基于化学荧光测定的板蓝根抗病毒效价检测方法的建立

Establishment of Bioassay Method for Antivirus Potency of Radix Isatidis Based on Chemical Fluorometric Determination

荧光测定法在化学分析和生物检测中已广泛应用,将该法引用到中药的生物效价检测中则是一种新的尝试。而生物效价检测与药效直接相关联,是研究中药质量控制与品质评价方法的新趋势。为了建立与抗病毒药效相关的板蓝根质量控制和评价方法,采用化学荧光测定法考察了板蓝根提取物对流感病毒神经氨酸酶(NA)的体外抑制活性,分析了其药理作用的量效曲线变化趋势和作用规律。在此基础上并根据生物检定的统计学原理,以"质反应平行线"法设计和优化检测条件,建立了板蓝根抗病毒生物效价荧光检测方法。实际测试结果能够灵敏、定量表征样品间的品质差异,方法的可靠性和可重复性较好。以此作为板蓝根质量生物控制和评价方法,能够体现中药药效特点。

In the present study, the in vitro inhibitory activity of extracts from radix isatidis on neuraminidase （NA） was investigated by the chemical fluoremetic determination to establish the quality control method for antivirus action of radix isatidis. The initial study indicated that radix isatidis had obvious in vitro inhibitory activity on NA with IC_50 = （0. 90±0. 20） mg（herb）·mL^-1. The correlation between logarithmic dose and reaction rate showed a ＂S＂ shape and a linear curve （linear equation, y= 8. 725 9±1. 216 9×og（D）, R= 0. 999 2） when the reaction rate was converted to probit-quite similar to Tamiflu＇s reaction curve, which hinted that radix isatidis had the same inhibitory function on NA as Tamiflu. According to the reaction type and the regularity of ＂parallel lines of qualitative effect＂, the experimental condition was optimized and a statistic method was confirmed based on the principle of bioassay statistic. Then the bioassay method for antivirus potency of radix isatidis based on fluorometric determination was established. The results of bio-potency assay showed the qualitative differences of radix isatidis samples sensitively and quantitatively. Meanwhile, this method has good reproducibility with RSD=5. 78% and reliability. The quality bioassay control method based on chemical fluorometric determination can reflect the pharmaco-dynamic features of Chinese medicine herb.