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.
Course of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan.
Kaname
K
Ohyama
Naoya
N
Kishikawa
Aya
A
Matsuo
Takahiro
T
Imazato
Yukitaka
Y
Ueki
Mitsuhiro
M
Wada
Kenichiro
K
Nakashima
Naotaka
N
Kuroda
D000328:Adult; D000418:Albumins / Q000737:chemistry*; D001565:Benzoates / Q000737:chemistry; D001774:Blood Chemical Analysis; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003920:Diabetes Mellitus / Q000097:blood*; D005260:Female; D005456:Fluorescent Dyes / Q000737:chemistry; D064368:Healthy Volunteers; D006801:Humans; D007093:Imidazoles / Q000737:chemistry; D008297:Male; D008875:Middle Aged; D010084:Oxidation-Reduction; D010100:Oxygen / Q000737:chemistry; D011485:Protein Binding; D015203:Reproducibility of Results; D012709:Serum Albumin / Q000032:analysis* / Q000737:chemistry; D000075462:Serum Albumin, Human
C584462:4-(4-(4-dimethylaminophenyl)-5-phenyl-1H-imidazol-2-yl)benzoic acid methyl ester; D000418:Albumins; D001565:Benzoates; D005456:Fluorescent Dyes; D007093:Imidazoles; D012709:Serum Albumin; C001702:mercaptoalbumin; C079732:nonmercaptalbumin; D010100:Oxygen; D000075462:Serum Albumin, Human
4-[4-(4-dimethylaminophenyl)-5-phenyl-1H-imidazol-2-yl]Benzoic acid methyl ester; Fluorescence probe; Mercaptalbumin; Nonmercaptalbumin; Redox state