Capillary electrophoretic method for measuring polymyxins antibacterial peptide and running buffer solution thereof

A technology of capillary electrophoresis and polymyxin, which is applied in the field of analytical chemistry, can solve the problems of low column efficiency and poor separation effect of polymyxin antimicrobial peptides, and achieve low cost, low consumption of samples and reagents, and easy The effect of using less reagent

Inactive Publication Date: 2018-12-18
SPH NO 1 BIOCHEM & PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to overcome the problems of poor separation effect and low column efficiency of polymyxin antimicrobial peptides in the prior art, the present invention provides a simple, sensitive capillary electrophoresis method suitable for analysis of polymyxin antimicrobial peptide components and related substances

Method used

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  • Capillary electrophoretic method for measuring polymyxins antibacterial peptide and running buffer solution thereof
  • Capillary electrophoretic method for measuring polymyxins antibacterial peptide and running buffer solution thereof
  • Capillary electrophoretic method for measuring polymyxins antibacterial peptide and running buffer solution thereof

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Embodiment 1

[0046] This example is a preferred running buffer for the determination of polymyxin antimicrobial peptides based on capillary electrophoresis in the present invention.

[0047] In this embodiment, the running buffer is a triethanolamine solution containing hydroxypropyl-β-cyclodextrin and isopropanol; wherein, in the buffer, the concentration of hydroxypropyl-β-cyclodextrin is 30 mmol / L, the content of isopropanol is 5% (volume fraction), the concentration of triethanolamine is 130mmol / L, and the pH is adjusted to 2.5 through phosphoric acid.

[0048] The experimental conditions of the capillary electrophoresis of the running buffer are: non-coated fused silica capillary, the column length is 60cm, the inner diameter is 50μm; the working voltage is 24kV; the column temperature is 25°C; the detection wavelength is 192nm; the injection pressure is 50mbar, and the injection time is 3s.

Embodiment 2

[0050] This example is a preferred running buffer for the determination of polymyxin antimicrobial peptides based on capillary electrophoresis in the present invention.

[0051] In this embodiment, the running buffer is a triethanolamine solution containing hydroxypropyl-β-cyclodextrin and isopropanol; wherein, in the buffer, the concentration of hydroxypropyl-β-cyclodextrin is 50 mmol / L, the content of isopropanol is 10% (volume fraction), the concentration of triethanolamine is 100mmol / L, and the pH is adjusted to 3 through sulfuric acid.

[0052] The experimental conditions of capillary electrophoresis for the running buffer are: cationic coated capillary, column length 100cm, inner diameter 100μm; working voltage 20kV; column temperature 30°C; detection wavelength 180nm; injection pressure 30mbar, injection time 5s.

Embodiment 3

[0054] This example is a preferred running buffer for the determination of polymyxin antimicrobial peptides based on capillary electrophoresis in the present invention.

[0055] In this embodiment, the running buffer is triethanolamine solution containing hydroxypropyl-α-cyclodextrin and acetonitrile; wherein, in the buffer, the concentration of hydroxypropyl-α-cyclodextrin is 5mmol / L , the content of acetonitrile is 8% (volume fraction), the concentration of triethanolamine is 80mmol / L, and the pH is adjusted to 2 by hydrochloric acid.

[0056] The experimental conditions of capillary electrophoresis for the running buffer are: anion-coated capillary, column length 30cm, inner diameter 25μm; working voltage 10kV; column temperature 40°C; detection wavelength 380nm; injection pressure 40mbar, injection time 10s.

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Abstract

The invention relates to a capillary electrophoretic method for measuring polymyxins antibacterial peptide. The method comprises the following steps: dissolving a polymyxins antibacterial peptide preparation in a solvent, directly measuring polymyxins antibacterial peptide through capillary electrophoresis in a running buffer solution, wherein the running buffer solution is a triethanolamine solution or triethylamine solution containing hydroxypropyls cyclodextrin and organic solvent, and the hydroxypropyls cyclodextrin comprises hydroxypropyl-beta-cyclodextrin, hydroxypropyl-alpha-cyclodextrin, hydroxypropyl-gamma-cyclodextrin, and / or sulfated / sulfonated cyclodextrin. The polymyxins antibacterial peptide is analyzed by firstly adopting the triethanolamine solution or triethylamine solution containing hydroxypropyls cyclodextrin and organic solvent in the capillary electrophoresis method, the component and the related substance are good in separation, especially the polymyxin B and E measurement; compared with the liquid phase method, the column efficiency is high, the operation is simple, sample and reagent consumption are less, and the cost is low; the method repeatability is good, and measurement results on samples of different manufacturers are stable and reliable.

Description

technical field [0001] The invention belongs to the field of analytical chemistry, and in particular relates to a capillary electrophoresis method for determining polymyxin antibacterial peptides and a running buffer used in the capillary electrophoresis method. Background technique [0002] Polymyxin is a lipopeptide antibacterial peptide, which has a strong bactericidal effect on Gram-negative bacilli, and because polymyxin antibacterial peptide has relatively large nephrotoxicity and neurotoxicity, it is currently used in most of the clinical trials. It is polymyxin B and E, which are considered to be the "last line of defense" in the treatment of multidrug-resistant Gram-negative bacilli infections. Because it is a fermentation product, polymyxin B or E is a mixture of a series of polypeptides with similar structures. The main components of polymyxin B are polymyxin B1, B2, B3 and B1-I. The structural formula is shown in the appendix figure 1 , the main components of po...

Claims

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Application Information

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IPC IPC(8): G01N27/447
CPCG01N27/44747G01N27/44791
Inventor 张含智刘浩秦峰闻宏亮赵敬丹裘亚徐晓曦吕丹丹
Owner SPH NO 1 BIOCHEM & PHARMA CO LTD
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