Application Note: Extraction of vitamins B1 and B6 from whole blood using ISOLUTE PLD+ Protein and Phospholipid Removal Plate
21 March 2023
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. In this application note, Biotage describes a
simple ISOLUTE®
PLD+ protocol for the extraction of vitamin B1
and B6 (TDP/PLP) from whole blood and LC-MS/MS analysis.
High reproducible recoveries and low protein and phospholipid
coextraction are demonstrated. ISOLUTE PLD+ Protein and
Phospholipid Removal plates and columns offer a fast and
extremely simple sample preparation approach. Proteins
are efficiently removed from the sample using an in-plate,
‘solvent first’, protein crash and filter procedure, which is
easily automated.