HPLC-MS/MS检测中药保健品中非法添加成分的研究

Simultaneous Determination of Four Antidiabetics Added to a Health-Care Food Illegally by HPLC-MS/MS

目的采用高效液相色谱-串联质谱技术（HPLC-MS/MS）,建立同时测定某“中药保健品”中非法添加的4种降糖药二甲双胍、苯乙双胍、罗格列酮、格列苯脲的方法。方法采用Thermo BDS Hypersil C8（2.1mm×100 mm,2.4μm）色谱柱,0.2%甲酸水溶液（A）和乙腈-甲醇（6∶4,v/v）（B）为流动相进行梯度洗脱,流速0.3 mL·min^-1,在电喷雾正离子模式下,以多反应监测（MRM）模式测定,定量离子对为m/z 129.9→70.9（二甲双胍）、m/z 205.9→59.9（苯乙双胍）、m/z 357.9→134.9（罗格列酮）、m/z 494.0→169.0（格列苯脲）。结果同时检出了4种降血糖化学药物。检出限为0.06～0.5 ng·mL^-1,定量限为0.21～1.62 ng·mL^-1,在3个不同浓度的添加水平下,4种化学药物的平均回收率为66.3%～110.2%,相对标准偏差（RSD）为2.3%～12.2%。结论该方法可快速、准确地检测保健食品中非法添加二甲双胍、苯乙双胍、罗格列酮、格列苯脲4种降血糖化学药,具有专属性强、灵敏度高的特点。

Objective To develop a HPLC-MS/MS method for simultaneous quantification of mefformin, phenformin, rosiglitazone, and glibenclamide, four antidiabetics which were illegally added into a health-care food. Methods The separation was performed on a Thermo BDS Hypersil C8 column（2.1 mm× 100 mm, 2.4 μm） by gradient elution with the mixture of 0.2 % aqueous formic acid（A） and acetonitrile-methanol（6 ： 4, v/v）（B） as mobile phase. The flow rate was set at 0.3 mL·min^-1. The detection of 4 target chemicals was performed by positive-ion electrospray ionization mass spectrometry（ESI-MS） in multiple-reaction monitoring（MRM） mode. The transitions of four antidiabetics executed as following： m/z 129.9→70.9 for metformin, m/z 205.9→9.9 for phenformin, m/z 357.9→134.9 for rosiglitazone, m/z 494.0→368.8 for glibenclamide. Results Four antidiabetics had been simultaneously identified from the health-care food by the established method. The limits of detection（LOD, S/N ≥ 3） and quantitation（LOQ, S/N 1〉 10） were 0.06～0.5 ng·mL^-1 and 0.21～1.62 ng·mL^-1, respectively. The average recoveries of the four analytes at three spiked concentration levels were 66.3 % to 110.2 % with relative standard deviations （RSDs） at 2.3 %-12.2 %. Conclusion The established method is convenient, rapid and accurate, and can be used for simultaneous determination of multiple antidiabetics in a health-care food which is labeled as a product of pure medicinal herbs.