Determination of the ratio between mercaptalbumin and nonmercaptalbumin by HPLC with fluorescence probe specifically binding to albumin

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• 作者：Kaname Ohyama ; Naoya Kishikawa ; Aya Matsuo ; Takahiro Imazato ; Yukitaka Ueki ; Mitsuhiro Wada ; Kenichiro Nakashima ; Naotaka Kuroda
• 关键词：4-[4-(4-dimethylaminophenyl)-5-phenyl-1H-imidazol-2-yl]Benzoic acid methyl ester ; Fluorescence probe ; Mercaptalbumin ; Nonmercaptalbumin ; Redox state
• 刊名：Journal of Pharmaceutical and Biomedical Analysis
• 出版年：25 January, 2014
• 年：2014
• 卷：88
• 期：Complete
• 页码：170-173
• 全文大小：539 K

文摘

A simple and selective HPLC-fluorescence (FL) method with FL probe, 4-[4-(4-dimethylaminophenyl)-5-phenyl-1H-imidazol-2-yl]benzoic acid methyl ester (DAPIM), for simultaneous determination of mercaptalbumin (HMA) and nonmercaptalbumin (HNA1) was developed. After HMA and HNA1 were separated on an ion-exchange column, they were on-line and post-column mixed with DAPIM. The DAPIM-albumin complex produces FL (位ex 370 nm and 位em 510 nm); however, DAPIM solution never gives the FL. Based on this mechanism, selective determination of HMA and HNA1 were achieved without any pretreatment and interfering peak. The proposed method was applied to the measurement of HMA and HNA1 in human serum of healthy volunteers and diabetes mellitus patients