• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    Zur Beseitigung von Trübungen in durch Probenmaterial getrübten Reaktionsgemischen, insbesondere für photometrische Messung werden dem Reaktionsgemisch a) eine oder mehrere niedermolekulare organische Verbindungen wie aromatische Alkohole oder Amine, die elektro-negative Substituenten tragen, z.B. Phenole oder Naphthole, welche mit ein oder mehreren Halogenatomen oder/und Hydroxylgruppen substituiert sind, und b) ein oder mehrere ebenfalls für sich klar lösliche Detergentien, welche mit dem Zusatz a) einen weniger löslichen Komplex bilden, wie nichtionogene Polyäthylenoxidaddukte oder kationische Detergentien, insbesondere Alkyl-, Aralkyl- oder Alkylthioäther oder Alkyl- oder Aralkylester von Poiyäthytenoxidgiykoten mit 8 bis 22 C-Atomen in Alkylgruppen und 5 bis 25 Äthyienoxideinheiten oder Alkyldimethylbenzyl-ammoniumchloridverbindungen mit 8 bis 22 C-Atomen in der Alkylgruppe, in einer zum Auftreten einer Fällung des Komplexes ausreichenden Menge zugesetzt und anschließend wird weiteres Detergens zugesetzt, bis der ausgefallene Komplex wieder gelöst ist.
    公开号:SU1443816A3
    申请号:SU792746998
    申请日:1979-04-06
    公开日:1988-12-07
    发明作者:Клозе Зигмар；Бушек Херберт；Шлумбергер Хельмут
    申请人:Берингер Маннхайм,Гмбх (Фирма)；
    IPC主号:
    专利说明:

    WITH
    with
      with
    00

    cm
    The invention relates to biochemistry, in particular, to the method of clarifying reaction mixtures, the mutation of which is caused by the test material, of a biological nature, such as serum or plasma.
    The aim of the invention is to increase the degree of clarification of the reaction mixture.
    It is known that the turbidity of some test samples of serum or plasma (lipemic serum or plasma) adversely affects the accuracy of the measurement in photometric analysis.
    An increase in the degree of clarification is achieved by adding detergents to the reaction mixture, which are one or more low-molecular-weight organic compounds and one or more transparent-soluble detergents that, with the addition of these organic compounds, form a soluble complex in sufficient quantities to begin the precipitation of the complex. There are another detergent before
    for example, butanone-2, cyclohexanone and the like; aldehydes, such as cinnamon al-dehyd or benzaldehyde; carboxylic acids, for example octanoic acid; ethers, for example 2-methoxyphenol; benzene.
    Among those forming with these compounds are poorly soluble complexes.
    fQ detergents, which in turn are clear solutions in the reaction mixture, meet non-ionic, ionic and amphoteric detergents. Preferred are
    5 non-ionic adducts of polyethylene oxide. Alkyl, aralkyl and alkylthioesters, as well as alkyl and aralkyl compounds of polyethylene oxide glycols give particularly good results. In these
    The 20 detergents alkyl groups typically have from 8 to 22 carbon atoms, the aryl group is, predominantly, a phenyl residue, the glycol residue of these detergents having
    25 usually from 3 to 25 units of ethylene oxide in condensed form. Typical examples of these groups of detergents are tesit, ganapol, terg tol, triton XI00, lenzodel, and bri 35.
    as long as the precipitated precipitated complex is also applicable.
    va does not dissolve.
    The compounds used are: aromatic (condensed and non-condensed) alcohols and amen, preferably those that are strongly electronegatively substituted (preferred electronegative substituents are halogens), for example benzyl alcohol, phenol, mono-, di- and trichlorophenols, dibromo - and tri- 40 bromophenols, cresols, naphthols, dihydrooxynaphthols, aniline and chloraniline, methylaniline} esters of aryl acids and alkyl acids, for example, ethanol, p-hydroxybenzoic acid, 45 lots, ethyl ester of acetic acid acid, acetic acid butyl ester, propionic acid ethyl ester, trichloroacetic acid ethyl ester, etc., are straight and cyclic and branched aliphatic alcohols with more than 3 carbon atoms, for example amyl alcohol, octanol, decananol, cyclohexanol, and t .P.; halogenated aliphatic compounds with a short chain of atoms, for example, dichloroethane, trichloroethnene, tetrachloroethane, dichlorobutene tetrachlorofluoroethane and the like; aliphatic ketones,
    for example, butanone-2, cyclohexanone and the like; aldehydes, such as cinnamon al-dehyd or benzaldehyde; carboxylic acids, for example octanoic acid; ethers, for example 2-methoxyphenol; benzene.
    Among those forming with these compounds are poorly soluble complexes.
    Q detergents, which, in turn, are clear solutions in the reaction mixture, are found to be non-ionic, ionic and amphoteric detergents. Preferred are
    5 non-ionic adducts of polyethylene oxide. Alkyl, aralkyl and alkylthioesters, as well as alkyl and aralkyl compounds of polyethylene oxide glycols give particularly good results. In these
    0 detergents alkyl groups usually have from 8 to 22 carbon atoms, the aryl group represents mainly the phenyl residue. The glycol residue of these detergents usually has from 3 to 25 ethylene oxide units in condensed form. Typical examples of these groups of detergents are tesit, ganapol, tergitol, triton XI00, lansodel, and bree 35.
    ionic detergents, in particular alkyl dimethyl benzyl ammonium chlorides. Secondary alkyl sulfates, the alkyl groups of which correspond to the above definition, are also suitable. N-lauryl diethanolamide is suitable for short-circuiting amphoteric detergents.
    The method is carried out as follows.
    A low molecular weight compound is selected in an amount of 0.5-300 mM, preferably 0.3-3.3 g / l, dissolved in buffer or saline solution, a clear detergent solution is added to this solution in small portions in an amount of 0.05- i%, preferably 1-30 g / l of the reaction mixture, so that when processing the test low molecular weight compound of the detergent there is a turbidity in the solution.
    If turbidity has occurred (which indicates that both substances can be used), then the resulting mixture is added to the sample to be analyzed in the indicated amounts so that additional turbidity is noticeable. Then add the same or different detergent portions to those
    ten
    15
    op, until the turbidity disappears with this. A test sample of serum or plasma treated in this way is then analyzed.
    It is preferable to use smaller amounts of detergents because of their possible negative effect on the activity of the enzymes, while at least one or two different detergents are used.
    The method is applicable to types of measurements in which turbidity can lead to distortion or interference of measurement, in particular in reaction mixtures that are intended for photometric measurements. Such reaction mixtures are known (Bergmeier G., W. Methods of enzyme analysis. - Chemie, Weine- „game / Bergstrasse).
    Tables 1 and 2 (examples 1-20) show the types of low-molecular compounds, their respective concentrations, tesit concentration (ThiSit) detergent and pH value ,.
    In Examples 21-36, in a solution that contains a low molecular weight compound (2,4-dichlorophenol) at a certain concentration, the personal detergents vary.
    In Examples 37-39, concentrations of reagents of a specific photometric test are given, in which the turbidity strongly interferes and is therefore eliminated by the additives contained in the reagent.
    The essence of the method is.
    25
    35
    To remove turbidity, the following additives are introduced,%: butyl ester — acetic acid 2, tesit (additive A) 1.5; dichloroethane 0.5; hexanol 0.5;
    that the poorly soluble complex, which is formed first, again the solution is the genepol X-0 (isotridecanol polyglycide by adding another detergent ester) 5; genapol X-100 (isotrite and at the same time also dissolves the turbidity to be eliminated in the solution of the sample being analyzed. Moreover,
    decanol polyglycol ether) 9; zit (additive B) 6.
    Test conditions: 2 ml of reagent
    those (with
    the use of a detergent characterized by additives A or B) ,. 0.1 ml of sample.
    from the first, it is used less.
    Examples 1-36. Brightening systems.
    The proposed combinations remove the dregs in the system within .1 minutes to a maximum of 2 hours: 2 ml of the test mixture 20 µl of milk with a fat content of 3.5% at a layer thickness of 1 cm, for a wave 546 by them, at 25 ° C.
    Adjustment options. Use 0.1 M Tris-HC1 buffer solution with 2% 7H-O.
    Variations of detergents. 0.1 M Tris-HC1 buffer solution (pH 8.0)
    50
    Starting from 0.6 V EU uricase J.7,3.3.
    The measurement is carried out at a pressure of 340 nm and at the end point.
    These additives contribute to the fact that significantly elevated lipidously turbid material under test, the initial extinctions rapidly decrease. The turbidity from extinction 2.0 in 10 minutes decreases to a stable extinction level of about 2, relative to the indicators of idle experience,
    Prev 38, Reagents for determining cholesterol in a short.
    0
    five

    d
    with 2% MgS04 7HiO and 8 mm / l of 2,4-dichlorophenol as an additive using the detergents shown in Table 2, a good bleaching of the mixture is obtained.
    The following names were used in Table 2: tesit - hydroxypolyethoxydodecane, tergitol NPX - 10.5-ethoxy-phenylphenol, triton X100 - 9.10-ethoxy-octylphenol, pegosperse - polyistilylene glycol luryl ether, Tepol 7 O and lenzodelle AB6 - mixed secondary alkyl sulfates (from Cg to C – C), tween 20-20-ethoxy-sorbitan-iolaurate, .briy 35 — polyoxyethylene-lauryl ether, gena-ol 0x80 and genapol XI00 — polyglycol ether of fatty alcohols (straight chain with C j; to.
    Example 37. Reagents for the determination of uric acid in serum. Formulation: 0.05 mol / l 10 vol.% Ethanol; HC1 is set at pH 8.5; 1 mg / ml of TPN (triphosphopyridine - 5 nucleotides); I v / ml of alcohol dehydration in EU- 1.2.1.5; 1000 v / ml of EC catalase 1.11.1.6; 0.05 mol / l Na oxalic acid; 0.05 mol / l pyrazole.
    The prescribed measurement wavelength and the relatively large required sample flow are due to the fact that with a hyperpathic turbid test material the measurement is difficult or impossible.
    To remove turbidity, the following additives are introduced,%: butyl ester — acetic acid 2, tesit (additive A) 1.5; dichloroethane 0.5; hexanol 0.5;
    35
     genapol X-0 (isotridecanol polyglycol ether) 5; genapol X-100 (isotrigenopol X-0 (isotridecanol polyglycol ether) 5; genapol X-100 (isotry
    decanol polyglycol ether) 9; zit (additive B) 6.
    Test conditions: 2 ml of reagent
    those (with
    additives A or B) ,. 0.1 ml of sample.
    Starting from 0.6 V EU uricase J.7,3.3.
    The measurement is carried out at a pressure of 340 nm and at the end point.
    These additives contribute to the fact that significantly elevated lipidously turbid material under test, the initial extinctions rapidly decrease. The turbidity from extinction 2.0 in 10 min is reduced to a stable level of extinction on the order of O, 2 relative to the indicators of idle experience,
    Prev 38, Reagents for determining cholesterol in a short.
    Recipe; 2.54 g / l 31.60 g / l K2HP04S 8 g / l 200 mg / l 4-aMHHoaHTHnHpHHaj 282 mg / l phenol; pH 7.90, 3 v / ml cholesterol teraz EC 3.1.1.13; 0.3 v / ml of cholesterol oxidase EC 1.1,3.6,
    This reagent is a mixture obtained in a single unit with a single-stage mixture, and therefore can be used with no change in the blank test. The fulfillment of this requirement is provided by the following additives (and for lipemically very turbid test material). Brightening agents: 815 mg / l 3,4-dichlorophenol, 4 g / l Oh-100 genapol and 3 g / l sodium deoxycholate; 2 g / l of eptanol and 8 g / l of tesit; I g / l of ethyl acetate and 7 g / l of tesite; at pH 7.0 in 80 mM K / Na-phosphate buffer; 1.5 g / l of parahydroxybenzoic acid ethyl ester and 4 g / l of tesite; at pH 7.50 in 0.2 M potassium phosphate buffer.
    In addition to the 3 mmol / l phenol required for the formation of color, 35 mmol / l phenol is used; 1.5 g / l of Oh-100 genapola; 1.5 g / l Tween 20; 1.0 g / l sodium cholate.
    Test conditions: 2 ml of reagent + 20 l of sample, 546 nm,.
    The extinction turbidity 1 1.5 caused by the material under test is removed completely or to a degree that can be neglected during the period required for the reaction,
    Example 39. Reagent for determination of serum glucose.
    Formulation: 14, 9 g / l Na2HP04; 13.2 g / l KHjPO; 8.0 g / l Na2SO4, 30 v / ml EU glucose oxidase 1.1.3.45 1.8 v / ml EU peroxidase 1.11,1.7; 0.2 v / ml EC esterase 3.1.1.13; 470 mg / l phenol; 156 mg / l 4-amino yntipirin, at pH 7.0,
    Brightening agents: 814 mg / l 3,4-dichlorophenol9 3.6 g / L of the genapole Ox-100 and 3.0 g / l of the tauroglycocholic ACID} 814 mg / l of the 3,4-dichlorophenol, 2.5 g / l of tesite and 3.9 g / l of tauroglycocholic acid, In 0.2 M potassium phosphate buffer, pH 7.0, in addition, 35 G / l of phenol, 489 mg / l 3,4-dichlor
    0
    five

    0
    five
    0
    five
    0
    phenol, 3.0 g / l of the Oxapol Oh-100 and 2.8 g / l of sodium cholate. ,
    The measurement is carried out at a pressure of 578 nm. Analogously to example 38, the additives used contribute to the fact that the turbidity caused by the material being used is reduced by the shortest in {5em.
    Examples include those for which one is used: one or more transparent-soluble detergents, which with an additive can form a weaker soluble complex, and those in which at least one detergent that cannot be added to the reaction mixture can be used in addition mixtures form a weaker soluble complex (marked).
    权利要求:
    Claims (1)
    [1]
    Invention Formula
    The method of clarifying the reaction mixture for photometric measurement of biological samples by introducing detergents into the sample, in contrast to the fact that, in order to increase the degree of clarification, a low molecular weight compound selected from the group of: phenol-25.3 or 2.4- Dichlorophenol, or 2,4,6-trichlorophenol, 2,4-dibromophenol, 1,5- or 1,7-, or 2,7-dihydroxy 1 naphthol, hydroquinone, aniline, 2-chloroaniline, o-toluidine, chloroform , benzene, gwa col, benzaldehyde, cinnamic aldehyde, cyclohexanone, at pH 7.1-8.0 and sodium salt of octanoic acid at pH 5.0, cat They are added in an amount of 0.3-3.3 g / l of the reaction mixture, then one or two detergents are selected, selected from the group: hydroxypolyethoxydodecane, 0.5-ethoxynonylphenol, polyglycol ether of aliphatic alcohols with the number carbon atoms in the direct chain from 12 to 15, 9.1 O-ethoxyoctylphenol, polyethylene glycol luryl ether, a mixture of secondary alkyl sulfates with the number of carbon atoms in the alkyl moiety from 8 to 18, 20-ethoxy-sorbitan monolaurate, polyoxyethylene llauryl ether, which use in the amount of 1.2-30 g / l of the reaction mixture.
    Table j
    31 Twin 20
    32 Bray 35
    , 33 Genapol 0x80
    34
    Twin 20
    14A381610
    Continuation of table 2
    ammonium bromide
    bis- (2-gyroxyethyl) -amine lauric acid
    bis- (2-hydroxyethyl) -amine lauric acid
    Dodecylsulfate sodium salt (in the presence of cations)
    Cetyltrimethylbromide am
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    法律状态:
    优先权:
    申请号 | 申请日 | 专利标题
    DE19782816229|DE2816229C2|1978-04-14|1978-04-14|Methods and means for removing opacities|
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