Determination of the ratio between mercaptalbumin and nonmercaptalbumin by HPLC with fluorescence probe specifically binding to albumin.
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 (λex 370nm and λem 510nm); 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.
['Ohyama|Kaname|K|', 'Kishikawa|Naoya|N|', 'Matsuo|Aya|A|', 'Imazato|Takahiro|T|', 'Ueki|Yukitaka|Y|', 'Wada|Mitsuhiro|M|', 'Nakashima|Kenichiro|K|', 'Kuroda|Naotaka|N|']
D000328:Adult D000418:Albumins D001565:Benzoates D001774:Blood Chemical Analysis D002851:Chromatography, High Pressure Liquid D002852:Chromatography, Ion Exchange D003920:Diabetes Mellitus D005260:Female D005456:Fluorescent Dyes D064368:Healthy Volunteers D006801:Humans D007093:Imidazoles D008297:Male D008875:Middle Aged D010084:Oxidation-Reduction D010100:Oxygen D011485:Protein Binding D015203:Reproducibility of Results D012709:Serum Albumin D000075462:Serum Albumin, Human
Mercaptalbumin Nonmercaptalbumin Fluorescence probe 4-[4-(4-dimethylaminophenyl)-5-phenyl-1H-imidazol-2-yl]Benzoic acid methyl ester Redox state