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Application of Monodisperse Polymethacrylate Microspheres in Purification of Vancomycin by Column Chromatography

A polymethacrylate, monodisperse technology, applied in the field of monodisperse polymethacrylate microspheres, can solve the problems of high pressure resistance, low group density, small specific surface area, etc., and achieve mobile phase pH resistance Change, high purity recovery, high recovery effect

Inactive Publication Date: 2011-12-14
SUZHOU NANOMICRO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the separation medium used in this method is a dextran matrix, and its bed volume will change significantly under different salt concentrations. At the same time, the dextran matrix is ​​soft and not resistant to high pressure, which has great advantages in rapid separation. big limitations
[0006] Wei et al. (Journal of Separation Science, 2006, 29, 5–13) synthesized a weak cationic column by copolymerizing glycidyl methacrylate and ethylene glycol dimethacrylate in a stainless steel column, but The separation medium is block-shaped, with a small specific surface area and a low group density
[0007] Lu Miaolong et al. (Chemical World, 1984, 10, 369-373) synthesized a weak cationic chromatographic medium based on polystyrene / divinylbenzene polymer. This medium has strong hydrophobicity, and the chloromethyl Chloromethyl ether is a strong carcinogenic reagent used in the chemicalization step, which has limitations in production

Method used

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  • Application of Monodisperse Polymethacrylate Microspheres in Purification of Vancomycin by Column Chromatography
  • Application of Monodisperse Polymethacrylate Microspheres in Purification of Vancomycin by Column Chromatography
  • Application of Monodisperse Polymethacrylate Microspheres in Purification of Vancomycin by Column Chromatography

Examples

Experimental program
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Effect test

Embodiment 1

[0016] Purification of vancomycin: using a 10×300 mm adjustable glass column at one end, Uni-PGM-50 resin (monodisperse polymethacrylate microspheres, 50 microns in diameter, Suzhou Nano Microbe Technology Co., Ltd.) as chromatography Packing, column height 20.5cm, column volume 16mL, first activate the chromatographic column with 3% NaCl in 3 column volumes (CV), and then equilibrate the chromatographic column with 5CV ultrapure water at a flow rate of 1mL / min. Vancomycin sample (purity 89%) was dissolved in ultrapure water, adjusted to pH 5.0 with dilute hydrochloric acid and then filtered. 10CV of 0.2%NH 4 HCO 3 Prewash at a flow rate of 1mL / min, then wash with 1% NH 4 HCO 3 It was eluted at a flow rate of 0.5mL / min and collected in separate tubes. During the elution process, the height of the column bed was 20.2cm, the volume of the packing was 15.9mL, and the volume of the packing bed remained basically unchanged. After testing the collected solution, the yield of van...

Embodiment 2

[0018] Purification of vancomycin: A 10×300 mm one-end adjustable glass column was used, CM Sephadex C-25 (GE Healthcare, particle size range 40-125 microns) was used as the chromatography packing material, the column height was 20.5 cm, and the column volume was 16 mL. Equilibrate the chromatographic column with 5CV of ultrapure water at a flow rate of 1mL / min. Vancomycin sample (purity 89%) was dissolved in ultrapure water, adjusted to pH 5.0 with dilute hydrochloric acid and then filtered. and 8CV of 0.4% NH 4 HCO 3 Prewash at a flow rate of 1 mL / min, then wash with 4% NH 4 HCO 3 Elute at a flow rate of 0.5mL / min and collect in separate tubes. During the elution process, the volume of the packed bed decreases significantly. After testing the collected solution, the yield of vancomycin with a purity greater than 95% was 68%.

Embodiment 3

[0020] Purification of vancomycin: A 10×300 mm one-end adjustable glass column was used, CM Sephadex C-25 (GE Healthcare, particle size range 40-125 microns) was used as the chromatography packing material, the column height was 20.5 cm, and the column volume was 16 mL. First activate the column with 3% NaCl for 3 column volumes (CV), and then equilibrate the column with 5CV of ultrapure water at a flow rate of 1 mL / min. Vancomycin sample (purity 89%) was dissolved in ultrapure water, adjusted to pH 5.0 with dilute hydrochloric acid and then filtered. 10CV of 0.2%NH 4 HCO 3 Prewash at a flow rate of 1mL / min, then wash with 1% NH 4 HCO 3 Elute at a flow rate of 0.5 mL / min for 5CV, and the volume of the packed bed decreases significantly during the elution process. Vancomycin was not detected in the collected fluid.

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Abstract

The invention discloses the application of monodisperse polymethacrylate microspheres in the column chromatography purification of vancomycin. Column chromatography was performed on a column packed with a packing material called Uni-PGM-50. When Uni-PGM-50 is used as a column chromatography filler, it has good hydrophilicity, high mechanical strength, acid and alkali resistance, and good separation effect. The salt concentration of the mobile phase changes, and at the same time when purifying and separating vancomycin, only a low-concentration saline solution is needed to elute the vancomycin adsorbed on the filler to achieve high purity and high recovery.

Description

technical field [0001] The invention relates to the application of monodisperse polymethacrylate microspheres, in particular to the application of monodisperse polymethacrylate microspheres in the extraction and purification of antibiotic vancomycin. Background technique [0002] Vancomycin is a glycopeptide antibiotic produced by microorganisms under controlled fermentation conditions. It is known to inhibit the synthesis of bacterial cell walls and alter the permeability of cell membranes. Its main use is to treat infections caused by Gram-positive bacteria. It has obvious curative effect on drug-resistant Staphylococcus aureus, and it is also used to treat patients who are allergic to penicillin or have no effect on penicillin and cephalosporins. [0003] In the separation and purification of antibiotics, amino acids, peptides, proteins and natural products, ion exchange chromatography is one of the most commonly used methods. The chromatographic medium is mainly compose...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K9/00C07K1/22
Inventor 江必旺陈荣姬吴俊成林生跃裘金明
Owner SUZHOU NANOMICRO TECH CO LTD