European Journal of Clinical Pharmacology

, Volume 28, Issue 2, pp 213–219 | Cite as

Determination of thiamine in human plasma and its pharmacokinetics

  • W. Weber
  • H. Kewitz
Originals

Summary

A sensitive assay for thiamine suitable for clinical use has been developed. It is based on precolumn oxidation of thiamine to thiochrome followed by HPLC-separation and fluorescence detection. The assay is applicable to various biological materials, including human plasma.

The minimum amount detectable was 5 fmol, minimum plasma concentration 0.5 nmol/l and minimum sample volume 0.3 ml plasma. Each chromatographic run took 3 min.

Inter- and intra-assay relative standard deviations (RSD) were 8.3% and 6.3%, respectively, at a stock plasma concentration of 10.8 nmol/l. At 38.8 nmol/l, interassay RSD was reduced to 3.4%. The recovery of 5 nmol/l added thiamine was 102 (SD±17)%, that of 30 nmol/l was 94±5%.

Plasma levels in 91 volunteers ranged from 6.6 to 43 nmol/l, showing a log normal distribution with a median of 11.6 nmol/l.

Thiamine kinetics were studied in plasma and urine from 8 men after intravenous and oral doses of 50, 100 and 200 mg thiamine hydrochloride. In all individuals, nonlinear renal elimination kinetics were demonstrated by plotting the fractional amount of thiamine excreted unchanged in urine against the corresponding area under the plasma concentration — time curve.

Key words

thiamine plasma level pharmacokinetics nonlinear renal elimination assay for clinical use 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Ellinger P, Holden M (1944) Quenching effect of electrolytes on the fluorescence intensity of riboflavin and thiochrome. Biochem J 38: 147–150Google Scholar
  2. 2.
    Kimura M, Fuyita T, Itokawa Y (1982) Liquid chromatographic determination of the total thiamin content of blood. Clin Chem 28 [1]: 29–31Google Scholar
  3. 3.
    Schrijver J, Speek A, Klosse J, van Rijn HJM, Schreurs WHP (1982) A reliable semiautomated method for the determination of total thiamine in whole blood by the thiochrome method with high-performance liquid chromatography. Ann Clin Biochem 19: 52–56Google Scholar
  4. 4.
    Warnock LG (1982) The measurement of erythrocyte thiamin pyrophosphate by high-performance liquid chromatography. Anal Biochem 126: 394–397Google Scholar
  5. 5.
    Baker H, Frank O, Pasher I, Sobotka H, Hutner S (1961) Vitamin-levels in blood and serum. Nature 191: 78–82Google Scholar
  6. 6.
    Baker H, Frank O, Fennelly JJ, Leevy CM (1964) A method for assaying thiamine status in man and animals. Am J Clin Nutr 14: 197–201Google Scholar
  7. 7.
    Roser RL, Andrist AH, Harrington WH (1978) Determination of urinary thiamine by high-pressure liquid chromatography utilizing the thiochrome fluorescent method. J Chromatogr 146: 43–53Google Scholar
  8. 8.
    Burch HB, Bessey O, Love RH, Lowry OH (1952) The determination of thiamine and thiamine phosphates in small quantities of blood and blood cells. J Biol Chem 198: 477–490Google Scholar
  9. 9.
    Mickelson O, Yamamoto RS (1958) In: Glick D (ed) Methods of biochemical analysis, Vol.6. Academic Press, New York, pp 191–257Google Scholar
  10. 10.
    Weber W, Kewitz H (1983) HPLC-procedure for thiamin determination to study kinetics in humans. Arch Pharmacol 622 [Suppl R2]Google Scholar
  11. 11.
    Myint T, Houser HB (1965) The determination of thiamine in small amounts of whole blood and serum by a simplified thiochrome method. Clin Chem 2: 617–623Google Scholar
  12. 12.
    Wielders JPM, Mink CHR JK (1983) Quantitative analysis of total thiamine in human blood, milk and cerebrospinal fluid by reversed phase ion-pair high performance liquid chromatography. J Chromatogr 277: 145–156Google Scholar
  13. 13.
    Dong MH, Green MD, Sauberlich HE (1981) Determination of urinary thiamin by the thiochrome method. Clin Biochem 14 [1]: 16–18Google Scholar
  14. 14.
    Heesen H (1975) Pharmacokinetics in normal persons and obese patients after 14 days of starvation. Inn Med 2: 89–96Google Scholar

Copyright information

© Springer-Verlag 1985

Authors and Affiliations

  • W. Weber
    • 1
  • H. Kewitz
    • 1
  1. 1.Department of Clinical Pharmacology, Klinikum SteglitzFree University of BerlinBerlinFederal Republic of Germany

Personalised recommendations