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

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.

Oral status of a noble European couple from the 16th century: A morphologic analysis of the teeth of Alessandro Farnese and his wife Maria D'Aviz.

Objective. To report the results of dental morphological analysis performed on the remains of two European nobles (Duke Alessandro Farnese and his wife Maria D'Aviz) who lived in the 16th century, together with hypotheses on their nutrition and oral hygiene habits. Design and results. The remains of Alessandro Farnese (1545-1592) and Princess Maria D'Aviz (1538-1577) were exhumed in Parma, Italy, in the context of an historical investigation into their possible causes of death. The skulls were examined and analysed through direct inspection, high-detailed photographs and radiographs. Ante mortem tooth loss (AMTL), postmortem tooth loss (PMTL), Scott and Smith dental wear indexes and the Kerr periodontal index were used to assess the dental and periodontal status of the couple. Alessandro Farnese suffered from severe dental wear while it was presumed that Maria D'Aviz was affected by periodontal disease and tooth decay. Conclusions. Based on the findings of the present analysis, we hypothesise that Alessandro Farnese's diet was mainly based on hard and unrefined foods, also suggested by limited historical reports. It is likely that Maria D'Aviz's nutrition was based on sugar-enriched foods.