Determination of plasma thiol bound to albumin using affinity chromatography and high-performance liquid chromatography with fluorescence detection: ratio of cysteinyl albumin as a possible biomarker of oxidative stress.
We examined the influence of oxidative stress on the relative amounts of various albumin-bound thiols in human plasma. To determine the ratio of thiols existing as mixed disulfides following oxidation, we developed a method combining fast purification of albumin using affinity columns and high-performance liquid chromatography (HPLC) with fluorescence detection for low molecular weight thiols which were labeled after reduction. When the effect of exposure of plasma to radical oxygen species on binding of thiols to albumin was determined by the present method, significant increases in the ratio of cysteine bound to albumin (Alb-Cys) to total cysteine were clearly demonstrated.
Ogasawara
Yuki
Y
Departments of Environmental Biology, Meiji Pharmaceutical University, 2-522-1 Noshio, Kiyose, Tokyo 204-8588, Japan. yo@my-pharm.ac.jp
Mukai
Yoshio
Y
Togawa
Tadayasu
T
Suzuki
Toshihiro
T
Tanabe
Shinzo
S
Ishii
Kazuyuki
K
D000328:Adult; D015415:Biomarkers / Q000032:analysis*; D002846:Chromatography, Affinity / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003545:Cysteine / Q000378:metabolism*; D005453:Fluorescence; D006801:Humans; D008875:Middle Aged; D018384:Oxidative Stress / Q000502:physiology; D015203:Reproducibility of Results; D012709:Serum Albumin / Q000737:chemistry*; D013438:Sulfhydryl Compounds / Q000097:blood*; D020122:tert-Butylhydroperoxide / Q000494:pharmacology
D015415:Biomarkers; D012709:Serum Albumin; D013438:Sulfhydryl Compounds; D020122:tert-Butylhydroperoxide; D003545:Cysteine