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Clinical Chemistry 29: 2073-2075, 1983;
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Clinical Chemistry, Vol 29, 2073-2075, Copyright © 1983 by American Association for Clinical Chemistry

Determination of thiamin and thiamin phosphate esters in blood by liquid chromatography with post-column derivatization

M Kimura and Y Itokawa

We demonstrate a liquid-chromatographic method involving post-column derivatization for determining the concentration of thiamin and its phosphate esters in human blood. Blood, erythrocytes, or plasma is deproteinized and centrifuged. Aliquots of the samples are applied to a mu Bondapak C18 column attached to a "high-performance" liquid chromatograph. Addition of potassium ferricyanide/sodium hydroxide solution to the column effluent with a proportioning pump converts thiamin phosphates into fluorophores, the intensities of which are measured with a spectrofluorophotometer. Thiamin, thiamin monophosphate, thiamin pyrophosphate, and thiamin triphosphate eluted as single peaks; no coeluting substances were detected. Thiamin pyrophosphate was the ester present in greatest concentration, followed by thiamin triphosphate; thiamin monophosphate and thiamin were present in slight amounts. This method allows easy determination of thiamin and its phosphate esters in 0.1 mL of blood.


The following articles in journals at HighWire Press have cited this article:


Home page
 Clin. Chem.Home page
D. Talwar, H. Davidson, J. Cooney, and D. St. JO'Reilly
Vitamin B1 Status Assessed by Direct Measurement of Thiamin Pyrophosphate in Erythrocytes or Whole Blood by HPLC: Comparison with Erythrocyte Transketolase Activation Assay
Clin. Chem., May 1, 2000; 46(5): 704 - 710.
[Abstract] [Full Text] [PDF]


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Copyright © 1983 by the American Association for Clinical Chemistry.