Abstract
Publisher Summary This chapter discusses the determination of total pyridoxal in human plasma using high-performance liquid chromatography. After cleavage of the pyridoxal (PL) phosphate (PLP) by acid phosphatase the total PL is determined. Reversed-phase high-performance liquid chromatography (HPLC) after protein precipitation is used with postcolumn on-line derivatization with semicarbazide. The effectiveness of phosphate cleavage was examined by hydrolysis of PLP in plasma. Three suitable enzymes are described along with their cleavage conditions: alkaline phosphatase, 1 hr at 37°; acid phosphatase, 1 hr at 37°; and Clara-Diastase, 2 hr at 45°. Several investigations demonstrated that 16 hr at 40° with acid phosphatase was sufficient to cleave all the PLP and leave only PL in plasma. The progress of cleavage was monitored at 0, 6, 12, and 16 hr, with regard to the build-up of pyridoxal and to the disappearance of the PLP in the plasma.
Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
