Investigation of volatile organic compounds in sweat samples as biomarkers in cancer diagnosis.

Author(s)

Fernanda Ferreira da Silva Souza Monedeiro

• Document type: Doctoral Thesis
• Press: Ribeirão Preto.
• Institution: Universidade de São Paulo (USP). Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (PCARP/BC)
• Defense date: 2018-05-23

Examining board members

Bruno Spinosa de Martinis; Celso Teixeira Mendes Junior; Fernanda Maris Peria

• Advisor: Bruno Spinosa de Martinis

Abstract

Natural metabolic processes in the human body lead to the formation of substances such as volatile organic compounds (VOCs), so that, in a pathological context, differentiated processes can occur in the cells, causing a different set of compounds to be produced. With this hypothesis, VOCs can be analyzed in biological samples with the intention to verify changes in their profiles that are indicative of certain pathologies. In the present study, sweat was selected as the matrix, due simple and non-invasive collection, with lower complexity composition and related to blood levels and skin emanations. In addition, urine samples were also analyzed to obtain comparative data. The present study comprised samples from healthy volunteers (control-C group) and individuals with confirmed cancer diagnosis (positive-P group). The sweat samples were collected with PharmChek® device, next, the patch was removed, inserted in a vial and the VOCs were isolated using an optimized Static Headspace (HS) technique. For urine samples, these were analyzed with and without ?-glucuronidase treatment. VOC profiles were obtained by gas chromatography coupled to mass spectrometry (GC-MS) for all samples. The attempt to identify the detected compounds was made by searching the NIST08 library and using the AMDIS32 software. Qualitative differences (chi-square test, p << 0.01) and quantitative tests (Mann-Whitney U test, p << 0.01) were evaluated between the profiles of the control and positive groups. For the sweat, the potential biomarkers pentanal, hexanal, heptanal, octanal, limonene, 2-ethyl-1-hexanol, 1-undecene, phenol, 2,6-dimethyl-7-octen-2-ol (DMOL), nonanal, decanal and tridecane; for urine, phenol and DMOL, both hydrolysis-dependent, were selected. HS-GC-FID (coupled to flame ionization detector) method was developed and validated according to RDC 27/2012- ANVISA, for both samples. In sweat, the analytes presented limits of quantification (LOQ) of 1 ng/patch, 5 ng/patch for phenol; in urine were 2 ng mL-1 for DMOL and 10 ng mL-1 for phenol. Linearity was observed for the range of 2 to 150 ng/patch and, 2 and 5 to 400 ng mL-1 in urine. In sweat, the precision ranged from 0.08 to 12.35% and the analytes were shown to be stable for the assays performed. Receiver Operating Characteristic (ROC) curves were evaluated and areas under the curve were all near to 1, with cut-off values of 1.71 to 35.44 ng/patch in sweat and 8.71 and 52.86 ng mL-1 in urine. 2-ethyl-1-hexanol was shown to be negatively correlated with the clinical stage in adenocarcinomas (rho= -0.527) and DMOL, in sweat, and C5-C8 aldehydes sum, positively related to the stage of prostate cancer (rho= 0.779 and 0.684, respectively). It was concluded, therefore, that the method presented proved to be efficient, however, practical and low cost, and the results corroborate to the idea of VOCs determination as a promising diagnostic tool for cancer diagnosis. (AU)

• FAPESP's process: 13/19653-4 - Investigation of volatile organic compounds in sweat samples as biomarkers for cancer diagnosis
• Grantee: Fernanda Ferreira da Silva Souza Monedeiro
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)