黄连抑菌效力生物检定方法的研究

Study on biological assay method of Coptis Chinensis bacteriostatic efficacy

目的探讨管碟法用于测定黄连抑菌效力的可行性。方法以盐酸小檗碱为工作参照物、抑菌圈直径(D)为反应值,按《中国药典》2010年版二部(附录Ⅺ)抗生素生物检定法(管碟法)的三剂量法设计试验,对黄连样品的抑菌效力进行测定。结果盐酸小檗碱在4.00～12.50μg/mL、黄连样品溶液在66.0～200.0μg/mL时,其相应浓度的对数值与抑菌圈D呈现良好的线性关系(r=0.998 9、0.999 0)。三剂量法实验剂间变异分析及可靠性测验结果表明回归非常显著(P<0.01),偏离平行、二次曲线、反二次曲线均不显著(P>0.05),组内(碟间)差异有统计学意义(P<0.01);可信限为1.884 5%。1 mg盐酸小檗碱抑菌效力相当于15.98 mg的黄连。结论管碟法能用于黄连抑菌效力的检测,可作为黄连品质评价的一种新方法。

Objective To study the feasibility of the cup-plate method used in determining the bacteriostatic efficacy of Coptis Chinensis. Methods In accordance with the experimental design of 3 doses of the cup-plate method in the antibiotics bioassay method in the appendixXI of the Chinese Pharmacopoeia （edition 2010, Part Ⅱ ）,with berherine hydroehlofide as the work reference substance and the diameter of inhibiting bacteria circle （D） as the reaction value, the bacteriostatic efficacy of Cop- tis Chinensis was detected. Results For berberine hydrochloride within the concentration range of 4.00-12.5 μg/mL and the sample solution of Coptis Chinensis within the concentration range of 66.0-200.0 μg/mL, the dose log-value showed the good lin- ear relation with the D value（r=0.998 9,0.999 0）. The vafiatioa analysis between doses showed that the regression was very signif- icant（P〈0.01 ）, and the deviations of parallelism, second degree curve and anti-second degree curve were not significant （P〉0.05）, the difference within groups had statistical significance （P〈0.01） and the average confidence limit was 1.884 5%. The bacteriostat- ic efficacy in 1 g of berberine hydrochloride was same to that of 15.98 mg Coptis Chinensiss. Conclusion The cup-plate method can be used for testing the bacteriostatic efficacy of Coptis Chinensiss and as a new method for the quality evaluation of Coptis Chinensis.