PVDF膜吸附染色法检测中药注射剂微量蛋白

Detection of Trace Protein in Chinese Materia Medica Injections by Dot Blotting

目的:建立一种快速、灵敏的中药注射剂微量蛋白质检测方法。方法:将样品点在PVDF膜上,用甲醇脱去杂色,经0.25%考马斯亮蓝染色液染色,脱色液甲醇-水-冰醋酸(4.5∶4.5∶1)脱去背景色,通过斑点颜色深浅判断蛋白质限量。结果:当上样量为1μL时,可以检测出浓度为12μg/mL(12 ng)的蛋白质,灵敏度高于药典磺基水杨酸法。结论:本法操作简单、具有较强的抗干扰能力,灵敏度高于现行《中国药典》磺基水杨酸比浊法,可以用于中药注射剂蛋白质限量的控制。

The main aim is to develop a sensitive method to detect trace protein in Chinese Materia Medica injections.A sample of 1μL was dotted on a PVDF membrane,and the membrane was washed with methanol to remove the color of the dot.Then,the membrane was stained with 0.25% Coomassie Brilliant Blue solution for at least 30 sec.After the stained membrane was washed with a mixed solution with methanol-water-acetic acid（4.5∶4.5∶1） to lower the background,a blot containing trace protein would appear.The color intensity of the blot was related to the protein content.This method was able to detect protein（〉12μg/mL） in 1μL sample,with high sensitivity and few interferences.Compared with the method of sulfosalicylic precipitation adopted by China Pharmacopoeia of 2010 edition,this method is more feasible.Also,the method is helpful to control protein content in Chinese Materia Medica injections.