Fluoroacetylation/fluoroethylesterification as a derivatization approach for gas chromatography–mass spectrometry in metabolomics: Preliminary study of lymphohyperplastic diseases

• Published in: Journal of Chromatography A 2013-08, Vol.1302, p.125-132
• Main Authors: Karamani, Anna A., Fiamegos, Yiannis Ch, Vartholomatos, George, Stalikas, Constantine D.
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; biomarkers; blood; cluster analysis; Derivatization; Fluoroacetylation; Fluoroethylesterification; Gas chromatography–mass spectrometry; General aspects; Investigative techniques, diagnostic techniques (general aspects); Lymphohyperplastic diseases; Medical sciences; metabolites; Metabolomic fingerprinting; metabolomics; Miscellaneous. Technology; multivariate analysis; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques; patients; saliva; Tumors; urine
• ISSN: 0021-9673
• DOI: 10.1016/j.chroma.2013.05.080

•Development of a derivatization procedure for hydroxyl, carboxyl and amino groups-bearing metabolites.•Application in human urine, whole blood and saliva.•Metabolomic fingerprinting and biomarker(s) discovery in lymphohyperplastic diseases using cluster analysis. Metabolic fingerprinting in combination with gas chromatography and multivariate analysis is being extensively employed for the improved understanding of biological changes induced by endogenous or exogenous factors. Chemical derivatization increases the sensitivity and specificity of gas chromatography–mass spectrometry (GC–MS) for polar or thermally labile biological compounds, which bear derivatizable groups. Thus, there is a constant demand for simple methods of derivatization and separation that satisfy the need for metabolite analysis, identifying as many chemical classes of compounds as possible. In this study, an optimized protocol of extraction and derivatization is established as a generally applicable method for the analysis of a wide range of classes of metabolites in urine, whole blood and saliva. Compounds of biological relevance bearing hydroxyl- carboxyl- and amino-groups are derivatized using single-step fluoroacetylation/fluoroethylesterification after proper optimization of the protocol. Subsequently, the developed derivatization procedure is engaged in finding blood metabolic biomarkers, induced by lymphohyperplastic disease, through the metabolomic fingerprinting approach, the multivariate modeling (hierarchical cluster analysis) and GC–MS. Our preliminary, GC–MS-based metabolomic fingerprinting study underlines the contribution of certain metabolites to the discrimination of patients with lymphohyperplastic diseases.