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题名: Binding of metal ions with protein studied by a combined technique of microdialysis with liquid chromatography
作者: Guo, M;  Zou, HF;  Wang, HL;  Kong, L;  Ni, JY
关键词: binding interaction ;  metal ions ;  human serum albumin ;  microdialysis ;  HPLC
刊名: ANALYTICA CHIMICA ACTA
发表日期: 2001-09-12
卷: 443, 期:1, 页:91-99
收录类别: SCI
文章类型: Article
部门归属: 18068
项目归属: 1809
WOS标题词: Science & Technology ;  Physical Sciences
类目[WOS]: Chemistry, Analytical
研究领域[WOS]: Chemistry
英文摘要: A method has been developed for the determination of interactions of metal ions and protein by using microdialysis sampling technique combined with pre-column derivation and reversed-phase ion-pair liquid chromatographic (HPLC analysis. Cu(II), Zn(II) and human serum albumin (HSA) were chosen as model metal ions and protein, respectively. The mixed solutions of metal ions and HSA with different molar ratios buffered with 0.1 M Tris-HCl containing 0.1 M NaCl at pH 7.43 were sampled with a mirodialysis probe by keeping perfusion rate at 1 mul/min and the temperature at 37 degreesC. The free concentrations of metal ions in microdialysates were assayed by precolumn derivatization with meso-tetra(4-sulfophenyl)-porphyrin (TPPS4) followed ion-pair HPLC analysis. The recovery (R) of microdialysis sampling was measured in vitro under similar conditions as 65.74% for Cu(II), 70.45% for Zn(II) with R.S.D. below 3.2%. The primary binding constants and number of binding site estimated by the Scatchard plot analysis are 5.04 x 10(6) M-1 and 0.85 for Cu(II), and 9.87 x 10(6) M-1 and 1.10 for Zn(II), respectively. The competition of Cu(II) and Zn(II) at the second binding site on HSA was investigated, and it was observed that there is a second site on HSA to bind Cu(II) and Zn(II), the affinity of Cu(II) is stronger than that of Zn(II) to this second site of HSA. (C) 2001 Elsevier Science B.V. All rights reserved.
关键词[WOS]: SERUM-ALBUMIN ;  SITE ;  EQUILIBRIUM ;  COPPER(II) ;  CONSTANTS ;  COMPLEX
语种: 英语
原文出处: 查看原文
WOS记录号: WOS:000170860300010
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/83721
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, Dalian 116011, Peoples R China

Recommended Citation:
Guo, M,Zou, HF,Wang, HL,et al. Binding of metal ions with protein studied by a combined technique of microdialysis with liquid chromatography[J]. ANALYTICA CHIMICA ACTA,2001,443(1):91-99.
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