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Published in: Forensic Toxicology 1/2020

01-01-2020 | Original Article

Pharmacokinetic study of xylazine in a zebrafish water tank, a human-like surrogate, by liquid chromatography Q-Orbitrap mass spectrometry

Authors: Rebecca Rodrigues Matos, Maria Elvira Poleti Martucci, Carina Souza de Anselmo, Francisco Radler Alquino Neto, Henrique Marcelo Gualberto Pereira, Vinícius Figueiredo Sardela

Published in: Forensic Toxicology | Issue 1/2020

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Abstract

Purpose

This study aims to investigate a zebrafish (Danio Rerio) water tank (ZWT) as an alternative model for the study of the metabolism of xylazine.

Methods

The ZWT approach for the study of metabolism consisted of two aquariums, where 18 fish and xylazine were added into the first tank. The second one, with 18 fish without drug, served as a negative control. The samples were submitted to a comprehensive analytical method developed for doping control purposes by liquid chromatography (LC) coupled with high-resolution mass spectrometry (HRMS) operating in five different acquisition modes. Glycoconjugate metabolites were evaluated indirectly by extracting the samples with and without the enzymatic hydrolysis step using β-glucuronidase.

Results

In total, 11 phase I and II metabolites were detected and characterized, of which four were previously described for humans and two for horses, and five metabolites were described for the first time. The main metabolites were 4-hydroxylated (M2) and oxygenated (M1) derivatives. Both metabolites were suggested as analytical targets for xylazine misuse. Around 79% of para- and meta-hydroxylated derivatives were in glycoconjugate form, whereas for oxo-hydroxylated and sulfone-hydroxylated derivatives of xylazine, around 83% and 70% were metabolized to the glycoconjugate form, respectively.

Conclusions

Xylazine was the subject of extensive metabolism in zebrafish. 4-Hydroxylated (M2) and oxygenated (M1) derivatives were the most abundant phase I metabolites as the main targets for doping control.
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Literature
5.
go back to reference Miksa IR, Cummings MR, Poppenga RH (2005) Determination of acepromazine, ketamine, medetomidine, and xylazine in serum: multi-residue screening by liquid chromatography-mass spectrometry. J Anal Toxicol 29(6):544–551 (PMID: 16168177) CrossRef Miksa IR, Cummings MR, Poppenga RH (2005) Determination of acepromazine, ketamine, medetomidine, and xylazine in serum: multi-residue screening by liquid chromatography-mass spectrometry. J Anal Toxicol 29(6):544–551 (PMID: 16168177) CrossRef
7.
go back to reference Se Hun K, Hyun Woo K, Chun S, Jinwoong N, Se Woong P, Min Jae K, Choong HL, Wan In L, Jae Pil K (2011) Effect of five-membered heteroaromatic linkers to the performance of phenothiazine-based dye-sensitized solar cells. Org Lett 13(21):5784–5787. https://doi.org/10.1021/ol2023517 CrossRef Se Hun K, Hyun Woo K, Chun S, Jinwoong N, Se Woong P, Min Jae K, Choong HL, Wan In L, Jae Pil K (2011) Effect of five-membered heteroaromatic linkers to the performance of phenothiazine-based dye-sensitized solar cells. Org Lett 13(21):5784–5787. https://​doi.​org/​10.​1021/​ol2023517 CrossRef
8.
go back to reference Carpy A, Leger JM, Leclerc G, Decker N, Rouot B, Wermuth CG (1982) Comparison of crystallographic and quantum mechanical analysiswith biological data on clonidine and some related analogues. Mol Pharmacol 21(2):400–408PubMed Carpy A, Leger JM, Leclerc G, Decker N, Rouot B, Wermuth CG (1982) Comparison of crystallographic and quantum mechanical analysiswith biological data on clonidine and some related analogues. Mol Pharmacol 21(2):400–408PubMed
10.
go back to reference Hoffmann U, Meister CM, Golle K, Zschiesche M (2001) Severe intoxication with the veterinary tranquilizer xylazine in humans. J Anal Toxicol 25(4):245–249CrossRef Hoffmann U, Meister CM, Golle K, Zschiesche M (2001) Severe intoxication with the veterinary tranquilizer xylazine in humans. J Anal Toxicol 25(4):245–249CrossRef
11.
go back to reference Gao X, Guo H, Du Y, Gu C (2015) Simultaneous determination of xylazine and 2,6-xylidine in blood and urine by auto solid-phase extraction and ultra high performance liquid chromatography coupled with quadrupole-time of flight mass spectrometry. J Anal Toxicol 39(6):444–450. https://doi.org/10.1093/jat/bkv040 CrossRefPubMed Gao X, Guo H, Du Y, Gu C (2015) Simultaneous determination of xylazine and 2,6-xylidine in blood and urine by auto solid-phase extraction and ultra high performance liquid chromatography coupled with quadrupole-time of flight mass spectrometry. J Anal Toxicol 39(6):444–450. https://​doi.​org/​10.​1093/​jat/​bkv040 CrossRefPubMed
18.
go back to reference Mutlib AE, Chui YC, Young LM, Abbott FS (1992) Characterization of metabolites of xylazine produced in vivo and in vitro by LC/MS/MS and by GC/MS. Drug Metabol Dispos 20(6):840–848 Mutlib AE, Chui YC, Young LM, Abbott FS (1992) Characterization of metabolites of xylazine produced in vivo and in vitro by LC/MS/MS and by GC/MS. Drug Metabol Dispos 20(6):840–848
21.
go back to reference Parker RJ, Collins JM, Strong JM (1996) Identification of 2,6-xylidine as a major lidocaine metabolite in human liver slices. Drug Metabol Dispos 24(11):1167–1173 Parker RJ, Collins JM, Strong JM (1996) Identification of 2,6-xylidine as a major lidocaine metabolite in human liver slices. Drug Metabol Dispos 24(11):1167–1173
23.
go back to reference Sardela VF, Anselmo CDS, Nunes IKC, Carneiro GRA, dos Santos GRC, de Carvalho AR, LaBanca BJ, Silva D, Ribeiro WR, de Araújo ALD, Padilha MC, de Lima CKF, de Souza VP, Aquino Neto FR, Pereira HMG (2018) Zebrafish (Danio rerio) water tank model for the investigation of drug metabolism: progress, outlook, and challenges. Drug Test Anal. https://doi.org/10.1002/dta.2523 CrossRefPubMed Sardela VF, Anselmo CDS, Nunes IKC, Carneiro GRA, dos Santos GRC, de Carvalho AR, LaBanca BJ, Silva D, Ribeiro WR, de Araújo ALD, Padilha MC, de Lima CKF, de Souza VP, Aquino Neto FR, Pereira HMG (2018) Zebrafish (Danio rerio) water tank model for the investigation of drug metabolism: progress, outlook, and challenges. Drug Test Anal. https://​doi.​org/​10.​1002/​dta.​2523 CrossRefPubMed
24.
go back to reference Howe K, Clark MD, Torroja CF, Torrance J, Berthelot C, Muffato M, Collins JE, Humphray S, McLaren K, Matthews L, McLaren S, Sealy I, Caccamo M, Churcher C, Scott C, Barrett JC, Koch R, Rauch GJ, White S, Chow W, Kilian B, Quintais LT, Guerra-Assunção JA, Zhou Y, Gu Y, Yen J, Vogel JH, Eyre T, Redmond S, Banerjee R, Chi J, Fu B, Langley E, Maguire SF, Laird GK, Lloyd D, Kenyon E, Donaldson S, Sehra H, Almeida-King J, Loveland J, Trevanion S, Jones M, Quail M, Willey D, Hunt A, Burton J, Sims S, McLay K, Plumb B, Davis J, Clee C, Oliver K, Clark R, Riddle C, Eliott D, Threadgold G, Harden G, Ware D, Mortimer B, Kerry G, Heath P, Phillimore B, Tracey A, Corby N, Dunn M, Johnson C, Wood J, Clark S, Pelan S, Griffiths G, Smith M, Glithero R, Howden P, Barker N, Stevens C, Harley J, Holt K, Panagiotidis G, Lovell J, Beasley H, Henderson C, Gordon D, Auger K, Wright D, Collins J, Raisen C, Dyer L, Leung K, Robertson L, Ambridge K, Leongamornlert D, McGuire S, Gilderthorp R, Griffiths C, Manthravadi D, Nichol S, Barker G, Whitehead S, Kay M, Brown J, Murnane C, Gray E, Humphries M, Sycamore N, Barker D, Saunders D, Wallis J, Babbage A, Hammond S, Mashreghi-Mohammadi M, Barr L, Martin S, Wray P, Ellington A, Matthews N, Ellwood M, Woodmansey R, Clark G, Cooper J, Tromans A, Grafham D, Skuce C, Pandian R, Andrews R, Harrison E, Kimberley A, Garnett J, Fosker N, Hall R, Garner P, Kelly D, Bird C, Palmer S, Gehring I, Berger A, Dooley CM, Ersan-Ürün Z, Eser C, Geiger H, Geisler M, Karotki L, Kirn A, Konantz J, Konantz M, Oberländer M, Rudolph-Geiger S, Teucke M, Osoegawa K, Zhu B, Rapp A, Widaa S, Langford C, Yang F, Carter NP, Harrow J, Ning Z, Herrero J, Searle SMJ, Enright A, Geisler R, Plasterk RHA, Lee C, Westerfield M, De Jong PJ, Zon LI, Postlethwait JH, NüssleinVolhard C, Hubbard TJP, Crollius HR, Rogers J, Stemple DL (2013) The zebrafish reference genome sequence and its relationship to the human genome. Nature 496:498–503. https://doi.org/10.1038/nature12111 CrossRefPubMedPubMedCentral Howe K, Clark MD, Torroja CF, Torrance J, Berthelot C, Muffato M, Collins JE, Humphray S, McLaren K, Matthews L, McLaren S, Sealy I, Caccamo M, Churcher C, Scott C, Barrett JC, Koch R, Rauch GJ, White S, Chow W, Kilian B, Quintais LT, Guerra-Assunção JA, Zhou Y, Gu Y, Yen J, Vogel JH, Eyre T, Redmond S, Banerjee R, Chi J, Fu B, Langley E, Maguire SF, Laird GK, Lloyd D, Kenyon E, Donaldson S, Sehra H, Almeida-King J, Loveland J, Trevanion S, Jones M, Quail M, Willey D, Hunt A, Burton J, Sims S, McLay K, Plumb B, Davis J, Clee C, Oliver K, Clark R, Riddle C, Eliott D, Threadgold G, Harden G, Ware D, Mortimer B, Kerry G, Heath P, Phillimore B, Tracey A, Corby N, Dunn M, Johnson C, Wood J, Clark S, Pelan S, Griffiths G, Smith M, Glithero R, Howden P, Barker N, Stevens C, Harley J, Holt K, Panagiotidis G, Lovell J, Beasley H, Henderson C, Gordon D, Auger K, Wright D, Collins J, Raisen C, Dyer L, Leung K, Robertson L, Ambridge K, Leongamornlert D, McGuire S, Gilderthorp R, Griffiths C, Manthravadi D, Nichol S, Barker G, Whitehead S, Kay M, Brown J, Murnane C, Gray E, Humphries M, Sycamore N, Barker D, Saunders D, Wallis J, Babbage A, Hammond S, Mashreghi-Mohammadi M, Barr L, Martin S, Wray P, Ellington A, Matthews N, Ellwood M, Woodmansey R, Clark G, Cooper J, Tromans A, Grafham D, Skuce C, Pandian R, Andrews R, Harrison E, Kimberley A, Garnett J, Fosker N, Hall R, Garner P, Kelly D, Bird C, Palmer S, Gehring I, Berger A, Dooley CM, Ersan-Ürün Z, Eser C, Geiger H, Geisler M, Karotki L, Kirn A, Konantz J, Konantz M, Oberländer M, Rudolph-Geiger S, Teucke M, Osoegawa K, Zhu B, Rapp A, Widaa S, Langford C, Yang F, Carter NP, Harrow J, Ning Z, Herrero J, Searle SMJ, Enright A, Geisler R, Plasterk RHA, Lee C, Westerfield M, De Jong PJ, Zon LI, Postlethwait JH, NüssleinVolhard C, Hubbard TJP, Crollius HR, Rogers J, Stemple DL (2013) The zebrafish reference genome sequence and its relationship to the human genome. Nature 496:498–503. https://​doi.​org/​10.​1038/​nature12111 CrossRefPubMedPubMedCentral
34.
go back to reference Grigoryev A, Savchuk S, Melnik A, Moskaleva N, Dzhurko J, Ershov M, Nosyrev A, Vedenin A, Izotov B, Zabirova I, Rozhanets V (2001) Chromatography mass spectrometry studies on the metabolism of synthetic cannabinoids JWH-018 and JWH-073, psychoactive components of smoking mixtures. J Chromatogr B 879:1126–1136. https://doi.org/10.1016/j.jchromb.2011.03.034 CrossRef Grigoryev A, Savchuk S, Melnik A, Moskaleva N, Dzhurko J, Ershov M, Nosyrev A, Vedenin A, Izotov B, Zabirova I, Rozhanets V (2001) Chromatography mass spectrometry studies on the metabolism of synthetic cannabinoids JWH-018 and JWH-073, psychoactive components of smoking mixtures. J Chromatogr B 879:1126–1136. https://​doi.​org/​10.​1016/​j.​jchromb.​2011.​03.​034 CrossRef
35.
go back to reference Sardela VF, Martucci MEP, de Araújo ALD, Leal ES, Oliveira DS, Carneiro GR, Deventer K, Van Eenoo P, Pereira HMG, Aquino Neto FR (2018) Comprehensive analysis by liquid chromatography-Q-Orbitrap mass spectrometry: fast screening of peptides and organic molecules. J Mass Spectrom 53(6):476–503. https://doi.org/10.1002/jms.4077 CrossRefPubMed Sardela VF, Martucci MEP, de Araújo ALD, Leal ES, Oliveira DS, Carneiro GR, Deventer K, Van Eenoo P, Pereira HMG, Aquino Neto FR (2018) Comprehensive analysis by liquid chromatography-Q-Orbitrap mass spectrometry: fast screening of peptides and organic molecules. J Mass Spectrom 53(6):476–503. https://​doi.​org/​10.​1002/​jms.​4077 CrossRefPubMed
37.
go back to reference Testa D, Krämer SD (2007) The biochemistry of drug metabolism—an introduction part 2. Redox reactions and their enzymes. Chem Biodiv 4:257–405CrossRef Testa D, Krämer SD (2007) The biochemistry of drug metabolism—an introduction part 2. Redox reactions and their enzymes. Chem Biodiv 4:257–405CrossRef
Metadata
Title
Pharmacokinetic study of xylazine in a zebrafish water tank, a human-like surrogate, by liquid chromatography Q-Orbitrap mass spectrometry
Authors
Rebecca Rodrigues Matos
Maria Elvira Poleti Martucci
Carina Souza de Anselmo
Francisco Radler Alquino Neto
Henrique Marcelo Gualberto Pereira
Vinícius Figueiredo Sardela
Publication date
01-01-2020
Publisher
Springer Singapore
Published in
Forensic Toxicology / Issue 1/2020
Print ISSN: 1860-8965
Electronic ISSN: 1860-8973
DOI
https://doi.org/10.1007/s11419-019-00493-y

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