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Improved method for plasma ADMA, SDMA, and arginine quantification by field-amplified sample injection capillary electrophoresis UV detection

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Abstract

Here, we describe an easy field-amplified sample injection capillary electrophoresis method with UV detection for the separation and detection of free plasma arginine and dimethylated arginines. The analytes were baseline-separated within 22 min by using 50 mmol/L Tris phosphate pH 2.3 as running buffer. The plasma samples were treated with acetonitrile/ammonia for protein elimination, the supernatants were dried, re-swollen in water and directly injected in the capillary without complex cleanup by solid phase extraction and/or tedious sample derivatization procedures. Due to the stacking effects of the electrokinetic injection, it was possible to operate a consistent on-line pre-concentration of the analytes before running the electrophoresis. This procedure allowed to reach a detection limit in the real sample of 10 nmol/L for dimethylated arginines and 20 nmol/L for arginine, thus improving about threefold our previous method, that required a more complicated pre-analytical procedure to concentrate samples. The recovery of plasma ADMA was 99–104% and inter-day CV was less than 3%. The assay performance was evaluated measuring the levels of arginine and its dimethyl derivatives in 50 subjects. The statistical tests for the methods comparison suggest that the data obtained by our new method and by our previous CE assay are similar.

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Abbreviations

ADMA:

Asymmetric dimethylarginine

Arg:

Arginine

CE:

Capillary electrophoresis

FASI:

Field-amplified sample stacking

SDMA:

Symmetric dimethylarginine

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Acknowledgments

This study was supported by the “Fondazione Banco di Sardegna—Sassari—Italy” and by the “Ministero dell’Università e della Ricerca” Italy. Angelo Zinellu was supported by Regione Autonoma della Sardegna “Ricerca cofinanziata PROGRAMMA OPERATIVO FSE SARDEGNA 2007–2013-L.R.7/2007—Promozione della ricerca scientifica e dell’innovazione tecnologica in Sardegna”. The manuscript language revision by Mrs Maria Antonietta Meloni is greatly appreciated.

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Authors and Affiliations

  1. Porto Conte Ricerche Srl, Tramariglio, 07041, Alghero, Sassari, Italy

    Angelo Zinellu

  2. Department of Biomedical Sciences, University of Sassari, Viale San Pietro 43/B, 07100, Sassari, Italy

    Angelo Zinellu, Salvatore Sotgia, Gianfranco Pintus, Luca Deiana & Ciriaco Carru

  3. Department of Botanic, Ecology and Geology Science, University of Sassari, 07100, Sassari, Italy

    Maria Franca Usai

  4. Department of Zoology and Evolutionary Genetics, University of Sassari, 07100, Sassari, Italy

    Maria Franca Usai

  5. National Laboratory of the National Institute of Biostructures and Biosystems, 07033, Osilo, Italy

    Ciriaco Carru

Authors
  1. Angelo Zinellu
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  2. Salvatore Sotgia
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  3. Maria Franca Usai
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  4. Gianfranco Pintus
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  5. Luca Deiana
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  6. Ciriaco Carru
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Correspondence to Ciriaco Carru.

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Zinellu, A., Sotgia, S., Usai, M.F. et al. Improved method for plasma ADMA, SDMA, and arginine quantification by field-amplified sample injection capillary electrophoresis UV detection. Anal Bioanal Chem 399, 1815–1821 (2011). https://doi.org/10.1007/s00216-010-4580-0

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  • DOI: https://doi.org/10.1007/s00216-010-4580-0

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