Method for determining antibiotics in human urine through solid phase extraction-high performance liquid chromatography-tandem mass spectrometry

A technology of high performance liquid chromatography and tandem mass spectrometry, which is applied in the field of solid phase extraction-high performance liquid chromatography-tandem mass spectrometry for the determination of antibiotics in human urine, can solve the problems of difficult separation and low detection accuracy, and achieves good selectivity, A large number, the effect of reducing the effect of matrix interference

Active Publication Date: 2021-10-01
SOUTH CHINA NORMAL UNIVERSITY
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  • Claims
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Problems solved by technology

[0005] For above-mentioned deficiency, the object of the present invention is to provide a kind of solid-phase extraction-high performance liquid chromatography-tandem mass spectrometry method for the antibiotic in human urine, can effectively solve the existing human urine antibiotic detection technology due to inorganic salt and protein etc. Difficult separation of components causes the problem of low detection accuracy, and the present invention establishes a pretreatment analysis method of extraction, enrichment, and determination, which can simultaneously detect sulfonamides, quinolones, and tetracyclines in human urine in a short time 37 kinds of antibiotics in seven categories, macrolides, lincomycin, florfenicol and carbadol, including sulfathiazole, sulfidesoxine, marbofloxacin, beilimycin, omeprem and Kabado is the first report of its human urine detection method

Method used

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  • Method for determining antibiotics in human urine through solid phase extraction-high performance liquid chromatography-tandem mass spectrometry
  • Method for determining antibiotics in human urine through solid phase extraction-high performance liquid chromatography-tandem mass spectrometry
  • Method for determining antibiotics in human urine through solid phase extraction-high performance liquid chromatography-tandem mass spectrometry

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Embodiment 1

[0033]This example provides a method for the determination of antibiotics in human urine by solid phase extraction-high performance liquid chromatography-tandem mass spectrometry, which specifically includes the following steps:

[0034] Step (1) pretreatment: add internal standard, 1M acetic acid-ammonium acetate buffer and β-glucuronidase to the human urine sample, and incubate at 37°C for 12 hours; among them, the addition of acetic acid-ammonium acetate buffer The amount is 20% (v / v), the addition amount of β-glucuronidase is 2% (v / v), and the pH of the acetic acid-ammonium acetate buffer is 5;

[0035] Step (2) enrichment and purification: pass the human urine sample obtained in step (1) into the activated HLB column (the activation process of the solid phase extraction column is: activate the HLB column with methanol and ultrapure water in sequence), and then use the ultrapure Water and 30% (v%) aqueous methanol solution are rinsed successively, and are sucked dry under ...

experiment example

[0040] In this example, the same method as in Example 1 was used to detect 4 unknown human urine samples collected in Guangzhou, and the experimental results are shown in Table 1. A total of 11 sulfonamide antibiotics, 2 quinolone antibiotics, 4 tetracycline antibiotics, 3 macrolide antibiotics, lincomycin and florfenicol were detected; the total concentration ranged from 0.67 ng / mL to 5.70ng / mL, among which, the content of sulfamethoxazole was the highest.

[0041] Table 1 The detection results of antibiotics in 4 unknown human urine samples in Guangzhou (ng / mL)

[0042]

[0043]

[0044] It can be seen from the above results that the detection method of the present invention has good separation of 37 kinds of antibiotics, stable baseline, high precision, high sensitivity, high stability, high selectivity and low detection limit, and the advantages of more real and reliable detection results .

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Abstract

The invention discloses a method for determining antibiotics in human urine through solid phase extraction-high performance liquid chromatography-tandem mass spectrometry, and belongs to the technical field of detection analysis, and the method mainly comprises the following steps: carrying out solid phase extraction treatment on a pretreated human urine sample to obtain a human urine concentrated sample, and carrying out high performance liquid chromatography-tandem mass spectrometry detection. According to the invention, a solid-phase extraction method is adopted to extract and purify a detected sample at the same time, so that the interference of high-concentration inorganic salt, protein, urea and other components in urine can be effectively removed, the matrix interference effect is reduced, and the detection result is more accurate; meanwhile, the method can simultaneously detect the residual conditions of 37 antibiotics in human urine at one time, and has the advantages of high precision, high sensitivity, high stability, high selectivity, low detection limit, more real and reliable detection result and the like.

Description

technical field [0001] The invention relates to the technical field of detection and analysis, in particular to a solid-phase extraction-high performance liquid chromatography-tandem mass spectrometry method for determining antibiotics in human urine. Background technique [0002] Antibiotics are a large class of drugs with antibacterial or bactericidal activity, which can be produced by microorganisms (bacteria, fungi, and actinomycetes) or higher animals and plants, and can also be synthesized artificially. They mainly include sulfonamides, tetracyclines, quinolones, macrolides and β-lactams. Antibiotics are mainly used in the treatment of bacterial infections in humans and animals, and also as feed additives to promote animal growth. At present, China is the world's largest producer and user of antibiotics. It is estimated that in 2013 China's antibiotic consumption was as high as 162,000 tons, accounting for about half of the world's total, of which 48% was for human u...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/08G01N30/14G01N30/34G01N30/72G01N30/86
CPCG01N30/02G01N30/06G01N30/08G01N30/14G01N30/34G01N30/7266G01N30/8675G01N2030/045G01N2030/062Y02A50/30
Inventor 刘思思陈长二应光国杨愿愿
Owner SOUTH CHINA NORMAL UNIVERSITY
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