HPLC-MS/MS法同时测定止咳平喘类中成药中添加的7个化学抗菌类物质

Simultaneous determination of adulteration of seven antimicrobial agents in Chinese patent medicines for relieving cough and asthma by HPLC-MS/MS

目的:建立高效液相色谱-质谱联用法测定止咳平喘类中成药中非法添加的磺胺嘧啶、吡哌酸、甲氧苄啶、氧氟沙星、诺氟沙星、磺胺二甲嘧啶、磺胺甲噁唑7个化学抗菌类物质。方法:以Venusil MP C_(18)(4.6 mm×250 mm,5μm)为色谱柱,乙腈-甲醇(1∶1)-甲酸铵溶液为流动相,梯度洗脱,柱温为30℃,流速为1 mL·min^(-1);质谱配置电喷雾电离源(ESI),干燥气温度325℃,干燥气流速5 L·min^(-1),雾化压力103.4 kPa,扫描方式为正离子模式进行一级质谱全扫描、二级质谱全扫描,扫描范围为m/z 50~650。结果:7个化学抗菌药检测下限均小于0.04μg,回收率为94.4%~98.9%;在1.4~17.6μg·mL^(-1)范围内,线性关系良好(r>0.9996);3批阴性样品呈阴性,3批模拟阳性样品呈阳性,方法专属性强;用本文建立的方法对假药A~E进行了检验,检出假药A、C、D含甲氧苄啶、磺胺甲噁唑。结论:本方法专属性强,灵敏度高,可用于7个化学抗菌类非法添加物的验证试验。

Objective:To establish an HPLC-MS/MS method for simultaneous determination of seven antimicrobial agents(sulfadiazine,pipemidic acid,trimethoprim,sulfamethazine,ofloxacin,norfloxacin and sulfamethoxazole)in adulterated Chinese patent medicines for relieving cough and asthma.Methods:The analysis was performed on a Venusil MP C_(18)column(4.6 mm×250 mm,5μm)with a gradient elution of acetonitrilemethanol(1∶1)-0.3%ammonium formate solution at a flow rate of 1 mL·min^(-1).The column temperature was 30℃.The target compounds were analyzed with LC-MS in two scan modes including full scan MS and full scan MS2.The scan range was m/z 50~650.The drying gas temperature was 325℃,the drying gas flow rate was 5 L·min^(-1),and the drying gas pressure was 103.4 kPa.Results:Sulfadiazine,pipemidic acid,trimethoprim,sulfamethazine,ofloxacin,ofloxacin and sulfamethoxazole were all less than low 0.04μg and had good linearities in the range of 1.4-17.6μg·mL^(-1)(r>0.9996).The recoveries of high,median,and low concentrations control samples were 94.4%-98.9%.The test result of three batches of negative samples was negative,and the test result of three batches of analog positive samples was positive.Trimethoprim and sulfamethoxazole were found in three batches of counterfeit drugs.Conclusion:The method can be used for examining seven antimicrobial agents in adulterated Chinese patent medicines.