Aplicação de dried saliva spots na determinação de marcadores de consumo de tabaco

• Main Author: Marques, Hernani Fortunato
• Publication Date: 2021
• Format: Master thesis
• Language: eng
• Source: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
• Download full: http://hdl.handle.net/10400.6/12698
• topic: Biomarcadores de Tabaco; Cromatografia Gasosa Acoplado a Espetrometria de Massa em Tandem; Dried Saliva Spots; Fluido Oral; Nicotina; Tabaco; Domínio/Área Científica::Engenharia e Tecnologia::Bioquímica

Exposure to tobacco smoke is one of the most common causes of premature death worldwide and is the cause of 8 million deaths annually. For this purpose, it was developed, optimized, and validated a procedure for the detection of nicotine, cotinine and trans-3-hydroxycotinine (biomarkers of tobacco exposure) in oral fluid using the dried saliva spots sampling approach and gas chromatography coupled to tandem mass spectrometry, thus allowing the distinction between active and passive smokers. For optimization, 4 parameters were evaluated, namely extraction solvent, extraction solvent volume, extraction time and spots drying time. During method validation, the parameters selectivity, linearity, precision and accuracy, recovery, stability, and dilution factor were assessed. Linearity was obtained for all target analytes in the concentration range of 10-200 ng/mL allowing the quantification of compounds up to 1000 ng/mL considering the dilution factor. The method recoveries ranged from 29.2% to 43.30% for nicotine, 66.60% to 89.10% for cotinine and 80.30% to 92.80% for trans-3-hydroxycotinine, while achieving intra-day, inter-day and intermediate precision and accuracy values never greater than 10.37% and ± 6.50% respectively for all compounds. The herein described analytical method is the first that allows the determination of tobacco biomarkers in oral fluid using dried saliva spots, being considered a sensitive, simple, and low-cost alternative to conventional methods.