熊胆粉、猪胆粉、牛胆粉及鸡胆粉中总胆酸的含量测定及其抑菌作用

目的:建立用紫外分光光度计测定熊胆、猪胆粉、牛胆粉、鸡胆粉中总胆酸含量的方法并评价抑菌效果。方法:分别以胆酸和熊去氧胆酸为对照品,采用加入硫酸-水(50∶50),在75℃水浴加热18 min反应显色,于388 nm波长处测定熊胆粉等动物胆粉中总胆酸的含量;同时以金黄色葡萄球菌和大肠杆菌为实验菌种,在650 nm波长处测定OD值以考察体外抑菌效果。结果:胆酸在5.7～34.3μg·mL-1范围内有良好的线性关系,线性方程为Y=41.573 X-0.166 1,r=0.999 5;熊去氧胆酸在4.7～23.5μg.mL-1范围内有良好的线性关系,线性方程为Y=24.957 X-0.067 9,r=0.999 6;胆酸及熊去氧胆酸在熊胆粉、牛胆粉、猪胆粉、鸡胆粉中的平均回收率及RSD值分别为:98.9%,100.5%,1.59%,1.07%;97.7%,98.8%,1.08%,1.12%;104.1%,99.4%,2.00%,1.67%;102.9%,103.6%,1.22%,1.78%。熊胆粉等4种动物胆粉对金黄色葡萄球菌和大肠杆菌均有明显的抑菌作用,但差异无显著性。结论:本法准确,简便,重复性好,可用于熊胆等4种动物胆粉中总胆酸的含量测定,熊胆等4种动物胆粉均可抑制金黄色葡萄球菌和大肠杆菌的生长。

Determination of total cholic acid in bear bile powder,porcine gall powder,ox bile powder and chicken bile powder and study on the bacteriostatic action of these bile powder

OBJECTIVE To develop a method for determining the total cholic acid content in the bear bile powder,the porcine gall powder,the ox bile powder and the chicken bile powder,and to evaluate bacteriostatic action of these bile powder.METHODS Sulfuric acid-water(50∶50) solution was added in each bile powder sample,heated and displayed color in water bath for 18 min at 75 ℃,using cholic acid and ursodesoxycholic acid as reference substance,the total cholic content in each kind of bile powder was determined at wavelength 388 nm,at the same tine,using Staphylococcus aureus and Escherichia coli as experimental bacteria,by determining the OD value at wavelength 650 nm,the bacteriostatic action in vitro of each one was investigated.RESULTS The linear range of cholic acid was 5.7-34.3 μg·mL-1,the linear equations was Y=41.573X-0.166 1,r=0.999 5.The linear range of ursodeoxycholic acid was 4.7-23.5 μg·mL-1,the linear equations was Y=24.957X-0.067 9,r=0.999 6,the recovery of cholic acid in the bear bile powder,the ox bile powder,the porcine gall powder and the chicken bile powder was 98.9%,97.7%,104.1% and 102.9% respectively,RSD was 1.59%,1.08%,2.00%and 1.22% respectively,and the recovery of ursodeoxycholic acid in the bear bile powder,the ox bile powder,the porcine gall powder and the chicken bile powder was 100.5%,98.8%,99.4% and 103.6% respectively,RSD was 1.07%,1.12%,1.67% and 1.78% respectively.All the four kinds of bile powders had obvious bacteriostatic actions on Staphylococcus aureus and Escherichia coli,but but the difference was not significant.CONCLUSION The method is simple,accurate,reproducibility,and suitable for the determination of total cholic acid content in the bear bile powder and other bile powder,each of them has significant inhibitory effect on Staphylococcus aureus and Escherichia coli.