猪粪中21种常见抗生素的同步提取检测方法研究及应用

Research and application of detection method of simultaneous extraction and detection of 21 common antibiotics from pig manure

为实现猪粪中抗生素的有效削减及控制,迫切需要开发快速、同步兽用抗生素的检测分析方法,明确猪粪中多种类抗生素赋存特征.本研究建立并优化了猪粪中21种常见抗生素的同步分析方法.该方法采用甲醇和Na_(2)EDTA-McIlvaine缓冲溶液(1∶1,V/V)提取,提取液经固相萃取净化后高效液相色谱串联质谱检测.猪粪样品中5种四环素类、2种β-内酰胺类、6种大环内酯类、4种磺胺类和4种喹诺酮类抗生素的内标定量回收率29.53%—116.36%,检出限0.90—26μg·kg^(-1)DS,定量限2.91—85.20μg·kg^(-1)DS,相关系数R2均大于0.997.采用该方法对单个猪场不同畜舍样品加标回收率的单因素方差分析发现,除了四环素-D6(TC-D6)有显著性影响之外,其它内标回收率在不同样品中无显著性影响.分别采集不同地区2个猪场的猪粪样品进行验证分析,发现猪粪样品中单个抗生素含量范围在ND—99.37 mg·kg^(-1)DS.结果表明,该方法可用于猪粪中多种类抗生素的同步检测.

In order to achieve effective reduction and control of antibiotics in pig manure,there is an urgent need to develop rapid and simultaneous detection and analysis methods for veterinary antibiotics,and to clarify the characteristics of the occurrence of multiple antibiotics in pig manure.In this study,a simultaneous analytical method was developed and optimized for the determination of 21 common antibiotics in swine manure.The method used methanol and Na_(2)EDTA-McIlvaine buffer solution(1∶1,V/V)for extraction,and the extracts were cleaned up by solid phase extraction and detected by high performance liquid chromatography tandem mass spectrometry.The recoveries of the internal standard quantification of five tetracyclines,two β-lactams,six macrolides,four sulfonamides and four quinolones antibiotics in pig manure samples ranged from 29.53%to116.36%,the limits of detection(LODs)were 0.90—26μg·kg^(-1) DS,and the limits of quantification(LOQs)were 2.91—85.20μg·kg^(-1) DS,with the correlation coefficients R^(2) were greater than 0.997.Using this method to analyze the single-factor analysis of variance for the recovery rates of samples from different houses of a single pig farm,it was found that except for the significant effect of tetracycline-D6(TC-D6),the recovery rates of other internal standards were not significant in different samples.Pig manure samples from two pig farms in different regions were collected for verification and analysis.It was found that the content of a single antibiotic in the pig manure samples ranged from ND—99.37 mg·kg^(-1) DS.The results show that this method can be used for simultaneous detection of multiple antibiotics in pig manure.