Y. Kawamoto et al., DETERMINATION OF ATP AND ITS METABOLITES RELEASED FROM RAT CAUDAL ARTERY BY ISOCRATIC ION-PAIR REVERSED-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY, Analytical biochemistry (Print), 262(1), 1998, pp. 33-38
Citations number
19
Categorie Soggetti
Biology,"Biochemical Research Methods","Chemistry Analytical
A sensitive and selective assay method for adenine compounds (purines)
using high-performance liquid chromatography with fluorescent detecti
on was developed. The 1,N-6-ethenoderivatives of adenine, adenosine, A
MP, ADP, and ATP formed by reaction with chloroacetaldehyde at 80 degr
ees C were separated by ion-pair reversed-phase chromatography within
15 min under isocratic conditions, alpha,beta-Methylene adenosine 5'-d
iphosphate could be used as an internal standard for the determination
of purines. The calibration graphs constructed with peak area ratios
against amounts were linear between 0.1 and 10.0 pmol of each purine.
The repeatability and intermediate precision were less than 6% (RSD, n
= 5) and 8% (RSD, n = 3), respectively, at 0.5 pmol of each purine. A
method for calculation of each purine amount which considers hydrolys
is by derivatization is described. The optimized method was applied to
determine the purines released from the rat caudal artery stimulated
by noradrenaline. (C) 1998 Academic Press.