超高效液相色谱-四级杆/静电轨道阱高分辨质谱法快速定性和定量筛查抗风湿类中成药和保健食品中16种非法添加化学药物

Rapid screening and quantitative analysis of 16 illegally added drugs in anti-rheumatic Chinese patent drugs or dietary supplements by ultra high performance liquid chromatography-quadrupole/obitrap mass spectrometry

目的建立抗风湿类中成药和保健食品中16种非法添加化学药物的超高效液相色谱-四级杆/静电场轨道阱高分辨质谱(UPLC-Q Extractive)联用的测定方法。方法样品用0.1%甲酸-甲醇提取、过滤后上机进行分析。经BEH C18色谱柱(100 mm×2.1mm,1.7μm)分离,以含0.05%甲酸的2 mmol/L乙酸铵溶液-甲醇溶液作为流动相,梯度洗脱,流速0.3 mL/min,柱温35℃。质谱分析则采用电喷雾离子源进行正离子和负离子模式同时扫描,数据采集使用以母离子全扫描和数据依赖的二级子离子扫描(Full MS/dd-MS2)模式,以分析物的保留时间和一级母离子以及自动触发采集的二级碎片离子信息建立数据库,进行筛查和定量分析。结果16种化学药物在10.0μg/L~100.0μg/L,线性范围内呈良好的线性关系(r2≥0.992);检出限为0.3μg/L和1.5μg/L,定量限为0.9μg/L和5.0μg/L;加标水品为10μg/L~80μg/L时,回收率为82.1%~108.5%,RSD为0.3%~4.7%。用上述方法对30个样本进行筛查,检出1批阳性样品并进行定量分析。结论该方法具有快速准确,灵敏度高,能够满足抗风湿类中成药和保健食品中非法添加化学药物的筛查和定量。

Objective To develop a method for the determination of 16 illegally added drugs in anti-rheumatic Chinese patent drugs or dietary supplements by ultra-high-performance liquid chromatography-quadrupole/obitrap mass spectrometry(UPLCQ Extractive).Methods The samples were extracted with 1%acetic acid-methanol and filtrated before analysis,separated on a BEH C18 column(100 mm×2.1 mm,1.7μm)by gradient elution with a water containing 2 mmol/L ammonium acetate combined with 0.05%formic acid-methanol as mobile phase at a flow rate of 0.3 mL/min,and the column temperature was 35℃.The analyses were detected by both positive and negative ion ESI in Full MS/dd-MS2 mode.A database was set up depending on the retention time,accurate mass of parent ions and confirmation in two-stage threshold-triggered full mass scan mode.Results The linear relationship was good within 10.0μg/L~100.0μg/L(r2≥0.992)for the 16 illegally added drugs.The LOD was 0.3μg/L and 1.5μg/L;the LOQ was 0.9μg/L and 5μg/L.Recovery of this method was in range of 82.1%~108.5%with spike levels of 10~80μg/L,and the RSD was within 0.3%~4.7%.The method mentioned above was applied in the detection of 30 samples,and 1 positive sample was found and quantitatively analyzed.Conclusion The UPLC-Q Extractive method is rapid,accurate,and high sensitive,which can meet the rapid screening and quantitative analysis of 16 illegally added drugs in anti-rheumatic Chinese patent drugs or dietary supplements.