Determination of thiol by high-performance liquid chromatography and fluorescence detection with 5-methyl-(2-(m-iodoacetylaminophenyl)benzoxazole.
A new high-performance liquid chromatographic (HPLC) method for measuring low molecular weight (LMW) thiol-containing compounds, including cysteine (CysH), glutathione (GSH), N-acetylcysteine (Nac), penicillamine (PA), and 2-mercaptoethanol (2-ME), has been developed by using 5-methyl-(2-(m-iodoacetylaminophenyl)benzoxazole (MIPBO) as fluorescence-labeling reagent. The derivatization and separation conditions have been investigated in detail. Detection limits ranging from 3.5 to 15.0 fmol were achieved for the thiols investigated in a 16 min separation with detection wavelengths 310 and 375 nm for the excitation and emission, respectively. The utility of the proposed method has been validated by measuring CysH in human urine samples.
Liang
Shu-Cai
SC
Department of Chemistry, Wuhan University, Wuhan, 430072, China.
Wang
Hong
H
Zhang
Zhi-Min
ZM
Zhang
Hua-Shan
HS
D001583:Benzoxazoles / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D013438:Sulfhydryl Compounds / Q000032:analysis* / Q000652:urine
C500681:5-methyl-(2-(m-iodoacetylaminophenyl)benzoxazole); D001583:Benzoxazoles; D005456:Fluorescent Dyes; D013438:Sulfhydryl Compounds