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- Comparison of spectroscopic and molecular methods for estimating age and sex for medico-legal purposes: protocol for a systematic reviewPublication . Lopes Cardoso, Inês; Loibl, Beatriz; Rodrigues, Carolina; Moreira, Teresa; Guimarães, Maria InêsIntroduction: In forensic contexts, accurate estimation of age and sex is fundamental for the identification of human remains. Spectroscopic methods, offering non-destructive and precise data insights, have emerged as promising alternatives/complements to traditional morphological and genetic techniques. Objective: This protocol outlines a systematic review aimed at evaluating the effectiveness of spectroscopic and molecular methods for estimating age and/or sex based on dental samples in medicolegal settings. Methodology: This review will be conducted in accordance with the PRISMA 2020 guidelines. Databases to be searched include PubMed, Web of Science, ScienceDirect, LILACS (via BVS), Cochrane Library (Protocols), MEDLINE (via EBSCOhost), Academic Search Complete (via EBSCOhost), CINAHL (via EBSCOhost) and grey literature. The eligibility criteria follow the PICOS framework. Risk of bias will be assessed using the QUIN tool for in vitro studies. Data synthesis will involve both quantitative and qualitative approaches. The protocol of this review has been registered in PROSPERO (CRD420250650858). Results: This review is expected to identify a range of spectroscopic techniques applied to dental samples for age and sex estimation, including mass spectrometry, Raman spectroscopy, FTIR spectroscopy, and X-ray fluorescence, as well as molecular methods such as amelogenin analysis, DNA methylation, and telomere length assessment. The evidence is anticipated to vary regarding study design, sample characteristics, analytical protocols, and reported outcome measures. The review will synthesize the available evidence on the accuracy, applicability, and methodological characteristics of these approaches within medico-legal contexts. Conclusion: This systematic review will provide a comprehensive overview of the available evidence regarding the use of spectroscopic and molecular methods for age and sex estimation from dental samples. By identifying the methodologies currently employed, their reported outcomes, and existing research gaps, the review may contribute to future research directions and support evidence-based decision-making in forensic and medico-legal practice, particularly in contexts where non-destructive analytical approaches are desirable.