板蓝根类制剂抑菌生物效价测定方法的研究  

Determination of biological titers of Radix Isatidis preparations on bacteriostasis

作　　者：阮健 陈彤 

机构地区：[1]烟台市食品药品检验检测中心,烟台264670

出　　处：《药物分析杂志》2016年第2期261-266,共6页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的：研究板蓝根微生物检定法的适用剂量范围,建立其对敏感菌株的剂量效应曲线,并且根据板蓝根量效曲线测定不同厂家板蓝根片的效价,从而建立板蓝根和板蓝根片的质量标准。方法：采用菌敏试验（最小抑菌浓度和最小杀菌浓度的测定）考察板蓝根水提液对5种标准菌株（伤寒沙门氏菌、乙型副伤寒沙门氏菌、福氏志贺氏菌、大肠杆菌和枯草芽孢杆菌）的抑菌性。利用不同浓度板蓝根提取液对敏感菌的抑菌圈直径大小和浓度的关系建立标准曲线,构建适用于板蓝根的生物检定法,通过测定不同厂家产地的板蓝根和板蓝根片的抑菌圈直径,代入量效曲线计算它们的生物效价。结果：抑菌实验表明,板蓝根对伤寒沙门氏菌、乙型副伤寒沙门氏菌、福氏志贺氏菌、大肠杆菌及枯草芽孢杆菌等多种菌敏感,其中枯草芽孢杆菌的抑菌圈边缘清晰,敏感度较高。当板蓝根的质量浓度在0.029～0.136 g·mL^-1范围内时,对数浓度与反应效应呈线性关系,且相关性好（r=0.991 2）。板蓝根在高、中、低浓度的回收率在90.0%～110.0%范围内,可信限率均在10%以内,RSD为3.7%。结论：采用微生物检定法建立板蓝根、板蓝根片的质量控制方法是可行的,它可以有效配合传统检验方法对板蓝根药材及其制剂进行质量监控。Objective： To study the suitable dose range of microbiological test of Radix Isatidis, establish the dose-response curve for sensitive strains, and determine titer of Radix Isatidis preparations from different manufacturers according to the dose-response curve, so as to set up the quality standard of Radix Isatidis and Radix Isatidis tablets. Methods： Bacteria sensitive test （ determination of minimal inhibitory concentration （ MIC ） and minimal bactericidal concentration （ MBC ） ） were employed to inspect the antimlcrobial activity of the water-extraction of Radix Isatidis to 5 kinds of standard strains （ Salmonella typhi, type B paratyphoid salmonella, Shigellaflexneri, Escherichia coli and Bacillus subtilis ）. The standard curves were established according to the relationship between anti-bacterial ring diameter to sensitive bacterial of different concentration of Radix Isatidis extraction solution and the concentration of Radix Isatidis, and a suitable biological test was built for Radix Isatidis.Through measuring the antibacterial circle diameter of Radix Isatidis from different manufacturers, the data was input to the dose-response curve and their biological potency was calculated.Results： The antimicrobial test showed that Radix Isatidis had sensitivity to Salmonella typhi, type B paratyphoid salmonella, Shigella flexneri, Escherichia coli and Bacillus subtilis etc., among them the antibacterial circle edge of Bacillus subtilis was clear and had higher sensitivity.There was a good linear relationship between logarithmic dose and response effect in the concentration range of 0.029-0.136 g·mL^-1 （ r = 0.991 2 ） .The recovery rate of Radix Isatidis in high, medium and low concentration were in the range of 90% to 110%, and the reliability limit rate was less than 10%, and RSD was 3.7%.Conclusion： As a quality control method of Radix Isatidis, the bioassay is feasible, and can effectively assist the traditional test method for quality control of Radix Isatidis and its preparation.

关 键 词：板蓝根 抗菌活性 生物效价 微生物检定法 剂量效应曲线 菌敏试验 最小抑菌浓度 最小杀菌浓度 

分 类 号：R917[医药卫生—药物分析学]