Determination of ornithine conjugates of some carboxylic acids in birds by high-performance liquid chromatography.
A high-performance liquid chromatographic (HPLC) method for the determination of ornithine conjugation of some carboxylic acids in vitro has been developed. The ornithine conjugates of benzoic acid, p-nitrobenzoic acid, furancarboxylic acid and phenylacetic acid in an incubation mixture with kidney mitochondria were well separated on a reversed-phase C18 column using a mixture of 10 mM ammonium acetate buffer and methanol as the mobile phase. In addition, by varying the pH of the mobile phase and utilizing the absorption wavelengths (nm) of the conjugates it was possible to resolve and specifically detect each conjugate. The calibration curves were linear in the range of 0.2-16 micrograms/ml for all compounds and the detection limits were about 50 ng/ml except for the ornithine conjugate of phenyl acetic acid (S/N = 2). The ornithine conjugation of some carboxylic acids with chicken kidney mitochondria were determined by this assay method. The activity of ornithine conjugation of benzoic acid, furancarboxylic acid, p-nitrobenzoic acid and phenylacetic acid were 14.5, 5.5, 0.5 and 6.9 nmol/mg of protein, respectively. Moreover, the ornithine conjugation and the glycine conjugation of benzoic acid were examined in birds and rodents. The ornithine conjugation was observed only in chicken (14.5 nmol/mg of protein) and mallard (0.99 nmol/mg of protein).
Igarashi
K
K
Faculty of Pharmaceutical Sciences, Kobe-gakuin University, Japan.
Suzuki
R
R
Kasuya
F
F
Fukui
M
M
D000818:Animals; D001717:Birds / Q000378:metabolism*; D002264:Carboxylic Acids / Q000032:analysis / Q000378:metabolism*; D002645:Chickens; D002851:Chromatography, High Pressure Liquid; D003370:Coturnix; D004372:Ducks; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D009952:Ornithine / Q000032:analysis / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar
D002264:Carboxylic Acids; D009952:Ornithine