琼脂糖凝胶电泳法中染色剂对人血白蛋白纯度检测结果的影响  

Effect of Stains on the Purity Determination of Human Serum Albumin by Agarose Gel Electrophoresis

作　　者：穆矛 陈思榕 刘冰滢 吴一平 梁蔚阳[1] MU Mao;CHEN Si-rong;LIU Bing-ying;WU Yi-ping;LIANG Wei-yang(Guangdong Institute for Drug Control,Guangzhou 510663,China)

机构地区：[1]广东省药品检验所,广州510663

出　　处：《中国药学杂志》2020年第13期1098-1102,共5页Chinese Pharmaceutical Journal

基　　金：国家药典委员会药品标准修订研究课题资助(2018S016,2018S017);广东省医学科学技术研究基金项目资助(B2018130,B2018220);广东省食品药品管理局科技项目引导扶持基金资助(2018YDB02)。

摘　　要：目的研究染色剂对琼脂糖凝胶电泳法检测人血白蛋白纯度的影响,选择合适的染色剂使琼脂糖凝胶电泳法与《中国药典》2015年版中规定方法(醋酸纤维素薄膜电泳法)的检测结果基本一致。方法在琼脂糖凝胶电泳法中,使用酸蓝、氨基黑、丽春红3种染色剂,对30批人血白蛋白样品进行纯度检测,并与醋酸纤维素薄膜电泳法的检测结果进行比较。结果在琼脂糖凝胶电泳中,丽春红染色无法染出杂质条带;酸蓝染色的纯度结果偏低,与醋酸纤维素薄膜电泳法有显著差异(P<0.01);氨基黑染色的纯度结果与醋酸纤维素薄膜电泳法比较结果未显示统计学差异(P=0.37),检测的30批样品有25批样品的醋酸纤维素薄膜电泳法结果在氨基黑染色结果的95%置信区间内。结论在琼脂糖凝胶电泳法检测人血白蛋白纯度的过程中,染色剂的种类对实验结果有较大影响,使用氨基黑染色液对电泳结果进行染色可以获得与现行药典方法较为一致的结果。OBJECTIVE To study the effect of stains on the purity detection results of human serum albumin by agarose gel electrophoresis,thus to select appropriate stain so that the detection result of agarose gel electrophoresis method can be basically consistent with the method specified in the 2015 edition of Chinese Pharmacopoeia,cellulose acetate membrane electrophoresis method.METHODS Acid blue,amino black and ponceau were used in agarose gel electrophoresis to detect the purity of 30 batches of human serum albumin samples,and the results were compared with those obtained by cellulose acetate membrane electrophoresis.RESULTS Impurity bands could not be detected by ponceau stain;the results of amino black staining was significantly lower than that of cellulose acetate membrane electrophoresis.There was no statistical difference between the results of amino black staining and cellulose acetate membrane electrophoresis.The results of 25 batches of samples detected by cellulose acetate membrane electrophoresis were within the 95%confidence interval of the results detected by amino black staining.CONCLUSION The category of stain has great influence on the detected purity of human serum albumin when using agarose gel electrophoresis.The results of amino black staining is consistent with those of the method recorded in the Chinese Pharmacopoeia in use.

关 键 词：琼脂糖凝胶电泳 人血白蛋白 纯度 染色剂 

分 类 号：R917[医药卫生—药物分析学]