超高效液相色谱串联质谱法检测山药丸中4种黄曲霉毒素

Determination of Four Aflatoxins in Shanyao Pills by UPLC-MS/MS

目的建立同时测定山药丸中4种黄曲霉毒素含量的超高效液相色谱串联质谱法(UPLC-MS/MS)。方法样品经70%甲醇提取,再用QuEChERS净化管和免疫亲和柱净化,色谱柱为Acquity UPLC BEH C18柱(100 mm×2.1 mm,1.7μm),流动相为乙腈-10 mmol/L乙酸铵溶液(梯度洗脱),流速为0.20 mL/min,柱温为25℃,进样量为2μL,采用多反应监测、电喷雾电离、正离子扫描模式,毛细管电压为2.4 kV,脱溶剂气温度和流量分别为450℃、950 L/h。结果黄曲霉毒素B1,B2,G1,G2的质量浓度分别在0.108~2.160 ng/mL、0.033~0.660 ng/mL、0.101~2.020 ng/mL、0.030~0.600 ng/mL范围内与峰面积线性关系良好(r≥0.998,n=5);精密度、稳定性、重复性试验的RSD均小于4.0%(n=6);平均回收率分别为93.00%,87.22%,87.88%,87.44%,RSD分别为1.63%,3.05%,1.85%,3.26%(n=6)。6批样品检出黄曲霉毒素,检出量分别为0.6,0.5,1.4,0.9,0.9,0.8μg/kg。结论该方法操作简便、灵敏度高、快速准确、专属性强,可用于同时测定山药丸中4种黄曲霉毒素的含量。

Objective To establish an ultra-high performance liquid chromatography-mass spectrometry(UPLC-MS/MS)method for simultaneous determination of four aflatoxins in Shanyao Pills.Methods The sample was extracted with 70%methanol and purified by the QuEChERS purification tubes and immunoaffinity columns.The chromatographic column was Acquity UPLC BEH C18 column(100 mm×2.1 mm,1.7μm),the mobile phase was acetonitrile-10 mmol/L ammonium acetate solution(gradient elution),the flow rate was 0.20 mL/min,the column temperature was 25℃,and the injection volume was 2μL.Multi-reaction monitoring mode was adopted with electric spray ion and positive ion scanning,the capillary voltage was 2.4 kV,the desolvation gas temperature was 450℃,and the flow rate was 950 L/h.Results The linear ranges of aflatoxins B1,B2,G1,and G2 were 0.108-2.160 ng/mL,0.033-0.660 ng/mL,0.101-2.020 ng/mL,and 0.030-0.600 ng/mL(r≥0.998,n=5),respectively.The RSDs of precision,stability,and repeatability tests were all lower than 4.0%(n=6).The average recoveries of aflatoxins B1,B2,G1,and G2 were 93.00%,87.22%,87.88%,and 87.44%,with RSDs of 1.63%,3.05%,1.85%,and 3.26%(n=6),respectively.Six batches of samples were detected with aflatoxin contents of 0.6,0.5,1.4,0.9,0.9,and 0.8μg/kg,respectively.Conclusion The method is simple,highly sensitive,fast,accurate,and highly specific,which can be used for the simultaneously determination of four aflatoxins in Shanyao Pills.