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Comparison of two methods for dimethylarginines quantification.

Vendula Sudová1,2, Pavel Prokop1,2, Ladislav Trefil1,2

  • 1Biomedical Center, Faculty of Medicine in Pilsen, Charles University, Alej Svobody 1655/76, Pilsen, 32300, Czech Republic.

Practical Laboratory Medicine
|February 5, 2024
PubMed
Summary

This study validated a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for measuring dimethylarginines in chronic kidney disease (CKD). Immunoassay overestimated ADMA and underestimated SDMA, suggesting simple sample dilution could improve accuracy.

Keywords:
Asymmetric dimethylarginineCKDELISALC-MS/MSSymmetric dimethylarginine

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Clinical Chemistry

Background:

  • Dimethylarginines (ADMA and SDMA) are biomarkers associated with chronic kidney disease (CKD).
  • Accurate measurement of these biomarkers is crucial for patient management and research.
  • Existing immunoassay methods may have limitations in accuracy compared to advanced analytical techniques.

Purpose of the Study:

  • To validate a published liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for quantifying asymmetric dimethylarginine (ADMA) and symmetric dimethylarginine (SDMA).
  • To compare the results obtained from LC-MS/MS with those from a conventional immunoassay.
  • To assess the accuracy and reliability of both methods in healthy individuals and patients with varying stages of CKD.

Main Methods:

  • Analysis was performed using UHPLC-MS/MS on a Dionex UltiMate 3000 system with a Bruker amaZon SL ion trap.
  • Method comparison involved Passing Bablok regression and Bland Altman plots.
  • Validation and comparison were conducted using samples from 40 healthy volunteers and 40 patients with CKD.

Main Results:

  • LC-MS/MS and immunoassay showed strong correlations for both ADMA (Spearman R=0.858) and SDMA (Spearman R=0.895).
  • Immunoassay results for ADMA were overestimated by approximately 30%, while SDMA levels were underestimated by about 3%.
  • Mean ADMA levels in CKD patients were 0.82 ± 0.16 μmol/L (LC-MS/MS) vs. 1.06 ± 0.30 μmol/L (immunoassay).
  • Mean SDMA levels in CKD patients were 2.14 ± 0.88 μmol/L (LC-MS/MS) vs. 1.65 ± 0.52 μmol/L (immunoassay).

Conclusions:

  • The LC-MS/MS method provides a validated and reliable approach for measuring ADMA and SDMA.
  • Significant discrepancies exist between LC-MS/MS and immunoassay results, particularly for ADMA.
  • Simple sample dilution may improve the accuracy of immunoassay measurements for dimethylarginines.