title: An improved method for the electrophoretic separation and fluorometric determination of thiamine and its phosphates in animal tissues.
abstract: An improved method for electrophoretic separation and fluorometric (or radiometric) determination of thiamine and thiamine mono-, di- and tri-phosphates in animal tissues (liver, small intestine, kidney, heart, brain) is described. The amount of total thiamine handled was 1.5-3.5 microgram. The procedure includes: acid excretion of the compounds from the tissue, deproteinization with TCA, purification of the extract on partially deactivated charcoal, elution with 10% n-propanol in 0.1 N formic acid, lyophilization, electrophoresis on gelatinized cellulose acetate strips, elution of the thiaminic bands with 50% ethanol and finally oxidation to thiochromes and reading of eluate fluorescence in a spectrofluorophotometer. The mean internal recovery was 83.3% and the external one 83.5%. Dephosphorylation of the T phosphates was rather scanty and each thiamine compound was comparably recovered (82.0-85.7%), apparently TTP being the worst and T the best recovered compound.
mesh_terms: D000818:Animals; D001923:Brain Chemistry; D004586:Electrophoresis / Q000379:methods*; D004588:Electrophoresis, Cellulose Acetate / Q000379:methods*; D007421:Intestine, Small / Q000032:analysis; D007668:Kidney / Q000032:analysis; D008099:Liver / Q000032:analysis; D008297:Male; D009206:Myocardium / Q000032:analysis; D051381:Rats; D013050:Spectrometry, Fluorescence / Q000379:methods; D013831:Thiamine / Q000032:analysis*; D013833:Thiamine Monophosphate / Q000032:analysis; D013835:Thiamine Pyrophosphate / Q000032:analysis; D013838:Thiamine Triphosphate / Q000032:analysis
chemical_list: D013838:Thiamine Triphosphate; D013833:Thiamine Monophosphate; D013835:Thiamine Pyrophosphate; D013831:Thiamine