UPLC-MS/MS法测定保健食品中添加的114个化学药

目的：建立一种通过1次实验能同时完成保健食品中非法添加114个化学药的快速准确的筛查和定性分析的方法。方法：样品采用乙腈超声提取。采用UPLC-MS/MS法，以Atlantis?dC₁₈（2.1 mm×150 mm，3 μm）为色谱柱，0.1%甲酸水溶液（A）-乙腈（B）为流动相，梯度洗脱（0~3 min，90% A；3~15 min，90% A→60% A；15~28 min，60% A→0% A；28~29 min，0% A；29~30 min，0% A→90% A；30~31 min，90% A）；体积流量0.3 mL·min^-1，柱温40℃，进样体积5 μL。选择ESI离子源，正负离子扫描；多反应监测（MRM）模式，测定114个临床常用的化学药，通过比较MRM通道中样品峰与对照品峰的分子离子峰和二级碎片离子峰等信息，确定添加的化学药。结果：在上述色谱及质谱条件下，114个化学药的质谱检测的线性范围宽，相关性好，r≥0.992；方法精密度RSD为1.2%~6.5%（n=6）；方法回收率采用3个浓度（20、100、200 ng·mL^-1）进行加样回收试验，回收率范围在80.1%~120.0%；检测下限在0.006 6~7.210 8 μg·g^-1，定量下限在0.022 0~24.035 9 μg·g^-1。所建立的方法能够在1次检测中同时完成对保健食品中添加的114个化学药的筛查和定性分析。应用该方法对198批样品进行了检测，其中有34批次样品中检出盐酸西布曲明、N-单去甲基西布曲明、酚酞、盐酸苯乙双胍、盐酸二甲双胍和格列本脲等化学药。结论：研究结果表明，本法专属性强，灵敏度高，可作为保健食品中非法添加化学药的快速筛查和准确定性。

Objective: To establish a rapid and accurate method for the determination of 114 chemicals illegally added into the health foods in a single analysis. Methods: The UPLC-MS/MS method was adopted. The samples were extracted with acetonitrile by ultrasonic processing and separated on an Atlantis?dC₁₈ (2.1 mm×150 mm,3 μm) column with 0.1%formic acid water solution (A) and acetonitrile (B) as the mobile phase by gradient elution (0-3 min,90%A;3-15 min,90%A→60%A;15-28 min,60%A→0%A;28-29 min,0%A;29-30 min,0%A→90%A;30-31min,90%A) at a flow rate of 0.3 mL·min^-1. The column temperature was 40℃and the injection volume was 5 μL. Positive and negative-ion ESI sources with MRM mode were used to determine the chemicals.The obtained molecular ions,fragment ions and MRM channels were used to identify the 114 kinds of drugs by comparison with those of the reference substances. Results: 114 linear calibration curves were obtained with r ≥ 0.992. The RSD values (n=6) for precision ranged from 1.2% to 6.5%. The recoveries at three concentrations (20,100,200 ng·mL^-1) ranged from 80.1% to 120.0%. The limits of qualitative and limits of quantitative were in the range of 0.006 6-7.210 8 μg·g^-1 and 0.022 0-24.035 9 μg·g^-1,respectively. The established method could complete the screening and confirmation for 114 kinds of drugs in a single analysis. 198 batches of samples were tasted by this method. Sibutramine hydrochloride,N-monodesmethyl sibutramine hydrochloride,phenothalin,phenformin hydrochloride,metformin hydrochloride and glibenclamide were detected in 34 samples. Conclusion: The established method is specific and sensitive. The method can use in screening and confirmation of illegally added chemicals in health foods.

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