Microscale determination of purines in tissue samples by capillary liquid chromatography with electrochemical detection.
A microscale method for purines involved in intracellular signaling and energy metabolism, including ADP, ATP, cyclic-AMP, NADH and GTP, was developed. The analytes were separated on a fused-silica capillary liquid chromatography column (50 microm inner diameter by 25 cm long) packed with 7 microm reversed-phase particles and detected with a carbon fiber cylinder microelectrode at +1.50 V versus Ag/AgCl reference electrode. With an acetonitrile gradient, the separation was carried out within 15 min. With a 100 nl injection volume, the detection limits varied from 0.9 to 8 fmol depending upon the analyte. The low detection limits make the method suitable for analysis of small tissue samples. As a demonstration of the method, islets of Langerhans were analyzed for their adenosine-related messenger content.
Deng
Qing
Q
Department of Chemistry, University of Michigan, Ann Arbor, MI 48109-1055, USA.
Kauri
Lisa M
LM
Qian
Wei-Jun
WJ
Dahlgren
Gabriella M
GM
Kennedy
Robert T
RT
D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods; D046508:Culture Techniques; D007515:Islets of Langerhans / Q000737:chemistry; D051379:Mice; D008832:Microchemistry / Q000379:methods*; D011687:Purines / Q000032:analysis*; D051381:Rats; D017207:Rats, Sprague-Dawley
D011687:Purines