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Method development and validation for rat serum fingerprinting with CE–MS: application to ventilator-induced-lung-injury study

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Abstract

In the search for a noninvasive and reliable rapid screening method to detect biomarkers, a metabolomics fingerprinting approach was developed and applied to rat serum samples using capillary electrophoresis coupled to an electrospray ionization-time of flight-mass spectrometer (CE–TOF-MS). An ultrafiltration method was used for sample pretreatment. To evaluate performance the method was validated with carnitine, choline, ornithine, alanine, acetylcarnitine, betaine, and citrulline, covering the entire electropherogram of pool of rat serum. The linearity for all metabolites was >0.99, with good recovery and precision. Approximately 34 compounds were also confirmed in the pool of rat serum. The method was successfully applied to real serum samples from rats with ventilator-induced lung injury, an experimental rat model for acute lung injury (ALI), giving a total of 1163 molecular features. By use of univariate and multivariate statistics 18 significant compounds were found, of which five were confirmed. The involvement of arginase and nitric oxide synthase has been proved for other lung diseases, meaning the increase of asymmetric dimethyl arginine (ADMA) and ornithine and the decrease of arginine found were in accordance with published literature. Ultimately this fingerprinting approach offers the possibility of identifying biomarkers that could be regularly screened for as part of routine disease control. In this way it might be possible to prevent the development of ALI in patients in critical care units.

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Acknowledgements

Shama Naz is receiving funding from the European Union Seventh Framework Programme (FP7/2007-2013) under grant agreement no-264864, and a research grant from Ministerio de Economia y Competitividad (MINECO, CTQ2011-23562). The authors thank Jesús Ruíz-Cabello, Andrés Esteban, and José Angel Lorente from the Getafe Hospital, Madrid, Spain, for the serum samples of VILI and control, Dr Marcela Erazo, for helping with the biological interpretation of choline, and Dr Emily G. Armitage, for checking the grammar throughout the manuscript.

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Correspondence to Coral Barbas.

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Naz, S., Garcia, A., Rusak, M. et al. Method development and validation for rat serum fingerprinting with CE–MS: application to ventilator-induced-lung-injury study. Anal Bioanal Chem 405, 4849–4858 (2013). https://doi.org/10.1007/s00216-013-6882-5

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  • DOI: https://doi.org/10.1007/s00216-013-6882-5

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