实心核颗粒色谱分离-HPLC-MS/MS法同时测定性保健品中非法添加15种壮阳类化学成分

Detection of 15 afrodyn drugs illegally added in sex-enhancing product by HPLC-MS/MS with solid nuclear particle chromatographic column

目的建立了性保健品中非法添加15种壮阳类化学成分的实心核颗粒色谱分离-HPLC-MS/MS检测方法。方法以高色谱性能的实心核颗粒色谱柱CORTECS-C18(150 mm×4.6 mm,2.7μm)分离,串联四级杆质谱仪检测,MRM模式进行定性定量分析,样品以甲醇为溶剂超声提取,检测添加在性保健品中的甲磺酸酚妥拉明、那红地那非、红地那非、伐地那非、西地那非、羟基豪莫西地那非、豪莫西地那非、氨基他达拉非、他达拉非、硫代艾地那非、伪伐地那非、那莫西地那非、甲睾酮、司坦唑醇和达那唑共15种化学成分。结果 15种壮阳类化学成分质谱检测的分离度较好,线性范围宽,相关性好,r2≥0.995 4;方法精密度以6次测定值的RSD表示,为0.9%~3.3%;方法回收率用3个质量浓度进行添加实验,回收率为88.3%~109.5%,RSD为0.9~5.2%;定量限范围为0.015~0.30μg/m L;日内精密度和日间精密度的RSD分别为0.9%~4.2%和1.0%~4.5%;25批次性保健品中检出阳性样品20批次,检出成分均为西地那非,不合格率达80%。结论本方法通用性强,操作简单,可作为性保健品中非法添加15种壮阳类化学成分的有效检测方法。

Objective To develop an HPLC-MS/MS method with solid nuclear particle chromatographic column to determine 15 afrodyn drugs illegally added in sex-enhancing product. Methods The analysis was performed by an HPLC-MS/MS system, with Cortecs-C18（150 mm × 4.6 mm, 2.7 μm） column. Multiple-reaction monitoring（MRM） was performed to identify and quantify phentolamine mesylate, noracetildenafil, acetildenafil, vardenafil, sildenafil, hydroxyhomosildenafil, homosildenafil, aminotadalafil, tadalafil, thioaildenafil, pseudovardenafil, norneosildenafil, methyltestosterone, stanozolol and danazol, which were extracted with methanol by ultrasonic. Results The linear calibration curves of 15 chemical components mentioned above were obtained at r2 ≥0.995 4 with better separation degree and wide linear range. The precision of the method was shown by RSD（n = 6） from six estimated values ranged from 0.9% to 3.3%. The recovery rates determined at three adding concentrations were from 88.3% to 109.5% and the RSD was from 0.9% to 5.2%. The range of limit of quantitation（LOQ） were from 0.015 μg/m L to 0.30 μg/m L and The RSD of inter-day and intra-day precision were from 0.9% to 4.2% and from 1.0% to 4.5%, respectively; Twenty batches of positive samples of 25 batches of sex-enhancing products were detected to be added with sildenafil and with an unqualified rate of 80%. Conclusion The method is specific and simple to detect that 15 afrodyn drugs were added illegally in sex-enhancing product. At the same time, the high positive detection rate should attract the attention of the department concerned, and the corresponding policies should be issued to guarantee the health and safety of the people.