Skip to main content
Top
Published in: Forensic Toxicology 1/2017

01-01-2017 | Short Communication

Simple protein precipitation-based analysis of Δ9-tetrahydrocannabinol and its metabolites in human serum by liquid chromatography–tandem mass spectrometry

Authors: Marek Dziadosz, Michael Klintschar, Jörg Teske

Published in: Forensic Toxicology | Issue 1/2017

Login to get access

Abstract

A liquid chromatography–tandem mass spectrometry (LC–MS/MS) quantification method based on a direct protein precipitation with the mobile phase was developed, validated and applied for the analysis of Δ9-tetrahydrocannabinol (THC), 11-hydroxy-Δ9-tetrahydrocannabinol (11-OH-THC) and 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THC-COOH) in human serum for forensic purposes. Chromatographic separation was performed with a Luna 5 µm C18 (2) 100 A, 150 × 2 mm analytical column and the elution with a programmed flow of a mobile phase consisting of A (H2O/methanol, 95:5, v/v) and B (H2O/methanol, 3:97, v/v), both with 10 mM ammonium acetate and 0.1 % acetic acid. The results of the validation experiments demonstrated that the strategy presented can be applied successfully for a fast, simple and cost effective LC–MS/MS analysis of THC and its metabolites. A potential conversion of THC-COOH glucuronide to THC-COOH during sample preparation/analysis could be excluded.
Literature
1.
go back to reference Baselt RC (2011) Disposition of toxic drugs and chemicals in man. Biomedical Publications, Seal Beach Baselt RC (2011) Disposition of toxic drugs and chemicals in man. Biomedical Publications, Seal Beach
2.
go back to reference Dziadosz M, Weller JP, Klintschar M, Teske J (2013) Scheduled multiple reaction monitoring algorithm as a way to analyse new designer drugs combined with synthetic cannabinoids in human serum with liquid chromatography–tandem mass spectrometry. J Chromatogr B 929:84–89CrossRef Dziadosz M, Weller JP, Klintschar M, Teske J (2013) Scheduled multiple reaction monitoring algorithm as a way to analyse new designer drugs combined with synthetic cannabinoids in human serum with liquid chromatography–tandem mass spectrometry. J Chromatogr B 929:84–89CrossRef
3.
go back to reference Dziadosz M, Klintschar M, Teske J (2014) Mixed consumption of cannabis and “Spice”. Forensic Sci Int 235:e1–e2CrossRefPubMed Dziadosz M, Klintschar M, Teske J (2014) Mixed consumption of cannabis and “Spice”. Forensic Sci Int 235:e1–e2CrossRefPubMed
4.
go back to reference Gustafson RA, Moolchan ET, Barnes A, Levine B, Huestis MA (2003) Validated method for the simultaneous determination of Δ9-tetrahydrocannabinol (THC), 11-hydroxy-THC and 11-nor-9-carboxy-THC in human plasma using solid phase extraction and gas chromatography–mass spectrometry with positive chemical ionization. J Chromatogr B 798:145–154CrossRef Gustafson RA, Moolchan ET, Barnes A, Levine B, Huestis MA (2003) Validated method for the simultaneous determination of Δ9-tetrahydrocannabinol (THC), 11-hydroxy-THC and 11-nor-9-carboxy-THC in human plasma using solid phase extraction and gas chromatography–mass spectrometry with positive chemical ionization. J Chromatogr B 798:145–154CrossRef
5.
go back to reference Hidvégi E, Somogyi GP (2014) Determination of main tetrahydrocannabinoids by GC–MS: impact of protein precipitation by acetonitrile on solid phase extraction of cannabinoids from human serum. Pharmazie 69:417–419PubMed Hidvégi E, Somogyi GP (2014) Determination of main tetrahydrocannabinoids by GC–MS: impact of protein precipitation by acetonitrile on solid phase extraction of cannabinoids from human serum. Pharmazie 69:417–419PubMed
6.
go back to reference Lowe RH, Karschner EL, Schwilke EW, Barnes AJ, Huestis MA (2007) Simultaneous quantification of Δ9-tetrahydrocannabinol, 11-hydroxy-Δ9-tetrahydrocannabinol, and 11-nor-Δ9-tetrahydrocannabinol-9-carboxylic acid in human plasma using two-dimensional gas chromatography, cryofocusing, and electron impact-mass spectrometry. J Chromatogr A 1163:318–327CrossRefPubMedPubMedCentral Lowe RH, Karschner EL, Schwilke EW, Barnes AJ, Huestis MA (2007) Simultaneous quantification of Δ9-tetrahydrocannabinol, 11-hydroxy-Δ9-tetrahydrocannabinol, and 11-nor-Δ9-tetrahydrocannabinol-9-carboxylic acid in human plasma using two-dimensional gas chromatography, cryofocusing, and electron impact-mass spectrometry. J Chromatogr A 1163:318–327CrossRefPubMedPubMedCentral
7.
go back to reference Nadulski T, Sporkert F, Schnelle M, Stadelmann AM, Roser P, Schefter T, Pragst F (2005) Simultaneous and sensitive analysis of THC, 11-OH-THC, THC-COOH, CBD, and CBN by GC–MS in plasma after oral application of small doses of THC and cannabis extract. J Anal Toxicol 29:782–789CrossRefPubMed Nadulski T, Sporkert F, Schnelle M, Stadelmann AM, Roser P, Schefter T, Pragst F (2005) Simultaneous and sensitive analysis of THC, 11-OH-THC, THC-COOH, CBD, and CBN by GC–MS in plasma after oral application of small doses of THC and cannabis extract. J Anal Toxicol 29:782–789CrossRefPubMed
8.
go back to reference Purschke K, Heinl S, Lerch O, Erdmann F, Veit F (2016) Development and validation of an automated liquid-liquid extraction GC/MS method for the determination of THC, 11-OH-THC, and free THC-carboxylic acid (THC-COOH) from blood serum. Anal Bioanal Chem 408:4379–4388CrossRefPubMedPubMedCentral Purschke K, Heinl S, Lerch O, Erdmann F, Veit F (2016) Development and validation of an automated liquid-liquid extraction GC/MS method for the determination of THC, 11-OH-THC, and free THC-carboxylic acid (THC-COOH) from blood serum. Anal Bioanal Chem 408:4379–4388CrossRefPubMedPubMedCentral
9.
go back to reference Röhrich J, Schimmel I, Zörntlein S, Becker J, Drobnik S, Kaufmann T, Kuntz V, Urban R (2010) Concentrations of Δ9-tetrahydrocannabinol and 11-nor-9-carboxytetrahydrocannabinol in blood and urine after passive exposure to cannabis smoke in a coffee shop. J Anal Toxicol 34:196–203CrossRefPubMed Röhrich J, Schimmel I, Zörntlein S, Becker J, Drobnik S, Kaufmann T, Kuntz V, Urban R (2010) Concentrations of Δ9-tetrahydrocannabinol and 11-nor-9-carboxytetrahydrocannabinol in blood and urine after passive exposure to cannabis smoke in a coffee shop. J Anal Toxicol 34:196–203CrossRefPubMed
10.
go back to reference Scurlock RD, Ohlson GB, Worthen DK (2006) The detection of Δ9-tetrahydrocannabinol (THC) and 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THCA) in whole blood using two-dimensional gas chromatography and EI-mass spectrometry. J Anal Toxicol 30:262–266CrossRefPubMed Scurlock RD, Ohlson GB, Worthen DK (2006) The detection of Δ9-tetrahydrocannabinol (THC) and 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THCA) in whole blood using two-dimensional gas chromatography and EI-mass spectrometry. J Anal Toxicol 30:262–266CrossRefPubMed
11.
go back to reference Steinmeyer S, Bregel D, Warth S, Kraemer T, Moeller MR (2002) Improved and validated method for the determination of Δ9-tetrahydrocannabinol (THC), 11-hydroxy-THC and 11-nor-9-carboxy-THC in serum, and in human liver microsomal preparations using gas chromatography–mass spectrometry. J Chromatogr B 772:239–248CrossRef Steinmeyer S, Bregel D, Warth S, Kraemer T, Moeller MR (2002) Improved and validated method for the determination of Δ9-tetrahydrocannabinol (THC), 11-hydroxy-THC and 11-nor-9-carboxy-THC in serum, and in human liver microsomal preparations using gas chromatography–mass spectrometry. J Chromatogr B 772:239–248CrossRef
12.
go back to reference Thomas A, Widmer C, Hopfgartner G, Staub C (2007) Fast gas chromatography and negative-ion chemical ionization tandem mass spectrometry for forensic analysis of cannabinoids in whole blood. J Pharm Biomed Anal 45:495–503CrossRefPubMed Thomas A, Widmer C, Hopfgartner G, Staub C (2007) Fast gas chromatography and negative-ion chemical ionization tandem mass spectrometry for forensic analysis of cannabinoids in whole blood. J Pharm Biomed Anal 45:495–503CrossRefPubMed
13.
go back to reference Weller JP, Wolf M, Szidat S (2000) Enhanced selectivity in the determination of Δ9-tetrahydrocannabinol and two major metabolites in serum using ion-trap GC–MS-MS. J Anal Toxicol 24:359–364CrossRefPubMed Weller JP, Wolf M, Szidat S (2000) Enhanced selectivity in the determination of Δ9-tetrahydrocannabinol and two major metabolites in serum using ion-trap GC–MS-MS. J Anal Toxicol 24:359–364CrossRefPubMed
14.
go back to reference Coulter C, Miller E, Crompton K, Moore C (2008) Tetrahydrocannabinol and two of its metabolites in whole blood using liquid chromatography–tandem mass spectrometry. J Anal Toxicol 32:653–658CrossRefPubMed Coulter C, Miller E, Crompton K, Moore C (2008) Tetrahydrocannabinol and two of its metabolites in whole blood using liquid chromatography–tandem mass spectrometry. J Anal Toxicol 32:653–658CrossRefPubMed
15.
go back to reference Dubois N, Paccou AP, De Backer BG, Charlier CJ (2012) Validation of the quantitative determination of tetrahydrocannabinol and its two major metabolites in plasma by ultra-high-performance liquid chromatography–tandem mass spectrometry according to the total error approach. J Anal Toxicol 36:25–29CrossRefPubMed Dubois N, Paccou AP, De Backer BG, Charlier CJ (2012) Validation of the quantitative determination of tetrahydrocannabinol and its two major metabolites in plasma by ultra-high-performance liquid chromatography–tandem mass spectrometry according to the total error approach. J Anal Toxicol 36:25–29CrossRefPubMed
16.
go back to reference Elian AA, Hackett J (2009) Solid-phase extraction and analysis of THC and carboxy-THC from whole blood using a novel fluorinated solid phase extraction sorbent and fast liquid chromatography–tandem mass spectrometry. J Anal Toxicol 33:461–468CrossRefPubMed Elian AA, Hackett J (2009) Solid-phase extraction and analysis of THC and carboxy-THC from whole blood using a novel fluorinated solid phase extraction sorbent and fast liquid chromatography–tandem mass spectrometry. J Anal Toxicol 33:461–468CrossRefPubMed
17.
go back to reference Del Mar Ramirez Fernandez M, De Boeck G, Wood M, Lopez-Rivadulla M, Samyn N (2008) Simultaneous analysis of THC and its metabolites in blood using liquid chromatography–tandem mass spectrometry. J Chromatogr B 875:465–470CrossRef Del Mar Ramirez Fernandez M, De Boeck G, Wood M, Lopez-Rivadulla M, Samyn N (2008) Simultaneous analysis of THC and its metabolites in blood using liquid chromatography–tandem mass spectrometry. J Chromatogr B 875:465–470CrossRef
18.
go back to reference Jagerdeo E, Schaff JE, Montgomery MA, LeBeau MA (2009) A semi-automated solid-phase extraction liquid chromatography/tandem mass spectrometry method for the analysis of tetrahydrocannabinol and metabolites in whole blood. Rapid Commun Mass Spectrom 23:2697–2705CrossRefPubMed Jagerdeo E, Schaff JE, Montgomery MA, LeBeau MA (2009) A semi-automated solid-phase extraction liquid chromatography/tandem mass spectrometry method for the analysis of tetrahydrocannabinol and metabolites in whole blood. Rapid Commun Mass Spectrom 23:2697–2705CrossRefPubMed
19.
go back to reference Jamey C, Szwarc E, Tracqui A, Ludes B (2008) Determination of cannabinoids in whole blood by UPLC–MS–MS. J Anal Toxicol 32:349–354CrossRefPubMed Jamey C, Szwarc E, Tracqui A, Ludes B (2008) Determination of cannabinoids in whole blood by UPLC–MS–MS. J Anal Toxicol 32:349–354CrossRefPubMed
20.
go back to reference König S, Aebi B, Lanz S, Gasser M, Weinmann W (2011) On-line SPE LC–MS/MS for the quantification of Δ9-tetrahydrocannabinol (THC) and its two major metabolites in human peripheral blood by liquid chromatography tandem mass spectrometry. Anal Bioanal Chem 400:9–16CrossRefPubMed König S, Aebi B, Lanz S, Gasser M, Weinmann W (2011) On-line SPE LC–MS/MS for the quantification of Δ9-tetrahydrocannabinol (THC) and its two major metabolites in human peripheral blood by liquid chromatography tandem mass spectrometry. Anal Bioanal Chem 400:9–16CrossRefPubMed
21.
go back to reference Laloup M, Del Mar Ramirez Fernandez M, Wood M, De Boeck G, Maes V, Samyn N (2006) Correlation of Δ9-tetrahydrocannabinol concentrations determined by LC–MS–MS in oral fluid and plasma from impaired drivers and evaluation of the on-site Dräger DrugTest®. Forensic Sci Int 161:175–179CrossRefPubMed Laloup M, Del Mar Ramirez Fernandez M, Wood M, De Boeck G, Maes V, Samyn N (2006) Correlation of Δ9-tetrahydrocannabinol concentrations determined by LC–MS–MS in oral fluid and plasma from impaired drivers and evaluation of the on-site Dräger DrugTest®. Forensic Sci Int 161:175–179CrossRefPubMed
22.
go back to reference Moralikova B, Weinmann W (2004) Simultaneous determination of Δ9-tetrahydrocannabinol, 11-hydroxy-Δ9-tetrahydrocannabinol and 11-nor-9-carboxy-Δ9-tetrahydrocannabinol in human plasma by high-performance liquid chromatography/tandem mass spectrometry. J Mass Spectrom 39:526–531CrossRef Moralikova B, Weinmann W (2004) Simultaneous determination of Δ9-tetrahydrocannabinol, 11-hydroxy-Δ9-tetrahydrocannabinol and 11-nor-9-carboxy-Δ9-tetrahydrocannabinol in human plasma by high-performance liquid chromatography/tandem mass spectrometry. J Mass Spectrom 39:526–531CrossRef
23.
go back to reference Schwope DM, Scheidweiler KB, Huestis MA (2011) Direct quantification of cannabinoids and cannabinoid glucuronides in whole blood by liquid chromatography–tandem mass spectrometry. Anal Bioanal Chem 401:1273–1283CrossRefPubMedPubMedCentral Schwope DM, Scheidweiler KB, Huestis MA (2011) Direct quantification of cannabinoids and cannabinoid glucuronides in whole blood by liquid chromatography–tandem mass spectrometry. Anal Bioanal Chem 401:1273–1283CrossRefPubMedPubMedCentral
24.
go back to reference Peters FT, Hartung M, Herbold M, Schmitt G, Daldrup T, Mußhoff F (2009) Anhang B zur Richtlinie der GTFCh zur Qualitätssicherung bei forensisch-toxikologischen Untersuchungen. Anforderungen an die Validierung von Analysenmethoden. Toxichem Krimtech 76:185–208 Peters FT, Hartung M, Herbold M, Schmitt G, Daldrup T, Mußhoff F (2009) Anhang B zur Richtlinie der GTFCh zur Qualitätssicherung bei forensisch-toxikologischen Untersuchungen. Anforderungen an die Validierung von Analysenmethoden. Toxichem Krimtech 76:185–208
25.
go back to reference Matuszewski BK, Constanzer ML, Chavez-Eng CM (2003) Strategies for the assessment of matrix effect in quantitative bioanalytical methods based on HPLC–MS/MS. Anal Chem 75:3019–3030CrossRefPubMed Matuszewski BK, Constanzer ML, Chavez-Eng CM (2003) Strategies for the assessment of matrix effect in quantitative bioanalytical methods based on HPLC–MS/MS. Anal Chem 75:3019–3030CrossRefPubMed
26.
go back to reference Skopp G, Pötsch L, Mauden M, Richter B (2002) Partition coefficient, blood to plasma ratio, protein binding and short-term stability of 11-nor-Δ9-carboxy tetrahydrocannabinol glucuronide. Forensic Sci Int 126:17–23CrossRefPubMed Skopp G, Pötsch L, Mauden M, Richter B (2002) Partition coefficient, blood to plasma ratio, protein binding and short-term stability of 11-nor-Δ9-carboxy tetrahydrocannabinol glucuronide. Forensic Sci Int 126:17–23CrossRefPubMed
27.
go back to reference Skopp G, Pötsch L (2002) Stability of 11-nor-Δ9-carboxy-tetrahydrocannabinol glucuronide in plasma and urine assessed by liquid chromatography–tandem mass spectrometry. Clin Chem 48:301–306PubMed Skopp G, Pötsch L (2002) Stability of 11-nor-Δ9-carboxy-tetrahydrocannabinol glucuronide in plasma and urine assessed by liquid chromatography–tandem mass spectrometry. Clin Chem 48:301–306PubMed
28.
go back to reference Schwope DM, Karschner EL, Gorelick DA, Huestis MA (2011) Identification of recent cannabis use: whole-blood and plasma free and glucuronidated cannabinoid pharmacokinetics following controlled smoked cannabis administration. Clin Chem 57:1406–1414CrossRefPubMedPubMedCentral Schwope DM, Karschner EL, Gorelick DA, Huestis MA (2011) Identification of recent cannabis use: whole-blood and plasma free and glucuronidated cannabinoid pharmacokinetics following controlled smoked cannabis administration. Clin Chem 57:1406–1414CrossRefPubMedPubMedCentral
Metadata
Title
Simple protein precipitation-based analysis of Δ9-tetrahydrocannabinol and its metabolites in human serum by liquid chromatography–tandem mass spectrometry
Authors
Marek Dziadosz
Michael Klintschar
Jörg Teske
Publication date
01-01-2017
Publisher
Springer Japan
Published in
Forensic Toxicology / Issue 1/2017
Print ISSN: 1860-8965
Electronic ISSN: 1860-8973
DOI
https://doi.org/10.1007/s11419-016-0344-7

Other articles of this Issue 1/2017

Forensic Toxicology 1/2017 Go to the issue