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Development of analytical methods to study the salivary metabolome: impact of the sampling

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Abstract

Advances in metabolomics have allowed the identification and characterization of saliva metabolites that can be used as biomarkers. However, discrepancies can be noted with the content of the same biomarker being increased or decreased for a given disease. Differences in the way saliva is collected, stored, and/or treated could cause these discrepancies. Indeed, there is no standardized method for saliva sampling and analysis. In this work, two chromatographic modes were used, i.e., RP-LC and HILIC both coupled to MS used in positive and negative ionization modes. The analytical conditions were optimized with a mixture of 90 compounds naturally present in saliva, representative of the wide range of molecular mass and polarity of salivary metabolites and being described as having a differential expression in various pathologies. These four methods were applied to the analysis of saliva samples collected by spitting, aspiration, or Salivette® with or without prior rinsing of the mouth. Rinsing had an effect on some metabolite concentrations. As it can induce an additional parameter of variability to the sampling, it seems therefore preferable to use methods without rinsing while effects of these parameters on the metabolites are investigated. Saliva obtained by spitting and aspiration gave statistically equivalent results for 84% of the metabolites studied. Conversely, Salivette® gave different results since the majority of the metabolites chosen for the study were not quantified in the samples. The Salivette® does not seem therefore to be a suitable sampling method for an untargeted analysis of the salivary metabolome, unlike aspiration and spitting.

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Acknowledgements

We thank Drs Marion Florimond (URP 2496, UFR Odontologie, Université Paris Cité) and Dominique Hotton (UMRS 1138, INSERM Centre des Cordeliers) for their advice in sample staining and light microscopy.

Funding

This work was supported by the “Plateforme d’expertise pour le suivi de toxines à l’état de traces en milieu reel” project from DIM analytics from the Ile de France region.

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Authors and Affiliations

  1. Laboratoire Des Sciences Analytiques, Bioanalytiques Et Miniaturisation, UMR 8231 CBI CNRS, ESPCI Paris, PSL université, Paris, France

    Pauline Bosman, Valérie Pichon, Hélène Chardin & Audrey Combes

  2. Sorbonne Université, Paris, France

    Valérie Pichon

  3. Laboratory of Oral Histopathology, Health Sciences Faculty of Brasilia Campus, Universitario Darcy Ribeiro, Brasilia, Brazil

    Ana Carolina Acevedo

  4. Université Paris Cité, Paris, France

    Ana Carolina Acevedo & Hélène Chardin

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  1. Pauline Bosman
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  2. Valérie Pichon
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  3. Ana Carolina Acevedo
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  5. Audrey Combes
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Contributions

Conceptualization: V. Pichon, A. Combes, H. Chardin. Formal analysis and investigation: P. Bosman, A. Combes. Writing—original draft preparation: P. Bosman. Writing—review and editing: V. Pichon, A. Combes, H. Chardin, A. C. Acevedo. Funding acquisition: V. Pichon. All authors have read and agreed to the published version of the manuscript.

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Correspondence to Audrey Combes.

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This study on human saliva samples have been performed in accordance with the ethical standards of the French General Health Directorate. And the participant gave informed consent for participation.

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Bosman, P., Pichon, V., Acevedo, A.C. et al. Development of analytical methods to study the salivary metabolome: impact of the sampling. Anal Bioanal Chem 414, 6899–6909 (2022). https://doi.org/10.1007/s00216-022-04255-5

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