Background: Symmetric dimethylarginine (SDMA) and asymmetric dimethylarginine (ADMA) are naturally occurring amino acids classed as uraemic toxins by the European Uremic Toxins Work Group. SDMA is principally excreted through the kidneys and is a well-known renal function marker, and ADMA is a potent inhibitor of nitric oxide production. Here, we describe the development of a rapid and sensitive liquid chromatography tandem mass spectrometry method for simultaneous measurement of SDMA, ADMA and creatinine.
Method: Serum samples were prepared by protein precipitation and dilution with acetonitrile prior to injection onto a Waters TQS-Micro. SDMA, ADMA, creatinine and their corresponding internal standard transitions were detected using multiple reaction monitoring after separation with a hydrophilic interaction liquid chromatography analytical column. Sample stability and intra-individual variation studies were also assessed following ethical approval.
Results: The retention time for creatinine was 0.43, SDMA 1.10 and ADMA 1.14 min. Mean recovery for creatinine was 103%, SDMA was 100% and ADMA was 103%; matrix effects were minimal (<6%). Lower limit of quantitation for creatinine and SDMA/ADMA was 17.5 µmol/L and 0.1 µmol/L, respectively. Analytical imprecision showed a coefficient of variation <10% for all analytes across the working range of the assays. Intra-individual variation for creatinine was 4.7%, SDMA 7.5% and ADMA 7.6%.
Discussion: We have developed a rugged assay for measurement of SDMA, ADMA and creatinine by LC-MS/MS suitable for routine use. It is easy to perform owing to its simplicity and reproducibility. The stability of SDMA and ADMA pre- and post-centrifugation allows for their routine use without any special sample handling requirements.
Keywords: Mass spectrometry; analytes; clinical studies; creatinine; laboratory methods; renal disease.