Broadening the coverage of metabolites in faeces through advanced analytical strategies

The analysis of human faecal metabolites can provide an insight into metabolic interactions between gut microbiota and the host organism; furthermore, it can be instrumental to not only treat, but also intercept or even prevent diseases. The present project proposes the development, validation and application of two complementary chromatographic platforms (GC/MS and LC-IM-QTOF-MS) in combination with solid phase microextraction (SPME) as a greener, high throughput sample preparation method for faecal metabolites study, more specifically, volatile organic compounds (VOCs) and bile acids (BAs). As a proof-of-concept, faeces from patients with anorexia nervosa (AN) will be analyzed and compared to that obtained from health individuals (controls) aiming to identify possible biomarkers and/or associations between metabolites. Furthermore, patients with AN display nutritional, metabolic, hypothalamic, and endocrine abnormalities, which can lead to them presenting modified microbiota. Interestingly, eventhough AN patients often avoid cholesterol-rich food, hypercholesterolemia is commonly diagnosed in AN patients. In this context, the current project also aims to investigate possible correlations between short chain fatty acids and bile acids levels, BMI, and blood lipidic profile (cholesterol) for patients with anorexia nervosa (AN) and healthy individuals (controls) individuals that could shed light into the gut health, development of hypercholesterolemia, and establish the microbiota role in the evolution of the disease. Moreover, this project aims to develop simple to execute, greener, automated sample preparation for faecal samples that could be easily implemented in clinical settings.