高效凝胶排阻色谱法测定牛血清白蛋白相对分子质量及其分布  

Determination Method of Relative Molecular Mass and Distribution of Bovine Serum Albumin by High Performance Size Exclusion Chromatography

作　　者：于小焱[1] 罗嫣[1] 张晓慧[1] 

机构地区：[1]国家海洋标准计量中心,天津300112

出　　处：《化学分析计量》2013年第5期20-23,26,共5页Chemical Analysis And Meterage

基　　金：海洋公益性行业科研专项(201105025)

摘　　要：建立了一种高效凝胶排阻色谱快速测定牛血清白蛋白的相对分子质量及其分布的方法。使用凝胶色谱仪，TSKgelG3000SWXL型（300mm×7．8mrn）高效凝胶排阻色谱柱，以0．05mol／L磷酸盐缓冲溶液（pH6．8）-0.3mol／L氯化钠溶液为流动相，流速为1mL／min，DAD检测器，检测波长为220nm，选择300-330nm作为参比波长，信号采集时间为15min。测得质控样牛血清白蛋白相对分子质量标准物质峰值相对分子质量的均值为7．160×10^4Da，与参考值的相对误差为7．76％，组间相对标准偏差为0．49％。采用该法对市售牛血清白蛋白样品的相对分子质量分布进行测试，组内相对标准偏差为0．004％-0．014％，组间相对标准偏差为1．89％。该法可以满足一般实验室对牛血清白蛋白样品的分子量分布测定的要求。The determination method of relative molecular mass and its distribution of bovine serum albumin （BSA） by high performance size exclusion chromatography was established. Chromatographic experiments conditions were as the following： TSK gel G3000SWXL column （300 mm × 7.8 mm）, a mobile phase of phosphate buffer solution （0.05 mol/L, pH 6.8） with 0.3 mol/L sodium chloride, DAD detector with the detection wavelength of 220 nm, 300-330 nm as the reference wavelength, the flow rate was 1.0 mL/min, the testing interval was 15 min. Mp of the standard material for quality control was 7.160 × 10^4 Da, the relative deviation between measured value and certified value of the standard quality control sample was 7.76% in average with inter-assay RSD of 0.49%. The relative molecular mass and distribution of BSA samples were tested, the intra-assay RSD ware 0.004%-0.014%, and the inter-assay RSD was 1.89%, which could reach the requirement for the lab to test the relative molecular mass and distribution of the BSA samples.

关 键 词：高效凝胶排阻色谱 牛血清白蛋白 超滤膜 相对分子质量 相对分子质量分布 

分 类 号：O657.7[理学—分析化学]