高效液相色谱-串联质谱法检测中成药/保健食品中非法添加的49种化学药物

Determination of 49 kinds of illegally added chemical drugs in Chinese patent medicines and health foods by high performance liquid chromatography-tandem mass spectrometry

中文关键词:  高效液相色谱-串联质谱法  非法添加  中成药  保健食品  降压  降糖

英文关键词:high performance liquid chromatography-tandem mass spectrometry  illegal addition  Chinese patent medicines  health foods  antihypertensive drugs  anti-diabetic drugs

中文摘要:

      目的 建立高效液相色谱-串联质谱法测定降压降糖类中成药和保健食品中非法添加的49种化学药物的分析方法。方法 60%甲醇超声提取样品, 采用Agilent pursuit 5 PFP色谱柱(100 mm×2.0 mm, 5 μm), 以0.1%甲酸水-乙腈为流动相, 0.3 mL/min进行梯度洗脱; 柱温20 ℃, 进样体积5 μL; 采用电喷雾离子源, 正离子模式进行检测; 通过对照品保留时间、分子离子及二级碎片离子比对定性; 多反应监测(multiple reaction monitoring, MRM)下内标法定量计算含量。结果 49种化学药物在各自的考察范围内线性关系良好, 相关系数均大于0.99; 方法日内精密度的相对标准偏差(relative standard deviations, RSDs)为0.18%~9.92%; 日间精密度RSDs为4.29%~9.96%; 回收率采用3个浓度进行加样回收实验, 回收率范围为70.31%~129.35%; 检出限为0.1~5.0 ng/mL。应用该方法对20批样品进行了检测, 其中3批次样品中检出了格列本脲、可乐定、二甲双胍化学药物。结论 该方法专属性强、灵敏度高, 可用于降压降糖类中成药和保健食品中非法添加化学药物的快速筛查和定量检测。

英文摘要:

      Objective To establish a method for the determination of 49 kinds of illegally added chemical drugs in antihypertensive and anti-diabetic Chinese patent medicines and health foods by high performance liquid chromatography-tandem mass spectrometry. Methods The samples were extracted with 60% methanol by ultrasonic processing and separated on an Agilent pursuit 5 PFP column (100 mm×2.0 mm, 5 μm) with 0.1% formic acid water solution and acetonitrile as the mobile phase by gradient elution at a flow rate of 0.3 mL/min; the column temperature was 20 ℃ and the injection volume was 5 μL; those chemicals were determined by Electrospray ion source in positive mode. The retention time of the reference substance, molecular ions and secondary fragment ions were compared for qualitative analysis; the content was quantitatively calculated by internal standard method under multiple reaction monitoring (MRM). Results The method validation showed good linearity with correlation coefficients above 0.99 for all chemical drugs; the relative standard deviations (RSDs) values for intra-day precision ranged from 0.18% to 9.92% and the RSDs of inter-day precision ranged from 4.29% to 9.96%; the recoveries at 3 concentrations ranged from 70.31% to 129.35%; the limits of qualitative detection were 0.1 to 5.0 ng/mL. Twenty batches of samples were tested by this method, and glibenclamide, clonidine and metformin were detected in three batches of samples. Conclusion The method has high specificity and sensitivity, and can be used for rapid screening and quantitative detection of illegal chemical drugs in antihypertensive and anti-diabetic Chinese patent medicines and health foods.