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Forensic Toxicology

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01-01-2021 | Short Communication

Use of injection-port derivatization for the analysis of cocaine and its metabolites in urine by gas chromatography–tandem mass spectrometry

Authors: Kelly Francisco da Cunha, Rafael Lanaro, Aline Franco Martins, Karina Diniz Oliveira, Jose Luiz Costa

Published in: Forensic Toxicology | Issue 1/2021

Abstract

Purpose

We developed a fast and reliable method for the analysis of cocaine and three metabolites in human urine, using injection-port derivatization and gas chromatography–tandem mass spectrometry (IPD–GC–MS/MS), with a simplified sample preparation procedure. IPD is an interesting and fast procedure that simplifies sample preparation, where the reaction between the analytes and the derivatization reagent occurs instantly in the heated instrument’s injection port. The IPD eliminates the incubation step required in regular derivatization procedures and leads the analyst exposure to toxic reagents to a minimum.

Method

We present a fully validated IPD–GC–MS/MS method to quantify cocaine, ecgonine methyl ester, benzoylecgonine and cocaethylene in urine, using low-sample volume and liquid–liquid extraction (LLE). After sample extraction, cocaine metabolites were derivatized with N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) plus 1% trimethylchlorosilane (TCMS) directly at the injection port of the GC system.

Results

Calibration curves were linear over 50–2000 ng/mL, with intra-day and inter-day imprecision values (% RSD) and the analyte concentrations of bias (%) less than 8.8%. Extraction efficiencies greater than 50% were observed for all analytes. All analytes were stable at autosampler conditions (room temperature) for at least 24 h. The method results showed that the developed method has analytical performance enough to be used as confirmation procedure in cocaine exposure verification.

Conclusions

To the best of our knowledge, this is the first method to use IPD-GC–MS/MS and silylation reaction in forensic toxicology analysis. The method showed great sensitivity, imprecision and accuracy, using simple and less expensive LLE (compared to solid-phase extraction), with low sample and solvent volumes.

The United Nations Office on Drugs and Crime - UNODC (2019) World drug report 2019. https://www.unodc.org/wdr2019/. Accessed 14 Apr 2020

Smith ML, Shimomura E, Paul BD, Cone EJ, Darwin WD, Huestis MA (2010) Urinary excretion of ecgonine and five other cocaine metabolites following controlled oral, intravenous, intranasal, and smoked administration of cocaine. J Anal Toxicol 34:57–63. https://doi.org/10.1093/jat/34.2.57CrossRefPubMedPubMedCentral

Huestis MA, Darwin WD, Shimomura E, Lalani SA, Trinidad DV, Jenkins AJ, Cone EJ, Jacobs AJ, Smith ML, Paul BD (2007) Cocaine and metabolites urinary excretion after controlled smoked administration. J Anal Toxicol 31:462–468. https://doi.org/10.1093/jat/31.8.462CrossRefPubMedPubMedCentral

Skopp G, Klingmann A, Pötsch L, Mattern R (2001) In vitro stability of cocaine in whole blood and plasma including ecgonine as a target analyte. Ther Drug Monit 23:174–181. https://doi.org/10.1097/00007691-200104000-00014CrossRefPubMed

da Costa JL, Tonin FG, Zanolli LA, Chasin AA, Tavares MF (2009) Simple method for determination of cocaine and main metabolites in urine by CE coupled to MS. Electrophoresis 30:2238–2244. https://doi.org/10.1002/elps.200900032CrossRefPubMed

Cone EJ, Sampson-Cone AH, Darwin WD, Huestis MA, Oyler JM (2003) Urine testing for cocaine abuse: metabolic and excretion patterns following different routes of administration and methods for detection of false-negative results. J Anal Toxicol 27:386–401. https://doi.org/10.1093/jat/27.7.386CrossRefPubMed

Taskinen S, Beck O, Bosch T, Brcak M, Carmichael D, Fucci N, George C, Piper M, Salomone A, Schielen W, Steinmeyer S, Weinmann W (2017) European guidelines for workplace drug testing in urine. Drug Test Anal 9:853–865. https://doi.org/10.1002/dta.2178CrossRefPubMed

Drummer OH, Kourtis I, Beyer J, Tayler P, Boorman M, Gerostamoulos D (2012) The prevalence of drugs in injured drivers. Forensic Sci Int 215:14–17. https://doi.org/10.1016/j.forsciint.2011.01.040CrossRefPubMed

Penning R, Veldstra JL, Daamen AP, Olivier B, Verster JC (2010) Drugs of abuse, driving and traffic safety. Curr Drug Abuse Rev 3:23–32. https://doi.org/10.2174/1874473711003010023CrossRefPubMed

10.

Paul BD, Lalani S, Bosy T, Jacobs AJ, Huestis MA (2005) Concentration profiles of cocaine, pyrolytic methyl ecgonidine and thirteen metabolites in human blood and urine: determination by gas chromatorgraphy-mass spectrometry. Biomed Chromatogr 19:677–688. https://doi.org/10.1002/bmc.495CrossRefPubMed

11.

Wang Q, Ma L, Yin CR, Xu L (2013) Developments in injection port derivatization. J Chromatogr A 1296:25–35. https://doi.org/10.1016/j.chroma.2013.04.036CrossRefPubMed

12.

Marsol-Vall A, Balcells M, Eras J, Canela-Garayoa R (2017) Dispersive liquid–liquid microextraction and injection-port derivatization for the determination of free lipophilic compounds in fruit juices by gas chromatography-mass spectrometry. J Chromatogr A 1495:12–21. https://doi.org/10.1016/j.chroma.2017.03.027CrossRefPubMed

13.

Tzing SH, Ding WH (2010) Determination of melamine and cyanuric acid in powdered milk using injection-port derivatization and gas chromatography-tandem mass spectrometry with furan chemical ionization. J Chromatogr A 1217:6267–6273. https://doi.org/10.1016/j.chroma.2010.07.081CrossRefPubMed

14.

Mandrah K, Satyanarayana GNV, Roy SK (2017) A dispersive liquid-liquid microextraction based on solidification of floating organic droplet followed by injector port silylation coupled with gas chromatography–tandem mass spectrometry for the determination of nine bisphenols in bottled carbonated bev. J Chromatogr A 1528:10–17. https://doi.org/10.1016/j.chroma.2017.10.071CrossRefPubMed

15.

Makoś P, Fernandes A, Boczkaj G (2017) Method for the determination of carboxylic acids in industrial effluents using dispersive liquid-liquid microextraction with injection port derivatization gas chromatography–mass spectrometry. J Chromatogr A 1517:26–34. https://doi.org/10.1016/j.chroma.2017.08.045CrossRefPubMed

16.

González A, Clavijo S, Cerdà V (2019) Estrogens determination exploiting a SIA-LOV system prior in-port derivatization-large volume injection-programmable temperature vaporization-gas chromatography. Talanta 194:852–858. https://doi.org/10.1016/J.TALANTA.2018.10.105CrossRefPubMed

17.

Oliveira AFF, de Figueiredo EC, dos Santos-Neto ÁJ (2013) Analysis of fluoxetine and norfluoxetine in human plasma by liquid-phase microextraction and injection port derivatization GC-MS. J Pharm Biomed Anal 73:53–58. https://doi.org/10.1016/j.jpba.2012.04.006CrossRefPubMed

18.

Scientific Working Group for Forensic Toxicology - SWGTOX (2013) Standard practices for method validation in forensic toxicology. J Anal Toxicol 37:452–474. https://doi.org/10.1093/jat/bkt054CrossRef

19.

Wu J, Hu R, Yue J, Yang Z, Zhang L (2009) Determination of fecal sterols by gas chromatography-mass spectrometry with solid-phase extraction and injection-port derivatization. J Chromatogr A 1216:1053–1058. https://doi.org/10.1016/j.chroma.2008.12.054CrossRefPubMed

20.

Miki A, Katagi M, Zaitsu K, Nishioka H, Tsuchihashi H (2008) Development of a two-step injector for GC–MS with on-column derivatization, and its application to the determination of amphetamine-type stimulants (ATS) in biological specimens. J Chromatogr B 865:25–32. https://doi.org/10.1016/j.jchromb.2008.01.051CrossRef

21.

Namera A, Yashiki M, Liu J, Liu J, Okajima K, Hara K, Imamura T, Kojima T (2000) Simple and simultaneous analysis of fenfluramine, amphetamine and methamphetamine in whole blood by gas chromatography–mass spectrometry after headspace–solid phase microextraction and derivatization. Forensic Sci Int 109:215–223. https://doi.org/10.1016/S0379-0738(00)00145-6CrossRefPubMed

22.

Jain NC, Chinn DM, Budd RD, Sneath TS, Leung WJ (1977) Simultaneous determination of cocaine and benzoyl ecgonine in urine by gas chromatography with on-column alkylation. J Forensic Sci 22:7–16CrossRef

23.

Rafla FK, Epstein RL (1979) Identification of cocaine and its metabolites in human urine in the presence of ethyl alcohol. J Anal Toxicol 3:59–63. https://doi.org/10.1093/jat/3.2.59CrossRef

24.

Yonamine M, Silva OA (2002) Confirmation of cocaine exposure by gas chromatography-mass spectrometry of urine extracts after methylation of benzoylecgonine. J Chromatogr B Anal Technol Biomed Life Sci 773:83–87. https://doi.org/10.1016/S0378-4347(01)00516-3CrossRef

25.

Woźniak MK, Banaszkiewicz L, Wiergowski M, Tomczak E, Kata M, Szpiech B, Namieśnik J, Biziuk M (2020) Development and validation of a GC–MS/MS method for the determination of 11 amphetamines and 34 synthetic cathinones in whole blood. Forensic Toxicol 38:42–58. https://doi.org/10.1007/s11419-019-00485-yCrossRef

26.

The United Nations Office on Drugs and Crime - UNODC (2011) Guidelines for the forensic analysis of drugs facilitating sexual assault and other criminal acts. https://www.unodc.org/documents/scientific/forensic_analys_of_drugs_facilitating_sexual_assault_and_other_criminal_acts.pdf. Accessed 14 Apr 2020

Title: Use of injection-port derivatization for the analysis of cocaine and its metabolites in urine by gas chromatography–tandem mass spectrometry
Authors: Kelly Francisco da Cunha
Rafael Lanaro
Aline Franco Martins
Karina Diniz Oliveira
Jose Luiz Costa
Publication date: 01-01-2021
Publisher: Springer Singapore
Published in: Forensic Toxicology / Issue 1/2021
Print ISSN: 1860-8965
Electronic ISSN: 1860-8973
DOI: https://doi.org/10.1007/s11419-020-00545-8

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Use of injection-port derivatization for the analysis of cocaine and its metabolites in urine by gas chromatography–tandem mass spectrometry

Abstract

Purpose

Method

Results

Conclusions

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Purpose

Method

Results

Conclusions

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