抗感颗粒中混淆品单芽狗脊贯众投料的HPLC-MS/MS检测

Inspection of Illegal Use of Woodwardia unigemmata in Kanggan Granules by HPLC-MS/MS

目的建立检测抗感颗粒以绵马贯众易混淆品单芽狗脊贯众投料的高效液相色谱-串联质谱(HPLC-MS/MS)检测方法。方法单芽狗脊贯众药材中山柰苷含量远超过绵马贯众,通过测定抗感颗粒中山柰苷的含量,可以判断生产企业是否存在以单芽狗脊贯众投料的情况。具体测定条件为:以十八烷基硅烷键合硅胶为填充剂,流动相为含0.02%甲酸的乙腈-0.02%甲酸溶液,梯度洗脱;质谱采用电喷雾正离子模式(ESI^+),多反应监测(MRM)定量分析,以m/z 579.2→287.1作为定量离子对。结果山柰苷在272.46~5 449.25 pg内线性关系良好,回归方程为Y=139.217 2 X-2 901.93,相关系数r=0.999 6,平均回收率(n=6)为94.97%,RSD为0.85%。在检测收集的12批样品中,2家企业的7批样品山柰苷含量异常偏高,可能存在以单芽狗脊贯众投料的情况。结论建立的方法简便、准确、可靠,可用于检测抗感颗粒中单芽狗脊贯众的使用情况。

OBJECTIVE To establish an HPLC-MS/MS method for the inspection of illegal use of Woodwardia unigemmata, an adulterant of Dryopteris crassirhizoma, in Kanggan granules. METHODS The content of kaempferitrin in Woodwardia unigemmata is significantly higher than that in Dryopteris crassirhizoma. Therefore, the content of kaempferitrin can reflect the material usage （ Wood- wardia unigemmata or Dryopteris crassirhizoma） in Kanggan granules. C,s column was applied with a mobile phase consisting of 0. 02% formic acid acetonitrile（ V/V）-0. 02% formic acid H2O（V/V） in gradient elution mode. The mass spectral analysis was perionned in a positive electrospray ionization mode, and scanned by a multiple reaction monitoring mode. The precursor/product ion pair used for quantification was m/z 579.2→287. 1. RESULTS A good linear relationship was obtained in the range of 272.46 - 5 449.25 pg for kaempferitrin, with the linear regression equation of Y = 139. 217 2X - 2 901.93 （ r = 0. 999 6） ; the average recovery was 94. 97% （RSD =0. 85% , n = 6）. Among 12 batches of Kanggan granules, 7 batches from 2 enterprises showed abnormally high contents of kaempferitrin, which indicated illegal use of Woodwardia unigemmata, CONCLUSION The method is simple, accurate, and relia- ble, which can be applied to detect the use of Woodwardia unigemmata in Kanggan granules.