ICOPMAP SPECIAL EDITION Development of MDMA analysis method in dried blood spot using gas chromatography mass spectrometry Quadrupole Time-of-Flight
DOI:
https://doi.org/10.46542/pe.2025.252.6267Keywords:
Dried blood spot, Ecstasy, Ephedrine HCl, GC/MS Q-ToF, Liquid-liquid microextraction, ValidationAbstract
Background: 3,4-Methylenedioxymethamphetamine (MDMA) or ecstasy is a synthetic stimulant of the central nervous system (CNS) that induces hallucinogenic effects, addiction, and drowsiness. To minimise MDMA misuse, accurate testing is needed using the most reliable specimen, which is blood.
Objective: This study aims to develop an MDMA analysis method using dried blood spot (DBS) sampling, optimise the analysis conditions, prepare the DBS samples, and validate the analytical process.
Method: MDMA was analysed using gas chromatography-mass spectrometry, Quadrupole Time of-Flight, with ephedrine HCl as an internal standard. DBS preparation involved applying 30 μL of blood, drying for 3 hours, and extracting with methanol, followed by vortexing, sonication, and reconstitution with ethyl acetate.
Result: The method validation was performed based on the FDA 2022 M10 Bioanalytical Method Validation and Study Sample Analysis guidelines, showing linearity within the concentration range of 15.00 200 ppb.
Conclusion: The optimised method demonstrated reliable results for the analysis of MDMA in DBS samples, offering a simplified, less invasive blood collection technique with high analytical precision.
References
Baillargeon, K. R., Brooks, J. C., Miljanic, P. R., & Mace, C. R. (2022). Patterned dried blood spot cards for the improved sampling of whole blood. ACS measurement science Au, 2(1), 31–38. https://doi.org/10.1021/acsmeasuresciau.1c00031
Dunlap, L. E., Andrews, A. M., & Olson, D. E. (2018). Dark classics chemical neuroscience: 3,4 Methylenedioxymethamphetamine. ACS chemical neuroscience, 9(10), 2408–2427. https://doi.org/10.1021/acschemneuro.8b00155
European Medicines Agency. (2011). Guideline on bioanalytical method validation. https://www.ema.europa.eu/en/documents/scientific-guideline/guideline-bioanalytical- method-validation_en.pdf
Evans, C., Arnold, M., Bryan, P., Duggan, J., James, C. A., Li, W., Lowes, S., Matassa, L., Olah, T., Timmerman, P., Wang, X., Wickremsinhe, E., Williams, J., Woolf, E., & Zane, P. (2015). Implementing dried blood spot sampling for clinical pharmacokinetic determinations: Considerations from the IQ consortium microsampling working group. AAPS Journal, 17(2) 292–300. Springer New York LLC. https://doi.org/10.1208/s12248-014-9695-3
Gerostamoulos, D., & Schumann, J. (2023). Blood analysis for traditional drugs of abuse. In M. M. Houck (Ed.), Encyclopedia of Forensic Sciences, (3rd ed., Vol. 1, pp. 356-364). Elsevier. https://doi.org/10.1016/B978-0-12-823677-2.00270-1
Ghosh, P., Chowdhury, R., Rahat, Md. A., Hossain, F., Arpha, N. E., Kristan, M., Higgins, M., El Wahed, A. A., Goto, Y., Islam, M. M. T., Campino, S., Cameron, M., Duthie, M. S., Haque, R., & Mondal, D. (2023). Dried Blood Spots (DBS): A suitable alternative to using whole blood samples for diagnostic testing of visceral leishmaniasis in the post- elimination era. PLOS Neglected Tropical Diseases, 17(10), e0011680. https://doi.org/10.1371/journal.pntd.0011680
Harahap, Y., Irawan, H., & Kuswardani (2020). Development and validation of analytical method of 3, 4-methylenedioxy-n-ethylamphetamine in dried blood spot using gas chromatography-mass spectrometry. International Journal of Applied Pharmaceutics, 12(4), 94-99. https://doi.org/10.22159/ijap.2020v12i4.32304
Jantos, R., Veldstra, J., Mattern, R., Brookhuis, K., & Skopp, G. (2011). Analysis of 3,4- Methylenedioxymetamphetamine: Whole blood versus dried blood spots. Journal Of Analytical Toxicology, 35(5), 269–273.
Lacorte, S., & Fernandez‐Alba, A. R. (2006). Time of flight mass spectrometry applied to the liquid chromatographic analysis of pesticides in water and food. Mass Spectrometry Reviews, 25(6), 866–880. https://doi.org/10.1002/mas.20094
Lazarovits, J., Chen, Y. Y., Sykes, E. A., & Chan, W. C. W. (2015). Nanoparticle-blood interactions: The implications on solid tumour targeting. Chemical Communications, 51(14), 2756–2767. https://doi.org/10.1039/c4cc07644c
McNeil, S. E. (2023). Drug testing. StatPearls. NIDA. (2024, April 19). MDMA (Ecstasy/Molly).
National Narcotics Agency. (2023). Indonesia drug report 2023.
Sarah Ingvarsson. (2020). Characterization of hepatocyte derived metabolites of various New Psychoactive Substances using LC-QTOF-MS [16 hp]. Linköping University.
Schenk, S., & Highgate, Q. (2021). Methylenedioxymethamphetamine (MDMA): Serotonergic and dopaminergic mechanisms related to its use and misuse. Journal of Neurochemistry, 157(5), 1714–1724. https://doi.org/10.1111/jnc.15348
Seo, S.-H., & Batterman, S. (2024). Estimating blood volume on dried blood spots. Forensic Chemistry, 37, 100552. https://doi.org/10.1016/j.forc.2024.100552
United Nations. (2006). Multilingual dictionary of narcotic drugs and psychotropic substances under international control. https://www.unodc.org/documents/scientific/MLD-06-58676_Vol_1_ebook.pdf
United Nations. (2006). Recommended methods for the identification and analysis of amphetamine, methamphetamine and their ring-substituted analogues in seized materials (revised and updated). Manual for Use by National Drug Testing Laboratories. United Nations Office on Drugs and Crime. https://syntheticdrugs.unodc.org/uploads/syntheticdrugs/res/library/forensics_html/Recommended_Methods_for_the_Identification_and_Analysis_of_Amphetamine_Methamphetamine_and_their_Ring-substituted_Analogues_in_Seized_Materials_ST-NAR-34.pdf