MassChrom® Vitamins B1 and B6 in Whole Blood, with 96 Deep Well Plate

Clinical relevance

Vitamin B1 (thiamine) plays an important role in deriving energy from carbohydrates and fats. The active form is thiamine pyrophosphate (TPP), a coenzyme for some enzymatic reactions in carbohydrate metabolism and for pyruvate decarboxylase. TPP is also significantly involved in the oxidative degradation of glucose. The determination of the TPP concentration is more significant than measurements of total thiamine, as by this method only the physiologically active form is taken into account.

Vitamin B6 comprises pyridoxine, pyridoxamine and pyridoxal, as well as their phosphate derivatives. It is ingested with food and transferred via several enzymatic conversions into its active form, pyridoxal-5’-phosphate (PLP). PLP is involved in nearly one hundred different enzymatic reactions, among other things, as co-factor in amino acid metabolism, and in the formation of haemoglobin or neurotransmitters in the brain.

Product advantages

• Fast and easy sample preparation
• Robust method with high selectivity
• Short run time of 2.5 minutes
• Isotopically-labelled internal standard for each analyte

This Chromsystems assay enables combined determination of the active forms of vitamin B1 (thiamine pyrophosphate, TPP) and vitamin B6 (pyridoxal 5‘-phosphate, PLP) in whole blood by LC-MS/MS. Sample preparation consists of a simple protein precipitation step, with no need for derivatisation. This makes for a much faster sample preparation and high sample throughput. Analytes and potential interferences are effectively and swiftly separated within 2.5 minutes, indicating the method's high selectivity and speed. The chromatographic separation of the MassChrom® assay is fast enough to ensure a high throughput in the laboratory, but sufficiently resolving for a clear separation of the two analytes with minimal potential matrix effects. Isotope-labelled internal standards for both parameters as well as 6PLUS1® calibrators and MassCheck® controls ensure accurate and precise results. The assay is available in 2 configurations either with reaction vials or with 96 deep well plates.