吉林大学学报(理学版)

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偶氮氯膦-Ⅲ分光光度法测定蛋白质

李景梅, 郝凤奇, 翟庆洲, 张旭晨   

  1. 长春理工大学 纳米技术研究中心, 长春 130022
  • 收稿日期:2014-11-05 出版日期:2015-11-26 发布日期:2015-11-23
  • 通讯作者: 翟庆洲 E-mail:zhaiqingzhou@163.com

ChlorophosphonazoⅢ as Chromogenic Agent forthe Spectrophotometric Determination of Protein

LI Jingmei, HAO Fengqi, ZHAI Qingzhou, ZHANG Xuchen   

  1. Research Center for Nanotechnology, Changchun University of Science and Technology, Changchun 130022, China
  • Received:2014-11-05 Online:2015-11-26 Published:2015-11-23
  • Contact: ZHAI Qingzhou E-mail:zhaiqingzhou@163.com

摘要:

基于酸性介质中OP100存在条件下蛋白质与偶氮氯膦Ⅲ(CPA-Ⅲ)可迅速反应生成复合物(CPA-Ⅲ)BSA, 并产生特征吸收峰的特征, 结合分光光度法精确检测蛋白质的质量浓度. 结果表明: (CPA-Ⅲ)BSA最大吸收波长为674 nm; 蛋白质质量浓度在0~300 μg/mL内与吸光度差值(Δ A)呈良好的线性关系, 其线性回归方程为Δ A=0.003 1ρ+0.003 9 (ρ: μg/mL), 相关系数r=0.996 4; 检出限为1.55 μg/mL. 利用该方法测定人血清和鸡蛋清样品中蛋白质的质量浓度, 相对标准偏差为2.43%~4.35%, 加标回收率为97.97%~103.07%.

关键词: 偶氮氯膦Ⅲ, 分光光度法, 蛋白质

Abstract:

A novel spectrophotometric method for the determination of protein was proposed based on the fact that in the presence of OP100, chlorophosphonazoⅢ(CPAⅢ) and bovine serum albumin (BSA) can react quickly to generate a composite under acidic conditions that shows a characteristic absorption peak by which protein can be accurately detected  with spectrophotometry. At 674 nm, (CPAⅢ)BSA composite  has a maximum absorption. The absorbance difference (Δ A) is linearly related with the concentration of BSA over a range of 0—300 μg/mL and the fitted equation is Δ A=0.003 1ρ+0.003 9 (ρ: μg/mL), with a correlation coefficient r=0.996 4. The detection limit of the method was 1.55 μg/mL. The proteins in human serum and egg albumen samples were successfully determined by this method. The relative standard deviation was 2.43%—4.35%. The recovery of standard addition of the method was 97.97%—103.07%.

Key words: chlorophosphonazoⅢ, spectrophotometry, protein

中图分类号: 

  • O657.32