Untargeted Metabolomics via UPLC-HRMS and ASCA-Based Statistical Modeling to Characterize Metabolic Alterations Following Acute and Repeated Brorphine Exposure
6 affiliations
- 1Department of Chemistry, Sapienza University of Rome, 00185 Rome, Italy
- 2Department of Public Health and Infectious Disease, Sapienza University of Rome, 00185 Rome, Italy
- 3Department of Environmental Sciences and Prevention, University of Ferrara, 44121 Ferrara, Italy
- 4Department of Translational Medicine, Section of Legal Medicine and LTTA Centre, University of Ferrara, 44121 Ferrara, Italy
- 5Carabinieri, Department of Scientific Investigation (RIS), 00191 Rome, Italy
- 6Collaborative Center for the Italian National Early Warning System (NEWS-D), Department of Anti-Drug Policies and Other Addictions, Presidency of the Council of Ministers, 00184 Rome, Italy
Abstract
Background: Novel Synthetic Opioids (NSOs) currently represent the third most prevalent group among New Psychoactive Substances (NPS), yet they represent the category associated with the highest health risk. Their extreme pharmacological potency compared to traditional opioids, coupled with structural heterogeneity and low concentrations in biological matrices, poses significant analytical challenges for detection. Methods: This study proposes an indirect screening approach based on untargeted metabolomics to monitor systemic responses following brorphine administration. Urinary metabolic profiling was conducted using a sex-balanced murine cohort (n = 10); metabolic profile variations were analyzed following acute and repeated administrations. Samples were collected at 0–12 h and 12–24 h intervals and analyzed by liquid chromatography coupled with high resolution mass spectrometry (LC-HRMS); data were processed using ANOVA–Simultaneous Component Analysis (ASCA), for the isolation of metabolic fingerprints associated with distinct experimental factors (sex, treatment, sampling time). Results: The results revealed a notable metabolic divergence upon repeated administration, linked to sexual diergism: while females exhibited a high recovery capacity supported by a more efficient metabolic biotransformation, males showed a stronger energetic deficit, likely due to greater substance persistence and accumulation. Male vulnerability is characterized by neuro-metabolic dysfunction, inhibition of γ-aminobutyric acid (GABA) release and altered dopaminergic turnover. This systemic imbalance is further defined by massive amino acid and nucleotide catabolism, alongside an energetic deficit exacerbated by inefficient β-oxidation. Conclusions: This study demonstrates that metabolomics may provide interesting insights for the study of NPS by recognizing the biochemical signatures of drug consumption and it is essential for developing sex-specific strategies within the broader context of opioid pharmacology.
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BibTeX
@article{bracaglia2026untargeted,
title = {{Untargeted Metabolomics via UPLC-HRMS and ASCA-Based Statistical Modeling to Characterize Metabolic Alterations Following Acute and Repeated Brorphine Exposure}},
author = {Ilenia Bracaglia and Ludovica Chiodo and Camilla Montesano and Sabrine Bilel and Marta Bassi and Nicole Cocita and Fabiana Di Rosa and Adolfo Gregori and Matteo Marti and Federico Marini and Manuel Sergi},
journal = {Metabolites},
year = {2026},
month = oct,
doi = {10.3390/metabo16100770},
url = {https://www.mdpi.com/2218-1989/16/10/770},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Untargeted Metabolomics via UPLC-HRMS and ASCA-Based Statistical Modeling to Characterize Metabolic Alterations Following Acute and Repeated Brorphine Exposure
AU - Ilenia Bracaglia
AU - Ludovica Chiodo
AU - Camilla Montesano
AU - Sabrine Bilel
AU - Marta Bassi
AU - Nicole Cocita
AU - Fabiana Di Rosa
AU - Adolfo Gregori
AU - Matteo Marti
AU - Federico Marini
AU - Manuel Sergi
PY - 2026
DA - 2026/10/01
JO - Metabolites
DO - 10.3390/metabo16100770
UR - https://www.mdpi.com/2218-1989/16/10/770
AB - Background: Novel Synthetic Opioids (NSOs) currently represent the third most prevalent group among New Psychoactive Substances (NPS), yet they represent the category associated with the highest health risk. Their extreme pharmacological potency compared to traditional opioids, coupled with structural heterogeneity and low concentrations in biological matrices, poses significant analytical challenges for detection. Methods: This study proposes an indirect screening approach based on untargeted metabolomics to monitor systemic responses following brorphine administration. Urinary metabolic profiling was conducted using a sex-balanced murine cohort (n = 10); metabolic profile variations were analyzed following acute and repeated administrations. Samples were collected at 0–12 h and 12–24 h intervals and analyzed by liquid chromatography coupled with high resolution mass spectrometry (LC-HRMS); data were processed using ANOVA–Simultaneous Component Analysis (ASCA), for the isolation of metabolic fingerprints associated with distinct experimental factors (sex, treatment, sampling time). Results: The results revealed a notable metabolic divergence upon repeated administration, linked to sexual diergism: while females exhibited a high recovery capacity supported by a more efficient metabolic biotransformation, males showed a stronger energetic deficit, likely due to greater substance persistence and accumulation. Male vulnerability is characterized by neuro-metabolic dysfunction, inhibition of γ-aminobutyric acid (GABA) release and altered dopaminergic turnover. This systemic imbalance is further defined by massive amino acid and nucleotide catabolism, alongside an energetic deficit exacerbated by inefficient β-oxidation. Conclusions: This study demonstrates that metabolomics may provide interesting insights for the study of NPS by recognizing the biochemical signatures of drug consumption and it is essential for developing sex-specific strategies within the broader context of opioid pharmacology.
SN - 2218-1989
ER -