基于生物抑菌活性的金银花颗粒效价测定方法

Potency Determination of Lonicera Granule Based on Bacterial Inhibitory Activity

目的:探讨符合中医药特点的中药固体制剂质量控制与评价的化学-生物联合评价模式。方法:以金银花颗粒为模型药,采用菌敏试验(最小抑菌浓度和最小杀菌浓度的测定)考察金银花提取液对4种标准菌株(金黄色葡萄球菌、变形链球菌、大肠埃希菌、铜绿假单胞菌)的抑菌性。建立敏感菌抑菌圈直径与不同浓度金银花提取液之间的相关曲线,计算不同厂家金银花颗粒的生物效价,并将测定结果与HPLC法测定(以绿原酸计)的结果相比较。结果:抑菌实验表明,金银花对金黄色葡萄球菌、变形链球菌、大肠埃希菌、铜绿假单胞菌敏感,其中金黄色葡萄球菌的抑菌圈边缘清晰、敏感度较高。当金银花的质量浓度在0.0142~0.0680 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=0.990 9)。结论:以生物活性为导向的效价评价方法有望成为评价中药固体制剂有效成分含量的手段之一,可配合传统检验方法对中药材及其制剂进行质量监控。

Objective： To explore the chemical-biological evaluation pattern for the quality control and evaluation of Chinese materia medica solid preparations（CMMSP） according with the characteristics of Chinese traditional medicines. Methods： Lonicera granule as the model drug, the bacteria sensitive tests [ minimal inhibitory concentration （MIC） and minimal bactericidal concentration （MBC） ] were applied to study the antimicrobial effect of Lonicera extract on 4 kinds of standard strains （Staphylococcus aureus, Streptococcus pneumoniae, Escherichia coli and Pseudomonas aeruginosa）. The standard curves of the relationship between the anti-bacterial ring diameter for sensitive bacterial and the concentration of Lonicera extracts were established, and the biological potency of Lonicera granule from differ- ent manufacturers was calculated and compared with the result of the HPLC method （measured by chlorogenic acid）. Results： The anti- microbial test showed that Staphylococcus aureus , Streptococcus pneumoniae , Escherichia coli and Pseudomonas aeruginosa were susceptible to Lonicera, and the antibacterial circle edge for Staphylococcus aureus was clear with higher sensitivity. There was a good linear relationship between the logarithmic dose and the response effect when the concentration of I^nicera was within the range of 0.014 2-0.068 0 g · ml - 1 （r = 0. 990 9）. Conclusion： As a quality control method for Lonicera and Lonicera granules, the bioassay is feasible, which can ef- fectively cooperate with the traditional test method for the quality control of Lonicera and its preparations.