Improved high-performance liquid chromatographic determination of thiamine and its phosphate esters in animal tissues

An improved method for the determination of thiamine and its phosphate esters in animal tissues using reversed-phase high-performance liquid chromatography with precolumn derivatization is described. Thiamine and its phosphate esters were converted into fluorophores by alkaline cyanogen bromide, and...

Full description

Saved in:
Bibliographic Details
Published in:Journal of Chromatography A 1988, Vol.450 (3), p.317-323
Main Authors: Iwata, Heitaroh, Matsuda, Toshio, Tonomura, Hideo
Format: Article
Language:English
Subjects:
Analysis
Biological and medical sciences
General pharmacology
Medical sciences
Pharmacology. Drug treatments
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:An improved method for the determination of thiamine and its phosphate esters in animal tissues using reversed-phase high-performance liquid chromatography with precolumn derivatization is described. Thiamine and its phosphate esters were converted into fluorophores by alkaline cyanogen bromide, and the derivatives were applied to an ODS packed column. Then the effluent obtained by an acidic mobile phase was mixed with an alkaline methanol solution to increase the fluorescence intensity of the derivatives which was determined spectrofluorometrically. A complete, rapid and quantitative separation of thiamin and its phosphate esters was achieved and the use of the acidic buffer as a mobile phase improved the column stability. The fluorophores of thiochrome ester peaks on the chromatogram were sensitive to pretreatment with thiamine triphosphatase or acid phosphatase. The applicability of the mehod to the determination of the form of thiamin in various tissues of rat is demonstrated.
ISSN:0021-9673
DOI:10.1016/S0021-9673(01)83586-X