基于化学荧光测定的板蓝根抗病毒效价检测方法的建立

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。

Abstract

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 IC50=(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 log(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.

参考文献

[1] XIAO Xiao-he, JIN Cheng, ZHAO Zhong-zhen, et al(肖小河, 金城, 赵中振, 等).China Journal of Chinese Material Medicine(中国中药杂志), 2007, 32(14)： 1377.

[2] WANG Ping, QIAN Zhong-zhi(王平, 钱忠直). Chinese Journal of Pharmacentical Analysis(药物分析杂志), 2008, 28(2): 337.

[3] YANG Ming-hua, YANG Su-bei, JIN Zu-han(杨明华, 杨苏蓓, 金祖汉). Journal of Chinese Medicinal Materials(中药材), 2002, 6(6): 410.

[4] ZHANG Gui-jun(张贵君). Bioassay of General Chinese Medicines(常用中药生物鉴定). Beijing： Chemical Industry Press(北京：化学工业出版社), 2006. 179.

[5] Khare M D, Sharland M. Exp. Opin. Pharmacother, 2000, 1(3): 367.

[6] Cox N J, Subbarao K. Lancet., 1999, 354(9186): 1277.

[7] PENG Yong, LI Wen-kui(彭勇, 李文魁). World Phytomedicines(国外医药·植物药分册), 1999, 14(1): 7.

[8] LI Yan, WU Ran-ran, YU Bai-hua, et al(李燕, 吴然然, 于佰华, 等). Spectroscopy and Spectral Analysis(光谱学与光谱分析), 2007, 27(6): 1081.

[9] HU Xing-chang, ZHENG Wei-qiang(胡兴昌, 郑伟强). Journal of Shanghai Teachers University(上海师范大学学报), 2003, 32(1): 62.

[10] LIU Sheng, CHEN Wan-sheng, QIAO Chuan-zhuo, et al(刘盛, 陈万生, 乔传卓, 等). Acad. J. Sec. Mil. Med. Univ.(第二军医大学学报), 2000, 21(3): 204.

[11] XIAO Shan-shan, JIN Yu, SUN Yu-qing(肖珊珊, 金郁, 孙毓庆). Journal of Shenyang Pharmaceutical University(沈阳药科大学学报), 2003, 20(6): 455.

[12] CANN A J. Virus Culture A Practical Approach. New York: Oxford University Press. 1999.

[13] Deba Prasad Nayaka, Udo Reichl. Journal of Virological Methods, 2004, 122: 9.

[14] Potier M, Mameli L. Anal. Biochem., 1979, 94: 287.

[15] ZHOU Hai-jun(周海钧). Drug Bioassary(药品生物检定). Beijing: People’s Medical Press(北京：人民卫生出版社), 2005. 87, 127.

[16] XUE Bing-chun, WANG Tao, LIU Er-bao(薛冰纯, 王滔, 刘二保). Spectroscopy and Spectral Analysis(光谱学与光谱分析), 2006, 26(5): 816.

[17] WU Cheng-ke, FENG Su-ling, FAN Jing(吴呈珂, 冯素玲, 樊静). Spectroscopy and Spectral Analysis(光谱学与光谱分析), 2007, 27(12): 2490.

[18] LI Han-bing, YAN Dan, WANG Jia-bo, et al(李寒冰, 鄢丹, 王伽伯, 等). Acta Pharmaceutica Sinica(药学学报), 2009, 44(2)： 162.

李寒冰, 鄢丹, 金城, 王伽伯, 魏丽, 肖小河, 曹俊岭. 基于化学荧光测定的板蓝根抗病毒效价检测方法的建立[J]. 光谱学与光谱分析, 2009, 29(4): 908. LI Han-bing, YAN Dan, JIN Cheng, WANG Jia-bo, WEI Li, XIAO Xiao-he, CAO Jun-ling. Establishment of Bioassay Method for Antivirus Potency of Radix Isatidis Based on Chemical Fluorometric Determination[J]. Spectroscopy and Spectral Analysis, 2009, 29(4): 908.

基于化学荧光测定的板蓝根抗病毒效价检测方法的建立

关于本站 Cookie 的使用提示

全站搜索

基于化学荧光测定的板蓝根抗病毒效价检测方法的建立

相关论文

相关资讯

关于本站 Cookie 的使用提示

全站搜索