[Liquid chromatographic determination of free thiamine and its esters in whole blood].
BACKGROUND
To appreciate vitamin B1 status in normal and pathophysiological states in local ivorian populations, a high-performance liquid chromatographic method for the simultaneous determination of thiamine and its phosphate esters: thiamine monophosphate and thiamine diphsophate or cocarboxylase in whole blood has been developed.


METHODS
The method involves extraction with diethylether, followed by pre-column alkaline derivatization by bromide cyanogen. The extract was analysed by isocratic reversed-phase isocratic high performance liquid chromatography with fluorescent detection. System suitability tests were applied to assess the method's continuing suitability for use. The main tests of method validation: linearity, precision, accuracy and sensibility were applied to the analytical procedure.


RESULTS
Thiamine and its esters were eluted within 10 minutes with a good resolution. System suitability tests showed that the chromatographic system was suitable for continuing use. Results of validation tests show the reliability of the method : linearity domain, satisfying precision and accuracy, detection limits between 0.067 pg/l et 0.92 pg/l (0.160 fmol/l et 1.822 fmol/l).


CONCLUSIONS
The proposed method is suitable for determination of thiamine and its phosphate esters in whole blood.
Ake
M
M
Laboratoire de Chimie Analytique, UFR Sciences Pharmaceutiques et Biologiques, Abidjan, Côte d'Ivoire. mdake@aviso.ci
Soko
Y N
YN
Malan
K A
KA
D017766:Acid Anhydride Hydrolases / Q000097:blood; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D010744:Phosphoric Monoester Hydrolases / Q000097:blood; D013831:Thiamine / Q000097:blood*; D013835:Thiamine Pyrophosphate / Q000097:blood
C022734:thiamine monophosphatase; D010744:Phosphoric Monoester Hydrolases; D017766:Acid Anhydride Hydrolases; C023837:nucleoside-diphosphatase; D013835:Thiamine Pyrophosphate; D013831:Thiamine