HPLC-MS/MS法检查止咳平喘类中药制剂非法添加研究

Study on HPLC-MS/MS Method in Inspection of Illegal Additions in Traditional Chinese Medicine Relieving Cough and Asthma Preparations

目的建立高效液相色谱-串联质谱(HPLC-MS/MS)法检查平喘止咳类中药制剂中非法添加的磺胺甲二唑、茶碱、磷酸可待因、枸橼酸喷托维林等10种化学药物。方法采用Kromasil 100-5C18色谱柱(250 mm×4.6 mm, 5μm)。流动相A、B依次为甲醇、0.1%甲酸,梯度洗脱,体积流量0.3 mL/min,柱温40°,进样量5μL。电喷雾离子源(ESI)行正离子模式扫描。结果 10种化学药物峰面积和浓度线性系数均≥0.9980,呈良好线性关系,检测限、定量限范围依次为(0.03~3.19)mg/kg、(0.05~9.32)mg/kg, RSD均≤4.64%,回收率范围74.63%~102.11%,7批次样品中检测出1批次样品存在非法添加情况,添加物为磷酸可待因,其他9种化学药物未被检出。结论 HPLC-MS/MS法,精密度、准确性高,回收率较好,适用于医疗行业中平喘止咳类中药制剂中非法添加化学药物的有效督查监管。

Objective To establish a high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS) method to inspect the illegal additions of 10 kinds of chemical drugs such as sulfamethizole, theophylline, codeine phosphate and pentoverine citrate in traditional Chinese medicine relieving cough and asthma preparations.Methods Kromasil 100-5 C18 chromatographic column(250 mm×4.6 mm, 5 μm) was performed. The mobile phases A and B were methanol and 0.1% formic acid, with gradient elution, volume flow rate of 0.3 mL/min, column temperature of 40℃ and sample size of 5 μL. Electrospray ionization(ESI) was used for positive ion mode scanning.Results The linear coefficients of peak area and concentration of the 10 chemical drugs were all more than 0.9980, showing a good linear relationship. The limit of quantification and limit of range were(0.03-3.19) mg/kg and(0.05-9.32) mg/kg, and the RSD were all less than 4.64%, and the recovery rate ranged from 74.63% to 102.11%. Among 7 batches of samples, 1 batch of samples was detected with illegal additions, and the additive was codeine phosphate and the other 9 chemical drugs were not detected.Conclusion HPLC-MS/MS method has high precision, high accuracy and good recovery rate, and it is suitable for effective supervision and inspection of illegal additions of chemical drugs in traditional Chinese medicine relieving cough and asthma preparations in the medical industry.