广西师范大学学报（自然科学版） ›› 2012, Vol. 30 ›› Issue (2): 99-105.

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5-溴-4-氯-3-吲哚基磷酸盐共振散射光谱法检测碱性磷酸酯酶

梁爱惠, 刘高伞, 蒋治良   

  1. 广西师范大学环境与资源学院,珍稀濒危动植物生态与环境保护省部共建教育部重点实验室,广西桂林541004
  • 收稿日期:2012-04-16 出版日期:2012-06-20 发布日期:2018-12-03
  • 通讯作者: 梁爱惠(1965—),女,广西岑溪人,广西师范大学研究员,硕士生导师。E-mail:ahliang2008@163.com
  • 基金资助:
    国家自然科学基金资助项目(21165005,21075023);广西研究生科研创新项目资助(2011106020830M62)

Resonance Scattering Spectral Assay for Alkaline Phosphatase Using BCIP as Substrate

LIANG Ai-hui, LIU Gao-san, JIANG Zhi-liang   

  1. Key Laboratory of Ecology of Rare and Endangered Species and Environmental Protection,Ministry of Education, School of Environment and Resource,Guangxi Normal University,Guilin Guangxi 541004,China
  • Received:2012-04-16 Online:2012-06-20 Published:2018-12-03

摘要： 在pH8.0的Tris-HCl缓冲溶液及32 mg/L PEG20000的存在下,碱性磷酸酯酶(ALP)催化底物5-溴-4-氯-3-吲哚基磷酸盐(BCIP)水解生成不溶性的蓝色BCIP二聚体微粒,该微粒在710 nm波长处产生一个共振散射峰。在选定条件下,随着ALP活性增大,710 nm波长处的共振散射峰强度线性增大,碱性磷酸酯酶浓度在0.078～1.25 U/L与共振散射强度增大值ΔI呈良好线性关系,其回归方程为:ΔI=512.6C+17.8,检出限为9.6×10-3 U/L。该法用于合成样品中碱性磷酸酯酶的测定,结果满意。

关键词: 5-溴-4-氯-3-吲哚基磷酸盐, 碱性磷酸酯酶, 共振散射光谱法

Abstract: In pH 8.0 Tris-HCl buffer solutions and in the presence of 32 mg/L PEG20000,using EDTANa2 as terminated agent to stop the enzymatic reaction,alkaline phosphatase (ALP) catalyzed the hydrolysis of 5-bromo-4-Chloro-3-indolylphosphat (BCIP) substrate to form blue BCIP dimer particles,which exhibited a strongresonance scattering (RS) peak at 710 nm.In the selected conditions,with theALP activity increased,the RS intensity (ΔI) linearly increased.ALP activitylinear range was 0.078～1.25 U/L,with a regression equation of ΔI=512.6C+17.8,and a detection limit of 9.6×10-3 U/L.

Key words: 5-bromo-4-chloro-3-indolyl phosphate, alkaline phosphatase, resonance scattering spectroscopy

中图分类号: 

  • O657.3
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