A candidate reference method for the determination of uric acid in serum based on high performance liquid chromatography, compared with an isotope dilution-gas chromatography-mass spectrometer method.
A method based on isocratic high performance liquid chromatography (HPLC) with UV detection at 292 nm is proposed as a candidate reference method for the determination of uric acid. Data obtained by this method are compared with those from an isotope dilution-gas chromatography-mass spectrometric method (ID-GC-MS), using [1,3-15N2]uric acid as internal standard and selected mass detection at m/z = 456 and m/z = 458. The inaccuracy of the ID-GC-MS method is maximally 0.4% for NBS-SRM-909 control sera with a concentration of 483 mumol/l. The coefficient of variation between days is 0.26%-0.80% and 0.37-0.90% for 14 control sera from other suppliers. The maximum bias of the HPLC method is 0.6%, and the coefficient of variation between days is 0.31%-0.65% for NBS-SRM-909 control sera. The coefficient of variation between days for the other 14 control sera tested is 0.35%-0.66%. Comparison of the HPLC method with the reference ID-GC-MS method resulted in a coefficient of correlation of r = 0.9998 (n = 14). The concentration of uric acid in the tested control sera ranged from 160 to 624 mumol/l.
Kock
R
R
Institut für Klinische Chemie und Pathobiochemie, Medizinischen Fakultät der RWTH Aachen.
Delvoux
B
B
Tillmanns
U
U
Greiling
H
H
D002851:Chromatography, High Pressure Liquid; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D007201:Indicator Dilution Techniques; D012016:Reference Values; D013056:Spectrophotometry, Ultraviolet; D014527:Uric Acid / Q000097:blood*
D014527:Uric Acid