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目的建立基于肝细胞毒价检测的雷公藤药材质量评价方法。方法以正常人肝细胞(L02细胞系)为研究载体,以细胞抑制率为检测指标,优化建立肝细胞毒价检测方法 ,评价不同产地雷公藤及其地方习用品昆明山海棠药材的质量。结果以对乙酰氨基酚(毒价定义为400 U/g)为阳性对照品,50%乙醇超声提取雷公藤后真空干燥为干膏,用培养基配制生药量3 mg/m L的供试液,稀释比1∶0.65,量反应平行线法检测,18份雷公藤样品肝细胞毒价17.78~4 131.4 U/g(相差超过200倍),5份昆明山海棠样品肝细胞毒价209.42~7 422.2 U/g(相差超过30倍),不同产地雷公藤及昆明山海棠药材的肝细胞毒价差异显著;并且所检测样品鲜品较市售干品毒性大。结论初步建立了基于肝细胞毒价检测的雷公藤质量评价方法 ,直接关联其临床肝脏毒副反应,对从质量控制角度提高雷公藤临床用药安全具有参考价值,也为雷公藤肝毒性成分筛选提供准确定量的方法 。
Objective To establish a method for the quality evaluation of Tripterygium wilfordii based on the detection of hepatotoxicity. Methods Normal human hepatocytes (L02 cell line) was used as the research carrier, and the cell inhibition rate was taken as the detection index to establish the hepatotoxicity detection method and to evaluate the quality of Tripterygium wilfordii and its local habitat Kunming Begonia. Results Paracetamol (the drug price was defined as 400 U / g) was used as the positive control. The extract of Tripterygium wilfordii was extracted with 50% ethanol and then vacuum dried to dry paste. The test solution of crude drug amount 3 mg / m L , The dilution ratio of 1: 0.65, the parallel reaction of the amount of reaction detection, 18 Tripterygium sample liver cell drug price 17.78 ~ 4 131.4 U / g (difference of more than 200 times), 5 copies of Kunming Begonia sample liver cell drug price 209.42 ~ 7 422.2 U / g (a difference of more than 30 times). The difference of hepatotoxicity between medicinal materials of Tripterygium wilfordii and Kunming Begonia from different areas was significant. And the tested products were more toxic than the commercially available ones. Conclusion The method of evaluating the quality of Tripterygium wilfordii was established based on the detection of hepatotoxicity in liver cells. It is directly related to the clinical liver toxicity and toxicity. It is of reference value to improve the safety of Tripterygium wilfordii from the perspective of quality control. Accurate quantitative method.