HPLC同时测定中成药中13种非法添加的降糖类化学药物

Simultaneously Detecting of 13 Kinds of Illegal Hpyerglycemic Additives in Chinese Patent Drug by HPLC

关键词:  中成药  降糖作用  非法添加  高效液相色谱  超高效液相色谱-串联质谱

Key words:  Chinese patent drug  hpyoglycemic effect  illegal additive  HPLC  UPLC-MS

摘要:

目的 建立同时测定中成药中非法添加13种降糖化学药物的方法。方法 采用HPLC-DAD进行测定,CAPCELL PAK C₁₈色谱柱(4.6mm×250mm,5μm),以甲醇-十二烷基硫酸钠溶液(pH 3.5)为流动相进行梯度洗脱,DAD检测器扫描,提取波长235nm,片剂、颗粒剂、丸剂、胶囊剂、口服液等中成药样品经甲醇提取,通过保留时间和紫外吸收光谱进行定性,峰面积外标法定量。检测的阳性样品以UPLC-MS进行确证,ESI源,正离子模式,SRM扫描。结果 13种降糖类药物成分能完全分离,在0.5~20μg·mL^-1内线性关系良好,回收率为94.36%~104.30%,RSD为0.5%~3.2%;检出的1批阳性样品,通过UPLC-MS能够得到确证。结论 该方法操作省时、结果准确可靠,适用日常监管检验。

Abstract:

OBJECTIVE To construct an approach which can be used to assess the concentration of 13 illegal chemical hypoglycemic additives in Chinese patent drug simultaneously.METHODS HPLC-DAD was employed in this research.CAPCELL PAK C₁₈ column(4.6 mm×250 mm, 5 μm) was used. The samples were eluted by methanolsodium dodecyl sulfate solution with gradient concentration. DAD detector was used with wavelength of 235 nm. Different forms of Chinese patent drug were analyzed, which included tablets, granules, pills, capsules and oral solution. The samples were extracted using methanol.Qualitative results were determined using retention time and ultraviolet spectrum. Quantitative results were obtained via external standard method based on the area of absorption peak. The positive samples were verified using UPLC-MS(ES1, positive ion mode, SRM scanning).RESULTS The 13 hypoglycemic chemicals were completely separated. The linearity were good when the concentration in the range of 0.5-20 μg·mL^-1. The recoveries rate were 94.36%-104.30% with RSDs of 0.5%-3.2%. One batch of samples were tested positive, which was verified using UPLC-MS.CONCLUSION The method is timesaving and reliable, which is suitable for daily drug controlling supervision.