High Throughput UHPLC, combined Vitamin B1/B6 Analysis - UHPLC

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

• High throughput due to separation in less than 3 minutes
• Reliable and reproducible results with a specific internal standard for each parameter
• Easy sample preparation with Pre-mixed Tubes

This UHPLC assay facilitates the combined analysis of vitamin B₁ in whole blood and vitamin B₆ in whole blood or plasma. Due to faster analysis times in less than 3 minutes, this method fulfils the requirements of clinical laboratories with higher sample throughput. The easy and efficient sample preparation consists of a single precipitation step and a final derivatisation in a reaction vial. The internal standard for each of the two parameters and special quality controls ensure precise and reliable quantification.

Chromsystems also offers two more assays that feature determination using HPLC: a standard version as well as one with pre-mixed tubes.

An automated method using HPLC is also available for laboratories with higher sample throughput.

Content for 1000 analyses
Mobile Phase A, 2 x 1000 ml
Mobile Phase B, 1000 ml
UHPLC Column, 1 pc.
Whole Blood Calibration Standard (lyoph.), 5 x 5 x 1 ml
Internal Standard, 10 x 10 ml
Precipitation Reagent, 10 x 30 ml
Pre-mixed Neutralisation Tubes, 10 x 100 pcs.
Derivatisation Reagent 1 (lyoph.), 10 x 2 x 0.3 ml
Derivatisation Reagent 2 , 10 x 15 ml
Reaction Vials, amber coloured (light protection), 10 x 100 pcs.