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Determination method of histamine content in veterinary gentamicin sulfate

A technology of gentamicin sulfate and determination method, which is applied in the field of analytical chemistry, can solve the problems of sensitivity, accuracy, stability not meeting the requirements, incapable of accurate quantitative detection, no histamine content, etc., achieving low cost, eliminating Matrix interference, quantitative and accurate effects

Active Publication Date: 2022-02-25
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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  • Determination method of histamine content in veterinary gentamicin sulfate
  • Determination method of histamine content in veterinary gentamicin sulfate
  • Determination method of histamine content in veterinary gentamicin sulfate

Examples

Experimental program
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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 100 μL of acetic acid and 10 mL of acetonitrile to obtain the extract, take 0.1 mL of sample solution 1 and sample solution 2, add 0.9 mL of ...

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 a supernatant; wherein, the mass ratio of the C18 powder and anhydrous magnesium sulfate 1:1:4;

[0080] (3) Nitrog...

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Abstract

The invention discloses a method for measuring histamine content in veterinary gentamicin sulfate, which belongs to the technical field of analytical chemistry. The method uses hydrochloric acid-methanol solution or hydrochloric acid-acetonitrile solution to extract and dilute veterinary gentamicin sulfate The histamine in the plain sample was purified by C18 powder, and then redissolved by nitrogen blowing and acetonitrile formic acid solution for LC-MS / MS determination. The method for determining the content of histamine in veterinary gentamicin sulfate provided by the present invention enables the high-performance liquid phase mass spectrometry instrument to better eliminate the interference of the matrix through the pretreatment of the sample, and then utilizes LC-MS / MS to quickly and accurately The method can accurately detect and quantify histamine in veterinary gentamicin sulfate, overcome the problem of difficult extraction of histamine in veterinary gentamicin sulfate, and make up for the inability to extract veterinary gentamicin sulfate in the prior art. Quantitative detection of histamine content in the vacancy.

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 Patents(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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