Capillary electrophoretic measurement of tissue metabolites.
Dillon, Patrick F., and Patrick R. Sears.
Department of Physiology, Michigan State University, East Lansing,
MI 48824. Telephone: 517-337-7947, Fax: 517-355-5125, e-mail:
dillon@psl.msu.edu
APStracts 4:0326C, 1997.
A method for the measurement of tissue metabolites from rabbit urinary
bladder using capillary eletrophoresis (CE) has been developed. The
method generates a reproducible electropherogram containing more than
twenty peaks, including NAD, NADH, lactate, UDP-glucose,
phosphocreatine, creatine, ATP, ADP, GTP and UTP, from less than 20
nanoliters of extract solution generated from 1.1 nl (or
approximately 1.2 micrograms) of tissue in less than 40 minutes.
Multiple samples from the same bladder produce standard errors
comparable to enzymatic or NMR measurements of metabolites:
phosphorus NMR measurement requires 106 more tissue than CE;
individual enzymatic measurements using 100 SYMBOL 109 \f l per
sample require 2000 l, 105 greater volume than CE requires for the
same number of metabolites. CE detects about 3 times more peaks than
phosphorus NMR on a similar time scale. Comparable measurements using
enzymatic analysis would require approximately 10 times longer. The
combination of minimal tissue volume requirements, rapid measurement,
and reproducibility makes CE a valuable tool in the investigation of
simultaneous changes in multiple metabolites from minute tissue
samples.
Received 8 September 1997; accepted in final form 3 November
1997.
APS Manuscript Number C460-7.
Article publication pending Am. J. Physiol. (Cell Physiology).
ISSN 1080-4757 Copyright 1997 The American Physiological Society.
Published in APStracts on 14 November 1997