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Method for detecting content and components of gentamicin in fermentation liquor by HPLC-ELSD

A technology of gentamicin and fermentation broth, which is applied in the fields of detecting the content and components of gentamicin, and detecting the content and components of gentamicin in fermentation broth by HPLC-ELSD. Problems such as multi-batch processing and long processing time

Pending Publication Date: 2021-07-30
HEBEI SHENGXUE DACHENG PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This treatment method has a good purification effect, but the processing time is too long in the sample detection process, and it is not suitable for tracking detection and multi-batch processing of the fermentation process.

Method used

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  • Method for detecting content and components of gentamicin in fermentation liquor by HPLC-ELSD
  • Method for detecting content and components of gentamicin in fermentation liquor by HPLC-ELSD
  • Method for detecting content and components of gentamicin in fermentation liquor by HPLC-ELSD

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Chromatographic conditions

[0040] Chromatographic column: C18, 4.6*250mm, 5.0μm;

[0041] Column temperature: 35°C;

[0042] Injection volume: 25μL;

[0043]Flow rate: 0.5mL / min;

[0044] Drift tube temperature: 110°C;

[0045] gain factor: 1;

[0046] Gas flow rate: 2.6L / min;

[0047] Running time: 50min;

[0048] Mobile phase: 0.2mol / L trifluoroacetic acid:methanol=96:4

[0049] a. Preparation of gentamicin reference substance solution: accurately weigh 50 mg of gentamicin reference substance, dissolve and dilute to a mass concentration of 5.0 mg / mL with mobile phase, shake well, and use it as reference substance solution 1; 1 Dilute 2.5 times, 5 times and 10 times respectively with the mobile phase solution to obtain reference substance solutions 2, 3 and 4;

[0050] b. Preparation of gentamicin sample solution: take an appropriate amount of gentamicin seed bottle fermentation liquid sample, acidify it with 10mol / L hydrochloric acid to a pH value of 1.5, le...

Embodiment 2

[0057] Chromatographic conditions

[0058] Chromatographic column: C18, 4.6*250mm, 5.0μm;

[0059] Column temperature: 30°C;

[0060] Injection volume: 20μL;

[0061] Flow rate: 0.6mL / min;

[0062] Drift tube temperature: 105°C;

[0063] gain factor: 1;

[0064] Gas flow rate: 2.8L / min;

[0065] Running time: 40min;

[0066] Mobile phase: 0.2mol / L trifluoroacetic acid:methanol=95:5

[0067] a. Preparation of gentamicin reference substance solution: accurately weigh 40 mg of gentamicin reference substance, dissolve and dilute to a mass concentration of 4.0 mg / mL with mobile phase, shake well, and use it as reference substance solution 1; 1 Dilute 2 times and 4 times respectively with mobile phase solution to obtain reference substance solution 2 and reference substance solution 3;

[0068] b. Preparation of gentamicin sample solution: take an appropriate amount of gentamicin fermentation liquid sample, acidify it with solid oxalic acid to a pH value of 1.0, and after ul...

Embodiment 3

[0073] Chromatographic conditions

[0074] Chromatographic column: C18, 4.6*150mm, 5.0μm;

[0075] Column temperature: 28°C;

[0076] Injection volume: 10μL;

[0077] Flow rate: 0.3mL / min;

[0078] Drift tube temperature: 120°C;

[0079] gain factor: 1;

[0080] Gas flow rate: 3.0L / min;

[0081] Running time: 30min;

[0082] Mobile phase: 0.1mol / L trifluoroacetic acid:methanol=94:6

[0083] a. Preparation of gentamicin reference substance solution: accurately weigh 50 mg of gentamicin reference substance, dissolve and dilute to a mass concentration of 5.0 mg / mL with mobile phase, shake well, and use it as reference substance solution 1; 1 Dilute 1.5 times, 3.75 times and 6 times respectively with the mobile phase solution to obtain reference substance solutions 2, 3 and 4;

[0084] b. Preparation of gentamicin sample solution: take an appropriate amount of gentamicin fermentation liquid sample, acidify it to pH 2.0 with 5mol / L sulfuric acid, and after ultrasonication f...

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Abstract

The invention provides a method for detecting the content and components of gentamicin in fermentation liquor by using HPLC-ELSD. The method comprises the following steps: preparing a gentamicin reference substance solution, treating a fermentation liquor sample, drawing a standard curve, detecting by using HPLC-ELSD, and calculating the content and components of gentamicin in the sample by using a standard curve method. The method has the beneficial effects that sample treatment steps are simple, derivatization reaction or resin adsorption is not needed, and detection is more convenient and timely; the detection specificity is high, the separation effect is good, and the chromatographic peak of each component of gentamicin in a sample can be effectively identified; the accuracy of the detection result is high, and the average value of the standard recovery rate of each component is 99.3%-101.6%; the repeatability and the durability are good, and the relative standard deviation is smaller than 2%; the method can be used for simultaneously measuring the total content of gentamicin and the proportion of each component concerned in pharmacopeia.

Description

technical field [0001] The invention belongs to the field of component detection, and relates to a method for detecting the content and components of gentamicin, in particular to a method for detecting the content and components of gentamicin in a fermentation liquid by using HPLC-ELSD. Background technique [0002] Gentamicin [Gentamicin, gentamycin (gentamycin)] is a multi-component mixture extracted from the fermentation broth of Actinomycetes Monospore, including C 1 、C 2 、C 1a 、C 2a Four main components and C 2b , sisomicin, gentamicin B and other minor components are commonly used aminoglycoside antibiotics, mainly used to treat bacterial infections. Gentamicin is currently one of the main antibiotics clinically used to resist various Gram-negative bacterial infections, and it is one of the few heat-stable antibiotics, so it has been widely used at home and abroad. There are also many enterprises that ferment and produce gentamicin. [0003] Due to the many compo...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06G01N30/16G01N30/30G01N30/32G01N30/34G01N30/74G01N30/86
CPCG01N30/482G01N2030/484G01N30/02G01N30/34G01N30/32G01N30/06G01N30/16G01N30/30G01N30/74G01N30/8651G01N30/8679G01N2030/324G01N2030/062G01N2030/3007
Inventor 杜精精王绘砖徐珍
Owner HEBEI SHENGXUE DACHENG PHARMA
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