Simple cation-exchange high-performance liquid chromatography optimized to the measurement of metabolites in the effluents from perfused rat livers using refractive index and ultraviolet detectors.
A high-performance liquid chromatographic method designed to analyse effluents from perfused organs is described. In the case of rat liver, compounds released by the liver are readily separated and quantitated, using a strong cation exchanger (Aminex HPX 87H), two detectors connected in series (ultraviolet detector at 210 nm and refractive index detector), and by optimizing the concentration of sulphuric acid in the mobile phase. Chromatographic conditions described in the present work enable the quantitation, in a single run, of metabolites derived from the tricarboxylic acid cycle, glycolysis, ketogenesis, adenine nucleotides catabolism and ethanol oxidation. The advantage of this method stems from its ease of implementation, sensitivity and flexibility.
Masson
S
S
Centre de Résonance Magnétique Biologique et Médicale (URA CNRS 1186), Faculté de Médecine, Marseille, France.
Sciaky
M
M
Desmoulin
F
F
Fontanarava
E
E
Cozzone
P J
PJ
D000225:Adenine / Q000378:metabolism; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D000431:Ethanol / Q000378:metabolism; D006019:Glycolysis; D006863:Hydrogen-Ion Concentration; D007474:Ion Exchange; D007659:Ketones / Q000378:metabolism; D008099:Liver / Q000737:chemistry / Q000378:metabolism*; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains; D012031:Refractometry; D014233:Tricarboxylic Acids / Q000378:metabolism; D014466:Ultraviolet Rays
D007659:Ketones; D014233:Tricarboxylic Acids; D000431:Ethanol; D000225:Adenine