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Method for measuring histamine content in gentamicin sulfate for veterinary use

A technology of gentamicin sulfate and a determination method, applied in the field of analytical chemistry, can solve the problems of unsatisfactory sensitivity, accuracy, stability, inability to accurately quantitatively detect, no histamine content, etc., and achieves low cost, elimination of Matrix interference, quantitatively accurate effect

Active Publication Date: 2019-12-31
INSPECTION & QUARANTINE TECH CENT OF FUJIAN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In the second half of 2016, users of gentamicin sulfate in some EU countries found that they had adverse reactions after intravenous injection of gentamicin sulfate to horses. Animal experiments showed that the adverse reactions may be related to the histamines contained in the product , but there is no corresponding quantitative detection method for histamine content in aminoglycoside broad-spectrum antibiotics such as gentamicin sulfate in the world, and it cannot be accurately quantitatively detected, and there are no limit regulations and requirements
However, it was found through the standard addition test in the early stage that the sensitivity, accuracy, stability and other indicators of the results of the above methods did not meet the requirements, and the above methods were not suitable for the detection of histamine in gentamicin sulfate.

Method used

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  • Method for measuring histamine content in gentamicin sulfate for veterinary use
  • Method for measuring histamine content in gentamicin sulfate for veterinary use
  • Method for measuring histamine content in gentamicin sulfate for veterinary use

Examples

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Effect test

Embodiment 1

[0033] Determination of the pretreatment method for veterinary gentamicin sulfate samples:

[0034] (1) Histamine extraction

[0035] Take 2g of veterinary gentamicin sulfate, add water to make up to 20mL, divide into two tubes, each 10mL, add 100μg of standard histamine to one of the tubes, and obtain sample solution 1 and sample solution 2.

[0036] 1 Acetic acid-methanol extraction

[0037] Mix 100 μL of acetic acid and 10 mL of methanol to obtain the extract, take 0.1 mL of sample solution 1 and sample solution 2, add 0.9 mL of extract respectively, shake and mix for 10 min, centrifuge at 12000 r / min for 10 min, take the supernatant, and filter through a 0.22 μm organic filter Membrane filtration, adding 1% acetic acid solution to double dilution, take 1mL on the machine.

[0038] 2 Acetic acid-acetonitrile extraction

[0039] Mix 100μL acetic acid and 10mL acetonitrile to obtain the extract, take 0.1mL sample solution 1 and sample solution 2 respectively, add 0.9mL ext...

Embodiment 2

[0076] A method for assaying histamine content in veterinary gentamicin sulfate, specifically comprising the following steps:

[0077] (1) Dilution: Add 1% acetic acid solution and acetonitrile to veterinary gentamicin sulfate samples FG1406171 and FG1611323, ultrasonicate for 5 minutes, shake and mix for 3 minutes, then add 100 g / mL phosphatase solution, vortex for 1 minute, and Incubate in a constant temperature oven at 37°C for 2 hours to obtain a diluted veterinary gentamicin sulfate sample;

[0078] (2) Extraction: use hydrochloric acid-methanol solution or hydrochloric acid-acetonitrile solution to extract the histamine in the diluted veterinary gentamicin sulfate sample to obtain an extract;

[0079] (2) Purification: add C18 powder and anhydrous magnesium sulfate successively to the extract, shake for 1min, and centrifuge at 5000r / min for 5min to obtain the supernatant; wherein, the mass ratio of the C18 powder and anhydrous magnesium sulfate 1:1:4;

[0080] (3) Nitr...

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Abstract

The invention discloses a method for measuring histamine content in gentamicin sulfate for veterinary use, and belongs to the technical field of analytical chemistry. The measuring method uses hydrochloric acid-methanol solution or hydrochloric acid-acetonitrile solution to extract histamine of diluted gentamicin sulfate for veterinary use, purifies impurities in the extracting solution with C18 powder, re-dissolves by nitrogen blowing and acetonitrile formic acid solution, and then performs LC-MS / MS determination. The method for measuring histamine content in gentamicin sulfate for veterinaryuse provided by the invention enables the HPLC mass spectrometer to better exclude matrix interferences by pre-processing the sample, and then uses LC-MS / MS to quickly and accurately perform detection and quantification of histamine in gentamicin sulfate for veterinary use, which overcomes the difficulty in extracting histamine in gentamicin sulfate for veterinary use, and makes up for the inability to quantitative detection of histamine content in gentamicin sulfate for veterinary use in the prior art.

Description

technical field [0001] The invention belongs to the technical field of analytical chemistry, in particular to a method for determining the content of histamine in veterinary gentamicin sulfate. Background technique [0002] Gentamicin is a family of aminoglycoside antibiotics produced by the fermentation and metabolism of Micromons pra prppurea. It is mainly composed of three components of gentamicin C1, C2 and C1a. It is mainly used to treat bacterial infections, especially those caused by Gram-negative bacteria. Gentamicin can bind to the 30s subunit of bacterial ribosomes and block bacterial protein synthesis. Gentamicin is one of the few antibiotics with few side effects, strong bactericidal power and thermal stability, so it is widely used in culture medium configuration. However, due to the introduction of gentamicin in the fermentation process (the culture medium is peptone made from leftovers of fish, shrimp, beef and mutton, etc.) and bacterial fermentation, gent...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/14G01N30/72
CPCG01N30/02G01N30/06G01N30/14G01N30/72G01N2030/143
Inventor 郑腾张志灯刘正才张体银陈艳唐寰宇林杰
Owner INSPECTION & QUARANTINE TECH CENT OF FUJIAN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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