基于UPLC-MRM-IDA-EPI法高通量筛查保健食品中60种非法添加的化学药物

High-throughput screening and rapid quantification of 60 illegally added chemical drugs in dietary supplements by UPLC-MRM-IDA-EPI

目的基于UPLC-MRM-IDA-EPI技术建立60种非法添加化学药物的快速定量和定性分析方法。方法色谱分析采用UPLC-MS/MS法,以Phenomenex kinetex C_(18)(100 mm×3.0 mm,2.6μm)为色谱柱,0.1%甲酸水溶液(A)-甲醇(B)为流动相进行梯度洗脱,流速0.3 mL·min^(-1),柱温40℃,进样体积5μL。质谱分析采用电喷雾离子源(ESI)同时扫描正负离子,利用多反应监测(MRM)-信息依赖性采集(IDA)-增强子离子扫描(EPI)建立60种非法添加化学药物的EPI信息库;待测样本的分析采用MRM-IDA-EPI扫描监测疑似化合物并对其进行初步定量,同时将EPI碎片信息与EPI信息库进行对比,增加定性结果的可靠性。结果建立了60种化学药物的MS/MS谱库和定量方法,该方法定量分析的线性范围宽,相关性好(r≥0.99);方法精密度RSD为0.70%~7.0%(n=6);低、中、高浓度的回收率在60.1%~115.8%;检测限在2.5~1101 ng·g^(-1),满足检出非法添加的要求。对EPI数据库进行方法学验证,基质加标溶液中化合物与数据库中化合物的“匹配度”值均在68~95。应用该方法对30批样品进行筛查,共检出4批含有非法添加的样品,其添加物为硝西泮、他达拉非、西地那非。结论本研究建立的分析方法操作简单、专属性强,适用于高通量筛查及初步定量分析保健食品中非法添加的60种化学药物。

Objective To establish a high-throughput analytical method to monitor and quantify 60 illegally added chemical drugs by UPLC-MRM-IDA-EPI.Methods Samples were analyzed by a Phenomenex kinetex C_(18)(100 mm×3.0 mm,2.6μm)column with 0.1%formic acid water solution(A)and methanol(B)as the mobile phase with gradient elution at 0.3 mL·min^(-1).The column temperature was 40℃and the injection volume was 5μL.The electrospray ionization(ESI)-MS was used in both positive and negative ion mode with MRM-IDA-EPI to establish an EPI database of 60 standard chemical drugs.Suspected compounds were monitored by MRM scan mode and further determined by MRMIDA-EPI.Results We successfully established an EPI database for each chemical drug and applied the database for compound monitoring and quantification.All the linear calibration curves were obtained with r≥0.99 and the precision of the method was satisfactory with relative standard deviation(RSD)ranging from 0.70%to 7.0%(n=6).The recovery at low,middle,and high concentrations ranged from 60.1%to 115.8%.The limit of detection was at 2.5~1101 ng·g^(-1).The database used for identification was further verified by analyzing standards in the matrix,and the similarity value(“Purity”)of each compound ranged from 68 to 95.Totally 30 batches of samples were tested by this method.Nitrazepam,tadalafil and sildenafil were detected in 4 samples.Conclusion The method is efficient and easy to operate for the rapid monitoring of 60 illegally added chemical drugs in dietary supplements.