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。
Abstract
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(Woodwardia unigemmata or Dryopteris crassirhizoma) in Kanggan granules. C18 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 performed 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 reliable, which can be applied to detect the use of Woodwardia unigemmata in Kanggan granules.
关键词
高效液相色谱-串联质谱 /
抗感颗粒 /
绵马贯众 /
单芽狗脊贯众 /
山柰苷
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Key words
HPLC-MS/MS /
Kanggan granule /
Dryopteris crassirhizoma /
Woodwardia unigemmata /
kaempferitrin
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中图分类号：
R917
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参考文献
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[8] LAN Y L. Studies on the chemical constituents of Woodwardia unigemmata [D] . Wuhan: Hubei College Tradit Chinese Medicine, 2009.
[9] VALENTE L M M, BIZARRI C H B, LIECHOCKI S, et al. Kaempferitrin from Uncaria guianensis(Rubiaceae) and its potential as a chemical marker for the species [J] . J Braz Chem Soc, 2009, 20(6): 1041-1045.
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脚注
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基金
国家十二五重大新药创制专项课题—中药质量安全检测和风险控制技术平台资助项目资助(2014ZX09304307-002); 山东省自然科学基金资助项目(ZR2013HM074)
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