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Comparative Study
. 2015 Jul 24;10(7):e0133912.
doi: 10.1371/journal.pone.0133912. eCollection 2015.

LC-MS/MS Method for Serum Creatinine: Comparison with Enzymatic Method and Jaffe Method

Meixian Ou  1 , Yunxiao Song  2 , Shuijun Li  1 , Gangyi Liu  1 , Jingying Jia  1 , Menqi Zhang  1 , Haichen Zhang  2 , Chen Yu  1
Affiliations
Comparative Study

LC-MS/MS Method for Serum Creatinine: Comparison with Enzymatic Method and Jaffe Method

Meixian Ou et al. PLoS One. .

Abstract

Accurate quantification of creatinine (Cre) is important to estimate glomerular filtration rate (GFR). Differences among various methods of Cre quantification were previously noted. This study aims to develop a liquid chromatography tandem mass spectrometry (LC-MS/MS) method for serum Cre and compare this method with clinical routine methods. LC-MS/MS analysis was performed on API 4000 triple quadrupole mass spectrometer coupled with an Agilent 1200 liquid chromatography system. After adding isotope-labeled Cre-d3 as internal standard, serum samples were prepared via a one-step protein precipitation with methanol. The LC-MS/MS method was compared with frequently used enzymatic method and Jaffe method. This developed method, with a total run time of 3 min, had a lower limit of quantification of 4.4 μmol/L, a total imprecision of 1.15%-3.84%, and an average bias of 1.06%. No significant matrix effect, carryover, and interference were observed for the LC-MS/MS method. The reference intervals of serum Cre measured by LC-MS/MS assay were 41-79 μmol/L for adult women, and 46-101 μmol/L for adult men. Using LC-MS/MS as a reference, the enzymatic method showed an average bias of -2.1% and the Jaffe method showed a substantial average bias of 11.7%. Compared with the LC-MS/MS method, significant negative bias was observed for the enzymatic and Jaffe methods in hemolytic and lipimic samples. We developed a simple, specific, and accurate LC-MS/MS method to analyze serum Cre. Discordance existed among different methods.

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Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Fig 1
Fig 1. Chromatograms of (A) blank (20% methanol), (B) LLOQ (4.4 μmol/L, S/N = 921) and (C) a patient sample (43.9 μmol/L).
Fig 2
Fig 2. Chromatograms of post-column infusion of Cre-d3 (132.7 μmol/L in 20% methanol) with a patient sample.
Fig 3
Fig 3. Method comparison of the serum Cre with (1) Deming regression and (2) Bland-Altman analysis of (A) LC-MS/MS versus enzymatic method, (B) LC-MS/MS versus Jaffe method, and (C) enzymatic method versus Jaffe method.
Fig 4
Fig 4. Effects of lipemia and hemolysis on the mean difference between (A) LC-MS/MS and enzymatic methods, and (B) LC-MS/MS and Jaffe methods.
See this image and copyright information in PMC

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