液质联用法检测保健食品中非法添加的13种5型磷酸二酯酶抑制剂

Determination of 13 kinds of PDE-5 inhibitors illegally added in health food by liquid chromatography-tandem mass spectrometry

目的建立同时检测保健食品中非法添加的13种5型磷酸二酯酶(PDE-5)抑制剂的液相色谱-串联质谱分析方法,并用于实际检测。方法样品经乙腈超声提取后,以资生堂CAPCELL PAK C18MGⅢ色谱柱(2.1 mm×150 mm,5μm)进行分离,甲醇(含15%乙腈)和0.02 mol/L乙酸铵水溶液(含15%乙腈和0.1%乙酸)为流动相,0.2 ml/min梯度洗脱;采用电喷雾电离源正离子(ESI^+),多反应监测(MRM)模式进行检测。结果 13种5型磷酸二酯酶(PDE-5)抑制剂在20 min内即可完成检测,在10 ng/ml^500 ng/ml内线性相关系数(r)均> 0.998,定量限(LOQ)为0.10μg/g^0.75μg/g,在12.5μg/g、25μg/g、50μg/g 3个不同添加水平的加样回收率在76.5%~107.6%,相对标准偏差(RSD)在0.3%~6.0%。结论该方法操作简单,分析速度快,灵敏度高,可应用于补肾壮阳类保健食品中非法添加的13种PDE-5抑制剂的筛查及检测。

Objective To develop a method for the simultaneous determination of 13 kinds of PDE-5 inhibitors illegally added in health food by liquid chromatography-tandem mass spectrometry(LC-MS/MS).Methods The samples were extracted using acetonitrile.The target analyses were separated on CAPCELL PAK C18 MGⅢcolumn(2.1 mm×150 mm,5μm)with a mobile phase consisting of methanol containing 15%acetonitrile and 0.02 mol/L ammonium acetate containing 15%acetonitrile and 0.1%formic acid by gradient elution,the flow rate was 0.2 ml/min.The detection was performed in ESI+and MRM modes.Results 13 kinds of PDE-5 inhibitors were determined within 20 minutes,and the linear correlation coefficients(r)were greater than 0.998 in the range of 10 ng/ml-500 ng/ml.The limits of quantitation(LOQs)were in the range of 0.10μg/kg-0.75μg/kg.The recoveries at spiked levels of 12.5μg/g,25μg/g and 50μg/g were in the range of 76.5%-107.6%,with relative standard deviations(RSDs)of 0.3%-6.0%.Conclusion This method is simple,rapid and sensitive,and it can be applied to the screening and detection of 13 PDE-5 inhibitors illegally added in health foods.