Quantification of arginine and its methylated derivatives in plasma by high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS)

Faye B. Vicente, Gina Vespa, Alan Miller, Shannon Haymond

Research output: Chapter in Book/Report/Conference proceedingChapter

6 Scopus citations

Abstract

Arginine is the substrate for nitric oxide synthases (NOS), thus the production of nitric oxide (NO) is based on arginine availability. Arginine is methylated through the activity of protein arginine methyltransferases (PRMT1 and PRMT2), to form asymmetrical dimethylarginine (ADMA) and symmetrical dimethylarginine (SDMA). These compounds have gained interest in recent years due to their influence on NO production rates and association with cardiovascular and renal diseases. The accurate and precise measurement of arginine and its methylated derivatives is needed for research studies investigating their role(s) in NO bioavailability and development of disease. We describe a high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method for quantifying arginine, ADMA, and SDMA requiring only 50 μL of plasma. The sample preparation involves addition of internal standards (ADMA-d7 for ADMA and SDMA, and 13C6 -arginine for arginine) prior to protein precipitation with LCMS grade acetonitrile. Samples are centrifuged and supernatant is dried under nitrogen gas at 50 °C. Samples are reconstituted with mobile phase (ammonium acetate—formic acid—water). Arginine, ADMA, and SDMA are separated using an isocratic HPLC method on a 3 μM silica analytical column. MS/MS detection is performed in the multiple-reaction monitoring (MRM) mode and the transitions monitored are m/z 203 to m/z 70 for ADMA and SDMA, m/z 210 to m/z 77 for ADMA-d7, m/z 175 to m/z 70 for arginine, and m/z 181 to m/z 74 for 13C6-arginine.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc
Pages21-30
Number of pages10
DOIs
StatePublished - Jan 1 2016

Publication series

NameMethods in Molecular Biology
Volume1378
ISSN (Print)1064-3745

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Keywords

  • Arginine
  • Asymmetric dimethylarginine
  • Liquid chromatography
  • Mass spectrometry
  • Plasma
  • Quantification
  • Symmetric dimethylarginine

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Vicente, F. B., Vespa, G., Miller, A., & Haymond, S. (2016). Quantification of arginine and its methylated derivatives in plasma by high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS). In Methods in Molecular Biology (pp. 21-30). (Methods in Molecular Biology; Vol. 1378). Humana Press Inc. https://doi.org/10.1007/978-1-4939-3182-8_3