journal article·2022-03-10Open access

Automation System for the Flexible Sample Preparation for Quantification of Δ9-THC-D3, THC-OH and THC-COOH from Serum, Saliva and Urine

Anna Bach, Heidi Fleischer, Bhagya Wijayawardena, Kerstin Thurow

Applied Sciences· Volume 12· Issue 6

Abstract

In the life sciences, automation solutions are primarily established in the field of drug discovery. However, there is also an increasing need for automated solutions in the field of medical diagnostics, e.g., for the determination of vitamins, medication or drug abuse. While the actual metrological determination is highly automated today, the necessary sample preparation processes are still mainly carried out manually. In the laboratory, flexible solutions are required that can be used to determine different target substances in different matrices. A suitable system based on an automated liquid handler was implemented. It has been tested and validated for the determination of three cannabinoid metabolites in blood, urine and saliva. To extract Δ9-tetrahydrocannabinol-D3 (Δ9-THC-D3), 11-hydroxy-Δ9-tetrahydrocannabinol (THC-OH) and 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THC-COOH) from serum, urine and saliva both rapidly and cost-effectively, three sample preparation methods automated with a liquid handling robot are presented in this article, the basic framework of which is an identical SPE method so that they can be quickly exchanged against each other when the matrix is changed. If necessary, the three matrices could also be prepared in parallel. For the sensitive detection of analytes, protein precipitation is used when preparing serum before SPE and basic hydrolysis is used for urine to cleave the glucuronide conjugate. Recoveries of developed methods are >77%. Coefficients of variation are <4%. LODs are below 1 ng/mL and a comparison with the manual process shows a significant cost reduction.
Source: crossref · Quality: baseline

Research context

OpenAlex topics: Forensic Toxicology and Drug Analysis · Analytical Chemistry and Chromatography · Metabolomics and Mass Spectrometry Studies

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

Affiliations

Institute of Automation, Faculty of Computer Science and Electrical Engineering, University of Rostock, 18119 Rostock, Germany

Beckman Coulter Life Sciences, Indianapolis, IN 46268, USA

Center for Life Science Automation, University of Rostock, 18119 Rostock, Germany

References (72)

Show source-provided references
1. (2021). Europäischer Drogenbericht 2021: Trends und Entwicklungen, Europäische Beobachtungsstelle für Drogen und Drogensucht, Amt für Veröffentlichungen der Europäischen Union.
2. Cannabis sativa L. and Nonpsychoactive Cannabinoids: Their Chemistry and Role against Oxidative Stress, Inflammation, and Cancer10.1155/2018/1691428 ↗
3. Simultaneous GC-EI-MS determination of Delta9-tetrahydrocannabinol, 11-hydroxy-Delta9-tetrahydrocannabinol, and 11-nor-9-carboxy-Delta9-tetrahydrocannabinol in human urine following tandem enzyme-alkaline hydrolysis10.1093/jat/31.8.477 ↗
4. Meng, Q., Buchanan, B., Zuccolo, J., Poulin, M.M., Gabriele, J., and Baranowski, D.C. (2018). A reliable and validated LC-MS/MS method for the simultaneous quantification of 4 cannabinoids in 40 consumer products. PLoS ONE, 13.10.1371/journal.pone.0196396 ↗
5. Validated method for the simultaneous determination of Delta 9-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 ionization10.1016/j.jchromb.2003.09.022 ↗
6. Cytochrome P450 enzymes involved in the metabolism of tetrahydrocannabinols and cannabinol by human hepatic microsomes10.1016/j.lfs.2006.12.032 ↗
7. Sensitive Determination of Cannabinoids in Whole Blood by LC-MS-MS After Rapid Removal of Phospholipids by Filtration10.1093/jat/bkx030 ↗
8. Simultaneous and sensitive LC-MS/MS determination of tetrahydrocannabinol and metabolites in human plasma10.1007/s00216-012-6501-x ↗
9. Differentiating new marijuana use from residual drug excretion in occasional marijuana users10.1093/jat/22.6.445 ↗
10. Validated method for the simultaneous determination of Delta9-THC and Delta9-THC-COOH in oral fluid, urine and whole blood using solid-phase extraction and liquid chromatography-mass spectrometry with electrospray ionization10.1016/j.forsciint.2007.03.026 ↗
11. Current knowledge on cannabinoids in oral fluid10.1002/dta.1514 ↗
12. Clinical and diagnostic utility of saliva as a non-invasive diagnostic fluid: A systematic review10.11613/BM.2015.018 ↗
13. Review: The physiology of saliva and transfer of drugs into saliva10.1016/j.forsciint.2004.10.026 ↗
14. Marihuana: Studies on the disposition and metabolism of delta-9-tetrahydrocannabinol in man10.1126/science.170.3964.1320 ↗
15. Metabolism, disposition, and kinetics of delta-9-tetrahydrocannabinol in men and women10.1038/clpt.1983.179 ↗
16. Cannabinoids in humans. II. The influence of three methods of hydrolysis on the concentration of THC and two metabolites in urine10.1093/jat/19.5.292 ↗
17. Disposition of Cannabinoids in Oral Fluid after Controlled Around-the-Clock Oral THC Administration10.1373/clinchem.2009.141853 ↗
18. Identification in human urine of Δ9-tetrahydrocannabinol-11-oic acid glucuronide: A tetrahydrocannabinol metabolite10.1111/j.2042-7158.1980.tb12966.x ↗
19. Determination of 21 drugs in oral fluid using fully automated supported liquid extraction and UHPLC-MS/MS10.1002/dta.2045 ↗
20. Alternative specimens for workplace drug testing10.1093/jat/25.5.396 ↗
21. Validation of a fully automated robotic setup for preparation of whole blood samples for LC-MS toxicology analysis10.1093/jat/bks014 ↗
22. Simple protein precipitation-based analysis of Δ9-tetrahydrocannabinol and its metabolites in human serum by liquid chromatography–tandem mass spectrometry10.1007/s11419-016-0344-7 ↗
23. Using THC-COOH cut-off concentrations for assessing cannabis consumption frequency: A recently detected THC-COOH isomer poses an important analytical problem
24. Pitfall in cannabinoid analysis—Detection of a previously unrecognized interfering compound in human serum10.1007/s00216-014-8301-y ↗
25. Analysis of 30 synthetic cannabinoids in serum by liquid chromatography-electrospray ionization tandem mass spectrometry after liquid-liquid extraction10.1002/jms.3020 ↗
26. A semi-automated solid-phase extraction liquid chromatography/tandem mass spectrometry method for the analysis of tetrahydrocannabinol and metabolites in whole blood10.1002/rcm.4174 ↗
27. Direct quantification of cannabinoids and cannabinoid glucuronides in whole blood by liquid chromatography-tandem mass spectrometry10.1007/s00216-011-5197-7 ↗
28. Determination of 12 commonly found compounds in DUID cases in whole blood using fully automated supported liquid extraction and UHPLC-MS/MS10.1016/j.jchromb.2018.06.050 ↗
29. Validation of a liquid chromatography tandem mass spectrometry (LC-MS/MS) method to detect cannabinoids in whole blood and breath10.1515/cclm-2019-0600 ↗
30. An Efficient, Robust Method for the Determination of Cannabinoids in Whole Blood by LC-MS-MS10.1093/jat/bkw063 ↗
31. 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 serum10.1007/s00216-016-9537-5 ↗
32. Delta(9)-THC, 11-OH-Delta(9)-THC and Delta(9)-THCCOOH plasma or serum to whole blood concentrations distribution ratios in blood samples taken from living and dead people10.1016/S0379-0738(01)00538-2 ↗
33. Enhanced selectivity in the determination of delta9-tetrahydrocannabinol and two major metabolites in serum using ion-trap GC-MS-MS10.1093/jat/24.5.359 ↗
34. Simultaneous quantification of major cannabinoids and metabolites in human urine and plasma by HPLC-MS/MS and enzyme-alkaline hydrolysis10.1002/dta.1998 ↗
35. Screening, quantification, and confirmation of synthetic cannabinoid metabolites in urine by UHPLC-QTOF-MS10.1002/dta.2464 ↗
36. Simultaneous quantification of 37 synthetic cannabinoid metabolites in human urine by liquid chromatography-tandem mass spectrometry10.1007/s11419-015-0265-x ↗
37. Sensitive screening of synthetic cannabinoids using liquid chromatography quadrupole time-of-flight mass spectrometry after solid phase extraction10.1002/dta.3052 ↗
38. Determination of 11-nor-delta9-tetrahydrocannabinol-9-carboxylic acid in urine using high-performance liquid chromatography and electrospray ionization mass spectrometry10.1016/S0378-4347(99)00276-5 ↗
39. A rapid quantitative method for the analysis of synthetic cannabinoids by liquid chromatography-tandem mass spectrometry10.1093/jat/bku092 ↗
40. Optimization of recombinant β-glucuronidase hydrolysis and quantification of eight urinary cannabinoids and metabolites by liquid chromatography tandem mass spectrometry10.1002/dta.2230 ↗
41. (2013). Automated Extraction of Synthetic Cannabinoids (SPICE) from Urine Using ISOLUTE® SLE + Prior to LC-MS/MS, Biotage. AN793.
42. Simultaneous quantification of free and glucuronidated cannabinoids in human urine by liquid chromatography tandem mass spectrometry10.1016/j.cca.2012.06.034 ↗
43. (2014). Sample Preparation with the AC Extraction Plate: Extraction of Urinary THC Metabolites for LC-MSMS Quantification, Tecan Trading AG.
44. Fast confirmation of 11-nor-9-carboxy-Delta(9)-tetrahydrocannabinol (THC-COOH) in urine by LC/MS/MS using negative atmospheric-pressure chemical ionisation (APCI)10.1016/S0379-0738(01)00459-5 ↗
45. A μ-SPE procedure for the determination of cannabinoids and their metabolites in urine by LC-MS/MS10.1016/j.jpba.2013.12.035 ↗
46. Surface-activated chemical ionization combined with electrospray ionization and mass spectrometry for the analysis of cannabinoids in biological samples. Part I: Analysis of 11-nor-9-carboxytetrahydro-cannabinol10.1002/rcm.5029 ↗
47. Quantitative analysis of 11-nor-9-carboxy-tetrahydrocannbinol (THC–COOH) in urine by LC–MS/MS following a simple filtration10.1016/j.jchromb.2014.02.056 ↗
48. Automated solid-phase extraction-liquid chromatography-tandem mass spectrometry analysis of 11-nor-Delta9-tetrahydrocannabinol-9-carboxylic acid in human urine specimens: Application to a high-throughput urine analysis laboratory10.1093/jat/33.8.456 ↗
49. A fast method for screening and/or quantitation of tetrahydrocannabinol and metabolites in urine by automated SPE/LC/MS/MS10.1007/s00216-010-3918-y ↗
50. Novel solid-phase extraction protocol for 11-nor-9-carboxy-delta9-tetrahydrocannabinol from urine samples employing a polymeric mixed-mode cation-exchange resin, Strata-X-C, suitable for gas chromatography-mass spectrometry or liquid chromatography-mass spectrometry analysis10.1016/j.chroma.2004.10.052 ↗
51. Urine toxicology screening by liquid chromatography time-of-flight mass spectrometry in a quaternary hospital setting10.1016/j.clinbiochem.2021.05.004 ↗
52. LC-QTOF-MS as a superior strategy to immunoassay for the comprehensive analysis of synthetic cannabinoids in urine10.1007/s00216-013-7574-x ↗
53. Cabrices, O.G., Foster, F.D., Taylor, A.M., and Lee, L.A. (2014). “Prep-and-Shoot”: The Completely Automated Workflow for the Hydrolysis and Analysis of Urine Samples by LC/MS/MS, Gerstel.
54. Simultaneous screening and quantification of 29 drugs of abuse in oral fluid by solid-phase extraction and ultraperformance LC-MS/MS10.1373/clinchem.2008.122341 ↗
55. Determination of illicit and medicinal drugs and their metabolites in oral fluid and preserved oral fluid by liquid chromatography–tandem mass spectrometry10.1007/s00216-008-2135-4 ↗
56. Analysis of Tetrahydrocannabinol and its Metabolite, 11-Nor-Δ9-Tetrahydrocannabinol-9-Carboxylic Acid, in Oral Fluid using Liquid Chromatography with Tandem Mass Spectrometry10.1093/jat/bks039 ↗
57. A validated method for the detection of Delta 9-tetrahydrocannabinol and 11-nor-9-carboxy-Delta 9-tetrahydrocannabinol in oral fluid samples by liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry10.1093/jat/31.3.157 ↗
58. Drugs of abuse in oral fluid collected by two different sample kits—Stability testing and validation using ultra performance tandem mass spectrometry analysis10.1016/j.jchromb.2011.09.002 ↗
59. Oral Fluid to Blood Concentration Ratios of Different Psychoactive Drugs in Samples from Suspected Drugged Drivers10.1097/FTD.0000000000000760 ↗
60. Micro extraction by packed sorbent coupled to liquid chromatography tandem mass spectrometry for the rapid and sensitive determination of cannabinoids in oral fluids10.1016/j.chroma.2013.05.072 ↗
61. Cannabinoids and metabolites in expectorated oral fluid after 8 days of controlled around-the-clock oral THC administration10.1007/s00216-011-5066-4 ↗
62. Simultaneous determination of Δ9-tetrahydrocannabinol and 11-nor-9-carboxy-Δ9-tetrahydrocannabinol in oral fluid using isotope dilution liquid chromatography tandem mass spectrometry10.1007/s00216-011-5439-8 ↗
63. Multiclass analysis of illicit drugs in plasma and oral fluids by LC-MS/MS10.1007/s00216-008-2456-3 ↗
64. Analysis of cannabis in oral fluid specimens by GC-MS with automatic SPE10.1016/j.scijus.2009.09.015 ↗
65. (2022, March 08). Cerillant Corporation Certificate of Analysis (COA). Available online: https://www.cerilliant.com/products/searchCOA.aspx.
66. Optimization of Automated Sample Preparation for Vitamin D Determination on a Biomek i7 Workstation10.1177/24726303211030291 ↗
67. Fleischer, H., and Thurow, K. (2017). Automation Solutions for Analytical Measurements, Wiley-VCH.10.1002/9783527805297 ↗
68. Otto, M. (2019). Analytische Chemie, Wiley-VCH.
69. Tetrahydrocannabinol stability in whole blood: Plastic versus glass containers10.1093/jat/10.4.129 ↗
70. Stability of 11 prevalent synthetic cannabinoids in authentic neat oral fluid samples: Glass versus polypropylene containers at different temperatures10.1002/dta.1497 ↗
71. Application of Biofunctionalized Magnetic Nanoparticles Based-Sensing in Abused Drugs Diagnostics10.1021/acs.analchem.9b04025 ↗
72. Scurati, S., Brambilla, M., D’Amato, L., and Brambilla, P. (2011). THC-COOH Quantification in Urine Using Dilute and Shoot LC-MS/MS Method for Forensic Toxicology, Thermo Fisher Scientific.