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. 2024 Jan 17:39:e00359.
doi: 10.1016/j.plabm.2024.e00359. eCollection 2024 Mar.

Comparison of two methods for dimethylarginines quantification

Vendula Sudová  1   2 , Pavel Prokop  1   2 , Ladislav Trefil  1   2 , Jaroslav Racek  1   2 , Daniel Rajdl  1   2
Affiliations

Comparison of two methods for dimethylarginines quantification

Vendula Sudová et al. Pract Lab Med. .

Abstract

Objectives: Both dimethylarginines are widely bound to chronic kidney disease (CKD). This study was focused to validate published LC-MS/MS method and compared the measured data with an immunoassay.

Design and methods: The analysis was performed on a Dionex UltiMate 3000 UHPLC-Standard (Thermo Fisher Scientific, Waltham, Massachusetts, USA) with an amaZon SL ion trap (Bruker, Billerica, Massachusetts, USA). Comparison was evaluated by using Passing Bablok regression and Bland Altman plot. Healthy volunteers (n = 40) were used for validation and as control group to patients group (n = 40) with different stages of CKD.

Results: The results in healthy controls determined by the LC-MS/MS (ELISA) method were 0.52 ± 0.0892 with 95 % CI: 0.49-0.55 (0.61 ± 0.1213 with 95 % CI: 0.57-0.64) μmol/L for AD MA and 0.56 ± 0.0810 with 95 % CI: 0.53-0.58 (0.62 ± 0.0752 with 95 % CI: 0.57-0.65) μmol/L for SDMA. In the same way, the patient group values determined by the LC-MS/MS (ELISA) method were 0.82 ± 0.1604 with 95 % CI: 0.75-0.88 (1.06 ± 0.3002 with 95 % CI: 0.94-1.19) μmol/L and 2.14 ± 0.8778 with 95 % CI: 1.47-2.58 (1.65 ± 0.5160 with 95 % CI: 1.40-1.98) μmol/L for ADMA and SDMA, respectively. The correlation between the methods, expressed as the Spearman correlation coefficient (R), was 0.858 (0.8059) for ADMA (p < 0.0001) and 0.895 (0.9607) for SDMA (p < 0.0001).

Conclusions: ADMA levels determined by the immunoassay were almost 30 % overestimated, in contrast to SDMA levels, which were 3 % underestimated. According to our findings, a better correlation could be obtained by simple sample dilution.

Keywords: Asymmetric dimethylarginine; CKD; ELISA; LC-MS/MS; Symmetric dimethylarginine.

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Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

Fig. 1
Fig. 1
ADMA (Fig. 1A), ADMA internal standard (Fig. 1B), SDMA (Fig. 1C) and SDMA internal standard (Fig. 1D) chromatograms with their mass spectra.
Fig. 2
Fig. 2
ADMA (2A) and SDMA (2B) serum concentrations (μmol/L) determined by LC-MS/MS and ELISA method in all samples; ADMA (2C) and SDMA (2D) serum concentrations determined by the LC-MS/MS method only in a control group and in patients with different stages of chronic kidney disease.
Fig. 3
Fig. 3
Bland-Altman and Passing-Bablok plots for ADMA (3A, 3B) and SDMA (3C, 3D) quantification method comparison (LC-MS/MS and ELISA).
See this image and copyright information in PMC

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