Simultaneous liquid chromatographic assessment of thiamine, thiamine monophosphate and thiamine diphosphate in human erythrocytes: a study on alcoholics

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2003-06, Vol.789 (2), p.355-363
• Main Authors: Mancinelli, Rosanna, Ceccanti, Mauro, Guiducci, Maria Soccorsa, Sasso, Guido Francesco, Sebastiani, Gemma, Attilia, Maria Luisa, Allen, John Paul
• Format: Article
• Language: English
• Subjects: Adolescent; Alcoholism - blood; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Case-Control Studies; Coenzymes, vitamins; Erythrocytes - chemistry; Female; Fundamental and applied biological sciences. Psychology; Humans; Male; Middle Aged; Other biological molecules; Reproducibility of Results; Sensitivity and Specificity; Thiamine; Thiamine - blood
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/S1570-0232(03)00139-9

An isocratic HPLC procedure for the assessment of thiamine (T), thiamine monophosphate (TMP) and thiamine diphosphate (TDP) in human erythrocytes is described. Several aspects of the procedure make it suitable for both clinical and research purposes: limits of detection and quantification of 1 and 2.5 nmol/l, respectively, recovery of 102% on average (range 93–112%), intra- and inter-day precisions within 5 and 9%, respectively, total elution time 15 min. This analytical methodology was applied to a case-control study on erythrocyte samples from 103 healthy subjects and 36 alcohol-dependent patients at risk of thiamine deficiency. Mean control values obtained were: T=89.6±22.7 nmol/l, TMP=4.4±6.6 nmol/l and TDP=222.23±56.3 nmol/l. T and TDP mean values of alcoholics were significantly lower than those of control cases: T=69.4±35.9 nmol/l ( P