ORIGINAL RESEARCH article
Front. Anal. Sci.
Sec. Forensic Chemistry
This article is part of the Research TopicVibrational Spectroscopy Trends in Forensic ChemistryView all articles
Sex determination of White-Tailed-Deer (Odocoileus virginianus) from plasma and serum sample by using Raman spectroscopy and PLS-DA method: A Forensic Perspective
Provisionally accepted
- Texas Tech University, Lubbock, United States
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Species identification and sex determination have significant relevance in wildlife monitoring, conservation, and forensic investigations. This study explores the feasibility of using Raman spectroscopy coupled with Partial Least Squares Discriminant Analysis (PLS-DA) for sex determination of white-tailed deer (Odocoileus virginianus) based on serum and plasma samples. A total of 720 Raman spectra (360 serum, 360 plasma) were acquired from five male and five female deer. PLS-DA models were developed using full data and four calibration-to-validation splits (1:4, 2:3, 3:2, 4:1). The serum-based PLS-DA model achieved predictive accuracies of 79.2% (1:4), 99.1% (2:3), 99.3% (3:2), and 98.6% (4:1), with balanced calibration-to-validation splits (≥2:3) demonstrating excellent robustness and generalizability exceeding 98%. The plasma-based model showed high internal accuracy (~99.2%) but reduced external accuracy depending on the split: 85.8% (1:4), 82.4% (2:3), 86.1% (3:2), and 57.0% (4:1), reflecting greater variability and susceptibility to overfitting. Spectral comparison revealed consistent Raman bands in both matrices associated with proteins, lipids, and carbohydrates, with subtle but distinct sex-specific intensity differences. Males showed enhanced signals at ~1315 and ~1445 cm⁻¹ (lipid regions), while females displayed stronger signals at ~1220-1280 and ~1540-1630 cm⁻¹ (protein regions). These findings highlight the potential of Raman-PLSDA models—particularly with serum—as a reliable, rapid, and non-destructive method for sex classification in forensic wildlife contexts, especially when morphological features are absent or degraded. This approach may enable field-adaptable identification that enhance species conservation and forensic investigations involving incomplete or decomposed biological remains.
Keywords: Raman spectroscopy, PLS-DA, White-tailed deer, sex determination, chemometricclassification, Wildlife forensics, multivariate analysis, pattern recognition
Received: 17 Oct 2025; Accepted: 03 Dec 2025.
Copyright: © 2025 Majumder, Smith and Halamkova. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Lenka Halamkova
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