
104/118 concentration of unsaturated fatty acids, while the fatty acid content with the same number of carbons was practically the same. On the other hand, the fatty acids of the oil purified by distillation exhibited more transisomers of unsaturated fatty acids compared to fluid-fluid extraction, as well as a slightly lower total content of unsaturated fatty acids.
Example 4
Investigation of the ability of arqinina to dissolve larger amounts of carboxylic acids in crude and spent oils.
Crude palm oil has a non-esterified fatty acid content of up to 35%, and spent oils of up to 40%. For commercial use, non-esterified fatty acids must be removed. An aqueous extraction with an arginine solution was performed twice. 101 of each crude oil was mixed with 301 of 0.5 mol / l arginine solution in a 501 tank. The mixture is stirred for 15 minutes and allowed to separate for 5 h. The aqueous phase was drained. Then, the procedure was repeated with 101 µl of a 100 mmol / l arginine solution. Similar experiments were carried out with solutions of 1,1-dimethylbiguanide, Arg-Gly-Asp, NG, NG-dimethylarginine, poly-L-arginine. An analysis of fatty acids was performed according to example 2.
The non-esterified fatty acid content of crude palm oil and spent oil was 33% and 36%, respectively, which were reduced to a final concentration of 0.1 to 0.3%, using arginine for extraction. The other tested compounds showed reductions in final concentrations of 0.2 - 0.5, and 0.4 - 0.9%, respectively.
Conclusion: carboxylic acids can be separated from various oils through aqueous solutions of arginine and comparative compounds, even when present in concentrations.
Example 5
Investigation on the ability of arqinine and other solubilizing compounds to solve carboxylic acids and crude oil ingredients
Fractions of extractable oils from plants and vegetables contain non-triglycerides in various grades that need to be removed in order to purify the oil product. Non-triglycerides include carboxylic acids, pigments, sterols, phospholipids, glycolipids, among others. Of these, amphiphiles such as phospholipids, carboxylic acids and glycolipids are added to vesicles incorporating other non-triglycerides. Amphiphiles join blades and membranes. It was investigated whether the inventive solubilization effect of the compounds of the invention in vesicles of
105/118 amphiphiles containing carboxylic acids is capable of removing carboxylic acids together with amphiphilic compounds complexed with carboxylic acids.
Crude sunflower and corn oils were vigorously stirred with solutions of arginine, 4-guanidinobenzoic acid, cimetidine, polyhexanide and melamin (100 mmol / l, vol 1: 1), respectively. The phases separated spontaneously, resulting in cloudy yellow-olive oil and creamy yellow oil phases. The aqueous phases were acidified with HCI in order to reach a pH of 5. Carboxylic acids were extracted by mixing with n-hexane, which was removed and analyzed according to the methods described above. Results documented the presence of fatty acids in the analytes without a substantial difference in their content between the investigated substances. The remaining aqueous phases were subsequently analyzed by HP-TLC (LiCrosphere). Among the substances that could be identified were: phospholipids, green pigments, tocopherol, phytosterols. The total phosphate content in the oil phases was analyzed according to F-16 (99) DGF. The free acid in the oil was determined by titration with KOH ethanol solution. It was found that more than 90% of non-esterified fatty acids were removed after the first extraction with the investigated substances. The total amount of fatty acids extracted correlated with the calculated amount of fatty acids recovered from the extraction of n-hexane from the aqueous phases.
The phosphate contents of the crude oils were reduced by more than half through the aqueous extraction procedures with the tested substances.
Conclusion: The solubilization effect of the invention of arginine and other solubilizing compounds can be used to release complexed carboxylic acids in solutions or emulsions of oils and amphiphiles that allow their extraction in an aqueous medium. In addition, amphiphilic compounds complexed with carboxylic acids can be solubilized separately (to an aqueous phase), at the same time, to a great extent.
Example 6
Investigation of the capacity of arginine and other solubilizing compounds to release, solve and extract complexed carboxylic acids from solid biological materials.
Most biological materials contain non-esterified carboxylic acids usually complexed with other organic materials. One of these solid materials is rice bran, which contains fatty acids and oils
106/118 up to 35% of its dry weight. Superfine crushed bran was suspended in a 200 mmol / l arginine solution, Νω-nitro-L-arginine, octopine, 2-guanidinoglutaric acid, and agmantin, respectively. The solutions were continuously stirred for 24 hours. The solutions became gray and were very cloudy in all cases. Solid matter was removed by filtration. The aqueous phases that had a pH of between 8-10 with the substances used were extracted with diethyl ether. Then, the aqueous solutions were acidified with ascorbic acid in order to reach a pH of 6. Extractions of non-esterified carboxylic acids and their determination were carried out, as in example 5. The fractions extracted with diethyl ether were dried and weighed. Then, they were resolved and analyzed by HP-TLC, according to example 5. The same was done for the residual aqueous solutions.
Results: The diethyl ether fractions contained triglycerides typically found in rice oils, the content was not significantly different between the substances used. The weight of the triglycerides corresponded to about 10 15% of the dry weight of the rice bran. The n-hexane fractions contained carboxylic acids and in the amphiphilic residual aqueous phases such as phospholipids and glycolipids could be identified, as well as pigments.
Conclusion: carboxylic acids often remain in organic solids that arise during food processing, and which are usually complexed with phospholipids and other lipids. An aqueous solution of arginine or other solubilizing compounds is capable of releasing complexed carboxylic acids, thus reducing the adhesion of phospholipids and other lipids to the solid material, thus allowing its aqueous extraction.
Example 7
Investigation of the ability of arginine to solve and extract aggregated carboxylic acids in mineral oils.
Fossil oils contain significant amounts of carboxylic acids that cause corrosion during oil refining. Therefore, there is a demand to reduce the content of carboxylic acids. The most common carboxylic acid in mineral oils is naphthenic acid. The ability of solubilizing compounds to separate the carboxylic acid content of a crude oil sample was investigated. The oil was a gift from an oil processing company and had a density of 0.85 g / cm ³ . The TAN (total acid number) was determined by KOH titration.
107/118
100 ml of crude oil (containing approximately 40 mmol of carboxylic acids) were mixed with 200 ml of a solution of 300 mmol / l of arginine or L-2-amino-3-guanidinopropionic acid, or L-canavanine in water for 1 hour at 45 ° C. Phases of each of the three samples were separated by centrifugation. After that, the oil phase was mixed with 100 ml of a 100 mmol / l solution in water, again either arginine or L-2-amino-
3-guanidinopropionic, or L-canavanine used in the first stage, for 30 minutes at room temperature and the phases were separated, preferably by centrifugation. Then, the TAN of the oil phase was determined.
Results: The TAN was reduced in all three samples from 1.8 mg KOH / g to 0.16 - 0.3 mg KOH / g through aqueous extraction with the solubilizing compounds.
Conclusion: The TAN of crude oil can be reduced by solubilizing compounds. As naphthenic acid prevails in crude oils, a significant reduction is possible.
Example 8
Investigation on oil stability in the presence of arginine and other solubilizing compounds
Refined sunflower oil was mixed with arginine solutions with varying concentrations and duration. The solutions had concentrations of 100, 200, 300, and 500 mmol of arginine, histidine, H-Cit-OH citrulline, Ν-ω-hydroxy-Lnorarginine, and L-NIL. They were added in a 1: 1 volume ratio. All solutions were vigorously stirred for 60 minutes, and then allowed to separate by sedimentation. One sample for each concentration was analyzed after 3 hours, another after 7 days and yet another after 14 days. The concentrations of fatty acids were determined as described in example 2. At concentrations of 100 and 200 mmol of arginine, the concentrations of fatty acids were the same in all samples. At a concentration of 300 to 500 mmol there was an insignificant increase in concentrations of fatty acids, depending on the concentration of arginine and the duration of exposure.
Conclusion: arginine and other solubilizing compounds do not hydrolyze triglycerides in low or moderate concentrations. However, at higher concentrations, hydrolysis may occur to a small extent.
Example 9
108/118 investigation of the ability of solubilizing compounds to solve and extract oil from carboxylic acids during fuel production.
The production of biodiesel depends on the hydrolysis of esterified carboxylic acids. More generally, hydrolysis is carried out by hydrolases. However, these enzymes are inhibited by their reaction products. Therefore, it is necessary to remove glycerin and carboxylic acids from the active center of the enzymes.
The dialysis apparatus of example 1 was used to test the feasibility and effectiveness of a continuous removal of fatty acids while carrying out the hydrolysis of soy oil triglycerides. Lipase (Novozyme 435) was immobilized according to Lee's method (Lipase Immobilization on Silica Gel Using a Cross-linking Method, DH Lee, CH Park, JM Yeo, and SW Kim, J. Ind. Eng. Chem., Vol 12, No. 5, (2006) 777-782). 150 ml of refined soy oil and 50 ml of either arginine, H-homoarginine-OH, or polyhexanide solution (100 mmol / l) were vigorously stirred and placed inside the reaction chamber. The circulation system constantly circulated the emulsion at a speed that did not allow phase separation. A PTFE filter with an exclusion size of 0.4 pm was mounted on the funnel-shaped outlet of the donor / reaction chamber in order to retain silica granules in the chamber while circulating the reaction solution. The acceptor chamber and the circulation system were filled with a 200 mmol / l solution of the respective compound. A PTFE separation membrane mounted between the reaction / donor and acceptor chambers was used. DC was applied between the reaction chamber and the acceptor chamber during the hydrolysis process, as in example 1. The solution in the acceptor chamber was circulated continuously and samples were taken every 10 minutes. The process was stopped after 30 minutes. 82% of the calculated fatty acid content of triglycerides submitted to hydrolysis was present in the acceptor chamber. The acceptor chamber solution was separated and acidified with HCI to a pH of 4. The fatty acids were separated by extraction of n-hexane. The separated hexane phase (10 ml) was mixed with 2 ml of methanol. Novozyme 435 immobilized on silica as described above was added. The esterification reaction was stopped after 30 minutes by filtering the solution. The solution was stirred vigorously with a 50 mmol / l solution of the solubilizing substance used in the previous step. The organic phase was separated after phase separation and sent to a rotary evaporator for residual methanol, destill and hexane.
11/118
Results: There was a rapid increase in the concentration of fatty acids in the accepting solutions up to a plateau. No mono, di, or triglycerides or glycerin were found in the acceptor solutions. The fatty acids that had passed into the acceptor chamber can be purified from water-soluble anions that have also undergone acidification and solvent extraction.
The purified fatty acids were esterified in non-aqueous medium by immobilized esterases. Unconverted fatty acids were removed by aqueous extraction with the solubilizing compounds. Evaporation of the alcohol and solvent resulted in a highly purified solution of the fatty acid methyl ester 10 with a degree of purity> 98%.
Conclusions: Hydrolysis of fatty acids esterified in an arginine solution is feasible, thus improving the convection of free fatty acids and, thus, the process conditions. Hydrolyzed fatty acids can be further purified by means of solutions of the solubilizing compounds and conducted to a non-aqueous reaction medium for methylesterification (i.e., the formation of a methyl ester). In addition, non-reactive carboxylic acids can be removed in a final purification step using an aqueous solution of solubilizing compounds. Highly purified fatty acid methyl esters were produced after evaporation of the solvent, with no need for distillation of the fatty acid methyl esters.
Example 10
Investigation of the ability to solubilize compounds to resolve substances of low solubility in aqueous solutions.
Many reaction substrates or reaction components must be present in an aqueous medium in an uncomplexed form, especially biological systems. The nano emulsification of substances improved their accessibility to biological transport mechanisms and reactions. However, many amphiphilic carrier systems exhibit biological toxicity or low bio-compatibility. Many of the solubilizing compounds are biocompatible.
Therefore, the emulsifying capacity of micro or nanoemulsions of these compounds and carboxylic acids was investigated in poorly soluble substances (water-soluble mg), such as tetrafenylporphyrin (2 mg / l), sudan red (insoluble), azoxystrobin (6.7mg / l) , co-phthalocyanine (insoluble), chlorprofam (110 35 mg / l).
110/118
The investigated substances were first dissolved in an organic solvent: tetrafenilporphyrin in dichloromethane (50mg / ml), red Sudan in acetone (2mg / ml), azoxystrobin in toluol (50mg / ml), co-phthalocyanine in acetonitrile (2mg / ml), chlorprofame in ethanol (50mg / ml).
Nanoemulsions (50 ml) of oleic acid (50 mmol / l) and arginine (80 mmol), linoleic acid (50 mmol) and L-2-Amino-3-guanidinopropionic acid (100 mmol), and 12-hydroxy-9 acid -octadecen (50 mg) and Νω-nitro-L-arginine (130mmol) were dissolved in organic medium containing the substances completely solubilized. The mixture was vigorously stirred. The organic solvent was evaporated, while slowly stirring at room temperature or at temperatures up to 50 ° C. The respective nanoemulsions were added until the solution became clear or up to a volume of 100 ml. The solutions were analyzed for transparency and residual solids immediately and after 24 hours.
Results: the substances of low water solubility investigated could be emulsified in an aqueous medium due to nano-emulsions after pre-solution in an appropriate organic solvent. Solvent-free nano-emulsified substances resulted in transparent emulsions without residual solid formation.
Conclusion: micro and nanoemulsions of solubilizing compounds and carboxylic acids can establish aqueous micro or nano emulsification of substances with low water solubility, obtaining a biocompatible transport or carrier system for those substances in aqueous media.
Example 11
Investigation of micro or nano emulsification of carboxylic acids to allow alternative chemical reactions
Hydrophobic carboxylic acids need to be dissolved in solvents to allow for many chemical reactions, such as peroxidation or polymerization. As many of these reactions can be carried out in aqueous media, solvent-free procedures would be advantageous. Emulsified nano molecules bring reagents close enough to react. Nano carboxylic acids emulgated with arginine or other solubilizing substances can be used to allow chemical reactions generally carried out in organic solvents to be tested in two experiments.
To prove the ability of microemulsified carboxylic acids to react with peroxides by enzymatic esterification, 200 mmol of 2-ethyl acid
111/118 hexanoic and 4-guanidinobutyric acid (1.5 mol equivalents) were dissolved in a mixture of water and THF at room temperature. Terc. butylhydroperoxide (200 mmol / l) was added and stirred gently. The solution was heated to 45 ° C and a 40 mg suspension of Lipase PS was added to allow a condensation reaction of tert-butylperoxo-2-ethylhexanoate. The reaction was terminated after 1 h. The rate of transmission was calculated by the peroxide fraction still present, as calculated by iodometric analysis. Repeated investigations have produced a transmission rate of 65 - 72%. The experiments were repeated with agmantin and 6-guanidinohexanoic acid. The respective transmission rates on the rates varied between 55-78%.
The nanoemulsion of 50 mmol / l perilla acid and 60 mmol / l arginine in water / THF (9: 1 vol: vol) was mixed with 2. eq. m-CPBA in three aliquots at 25 ° C for 3h, and stirred for another 12h. The reaction mixture was acidified to a pH of 4, extracted 3 times with CHCl ₃ . The organic phase was dried over Na ₂ SO ₄ , evaporated to 10 ml and a GC was measured. GC yield 65%. This procedure was also applied to geranium acid, citronella acid, oleic acid, linoleic acid using arginine, 1,1-dimethylbiguanide and, Ν-ω-hydroxy-L-arginine, as solubilizing compounds. The yield varied between 45-85%.
Conclusion: emulsified nano carboxylic acids using the substances of the invention allow chemical solubilization reactions in aqueous media without the need for a solvent.
Example 12
Investigation of the solubility of various carboxylic acids in an aqueous medium using arginine and other solubilizing compounds.
Various aqueous solubilizing compounds for their potential to solubilize a reference fatty acid, that is, oleic acid.
In the beginning, the influence of pH on the solubility of oleic acid in aqueous medium was tested. These tests were carried out in order to exclude the possibility that the observed solubility of oleic acid was due to pH changes and not mainly due to the interaction of functional groups.
Tests have shown that there is no interaction between oleic acid and the solution in a pH range of 9-12, at room temperature. At pH 13, oleic acid begins to dissolve. At pH 14, the addition of oleic acid led to the formation of a solid precipitate.
In the following tests, oleic acid was mixed with the test substances in H ₂ O. Subsequently, logP and pH were measured and the solubility of the
112/118 test substance in H2O was estimated together with an estimate of the interaction between oleic acid and the test substance according to the scheme shown below.
Solutions of the water-solubilizing compounds were prepared at a concentration of 6 to 600 mmol / l, and preferably approximately 60 mmol / l. For aqueous solutions of the solubilizing compounds 0.833 mol equivalent of oleic acid or a corresponding carboxylic acid was added and mixed. The solutions were allowed to stand for 1 hour and the pH was measured. In the case of incomplete dissolution and a pH below 7, a 1M NaOH solution was added dropwise until the turbidity resolved. Then, the mixtures were stirred or shaken. In order to assess the stability of the micro or nanoemulsion obtained, the solution was heated for 1 hour at 60 ° C and cooled to room temperature. Another part of the clear solution was stored overnight at 4 ° C and then reheated to room temperature. Subsequently, the solutions were analyzed for solid formation, residual oil, viscosity, turbidity and DLS measurements were performed.
In emulsions both micelles and vesicles are formed. Its size and volume can be heterogeneous. In micro and nanoemulsions, only vesicle size visualized agglomerates can prevail. The respective distribution and their relative frequency can be measured by dynamic laser light scattering (DLS), which was carried out on these samples that showed a micro or nano emulsifying solubilization behavior for oleic acid. For DLS measurements a Zetasizer Nano S from Malvern (USA) was used. All measurements were repeated three times undiluted or diluted 1:10 and 1: 100 in water. For the measurements of water viscosity and refractory water index were used. It could be demonstrated that all the solubilizing compounds used caused solubilization of lipophilic carboxylic acids in water.
The Solubilizing Compound column in Table A indicates the form of the solubilizing compound dissolved in water. If, for example, hydrochloride is indicated, the hydrochloride salt has been dissolved in water. However, at the pH used for the solubilization of the carboxylic acid, the solubilizing compound may no longer be in the form of the hydrochloride salt. Thus, the Solubilizing Compound column indicates the starting material, not the active form of the solubilizing compound that is capable of emulsifying the carboxylic acid in the aqueous medium.
113/118
Peak peak by Intensity: the size is determined directly from the measurement without weighing the volume or number of particles.
Peaks are shown corresponding to their percentage in the sample.
Maximum peak proportion: percentage distribution of particle sizes.
Solubility estimate:
X = Compound is completely dissolved (X) = Compound is partially dissolved ((X)) = Compound is not dissolved (visual aspect), but there is no change in pH. Therefore, it is assumed that at least a small part is dissolved.
= Insoluble
Estimated interaction between oleic acid and the test substance:
X = Interaction between oleic acid and the test substance (X) = Interaction between oleic acid and the test substance. There is significantly less oleic acid than water in the solution.
E = Emulsion
S = Formation of a solid precipitate
E, S = The solution becomes cloudy, followed by the formation of a solid precipitate
n.d. = Not determined
114/118
Table A
Solubilizing compound Dissolved (H ₂ O) logP pH Interaction with Oleic Acid Average size per intensity [nm] Part [%] Amino Acids L- acid hydrochloride2-Amino-3guanidinopropionic X - 3,8094 11.05 X AND 113 87.2 L-Arginine X -3,517 10.20 X AND 145 99.0 L-Lysine X -3,424 9.56 X AND 133 95.6 L-NIL X -3,517 9.12 X AND 349 94.1 H-Homoarg-OH X - 4,264 8.66 X AND 107 68.5 Histidine hydrochloride monohydrate X - 4,367 9.30 X AND 647 81.9 Arginine derivatives Ν-ω-Nitro-L-arginine X - 5,2386 11.95 X AND 85.5 46.7 Ν-ω-Hydroxy-Lnorarginine X 3,98584 9.51 X, E, S 325 93.9 D-Arginine methyl ester dihydrochloride X -2,1082 11.35 x, s n.d. n.d. Ν-ω monomethyl-L-arginine acetate X - 4,044 8.49 X AND n.d. n.d. DihydrochlorideNG.NG-Dimethylarginine X - 3,752 9.45 X AND 22.9 87.4 D - (+) - Octopine- 4,0096 6.70 X AND 435 94.7 Argininosuclonic acid disodium salt hydrate X 2.50212 9.93 X AND 721 74.6 LCanavanine free base X 3,98584 8.679.04 (X), s, EX AND n.d n.d
115/118
Derivatives of guanidine carboxylic acid Creatine X - 0.8842 8.3210.01 (X AND(X AND n.d. n.d. Guanidine acetic acid ((x)) - 2,387 6.20 (X), I / O n.d. n.d. 3guanidinopropionic acid X - 3,765 11.71 X AND 153 64.7 4-Guanidinobutyric acid X - 3,443 10.41 X AND n.d. n.d. 4- (4,5-dihydro-1 Himidazol-2-ylamino) butyric acid X - 1,601 8.63 / 11.5 X AND 627 58.9 (S) - (-) - 2guanidinoglutaric acid X -3.1378 10.15 X AND n.d. n.d. 6guanidinohexanoic acid (X) - 3,604 9.01 (X AND n.d. n.d. 4guanidinobenzoic acid hydrochloride (X) 0.6036 11.19 X AND n.d. n.d. Guanidine derivatives Guanidinachlorhydrate X - 1.03 12.71 X AND n.d. n.d. Sulfaguanidine ((X)) - 1,242 8.25 X, S n.d. n.d. Agmatine sulfate X -1,811 10.53 X, S n.d. n.d. 1,3-Di-o-toiyl-guanidine ((X)) 3,008 n.b. X, S 723 73.5 Clothianidin (X) 2.02559 9.81 X AND 153 53.9 N-guanylurea sulfate salt hydrate X 3,541 8.93 X AND n.d. n.d. Cimetidine ((X)) 0.19022 9.54 X AND 239 67.2Derivatives of Biguanidlna 1- (o-Tolyl) biguanide (X) 1,414 11.53 X AND 850 95.5 Chlorhexidine diacetate (X) 6.18 6.51 x, s n.d. n.d. Chlorhexidine diacetate (X) 6.18 9.58 X, s n.d. n.d. 1,1dimethylbiguanide hydrochloride (97%) X -1,633 9.20 Χ.Ε 106 89.4
116/118
Proguanil hydrochloride (X) 2,532 7.66 X, E n.d. n.d. Polymers(** = P log values are calculated on the basis of monomer units) Polyhexanide -5% (2300 bis 3100 MW) ___________ X 2.78 ** 5.61 X, s n.d. n.d. Polyhexanide -5% (2300 to 3100 MW) X 2,798 ** 8.61 X, S n.d. n.d. Poly-L-arginine HCI(70,000-150,000MW) X -1.32 ** 11.37 X, S n.d. n.d. Other compounds Decreased acetate(basic) X 0.90535 8.07 X, s, (E) 426 60.7 Comparative compound DL-N-Acetylomocysteine thiolactone (basic) X - 0.4908 10.70 - n.d. n.d. Melamine (X) - 0.96075 8.25 X, s n.d. n.d. 4- (4,6-Diamino-2,2dimethyl-2H- [1,3,5] triazine1-yl (x) 0.2952 11.28 X, E, S n.d. n.d. Comparative compound urea X -3 8.59 RT: -60 ° C: ((X)) 341 96.3 Imidazole X -0.67 9.50 X, I / O 244 84.6 Methylimidazole X -0.43 8.90 X, I / O 224 96.7 Di, tri, and tetrapeptides Tyr-Arg (kiotorphin acetate) X - 4,0044 8.54 X AND n.d. n.d. Arg-GIn Hydrochloride X - 6,5634 8.15 X AND n.d. n.d. Gly-Arg __________________ X - 5.0644 9.18 X AND n.d. n.d. Arg-Phe X - 3.3374 8.39 X, I / O n.d. n.d. Arg-Glu X -7.9134 8.83 X AND 191 97.7 Lys-Arg acetate X - 5.0554 11.93 X AND 254 89.9 His-Arg X - 5.7384 8.39 X, I / O 265 69.0 Arg-Gly-Asp (RGD) X - 8.2394 6.04 (X AND n.d. n.d. Ara-GIv-Asp (RGD) - X - 8.2394 8.25 X AND 220 96.
117/118
basic9 Arg-Phe-Ala acetate X - 3,595 8.25 X AND 223 90.0 Thr-Lys-Pro-Arg (Tuftsina) X - 3,982 8.68 X AND 174 96.3 Gly-Gly-Tyr-Arg X - 6.77304 8.57 X AND n.d. n.d. Others Comparative compoundAminophenylacetic acid (X) 0.73 9.23 ((X)) n.d. n.d. Comparative compound L-Glutamine (X) -2.05 8.37 ((X)) n.d. n.d. Comparative compoundGly-Phe X -0.65 7.25 ((X)) 834 86.3 Comparative compound4-Aminobutyric acid X -0.82 8.57 ((X)) n.d. n.d. Gly-His X -2.42 12.1 X AND n.d. n.d.
The following solubilizing compounds (A) to (T).
L-arginine (A), L-2-amino-3-guanidinopropionic acid (B), L-NIL (C), Ν-ω-nitro-Larginine (D), NG.NG-dimethylarginine (E), agmatine ( F), 1,1-dimethylbiguanide (G), Lcanavanin (H), argininosuccinic acid (I), octopine (J), ηω-monomethyl-L-argenine (K), arginine methyl (L), Ν-ω-hydroxy -L-arginine (M), histidine (N), Hhomoarginine-OH (O), L-2-amino-3-guanidinopropionic (P), 6guanidinohexanoic acid (Q), Ν-ω-hydroxy-L-norarginine (R ), 4guanidinobenzoic acid (S) and polyhexanide (T)
They were each used to solubilize the following carboxylic acids (I) to ():
Hexadecaoic acid (I), eicosanoic acid (II), stearic acid (III), docosapentaenoic acid (IV), benzoic acid (V), caffeic acid (VI), terephthalic acid (VII), naphthenic acid (VIII), perfluorooctanoic acid (IX), eicosapentaenoic acid (20: 5) (X), linolenic acid (18: 3) (XI) and docosapentaenoic acid (22: 5) (XII).
All carboxylic acids could be solubilized with the respective solubilizing compound used in the 1.5 mol equivalents at pH values between 8 and 10 as shown in Table B below:
Table Β: X: compound completely dissolved (X): compound partially therefrom
(I) (II) í !!! l (IV) (V) IYÜ (VII) (saw) (IX) IX) (XI) XXHL
118/118
(THE) X X X X (X) X X X X X X X (B) X X (X) X X X X X X X X X (Ç) X X X (X) X X X X X X X X (D) X X X X X X X X X X X X (AND) X X X X X X X X X X X X (F) X X X X X X X X X X X X (G) X X X X X X X X X X X X (H) X X X X X X X X X X X X (I) X X X X X X X X X X X X (J) X X X X X X X X X X X X (K) X X X X X X X X X X X X (L) X X X X X X X X X X X X (M) X X X X X X (X) X X X X X (N) X X X (X) X X X X X X X X (0) X X X X X X X X X X X X (P) X X X X X X X X X X X X (Q) X X X X X X X X X (X) X X (R) X X X X X X X X X X X X (S) X X X X X X X X X X X XX X X X X X X X X X X X
1/5

权利要求:
Claims (5)
[1]
1. Method for solubilizing and separating carboxylic acids with a solubilizing compound from aqueous or organic solutions, emulsions, suspensions resulting from medical therapy, medical analysis, food analysis, food processing, oil processing, oil analysis, fuel processing, modulation of chemical or physicochemical reactions, solubilization of poorly solvable molecules, analyzes in the pharmaceutical or chemical industry or science, removal of carboxylic acids from private sewage, commercial or industrial cleaning, removal of carboxylic acids from biological reactor processes , organogelation or nanoemulsification of carboxylic acids, in which said solubilizing compound contains at least one amidino group and / or at least one guanidino group and in which the solubilizing compound has a division coefficient between n-octanol and Kow water <6.30 , the method characterized by the fact that it comprises the following steps:
i) Provide the solution, or emulsion or suspension containing the carboxylic acids;
ii) Addition of at least equimolar amounts of at least one solubilizing compound;
iii) Separate the solubilized carboxylic acids from the solution, or emulsion or suspension by phase separation, filtration, nanofiltration, dialysis, absorption, complexation, electrophoresis, evaporation, distillation and / or extraction.
[2]
2/5 electrical or a combination thereof, where the phase separation interface consists of a gel, an organogel or a solid material or a combination thereof; or filter the carboxylic acids using at least one solubilizing compound; or nanofiltrate the carboxylic acids using at least one
solubilizing compound; ordialysing carboxylic acids using at least one solubilizer; oradsorb carboxylic acids using at least one solubilizer; orcomplex carboxylic acids using at least one solubilizer; ordistill carboxylic acids using at least one solubilizer; orseparate the carboxylic acids using at least one solubilizer by extracting supercritical fluid. compoundcompound compound compound
2. Method according to claim 1, characterized by the fact that step iii) is achieved by means of one of the following separation methods or a combination of them:
pass the carboxylic acids separately or together with at least one solubilizing compound through a separation membrane, or through a hollow capillary tube or assembly by applying a concentration gradient, a thermal gradient, a physical-chemical gradient, a pneumatic gradient, a electric gradient or a combination thereof; or perform phase separation by combining two or more phase separations of media construction; or passing the carboxylic acids together with at least one solubilizing compound through a phase separation interface that allows the passage of said carboxylic acids and said at least one solubilizing compound by applying a concentration gradient, a thermal gradient, a physical gradient chemical, a pneumatic gradient, a gradient
Petition 870190048064, of 05/22/2019, p. 9/13
[3]
3/5
f) Add water and / or a co-solvent to the solution; and / or
g) Optimize the reaction conditions by heating and / or mixing the solution, thus generating an improved micro or nanoemulsion.
3. Method, according to claim 1, characterized by the fact that it comprises the following steps:
a) Prepare this solution by reducing ionic strength by means of complexation, adsorption, separation or dialysis of cations and anions bound and unalloyed;
b) Adjust the pH of the solution by adding an acid or a base;
c1) Adjust the molarity of the solubilizing compound to be in the range of 1:10 to 20: 1 compared to the estimated concentration of the carboxylic acids to be solubilized; and
d) Add said solubilizing compound in a solid form or in a solution to said aqueous or organic solution containing carboxylic acid to generate a micro or nanoemulsion;
optionally comprising any one of the steps of a1) release of linked carboxylic acids by complexation or covalent bonding c2) If the solubilizing compound is administered in a solution, adjust the pH of that solution to optimize compatibility and reaction conditions with the acids carboxylics to be solubilized by acidification or alkanization;
e) Add esterases, hydrolases or a complex builder;
Petition 870190048064, of 05/22/2019, p. 10/13
[4]
4/5
k) Pump the carboxylic acid acceptor solution from an acceptor solution storage container in said second chamber of the second dialyzer;
l) Remove the loaded carboxylic acid acceptor solution in a waste container; and
m) Bring the purified solution containing the solubilizing compound back from said first chamber of the second dialyzer to the entrance of said second chamber of the first dialyzer.
6. Method, according to claim 5, characterized by the fact that the effect is achieved by means of dialysis, hemofiltration, hemoperfusion, centrifugal plasma separation, plasmapheresis, cascade filtration and thermo filtration.
Method according to claim 6, characterized by the fact that the separation efficiency is increased by additional hydrolysis of esterified fatty acids and / or by reinforcement of lipolysis.
8. Method according to any one of claims 1 to 6, characterized in that the solution to be purified originates from plants, organisms, fossil materials, natural or synthetic reaction mixtures.
9. Method according to any one of claims 1 to 8, characterized by the fact that the use of said solubilizing compound leads to the micro or nanoemulsion of said carboxylic acids and allows their separation through complexation, adsorption, absorption, diffusion , osmosis, dialysis, filtration, nanofiltration, distillation, fluid-fluid extraction or supercritical fluid extraction using a concentration gradient, a thermal gradient, an electrical gradient, a physical chemical gradient or a combination thereof.
Method according to any one of claims 1 to 9, characterized in that the at least one solubilizing compound is added to an emulsion, solution or suspension containing the carboxylic acids in order to use said emulsion to release, decompress, remove, react, aggregate, complex, coagulate, flocculate, sediment or separate complexes containing carboxylic acid.
11. Method according to claim 9 or 10, characterized by the fact that micro or nanoemulsions are used to decrease a physical-chemical or chemical reaction, to allow, to increase the absorption and transport of reaction products or components in biological or chemical reaction processes, to remove, solubilize, release, convect, transport substances
Petition 870190048064, of 05/22/2019, p. 12/13
4. Method, according to claim 3, characterized by the fact that it additionally comprises the steps, after step g), of:
g2) Conduct the reactive solution from a first chamber to a second chamber through a separation panel using a nanofiltration technique applying a concentration gradient, a chemical gradient, a pneumatic gradient, an electrical gradient or a combination thereof;
optionally comprising the steps of
h) Remove the carboxylic acid associates and solubilize the compound from the filtered solution through the convection of an acceptor solution being carried through an inlet into said second chamber and allowed to flow through an outlet of said second chamber; and
i) Remove the purified solution from the second chamber via an additional outlet.
5. Method according to claim 3, characterized by the fact that the aqueous solution is a subject's ex vivo blood sample from which fatty acid micro and / or blood nanoemulsions must be removed, further comprising , the following steps g1) to m) after step g):
g1) Release fatty acids esterified in the blood of a subject by hydrolases immobilized in support materials within the said first chamber, thus generating a micro or nanoemulsion;
h) Pump the filtered solution from said second chamber to a first chamber of a second dialyzer;
i) Conduct the solution containing carboxylic acid from said first chamber of the second dialyzer to a second chamber of the second dialyzer through a second separation panel applying a concentration gradient, a chemical gradient, a pneumatic gradient, an electrical gradient or a combination thereof;
j) Remove the associates of carboxylic acid and solubilizing compound by passing through said second separation panel through a tertiary circulation;
Petition 870190048064, of 05/22/2019, p. 11/13
[5]
5/5 through the absorption of vesicles, or allow or increase the penetration of emulsified carboxylic acids through hydrophilic or amphiphilic media or solids.

类似技术:

---

公开号 | 公开日 | 专利标题

---

BR112012032341B1|2020-01-28|method for solubilizing and separating carboxylic acids with a solubilizing compound

---

Nacka et al.2001|Physical and chemical stability of marine lipid-based liposomes under acid conditions

---

EP3592360A1|2020-01-15|Acefapc for the treatment of acetylcholine-dependent diseases

---

Bordi et al.2003|Charged lipid monolayers at the air–solution interface: coupling to polyelectrolytes

---

OA16283A|2015-04-24|Device and method for solubilizing, separation, removing and reacting carboxylic acids in oils, fats, aqueous or organic solutions by means of micro- or nanoemulsification.

---

US20210128810A1|2021-05-06|Cleaning of biological fluid

---

FR2631236A1|1989-11-17|ACTIVE AGENT SYSTEM FOR LIPID EXCHANGE WITH TARGET STRUCTURES, PROCESS FOR PREPARING AND USING THE SAME, AND PRODUCTS CONTAINING THE SAME

---

JPWO2013047263A1|2015-03-26|Hemoglobin-containing liposome and method for producing the same

---

CA2123585A1|1993-05-27|Polar lipidic composition from plants

---

JP6117789B2|2017-04-19|Lipophilic dopamine derivatives and uses thereof

---

JP2007106679A|2007-04-26|Accelerator for blood coagulation reaction

---

JP6552526B2|2019-07-31|Homogeneous formulation for oral administration comprising omega-3 polyunsaturated fatty acid and resveratrol

---

Anton2006|Recent advances concerning the functional properties of egg yolk low-density lipoproteins

---

JP2007091599A|2007-04-12|Blood coagulation reaction promoter

---

BR112014029492B1|2020-12-08|method for solubilizing and separating one or a plurality of carboxylic acids and use of an immobilized solubilizing compound

---

Kaneda et al.2013|Liposome-Encapsulated Hemoglobin as an Artificial Oxygen Carrier: Technological Features, Manufacturing and Issues for Practical Application

---

JP2008024646A|2008-02-07|Liposome suspension and method for producing the same

同族专利:

---

公开号 | 公开日

---

NZ603821A|2014-10-31|

---

CY1117737T1|2017-05-17|

---

MX339922B|2016-06-17|

---

US20130090488A1|2013-04-11|

---

MY160966A|2017-03-31|

---

PT2585420T|2016-07-15|

---

HUE028700T2|2016-12-28|

---

EP2585420A2|2013-05-01|

---

EP3042718A1|2016-07-13|

---

MX2012014763A|2013-10-04|

---

AU2011269269A1|2012-12-13|

---

EP2399885A1|2011-12-28|

---

WO2011160857A3|2012-02-16|

---

UA113497C2|2017-02-10|

---

SG185563A1|2012-12-28|

---

PL2585420T3|2016-09-30|

---

EP2585420B1|2016-04-06|

---

AU2011269269B2|2016-10-13|

---

ZA201208641B|2014-04-30|

---

CN103038195A|2013-04-10|

---

US9127233B2|2015-09-08|

---

WO2011160857A4|2012-04-05|

---

JP6215051B2|2017-10-18|

---

JP2013538106A|2013-10-10|

---

DK2585420T3|2016-07-18|

---

ES2582094T3|2016-09-09|

---

CA2801624A1|2011-12-29|

---

AP3267A|2015-05-31|

---

IL223104D0|2013-02-03|

---

KR102002113B1|2019-07-19|

---

KR20130087395A|2013-08-06|

---

AP2013006668A0|2013-01-31|

---

CN103038195B|2016-04-13|

---

WO2011160857A2|2011-12-29|

---

BR112012032341A2|2016-11-08|

---

CL2012003567A1|2013-11-08|

---

US20160122686A1|2016-05-05|

---

TN2012000573A1|2014-04-01|

---

SMT201600201B|2016-08-31|

---

CA2801624C|2019-10-29|

---

RU2013102533A|2014-07-27|

---

RU2581368C2|2016-04-20|

引用文献:

---

公开号 | 申请日 | 公开日 | 申请人 | 专利标题

---

---

US2945049A|1959-05-25|1960-07-12|Gen Mills Inc|Salts of basic amino acids and linoleic acid|

---

JPS538769B2|1974-10-14|1978-03-31|

---

JP2532042B2|1988-10-20|1996-09-11|ダイセル化学工業株式会社|Organic acid recovery method|

---

EP0430947A1|1989-06-02|1991-06-12|Fina Research S.A.|Enzymes immobilised on microporous composite membranes, activated supports prepared therewith, and uses thereof|

---

DE19756276A1|1997-12-18|1999-06-24|Basf Ag|Ethylene homo- and co-polymers, stabilized by addition of quinoline or diphenylamine derivatives, useful for storage and transport of vegetable oil esters|

---

EP1362605A1|2002-05-14|2003-11-19|Bernhard Dr. Kreymann|Dialysis apparatus to remove protein bound substances|

---

US6777213B2|2002-10-29|2004-08-17|Cognis Corporation|Isolation of carboxylic acids from fermentation broth|

---

BRPI0616877A2|2005-10-05|2011-07-05|Tokyo Cro Inc|biocompatible block copolymer, micelle, particulate, composite, pharmaceutical composition, method for making a micelle, particulate or composite, and use of a biocompatible block copolymer|

---

JP2009530100A|2006-03-20|2009-08-27|ビー．ピー．ティー．−バイオピュアテクノロジエルティーディー．|Hybrid membrane module, system and process for treating industrial wastewater|DE102012104668A1|2012-05-30|2013-12-05|Gea Mechanical Equipment Gmbh|A method of solubilizing and separating one or more carboxylic acids and using a demobilized solubilizing compound|

---

CN103254225B|2013-05-08|2015-10-28|浙江大学|A kind of method adopting ion liquid abstraction separating and purifying phosphatidyl choline|

---

DE102013012671A1|2013-07-31|2015-02-05|Mann + Hummel Gmbh|Ceramic hollow fiber membranes with improved mechanical properties|

---

CA2950269A1|2014-05-28|2015-12-03|The Board Of Regents Of The University Of Texas System|Novel compounds supports hematopoietic stem cells and red blood cells|

---

AU2015265869B2|2014-05-28|2018-11-08|Drei Lilien Pvg Gmbh & Co. Kg|Method for refining lipid phases, and use|

---

PL3149133T3|2014-05-30|2019-08-30|Drei Lilien Pvg GmbH&Co. KG|Method for purifying refined lipid phases|

---

US9994464B2|2014-06-03|2018-06-12|Drei Lilien Pvg Gmbh & Co. Kg|Method and devices for de-emulsifying and complexing organic compounds in emulsions|

---

DE102014210662A1|2014-06-04|2015-12-17|Gea Westfalia Separator Group Gmbh|Apparatus and method for obtaining glycoglycerolipids and glycosphingolipids from lipoid phases|

---

EP2952565A1|2014-06-04|2015-12-09|Nanoscience for life GmbH & Co. KG|Device and method for recovering glycoglycerolipids and glycosphingolipids from lipid phases|

---

CN104497684B|2014-12-16|2016-08-31|中国海洋石油总公司|A kind of polyurea type anti-settling agent|

---

US20170056352A1|2015-08-25|2017-03-02|Rgenix, Inc.|PHARMACEUTICALLY ACCEPTABLE SALTS OF beta-GUANIDINOPROPIONIC ACID WITH IMPROVED PROPERTIES AND USES THEREOF|

---

WO2017173382A1|2016-03-31|2017-10-05|ImMutriX Therapeutics, Inc.|Selective adsorbent of disease mediators|

---

KR101742295B1|2016-05-26|2017-06-01|한국에너지기술연구원|Calcium removal method from hydrocarbon fractions using extraction comprising 2-oxopropanal or derivatives thereof|

---

KR101829051B1|2016-10-31|2018-02-13|주식회사 삼양사|Thick hydrogel sheet with high adhesive power|

---

CN106592010B|2016-12-16|2018-09-21|浙江华峰氨纶股份有限公司|A kind of preparation method and applications of polyurethane elastomeric fiber|

---

CN106823451B|2017-02-27|2020-12-18|山西省食品研究所|Increase of CO2Charging process for supercritical extraction yield|

---

WO2018160178A1|2017-03-01|2018-09-07|Rgenix, Inc.|Pharmaceutically acceptable salts of b-guanidinopropionic acid with improved properties and uses thereof|

---

US10967352B1|2017-04-07|2021-04-06|Triad National Security, Llc|Microfluidic liquid-liquid contactor|

---

CN108096236B|2018-01-04|2019-10-25|武汉大学|Application of the leonurine in the drug of preparation treatment urarthritis|

---

CN108226150A|2018-01-11|2018-06-29|常州市五洲化工有限公司|A kind of method of quick measure terephthaldehyde acid content|

---

CN110927261A|2018-09-19|2020-03-27|内蒙古联邦动保药品有限公司|Method for detecting residual quantity of amoxicillin and clavulanic acid in tissues|

---

US10685906B2|2018-11-13|2020-06-16|International Business Machines Corporation|Electrically conductive deterministic lateral displacement array in a semiconductor device|

---

CN109675491B|2018-12-27|2020-10-16|湖北理工学院|Method for preparing microvesicle based on non-covalent complex of myristic acid and imidazole derivatives|

---

CN109942729B|2019-02-27|2021-11-30|宁波大学|Amphiphilic acethydrazide initiator and preparation and use methods thereof|

---

CN109939623B|2019-04-01|2021-11-12|山东大学|Monoalkyl phosphate vesicle and preparation and application thereof|

---

CN110092447B|2019-05-06|2021-12-21|重庆工商大学|High COD emulsification wastewater breakdown of emulsion degradation device of high pressure discharge plasma|

---

CN110988175A|2019-12-17|2020-04-10|南京正济医药研究有限公司|Method for determining genotoxic impurity methylglyoxal in cimetidine|

---

CN111450553B|2020-03-27|2022-03-04|聊城鲁西甲胺化工有限公司|Dimethyl formamide evaporation system and production process|

法律状态:
2018-09-25| B07A| Application suspended after technical examination (opinion) [chapter 7.1 patent gazette]|

---

2019-02-26| B07A| Application suspended after technical examination (opinion) [chapter 7.1 patent gazette]|

---

2019-11-26| B09A| Decision: intention to grant [chapter 9.1 patent gazette]|

---

2020-01-28| B16A| Patent or certificate of addition of invention granted [chapter 16.1 patent gazette]|Free format text: PRAZO DE VALIDADE: 20 (VINTE) ANOS CONTADOS A PARTIR DE 22/06/2011, OBSERVADAS AS CONDICOES LEGAIS. |

优先权:

---

申请号 | 申请日 | 专利标题

---

EP10075274A|EP2399885A1|2010-06-22|2010-06-22|Device and method for solubilizing, separating, removing and reacting carboxylic acids in aqueous or organic solutions by means of micro- or nanoemulsification|

---

US34431110P| true| 2010-06-28|2010-06-28|

---

PCT/EP2011/003182|WO2011160857A2|2010-06-22|2011-06-22|Device and method for solubilizing, separating, removing and reacting carboxylic acids in oils, fats, aqueous or organic solutions by means of micro- or nanoemulsification|

---