Validação do teste lactulose: manitol através de hplc- ms/ms para análise de barreira morfofuncional gastrointestinal

• Ano de defesa: 2021
• Autor(a) principal: Magalhães, Lyvia Maria Vasconcelos Carneiro
• Tipo de documento: Tese
• Tipo de acesso: Acesso aberto
• Idioma: por
• Palavras-chave em Português: Biomarcadores; Dosagem; Espectrometria de Massas; Lactulose; Manitol
• Link de acesso: http://www.repositorio.ufc.br/handle/riufc/58630

Mass spectrometry associated with high performance liquid chromatography (MS / MS HPLC) promises many benefits over liquid chromatography in the dosage of excretion of lactulose and mannitol sugars for the evaluation of the gastrointestinal morphofunctional barrier. The technique has high analytical performances. The present work involves the development, optimization and validation of an analytical method for simultaneous determination of lactulose and mannitol in urine samples from children in the city of Fortaleza - Ceará using MS / MS- HPLC. The main objective is to validate the efficiency in an MS / MS- HPLC system: Ionization by Eletrospray (ESI), evaluating the sensitivity and the matrix effect of the compounds under study. The figures of merit such as selectivity, detection limits (LD) and quantification (LQ), linearity and accuracy were evaluated through tests of recovery of the method. The analytical curves were statistically validated, and showed good linearity, with determination coefficients above 0.99, in accordance with the standards adopted by ANVISA. LD values for lactulose and mannitol were respectively 0.0055 and 0.0003 ng / mL and LQ 0.0168 and 0.0010. The accuracy and precision tests were evaluated at three concentration levels (100, 500 and 1000 ng / mL). Matrix effect was present between 8-30% (ionization suppression). Thus, the method was applied to 41 urine samples to study the present study. Lactuloses and mannitol were quantified in all analyzed samples. The samples came from children living in neighborhoods with different IDH in Fortaleza-CE. Children living in low IDH showed a significantly higher L: M ratio than children living in neighborhoods with higher IDH (P <0.05) and significantly lower percentage of mannitol excretion (P <0.05). The present study demonstrated that the L: M test using MS / MS- HPLC is fast, simple and sensitive in detecting the biomarkers lactulose and mannitol in the urine to assess the impairment of the gastrointestinal morphofunctional barrier and environmental enteric dysfunction in children living in regions with lower IDH which have impaired gastrointestinal morphofunctional barrier.