Ultra-high performance liquid chromatography high-resolution mass spectrometry for metabolomic analysis of dental calculus from Duke Alessandro Farnese and Maria D’Aviz

Abstract

Dental calculus is a valuable resource for the reconstruction of dietary habits and oral microbiome of past populations. In 2020 the remains of Duke Alessandro Farnese and his wife Maria D’Aviz were exhumed to get novel insights into the causes of death. This study aimed to investigate the dental calculus metabolome of the noble couple by untargeted metabolomics. The pulverized samples were decalcified in a water-formic acid mixture, extracted using methanol/acetonitrile and analyzed by ultra-high performance liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-HRMS) using a reversed-phase separation followed by electrospray ionization and full scan in positive and negative ion mode. Waters Synapt-G2-Si High-Definition hybrid quadrupole time-of-flight mass spectrometer was used. Significant features were then identified using MSE acquisition mode, recording information on exact mass precursor and fragment ions within the same run. This approach, together with data pre-treatment and multivariate statistical analysis allowed for the identification of compounds able to differentiate between the investigated samples. More than 200 metabolites were identified, being fatty acids, alcohols, aldehydes, phosphatidylcholines, phosphatidylglycerols, ceramides and phosphatidylserines the most abundant classes. Metabolites deriving from food, bacteria and fungi were also determined, providing information on the habits and oral health status of the couple.

Ultra-high Performance Liquid Chromatography–Ion Mobility–High-Resolution Mass Spectrometry to Evaluate the Metabolomic Response of Durum Wheat to Sustainable Treatments

“…Advances in high-resolution mass spectrometry (HRMS) permit a comprehensive profiling of food samples, representing a valuable technique for studying the metabolic changes developed by organisms in the presence of variable environmental factors. Ultra-high performance liquid chromatography coupled to HRMS (UHPLC-HRMS) currently represents the best tool to face challenges related to the complexity of metabolome . UHPLC provides efficient chromatographic separation of compounds, whereas HRMS is characterized by unparalleled specificity, sensitivity, and availability of large spectral databases.…”

“…Ultra-high performance liquid chromatography coupled to HRMS (UHPLC-HRMS) currently represents the best tool to face challenges related to the complexity of metabolome. 2 UHPLC provides efficient chromatographic separation of compounds, whereas HRMS is characterized by unparalleled specificity, sensitivity, and availability of large spectral databases. In addition, ion mobility spectrometry (IMS) offers great potential for improving depth of coverage in metabolomic studies, separating ions according to their collisional cross-section (CCS) and providing unique features for metabolite identification.…”