Specific method for determining uric acid in serum using high-performance liquid chromatography and gas chromatography-mass spectropmetry.
A method using a combination of high-performance liquid chromatography and stable-isotope dilution-mass spectrometry is described for the specific quantitation of uric acid in serum. The procedure involves addition of a known amount of [1,3,9-15n]uric acid, as intenral standard, to the serum sample followed by equilibration with the endogenous analyte. After separation from serum proteins, cationic and neutral compounds by anion-exchange chromatography, the purified uric acid is converted into its tetraethyl derivatives. High-performance liquid chromatography is used to isolate the three major isomeric derivatives for measurement of the isotope ratio m/e 280 to m/e 283. This ratio gives the relative abundances of the molecular ions of natural and of labelled tetraethyluric acid, and from it the amount of uric acid in the original serum specimen is determined. Effective separation of tetraethyluric acid isomers can be achieved by adsorption or reversed-phase high-performance liquid chromatography using n-heptane-isopropanol (80:1, v/v) and methanol-water (3:2, v/v), respectively, as solvent systems.
D000327:Adsorption; D000818:Animals; D002417:Cattle; D002849:Chromatography, Gas / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D012997:Solvents; D014527:Uric Acid / Q000097:blood*
D012997:Solvents; D014527:Uric Acid