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Use of solid-phase microextraction coupled to gas chromatography–mass spectrometry for determination of urinary volatile organic compounds in autistic children compared with healthy controls

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Abstract

Autism spectrum disorders (ASDs) are a group of neurodevelopmental disorders which have a severe life-long effect on behavior and social functioning, and which are associated with metabolic abnormalities. Their diagnosis is on the basis of behavioral and developmental signs usually detected before three years of age, and there is no reliable biological marker. The objective of this study was to establish the volatile urinary metabolomic profiles of 24 autistic children and 21 healthy children (control group) to investigate volatile organic compounds (VOCs) as potential biomarkers for ASDs. Solid-phase microextraction (SPME) using DVB/CAR/PDMS sorbent coupled with gas chromatography–mass spectrometry was used to obtain the metabolomic information patterns. Urine samples were analyzed under both acid and alkaline pH, to profile a range of urinary components with different physicochemical properties. Multivariate statistics techniques were applied to bioanalytical data to visualize clusters of cases and to detect the VOCs able to differentiate autistic patients from healthy children. In particular, orthogonal projections to latent structures discriminant analysis (OPLS-DA) achieved very good separation between autistic and control groups under both acidic and alkaline pH, identifying discriminating metabolites. Among these, 3-methyl-cyclopentanone, 3-methyl-butanal, 2-methyl-butanal, and hexane under acid conditions, and 2-methyl-pyrazine, 2,3-dimethyl-pyrazine, and isoxazolo under alkaline pH had statistically higher levels in urine samples from autistic children than from the control group. Further investigation with a higher number of patients should be performed to outline the metabolic origins of these variables, define a possible association with ASDs, and verify the usefulness of these variables for early-stage diagnosis.

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Acknowledgment

We thank study subjects and their families for participating in this study, and Dr Spagnuolo, head of the no-profit association “Associazione Irpinia Pianeta Autismo (AIPA)” located in Avellino (Italy), for her valuable support. We would also like to acknowledge Christopher Nardone for reviewing the English of the manuscript.

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Authors and Affiliations

  1. Institute of Food Science, National Council of Research, via Roma 64, 83100, Avellino, Italy

    Rosaria Cozzolino, Paola Saggese, Antonella Martignetti, Michele Di Stasio & Floriana Boscaino

  2. CIRANAD, Second University of Naples, via Pansini 5, Building 3, 80131, Naples, Italy

    Laura De Magistris

  3. S-IN Soluzioni Informatiche Srl, via Ferrari 14, 36100, Vicenza, Italy

    Matteo Stocchero

  4. Department of Environmental, Biological and Pharmaceutical Sciences and Technologies, Second University of Naples, Via Vivaldi 43, 81100, Caserta, Italy

    Antonio Malorni

  5. Department of Psychiatry, The Magna Graecia University of Catanzaro, Via T. Campanella, 88100, Catanzaro, Italy

    Rosa Marotta

  6. Department of Experimental Medicine, Section of Hygiene, Occupational Medicine and Forensic Medicine, School of Medicine, Second University of Naples, Via L. De Crecchio 7, 80138, Naples, Italy

    Livia Malorni

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  1. Rosaria Cozzolino
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  2. Laura De Magistris
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Correspondence to Rosaria Cozzolino.

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Cozzolino, R., De Magistris, L., Saggese, P. et al. Use of solid-phase microextraction coupled to gas chromatography–mass spectrometry for determination of urinary volatile organic compounds in autistic children compared with healthy controls. Anal Bioanal Chem 406, 4649–4662 (2014). https://doi.org/10.1007/s00216-014-7855-z

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  • DOI: https://doi.org/10.1007/s00216-014-7855-z

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