Capillary electrophoretic measurement of tissue metabolites.
A method for the measurement of tissue metabolites from rabbit urinary bladder using capillary electrophoresis (CE) has been developed. The method generates a reproducible electropherogram containing > 20 peaks, including NAD, NADH, lactate, UDP-glucose, phosphocreatine, creatine, ATP, ADP, GTP, and UTP, from < 20 nl of extract solution generated from 1.1 nl (or approximately 1.2 micrograms) of tissue in < 40 min. Multiple samples from the same bladder produce SE comparable with enzymatic or nuclear magnetic resonance (NMR) measurements of metabolites: phosphorus-NMR measurement requires 10(6) more tissue than CE; individual enzymatic measurements using 100 microliters/sample require 2,000 microliters, a 10(5) greater volume than required by CE for the same number of metabolites. CE detects about three 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.
Dillon
P F
PF
Department of Physiology, Michigan State University, East Lansing 48824, USA.
Sears
P R
PR
D000818:Animals; D019075:Electrophoresis, Capillary / Q000379:methods*; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D009130:Muscle, Smooth / Q000737:chemistry / Q000378:metabolism; D011817:Rabbits; D014020:Tissue Extracts / Q000737:chemistry; D001743:Urinary Bladder / Q000737:chemistry* / Q000378:metabolism
D014020:Tissue Extracts