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Vol. 38 No. 7 (2026): Vol. 38, No 7 (2026)

Copyright (c) 2026 ETIENNE NDIKUMANA, ASTHA PANDEY, ANIRUDHA DIXIT, AAKANKSHA SONI, TULSIDAS R. BAGGI

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This work is licensed under a Creative Commons Attribution 4.0 International License.

Targeted LC-MS Quantification of Eight Urinary Opioids using Validated 10-Fold Enrichment: Forensic and De-addiction Applications in Casework

https://doi.org/10.14233/ajchem.2026.36136
Etienne Ndikumana
Centre of Excellence for Research and Analysis of Narcotic Drugs and Psychotropic Substances and School of Forensic Science, National Forensic Sciences University, Sector 9, Gandhinagar-382007, India
https://orcid.org/0009-0000-7878-1466
Astha Pandey
Centre of Excellence for Research and Analysis of Narcotic Drugs and Psychotropic Substances and School of Forensic Science, National Forensic Sciences University, Sector 9, Gandhinagar-382007, India
https://orcid.org/0000-0002-9096-2566
Anirudha Dixit
Centre of Excellence in Nutritional Supplements Testing for Sports Person and School of Forensic Science, National Forensic Sciences University, Sector 9, Gandhinagar-382007, India
https://orcid.org/0009-0001-3783-2973
Aakanksha Soni
Centre of Excellence in Nutritional Supplements Testing for Sports Person and School of Forensic Science, National Forensic Sciences University, Sector 9, Gandhinagar-382007, India
https://orcid.org/0009-0001-1368-2886
Tulsidas R. Baggi
Central Forensic Science Laboratory, Ministry of Home Affairs, Government of India, Ramanthapur, Hyderabad-500013, India; Department of Forensic Science, University College of Science, Osmania University, Hyderabad-500007, India
https://orcid.org/0000-0002-4310-4358

Corresponding Author(s) : Astha Pandey

astha.pandey@nfsu.ac.in

Asian Journal of Chemistry, Vol. 38 No. 7 (2026): Vol. 38, No 7 (2026)

Abstract

Limited access to liquid chromatography-tandem mass spectrometry (LC-MS/MS) can restrict targeted urinary opioid testing in some forensic laboratories. A fit-for-purpose single-quadrupole liquid chromatography-mass spectrometry (LC-MS) workflow was developed and validated for the quantification of urine morphine, codeine, 6-monoacetylmorphine (6-MAM), tramadol, 6-acetylcodeine (6-AC), heroin, fentanyl and methadone with 10-fold enrichment. The samples were subjected to passive hydrolysis, Oasis HLB solid-phase extraction (SPE) cleanup, enrichment, C18 separation, positive electrospray ionisation (ESI) full-scan acquisition and extracted-ion chromatogram (EIC)-based monitoring. This method was applied to 63 de-addiction and forensic casework specimens. The effective urine-equivalent limit of quantification (LOQ) was 50.0 ng/mL, corresponding to 0.5 µg/mL in the final enriched extract. Calibration was linear over 0.5-25.0 µg/mL with a correlation coefficient (R2) of ≥ 0.990. Precision varied from 0.2% to 16.8%, bias from -17.3% to 18.0%, matrix effect (ME) from 85.8% to 109.1% and extraction recovery (RE) from 82.4% to 114.3%. Codeine and morphine were the most frequently quantified analytes, followed by 6-MAM and 6-AC. The method provides targeted urinary opioid quantification within a defined validation and LOQ-based reporting scope, without claiming equivalence to LC-MS/MS/multiple reaction monitoring (MRM) confirmation.

Keywords

Targeted urinary opioids LC-MS 10-Fold enrichment De-addiction monitoring Forensic toxicology
(1)
Ndikumana, E.; Pandey, A.; Dixit, A.; Soni, A.; Baggi, T. R. Targeted LC-MS Quantification of Eight Urinary Opioids Using Validated 10-Fold Enrichment: Forensic and De-Addiction Applications in Casework. ajc 2026, 38, 1897-1911.
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