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5361620 
Journal Article 
Determination of Phenol Sulphotransferase Activity by High-Performance Liquid Chromatography 
Honkasalo, T; Nissinen, E 
1988 
1 
Journal of Chromatography
ISSN: 0021-9673 
424 
1 
A fast and sensitive assay for the thermolabile (TL) and the thermostable (TS) forms of phenol-sulfotransferase (PST) activity in tissues is described. The method is based on the separation and quantification of p-nitrophenylsulfate (p-NPS) derived from p-nitrophenol (pNP) by high performance liquid chromatography (HPLC) and UV detection at 300 nanometers. PST catalyzes the conjugation of phenols and catechols with sulfates in the presence of the cofactor 3'-phosphoadenosine-5'-phosphosulfate (PAPS). Chromatograms obtained with crude rat liver homogenates show that the enzyme was totally inactive in the absence of PAPS. The distinct patterns of the TS and TL forms were also observed. The rate of pNPS formation was linear for both forms of PST. The two forms of PST were different with respect to the Michaelisenten constant, and they also showed different sensitivities to inhibition by 2,4-dichloro-6-nitrophenol (DCNP), a specific PST inhibitor. It is concluded that the method is precise, sensitive and fast, allowing the handling of approximately 40 samples per day.