Extraction of Vitamins B1 and B6 from Whole Blood Using ISOLUTE® PLD+ Protein and Phospholipid Removal Plate

Vitamin B1 (Thiamine Diphosphate, TDP) and B6 (Pyridoxal-5-Phosphate, PLP) in whole blood is measured clinically to help measure various metabolic functions. TDP and PLP are highly water soluble (logP= -5.9, -2.2 respectively) and are extremely light-sensitive. Traditionally, TDP and PLP are measured through fluorescence detection, requiring the use of hazardous reagents for derivatization. This additional derivatization step is also time consuming. This application note describes a simple, ISOLUTE® PLD+ protocol for the extraction of vitamin B1 and B6 (TDP/PLP) from whole blood and LC-MS/MS analysis.