催化分光光度法测定血红蛋白
Catalytic Spectrophotometric Determination of Hemoglobin
摘 要
基于血红蛋白的过氧化物酶特性,建立了一种测定血红蛋白的催化动力学光度分析法.在pH 9.5的缓冲溶液中,血红蛋白对过氧化氢氧化酸性铬蓝K反应体系的褪色作用明显加速.在最佳试验条件下,由于褪色作用的加速所引起的吸光度下降(ΔA)与血红蛋白的浓度在7.0×10-10~9.0×10-8mol·L-1范围内呈线性关系.表观摩尔吸光率为3.9×108L·mol-1·cm-1,检出限为1.7×10-11mol·L-1.用于尿液中血红蛋白含量的测定,测定结果的相对标准偏差(n=5)在1.9%~4.2%之间,回收率在96.0%~102.0%之间.对此酶催化反应的机理也作了探讨.
血红蛋白
酸性铬蓝K
过氧化氢
Catalytic spectrophotometry
Hemoglobin
Acid chrome blue K
Hydrogen peroxide
Abstract
Based on its peroxidase property,a new method of catalytic spectrophotometric determination of hemoglobin (Hb) was proposed.In an ammoniacal buffer medium of pH 9.5,oxidation of acid chrome blue K (ACBK) by hydrogen peroxide was catalyzed by hemoglobin,leading to acceleration of color-fading of the reaction system.Under the optimized reaction condition,the magnitude of decrease in absorbance (ΔA) due to catalytic effect was found to keep a linear relationship with the concentration of Hb in the range of 7.0×10-10-9.0×10-8mol·L-1,with its detection limit (3Sb/k) of 1.7×10-11mol·L-1,and apparent molar absorptivity of 3.9×108L·mol-1·cm-1.In application of the proposed method to the determination of Hb in urine,values of RSD′s (n=5) obtained were in the range of 1.9%-4.2%,and values of recovery were in the range of 96.0%-102.0%.A brief discussion on the mechanism of the enzyme catalysis was also given.
中图分类号 O657.31
所属栏目 工作简报
基金项目 河南省教育厅自然科学基金资助项目(2007150048)
收稿日期 2007/5/15
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备注陈亚红(1975-),女,河南鲁山人,博士,副教授,从事分析化学的教学及分子光谱的科研工作.
引用该论文: CHEN Ya-hong,ZHANG Jian-fu,ZHAI Xiang-na. Catalytic Spectrophotometric Determination of Hemoglobin[J]. Physical Testing and Chemical Analysis part B:Chemical Analysis, 2008, 44(10): 957～959
陈亚红,张建夫,翟向娜. 催化分光光度法测定血红蛋白[J]. 理化检验－化学分册, 2008, 44(10): 957～959
被引情况:
【1】陈亚红,田丰收,冯莉莉,陈辉, "酶催化分光光度法测定盐酸甲氧氯普胺",理化检验－化学分册 47, 542-544(2011)
【2】陈亚红,田丰收,冯莉莉,陈辉, "酶催化分光光度法测定盐酸甲氧氯普胺",理化检验－化学分册 47, 542-544(2011)
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参考文献
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【9】林丽,仇佩虹,杨丽珠,等.纳米银粒子修饰电极法测定血红蛋白[J].分析化学,2006,34(1):31-34．
【2】Guibaut G G.Handbool of enzymatic methods of analysis[M].New York and Basel: Marcel Dekker,Inc,1976．
【3】Liu Z H,Cai R X.Highly sensitive spectrofluorimetric determination of hydrogen peroxide with β-cycpldextrin-hemin as catalyst[J].Analyst,1999,124(1):173-179．
【4】Zhang K,Cai R X,Chen D.Determination of hemoglobin based on its enzymatic activity for the oxidation of o-phenylenediamine with hydrogen peroxide[J].Anal Chim Acta,2000,413(1):109-113．
【5】李晓燕,张元勤.一种高灵敏度测定血红蛋白的荧光分析法[J].分析化学,2005,33(1):54-57．
【6】王全林,刘志洪,蔡汝秀,等.明胶增敏催化光度法测定血红蛋白[J].分析化学,2001,29(4):421-424．
【7】胡涌刚,庞志涛.一种高灵敏度测定血红蛋白的化学发光分析方法[J].华中科技大学学报:自然科学版,2006,34(1):114-117．
【8】莫小曼,黄春保,刘巧玲.反相流动注射化学发光法测定牛血红蛋白的研究和应用[J].分析科学学报,2004,20(5):528-530．
【9】林丽,仇佩虹,杨丽珠,等.纳米银粒子修饰电极法测定血红蛋白[J].分析化学,2006,34(1):31-34．
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