高效液相色谱串联质谱法检测饲料中7种青霉素类药物

Determination of seven penicillins in feed by HPLC-MS/MS

试验建立了高效液相色谱串联质谱法测定饲料中7种青霉素类药物的方法,根据饲料种类特点,采用不同方法进行前处理,对配合饲料、浓缩饲料、精料补充料和单一饲料,通过20%乙腈溶液提取,提取液直接过膜上机测定。对预混合饲料采取以Na2EDTA-Mcllvaine溶液(pH=8)作为提取溶液,通过HLB固相萃取小柱净化的方法。采用C18色谱柱分离,0.2%甲酸溶液+乙腈作为流动相进行梯度洗脱,通过多反应监测模式进行测定。在10~1 000μg/L时线性良好,相关系数均大于0.99。在0.1、0.2、0.4 mg/kg添加水平下,平均回收率为62.2%~102.5%,相对标准偏差均小于15%,检测限为0.05 mg/kg,定量限(LOQ)为0.1 mg/kg,该方法操作简便、结果准确、稳定性好。

A method of HPLC-MS/MS was developed to determine seven penicillins in feed. According to the kind and characteristic of feed, formula feed, concentrated feed,concentrate supplement and single feed were extracted by 20% of acetonitrile by different pretreatment methods, and the extracting solution was detected after filtering with filter membrance. Premixed feed was extracted by Na2EDTA-Mcllvaine(pH=8) and purified by HLB SPE. The sample solution was separated by C18 chromatographic column. Acetonitrile and 0.2% of formic acid were used as the moving phase to carry out gradient elution under the multiple reaction monitoring mode. Qualitative and quantitative analysis of seven penicillins were carried out in multiple reactions monitoring mode(MRM). The limits of detection(LODs) of seven penicillins were 0.05 mg/kg, and the limit of quantitation(LOQs) of seven penicillins was 0.1 mg/kg. The correlation coefficient of the seven penicillins ranged from 10.0~1 000 μg/L was greater than 0.99. The results showed that when the concentrations of seven penicillins were between 0.1 mg/kg and 0.4 mg/kg, the average recoveries ranged from 62.2% to 102.5%, with relative standard deviations less than 15%. The method is simple, accurate and stable.