A new HPLC-method for the simultaneous determination of B1-, B2- and B6-vitamers in serum and whole blood.

Considering that the B1-, B2- and B6-vitamers in whole blood and serum are partly protein bound and partly in phosphorylated form as co-enzyme, we developed a sample treatment which is able to transduce all three vitamers in their free forms. After cleaning the sample on solid-phase-extraction columns (SPE-column) we are able to separate all three vitamers in one run with HPLC and to quantify them with simultaneous detection (UV and Fluorescence). The following reference values were obtained: (ng/ml) Serum: Thiamin 6.55 +/- 2.38, Riboflavin 4.92 +/- 1.01, vitamin B6 11.21 +/- 3.17. Whole blood: Thiamin 62.3 +/- 20.8, riboflavin 69.8 +/- 20.9, vitamin B6 59.5 +/- 22.4.