Clinical Chemistry
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Clinical Chemistry 41: 995-1003, 1995;
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Clinical Chemistry, Vol 41, 995-1003, Copyright © 1995 by American Association for Clinical Chemistry

Candidate reference method for determining serum creatinine by isocratic HPLC: validation with isotope dilution gas chromatography- mass spectrometry and application for accuracy assessment of routine test kits

LM Thienpont, KG Van Landuyt, D Stockl and AP De Leenheer
Laboratoria voor Medische Biochemie en voor Klinische Analyse, University of Ghent, Belgium.

We present a candidate Reference Method for determining creatinine in serum, based on isocratic HPLC. The chromatographic column, with alkaline-treated aluminum oxide as stationary phase, is eluted with an equivolume mixture of methanol/acetonitrile containing 70 mL/L aqueous NaOH (10 mmol/L), and ultraviolet absorbance is detected at 240 nm. We investigated the ruggedness of the method and validated its performance with isotope dilution gas chromatography-mass spectrometry (ID GC-MS) for a set of 22 patients' sera and 10 commercially available lyophilized control materials. The mean deviation from ID GC-MS was +0.1% (range, -2.2% to +2.2%). The between-day CV, calculated from six independent measurements performed on three different days, was 0.9% (range 0.2% to 1.9%); the within-run CV was 0.8%. The total error of the method was < 3%. These performance characteristics make the method suitable for target-setting of quality-control materials and accuracy assessment of routine test kits. For the latter application, done with split-sample measurements of a panel of 83 patients' specimens, we used the Boehringer Mannheim enzymatic creatinine PAP test on the Hitachi 911, the Kodak Ektachem single-slide enzymatic creatinine test on the Ektachem 700, the Merck Jaffe kinetic assay on the Mega analyzer, and the Roche Jaffe kinetic test on the Cobas Mira.


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P. H. Petersen, D. Stockl, O. Blaabjerg, B. Pedersen, E. Birkemose, L. Thienpont, J. F. Lassen, and J. Kjeldsen
Graphical interpretation of analytical data from comparison of a field method with a Reference Method by use of difference plots
Clin. Chem., November 1, 1997; 43(11): 2039 - 2046.
[Abstract] [Full Text] [PDF]


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Copyright © 1995 by the American Association for Clinical Chemistry.