Optimisation of a gas chromatography-mass spectrometry method for the simultaneous determination of tetrahydrocannabinol and its metabolites in rat urine

Fuchs, Nino; Miljanić, Alena; Katić, Anja; Brajenović, Nataša; Micek, Vedran; Fuchs, Radovan; Karačonji, Irena Brčić

doi:10.2478/aiht-2019-70-3352

Abstract

In order to evaluate the effect of irinotecan (IRI) on urinary elimination of delta-9-tetrahydrocannabinol (THC) in a rat experimental model, we developed an analytical method for the determination of the mass concentration of THC and its metabolites [11-hydroxy-delta-9-tetrahydrocannabinol (THC-OH) and 11-nor-9-carboxy-delta-9-tetrahydrocannabinol (THC-COOH)] in the urine of rats treated only with THC and treated simultaneously with THC and irinotecan. For this purpose, hydrolysis and solid phase extraction conditions of the investigated analytes were optimised and a gas chromatography-mass spectrometry (GC-MS) method was developed to determine all three analytes in rat urine. The most effective hydrolysis method for THC, THC-OH, and THC-COOH conjugates was so-called tandem hydrolysis by the β-glucuronidase enzyme from Escherichia coli at 50 °C for 2 hours and followed by alkaline hydrolysis. The proposed method was then applied for determining concentrations of analytes in 24-hour rat urine. THC was not detected in either sample, THC-OH was detected in 50 % of samples, and THC-COOH in all of the samples. Enhanced urinary THC-COOH excretion was noted in rats administered combined treatment compared to single THC treatment. The method described herein was suitable for determining the mass concentration of THC metabolites in the rat urine due to its sensitivity (detection limits: 0.8-1.0 μg/L), accuracy (>96 %), and precision (RSD <6 %).

References

1. MacCallum C, Russo E. Practical considerations in medical cannabis administration and dosing. Eur J Intern Med 2018;49:12-9. doi: 10.1016/j.ejim.2018.01.00410.1016/j.ejim.2018.01.004
Open DOI Search in Google Scholar Back to article
2. Lucić Vrdoljak A, Fuchs N, Mikolić A, Žunec S, Brčić Karačonji I, Jurič A, Prester L, Micek V, Neuberg M, Čanović S, Mršić G, Kopjar N. Irinotecan and ∆⁹-tetrahydrocannabinol interactions in rat liver: a preliminary evaluation using biochemical and genotoxicity markers. Molecules 2018;23:1332. doi: 10.3390/molecules2306133210.3390/23061332
Open DOI Search in Google Scholar Back to article
3. Atakan Z. Cannabis, a complex plant: Different compounds and different effects on individuals. Ther Adv Psychopharmacol 2012;2:241-54. doi: 10.1177/204512531245758610.1177/2045125312457586
Open DOI Search in Google Scholar Back to article
4. Lowe RH, Abraham TT, Darwin WD, Herning R, Cadet JL, Huestis MA. Extended urinary Δ9-tetrahydrocannabinol excretion in chronic cannabis users precludes use as a biomarker of new drug exposure. Drug Alcohol Depend 2009;105:24-32. doi: 10.1016/j.drugalcdep.2009.05.02710.1016/j.drugalcdep.2009.05.027
Open DOI Search in Google Scholar Back to article
5. Đorđević S, Krstić N, Kilibarda V, Šulić K, Jovanović B. Prikaz metoda za dokazivanje i određivanje kanabinoida u biološkom materijalu [A review of methods for determination of cannabinoids in biological samples, in Serbian]. Med Rev 2011;3:063-8.
Search in Google Scholar Back to article
6. Musshoff F, Madea B. Review of biologic matrices (urine, blood, hair) as indicators of recent or ongoing cannabis use. Ther Drug Monit 2006;28:155-63. doi: 10.1097/01.ftd.0000197091.07807.2210.1097/01.ftd.0000197091.07807.22
Open DOI Search in Google Scholar Back to article
7. Grotenhermen F. Clinical pharmacokinetics of cannabinoids. J Cannabis Ther 2003;3:3-51. doi: 10.1300/J175v03n01_0210.1300/J175v03n01_02
Open DOI Search in Google Scholar Back to article
8. Abraham TT, Lowe RH, Pirnay SO, Darwin WD, Huestis MA. Simultaneous GC–EI-MS determination of Δ9-tetrahydrocannabinol, 11-hydroxy-Δ9-tetrahydrocannabinol, and 11-nor-9-carboxy-Δ9-tetrahydrocannabinol in human urine following tandem enzyme-alkaline hydrolysis. J Anal Toxicol 2007;31:477-85. doi: 10.1093/jat/31.8.47710.1093/jat/31.8.477
Open DOI Search in Google Scholar Back to article
9. Brenneisen R, Meyer P, Chtioui H, Saugy M, Kamber M. Plasma and urine profiles of Δ⁹-tetrahydrocannabinol and its metabolites 11-hydroxy-Δ⁹-tetrahydrocannabinol and 11-nor-9-carboxy-Δ⁹-tetrahydrocannabinol after cannabis smoking by male volunteers to estimate recent consumption by athletes. Anal Bioanal Chem 2010;396:2493-502. doi: 10.1007/s00216-009-3431-310.1007/s00216-009-3431-3
Open DOI Search in Google Scholar Back to article
10. Weinmann W, Vogt S, Goerke R, Muller C, Bromberger A. Simultaneous determination of THC-COOH and THC-COOH-glucuronide in urine samples by LC/MS/MS. Forensic Sci Int 2000;113:381-7. doi: 10.1016/S0379-0738(00)00210-310.1016/S0379-0738(00)00210-3
Search in Google Scholar Back to article
11. Felli M, Martello S, Chiarotti M. LC-MS-MS method for simultaneous determination of THCCOOH and THCCOOH-glucuronide in urine: Application to workplace confirmation tests. Forensic Sci Int 2011;204:67-73. doi: 10.1016/j.forsciint.2010.05.00410.1016/j.forsciint.2010.05.00420627630
Open DOI Search in Google Scholar Back to article
12. Schaefer N, Kettner M, Laschke MW, Schlote J, Peters B, Bregel D, Menger MD, Maurer HH, Ewald AH, Schmidt PH. Simultaneous LC-MS/MS determination of JWH-210, RCS-4, ∆(9)-tetrahydrocannabinol, and their main metabolites in pig and human serum, whole blood, and urine for comparing pharmacokinetic data. Anal Bioanal Chem 2015;407:3775-86. doi: 10.1007/s00216-015-8605-610.1007/s00216-015-8605-625772567
Open DOI Search in Google Scholar Back to article
13. Teixeira H, Verstraete A, Proença P, Corte-Real F, Monsanto P, Vieira DN. Validated method for the simultaneous determination of Δ⁹-THC and Δ⁹-THC-COOH in oral fluid, urine and whole blood using solid-phase extraction and liquid chromatography-mass spectrometry with electrospray ionization. Forensic Sci Int 2007;170:148-55. doi: 10.1016/j.forsciint.2007.03.02610.1016/j.forsciint.2007.03.02617614225
Open DOI Search in Google Scholar Back to article
14. Battista N, Sergi M, Montesano C, Napoletano S, Compagnone D, Maccarrone M. Analytical approaches for the determination of phytocannabinoids and endocannabinoids in human matrices. Drug Test Anal 2014;6:7-16. doi: 10.1002/dta.157410.1002/dta.157424218186
Open DOI Search in Google Scholar Back to article
15. Kemp PM, Abukhalaf IK, Manno JE, Manno BR, Alford DD, McWilliams ME, Nixon FE, Fitzgerald MJ, Reeves RR, Wood MJ. Cannabinoids in humans. II. The influence of three methods of hydrolysis on the conentration of THC and two metabolites in urine. J Anal Toxicol 1995;19:292-8. doi: 10.1093/jat/19.5.29210.1093/jat/19.5.2927500615
Search in Google Scholar Back to article
16. Bansal T, Mishra G, Jaggi M, Khar RK, Talegaonkar S. Effect of P-glycoprotein inhibitor, verapamil, on oral bioavailability and pharmacokinetics of irinotecan in rats. Eur J Pharm Sci 2009;36:580–90. doi:10.1016/j.ejps.2008.12.005.10.1016/j.ejps.2008.12.00519135530
Open DOI Search in Google Scholar Back to article
17. Böhmdorfer M, Maier-Salamon A, Riha J, Brenner S, Höferl M, Jäger W. Interplay of drug metabolizing enzymes with cellular transporters. Wien Med Wochenschr 2014;164:461-71. doi: 10.1007/s10354-014-0301-110.1007/s10354-014-0301-125271108
Open DOI Search in Google Scholar Back to article
18. Horikawa M, Kato Y, Sugiyama Y. Reduced gastrointestinal toxicity following inhibition of the biliary excretion of irinotecan and its metabolites by probenecid in rats. Pharm Res 2002;19:1345-53. doi: 10.1023/A:102035891049010.1023/A:1020358910490
Open DOI Search in Google Scholar Back to article
19. Li C, Li X, Choi J. Enhanced bioavailability of etoposide after oral or intravenous administration of etoposide with kaempferol in rats. Arch Pharm Res 2009;32:133-8. doi: 10.1007/s12272-009-1127-z10.1007/s12272-009-1127-z19183886
Search in Google Scholar Back to article
20. Yamamoto M, Kurita A, Asahara T, Takakura A, Katono K, Iwasaki M, Ryuge S, Wada M, Onoda S, Yanaihara T, Yokoba M, Mitsufuji H, Nishii Y, Fukui T, Masuda N. Metabolism of irinotecan and its active metabolite SN-38 by intestinal microflora in rats. Oncol Rep 2008;20:727-30. doi: 10.3892/or_0000006610.3892/or_00000066
Open DOI Search in Google Scholar Back to article
21. Karačić V, Skender Lj. Analysis of drugs of abuse in urine by gas chromatography/mass spectrometry: Experience and application. Arh Hig Rada Toksikol 2000;51:389-400. PMID: 11276966
Search in Google Scholar Back to article
22. Jung J, Meyer MR, Maurer HH, Neusüß C, Weinmann W, Auwärter V. Studies on the metabolism of the Δ9-tetrahydrocannabinol precursor Δ9-tetrahydrocannabinolic acid A (Δ9-THCA-A) in rat using LC-MS/MS, LC-QTOFMS and GC-MS techniques. J Mass Spectrom 2007;44:1423-33. doi: 10.1002/jms.162410.1002/jms.162419728318
Open DOI Search in Google Scholar Back to article
23. Martignoni M, Groothuis GMM, de Kanter R. Species differences between mouse, rat, dog, monkey and human CYP-mediated drug metabolism, inhibition and induction. Expert Opin Drug Metab Toxicol 2006;2:875-94. doi: 10.1517/17425255.2.6.87510.1517/17425255.2.6.87517125407
Open DOI Search in Google Scholar Back to article
24. Coskun ZM, Bolkent S. Evaluation of Δ⁹-tetrahydrocannabinol metabolites and oxidative stress in type 2 diabetic rats. Iran J Basic Med Sci 2016;19:154-8. PMCID: PMC4818362
Search in Google Scholar Back to article
25. Santos A, Zanetta S, Cresteil T, Deroussent A, Pein F, Raymond E, Vernillet L, Risse ML, Boige V, Gouyette A, Vassal G. Metabolism of irinotecan (CPT-11) by CYP3A4 and CYP3A5 in humans. Clin Cancer Res 2000;6:2012-20. PMID: 10815927
Search in Google Scholar Back to article
26. Mazur A, Lichti CF, Prather PL, Zielinska AK, Bratton SM, Gallus-Zawada A, Finel M, Miller GP, Radomińska-Pandya A, Moran JH. Characterization of human hepatic and extrahepatic UDP-glucuronosyltransferase enzymes involved in the metabolism of classic cannabinoids. Drug Metab Dispos 2009;37:1496-504. doi: 10.1124/dmd.109.02689810.1124/dmd.109.026898269894319339377
Open DOI Search in Google Scholar Back to article

Optimisation of a gas chromatography-mass spectrometry method for the simultaneous determination of tetrahydrocannabinol and its metabolites in rat urine

Abstract

Paradigm

My account