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
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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)
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[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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