• 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:
    The invention relates to a new polyacrylamide adhesive suitable for the fixing of fine-particle sorbent layers based on organic and/or inorganic compounds, used in overpressured one- and multilayer chromatographic procedures. The said adhesive is prepared by polymerizing the aqueous solutions of acrylamide and N,N'-methylene-bis-acrylamide in the presence of a catalyst pair at from 21 DEG to 30 DEG C., pulpifying the precipitate thus obtained and repeating the precipitation and the pulpification more times.
    公开号:SU1393316A3
    申请号:SU843786905
    申请日:1984-08-15
    公开日:1988-04-30
    发明作者:Тьихак Ерне；Минчович Эмиль；Золтан Шандор；Кемени Габор；Матуни Анико；Секели Тибор；Немет Шандор；Антал Жужанна；Фатер Жужанна
    申请人:Реанал Финомведьсердьяр И Лабор Мюсерипари Мювек (Инопредприятие)；
    IPC主号:
    专利说明:

    00
    with
    oo with
    Oi

    cm
    The invention relates to high molecular weight compounds for the preparation of acrylamide based gels and can be used to obtain chromatographic plates for thin layer and high performance chromatography, including for pressure chromatography.
    The aim of the invention is to provide an adhesive for fixing the sorbent layers on chromatographic plates.
    : PRI me R 1, A. Preparation of a polycrylamide adhesive.
    , | Dissolved in 1000 ml of distilled water of 20 g of acryl and 8 g of I, N-methylene-bis-acrylamide, is added to the resulting solution of 3 T of tetra-: ethylenediamine (TMEDA), mixed with 22.5 ° С in one direction and pour 0.8 g of | (NH4) SjOB dissolved in 3 ml of water into the center of the solution. After 4 minutes, the mixture begins to acquire an opal color, the temperature rises to, and after 30 minutes, the re-extraction ends.
    The resulting gel. Suspend 5 s. In a Waring blender and an ал 1000 ml of acetone is added to 11000 ml of an opal color solution. The solution is decanted three times after cooling, the cheesy sediment is collected together with the sediment, the S walls of the container adhere to it, for which 1000 ml of distilled water is added to it and suspension is carried out for 30 s., J (the resulting opal solution is again added 1000 ml of acetone and the operation described above is repeated, i.e. the precipitation is again carried out with acetone (solutions obtained after decanting are discarded).
    The polyacrylamide content in 1000 m of the opal color solution obtained in the last stage is 0.021 g / ml, the density of the solution is 1.004 g / ml, and the viscosity g is 4.65.
    B. Obtaining layers of adsorbent.
    To 44 g of silica gel (average particle size 6 µm, inner diameter 90 A) was added 57 mp of the solution of the β-adhesive, diluted with 87.5 ml of water (concentration: 2.7% based on silica gel). The resulting suspension is gently mixed, and then pulsified for 60 seconds in a Waryn mixer. The creamy consistency suspension prepared in this way is applied by continuous spraying onto an aluminum foil 0.1 mm thick with a layer 0.2 mm thick so that after drying the thickness of the applied layer is 0.16 mm. Cutting the edge of the foil, get 7 chromatographic layers with a length of 20 and a width of 20 cm.
    B. Single layer chromatography under pressure.
    On three sides of one of the chromatographic plates prepared according to Example 1B, a layer of sorbent 5 mm wide is removed, and from the polymer dispersion containing polyvinyl acetate and polyacrylamide, a polymer film is formed that is resistant to the action of water and solvents. Fifteen identical samples of the CAMA-11 color control reagent were applied at a distance of 3.0 cm from the impregnated end (at a distance of 9–9 mm from each other). After the spot of the printed samples was found, the chromatographic plate was placed in an ultra-microcamera. CHROMPRES-10 under pressure with a Teflon plate there, in which there are channels for moving the solvent. The pressure in the chamber is 1.2 MPa, the velocity of the solvent is 185 MP / h and the development time is 4.6 minutes. The solvent used is methylene chloride. In the chromatogram thus developed, the deviation for the sample of each color is i 1.5%.
    D. Many (four) layer chromatography under pressure.
    Four chromatographic plates with an adsorbent layer prepared according to Example 1B are impregnated on three sides with a dispersion of a polymer containing polyvinyl acetate and polyacrylamide, after which the plates thus treated are treated for 10 minutes at 100 ° C. In the top three of four curved plates with impregnated edges, a slot is made, 185 long and 2 mm wide, at a distance of 15 mm from the unpaired edge of the impregnated part of the adsorbent layer.
    Three perforated plates with a layer of adsorbent prepared in the manner described above, put aliquots of the drug from chamomile flowers (3 X 15 samples) at a distance of 15 mm from the slot and 9–9 mm from each other. After drying, the spot sample plate gently fits one over the other. The visas are laid with a non-perforated plate, the edges of which are also O1) opitima and on which 15 samples of the extract of the medicine extract from chamomile flowers are applied. All plates thus laid are placed in a CHRO # RES-10 ultra-chamber under pressure. Benzene is used as a solvent. The pressure in the chamber is 1.1 MPa, the time taken to be 16.2 minutes. After completion of the plate, the plates are disassembled, dried, sprayed with 90% sulfuric acid solution containing 0.2% vanillin, heated for 5 minutes at and assessed visually and by chromatographic spectrophotometer. The distance to spots and Rf are the same for all plates, the standard deviation is within the allowable interval.
    Example 2. Chromatographic plates with a layer of silica gel adsorbent prepared according to example 1 are cut into samples of size 20 X 40 cm. From the opposite sides of the plates remove the layer of adsorbent 5-5 mm wide and form a waterproof and solvent-resistant polymer film at these points . The chromatographic plates thus prepared can be used for two-dimensional thin layer chromatography under pressure. In the center of the adsorbent layer, a channel is formed with a width of 180 mm for the solvent, after which 15-15–15 samples of protein hydrolyzate from seeds of various wheat varieties are equivalent to the plate. Samples are applied on both sides of the channel, at a distance of 15 mm from it, with an interval of 9 mm. After drying the spot sample, the plates are placed in an ultra-microcamera CHROMPRES-10. The pressure in the chamber is 1.2 MPa. A mixture of n-butanol, glacial acetic acid and water in a ratio of 4: 1: 1 is used as a crucible. Time of occurrence 68 min.
    After the end of the development, the dry layer of the adsorbent is sprayed with a 0.2% solution of ninhydrin in a mixture of methanol and acetic acid saturated with sulfuric acid copper. The plates are then heated and the color chromatogram is analyzed with a spectrophotometer.
    Example 3. The next adsorbent layer is prepared using an adhesive prepared according to Example 1.
    Q with n
    5 ,, 5
    0
    40 g of talc with an average particle size of 3-4 μm are suspended in 40 ml of an adhesive solution diluted with 40 ml of distilled water for 75 seconds in a Waring mixer. A creamy consistency suspension prepared in this way is applied in a layer of 0.15 mm thick onto a polythale phthalate foil 0.15 mm thick so that after drying the thickness of the talcum layer is 0.1 mm. The resulting film is cut into 6 hours with a size of 20 x 20 cm. On three sides of the chromatographic plates prepared in this way, a layer of the adsorbent with a width of 5 is removed and a polymer film containing a polyvinyl acetate polymer dispersion is formed on these strips. and polyacrylamide. Digitalis extracts and authentic Digitylis cardiac glycosides dissolved in a mixture of methanol and chloroform are deposited in equivalent amounts on one of the prepared plates with impregnated edges, after which the plate is placed in an ultramicrocamera CHROMPRES-IO. A mixture of methyl ethyl ketone and water is used as eluent in a ratio of 18: 85. Time of occurrence 25 min.
    After drying, the adsorbent layer is sprayed with a 20% aqueous solution of phosphoric acid and the plate is heated for 20 minutes at 120 ° C. As a result, the spots turn yellow-green. Colored spots are subjected to qualitative and quantitative analysis.
    Example4. Using an adhesive solution prepared according to Example 1, an adsorbent layer with a concentration zone is formed, for which 33 g of silica gel suspension with an average particle size of 6 μm and an internal particle size are poured into a larger container with a special spray chamber (with a larger cutting surface). with a diameter of 90 A and 43 ml of diluted 66 ml of distilled water (concentration 2.7% based on silica gel) of the adhesive solution (after its corresponding pulpification). I pour into a smaller container (with a diffusion chamber of a smaller surface) 11 g of the slurry of silica with an average particle size of 15 microns and 15 ml of adhesive solution diluted to 21.5 ml
    five
    chops Both suspensions are applied on an aluminum foil with a thickness of 0.1 mm in a layer with a thickness of 0.1 mm so that after drying the thickness of the adsorbent layer with a concentration zone is 0.08 mm. The adsorbent layer thus prepared consists of a strip of inert silica 4 cm wide and a strip of active fine silica gel 16 cm wide.
    ; The resulting foil is cut into 10 I plates of 20 x 20 cm in size, the edges of which are passed through as described in Example 1. The prepared plates are used for single-layer chromatography under pressure. Samples of chamomile oil Raz-: of personal origin are separated on the prepared layer of adsorbent. Sample components (polyins, sesquiterpene alcohols) are divided into pronounced bands.
    Example 5. Using an adhesive solution prepared in Example 1, an adsorbent layer was prepared consisting of silica gel and a resin. For this, 28 g of silica gel with an average particle size of 6 μm and 10 g of a catnon-exchange resin with an average particle size of 5-8 μm are suspended for 60 s in 42.5+ +70 ml of distilled water. The prepared suspension is applied to aluminum foil with a thickness of 0.4 mm and a thickness of 0.2 mm. As a result, a smooth pale yellow color adsorbent with a thickness of 0.17 mm is formed. The foil is then cut into 7 parts of size 20 X 20 cm to give chromatographic plates.
    In order to use these plates for single and multilayer chromatography under pressure, their edges are impregnated with Example 1.
    To check the effectiveness of the obtained plates with a layer of adsorbent, samples of hydrochloric acid hydrolysates of peptides of various plants and artificially prepared identical mixtures of amino acids were applied on them. After drying, the applied samples were placed in a CHROMPRES-10 ultramicrocamera. Citrate buffer with a pH of 3.45 was used as the solvent and 0.2% ninhydrin solution in a mixture of methanol and acetic acid was used as a solvent.
    And p and mep 6 „20 g of acrylamide and 8 g of N, N-methyl-bis-acrylamide
    ABOUT
    from 0
    5 Q d
    0
    five
    P66
    dissolve n oo ml of distemped water, which is then added to the resulting solution of 2.8 mHg TMEDL. The temperature of the solution is maintained at 23 ° C. The solution is thoroughly mixed in one direction and pour into the center of the stirred solution 0.9 g of crystalline (III). After 5 minutes, the solution begins to acquire opal color, and after 30 minutes, the gelation is completely completed.
    The resulting gel is treated as described in Example 1. An opal adhesive solution with a density of 1.005 g / ml and a viscosity is obtained.
    using the resulting adhesive, prepare a layer of adsorbent as follows.
    40 g of silica gel with an average particle size of 3 μm and internal DNA
    60 A is suspended in 56 ml of an adhesive solution diluted with 90 ml of water in a Waring mixer. The prepared suspension is applied on aluminum foil O, 1 mm thick, O, 1 mm thick, resulting in a 0.06 mm adsorbent after drying. After cutting the edges of the foil, it is cut into several sections and the edges of the obtained chromatographic plates are drawn in Example 1. The prepared plates are used to check the colored reagents according to the method of single-layer chromatography under pressure, put on them rather small 5-10 mkg) samples.
    Example 7. Using an adhesive in accordance with Example 6, chromatographic plates with an adsorbent layer were prepared as follows.
    40 g of alumina with an average particle size of 10 µm are suspended in a mixture of 28 ml of adhesive solution and 39 ml of water for 225 seconds, after which the resulting suspension is applied in a layer of 0.2 mm in thickness onto glass plates measuring 20 x 20 cm and thickness 2 mm After drying, plates with a layer of alumina 0.6 mm thick are obtained.
    The adsorbent layer, activated when, is removed from three sides of the chromatographic plates to a width of 4 mm, and a polymer film is applied onto these strips. The thus obtained chromatographic plate was applied in aliquot amounts of peeled alkaloid extracts prepared from poppy and dissolved in a mixture of methanol and chloroform, taken in a 1: 1 ratio. After the spot sample dries, the plates provided with a protective plastic frame (to protect the shock absorbing water system in an ultramicrochamber of elevated pressure from the splitting action of the edges of the glass plates) are placed in an CHROMPRES 101g ultramicrocamera made of aluminum foil
    The pressure in the chamber is 1.2 MPa. A mixture of heptane, chloroform and ether, taken at a ratio of 40:50:10, is used as an eluent, and 20 Dragendorf reagent modified by Vaguyfalvi is used as a developer for the manifestation of alkaloids.
    EXAMPLE 8 Using an adhesive in accordance with Example 6 and a suspension, containing 1.8% zinc silicate and a UV indicator activated by manganese, as the starting materials, an adsorbent layer similar to the silica adsorbent of the example is prepared. 6.20
    PRI me R 9. Dissolve in 2000 ml of distilled water 36 g of acrylamide and 16.2 N, N-methylene bis-acrylamide, add to the resulting solution 4 ml of 3-dimethylaminopropionitrile, set the temperature of the solution to be equal and thoroughly mix it in one direction. 1.0 g of (NH) 2S20g was quickly dissolved in 3 ml of distilled water and the resulting solution was poured into the center of the stirred mixture. Opalescence begins after 8 minutes, and after 45 minutes, gelation is completely completed.
    The gel thus obtained is treated in accordance with Example 1. As a result, an opal-colored adhesive solution with a density of 1.006 g / mp is obtained, and in h - yl, 4.3.
    Using the obtained adhesive, the next sorbent layer is prepared. 30 g of high-purity cellulose with an average particle size of 5 µm is mixed with 30 ml of adhesive diluted with 30 ml of water, and the resulting suspension is applied in the usual way on aluminum 2 "1 foil 0 thickness , 9 mm layer thickness of 0.15 mm. Kra foil cut and cut it into separate
    35
    40
    45
    50
    55
    adhesive according to example 9.
    20 g of the reverse phase of the force powder Kage C-18 are suspended for 175 s in VO ml of polyacrylamide adhesion and 60 ml of isopropanol. The resulting suspension is applied to a sheet of aluminum foil and 0.12 mm thick, 0.18 mm thick. Chromatographic plates obtained by working a sheet of foil, impregnating the drying edge, are used to separate the reverse phase of poppy alkaloids with an CHROMPRES-10 ultramicrochamber. A mixture of a tonitrile and 0.005 is used as an eluent, and the Dragend reagent containing ethyl acetate is used as a product.
    Take p11. With the aid of admixture according to example 9, the next sorbent layer is prepared.
    88 g of silica gel with an average particle size of 5 μm and a pore diameter of 60 A are suspended for 85 s in 135 ml of adhesion diluted with 175 ml of water in a Waring's mixture. The resulting suspension was applied to a sheet of defatted aluminum foil with a thickness of 11 mm and a layer of 0.65 mm so as to obtain a layer of silica gel 0.5 mm.
    On the thus prepared adsorbent layer, N-methylated lysines, formed during spontaneous methylation and formylation at the interaction of L-lysine with formaldehyde, at a pressure of 1.3 MPa, are separated. Distilled water is used as eluent.
    权利要求:
    Claims (2)
    [1]
    Formula invented
    The method of obtaining polyacrylamide go gel by copolymerization of acrylics with N, N-methylene-bis-acyramide in a
    13933168
    pieces The edges are impregnated and the plates thus obtained are used for single-layer chromatography under the pressure of amino acids. A mixture of methyl ethyl ketone, acetonitrile, and acetic acid is used as eluent.
    acids and water, taken in a ratio of 3: 4: 1:
    [2]
    2. The pressure in the chamber is 1.5 MPa, the manifestation time is 59 min (during replenishment).
    Example 10. For. fixation -oil-kagel RP-18, chemically bound to the stationary phase, on the plates
    0
    50
    five
    0
    five
    0
    five
    adhesive according to example 9.
    20 g of the reverse phase of C-18 silica gel powder are suspended for 175 s in a mixture of VO ml of polyacrylamide adhesive and 60 ml of isopropanol. The resulting fine suspension is applied to a sheet of aluminum foil and a thickness of 0.12 mm with a layer of 0.18 mm. Chromatographic plates, obtained by cutting a sheet of foil, impregnating the edges and drying, are used to separate the reverse phase of the poppy alkaloids in an CHROMPRES-10 ultramicrochamber. A mixture of acetonitrile and 0.005 is used as the eluent, and the Dragendorff reagent containing ethyl acetate is used as the developer.
    Take p11. Using the adhesive of Example 9, the next layer of sorbent is prepared.
    88 g of silica gel with an average particle size of 5 μm and a pore diameter of 60 A are suspended for 85 s in 135 ml of adhesive diluted with 175 ml of water in a Waring mixer. The resulting suspension is applied to a sheet of defatted aluminum foil with a thickness of 11 mm and a layer of 0.65 mm so as to obtain a layer of silica gel with a thickness of 0.5 mm.
    On the thus prepared v layer of the adsorbent, the separation of N-methylated lysines, which are formed by spontaneous methylation and formylation by the interaction of L-lysine with formaldehyde, at a pressure of 1.3 MPa. Distilled water is used as eluent.
    Invention Formula
    The method of obtaining polyacrylamide gel by copolymerization of acrylamide. with N, N-methylene bis-akrashamidom in water 9
    In the presence of tertiary amine and ammonium persulphate, followed by washing the gel from low molecular weight impurities, characterized in that, in order to obtain an adheivant for: fixing Sorbent layers on chromatographic plates, 3-dimethylaminopropshonite- pl or N are used as tertiary amine NN, Y-tetramethylenediamine, and the copolymerization is carried out at a mass ratio of acrylic: amide and N, | hylen-bis-acrylamide (2.2-2.5); 1
    139331610Acrylamide 1.8-2.0 M, N-methylene-bis-acrylamide 0.8-0.81 3-dimethylaminopropylonitrile or N, NN, N -TeT-, ramethylenediamine 0.16-0 , 22 Ammonium persulphate
     0.05-0.09 with this, the mouse gel is carried out by repeated alternation with
    / ii the following content of reagents, May.%: | dirovani and processing precipitator.
    Acrylamide 1.8-2.0 M, N-methylene-bis-acrylamide 0.8-0.81 3-dimethylaminopropyl nitrile or N, NN, N -TeT-, ramethylenediamine 0.16-0 , 22 Ammonium persulphate
     0.05–0.09, while removing the gel from the gel by repeated alternation of suspensions.
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    同族专利:
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    引用文献:
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    法律状态:
    优先权:
    申请号 | 申请日 | 专利标题
    HU287583A|HU190910B|1983-08-16|1983-08-16|Method for producing glue suitable for fixing sorbent layer of pressurized one and multiple layer chromatography|
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