OILVEQ: an Italian external quality control scheme for cannabinoids analysis in galenic preparations of cannabis oil

Maria Concetta Rotolo; Silvia Graziano; Adele Minutillo; Maria Rosaria Varì; Simona Pichini; Emilia Marchei

doi:10.1515/cclm-2024-0311

Article

OILVEQ: an Italian external quality control scheme for cannabinoids analysis in galenic preparations of cannabis oil

Maria Concetta Rotolo , Silvia Graziano , Adele Minutillo , Maria Rosaria Varì , Simona Pichini and Emilia Marchei

Published/Copyright: May 13, 2024

Published by

Become an author with De Gruyter Brill

Submit Manuscript Author Information Explore this Subject

From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 62 Issue 11

Abstract

Objectives

Italy legalized cannabis oil for specific medical conditions (neuropathic pain, refractory epilepsy and other established pathologies) in 2015, but mandates titration of principal cannabinoids before marketing each batch using iphenated techniques coupled with mass spectrometry. To assess reliability of laboratories from the Italian National Health Service in charge of titrating the batches, the Italian National Institute of Health set up an quality control program on determination of Δ9-tetrahydrocannabinol l (THC), cannabidiol (CBD), Δ9-tetrahydrocannabinolic acid A (THCA-A) and cannabidiolic acid (CBDA) in cannabis oil preparations.

Methods

Two rounds of exercises have been carried out since 2019, involving sixteen Italian laboratories. Five different cannabis oil samples (19-1A and 19-1B for the first round and 22-1A, 22-1B and 22-1C for the second one were prepared and 1 mL amount of each sample was sent to the laboratories. The quantitative performance of each laboratory was assessed calculating the Z-score value, a statistical measurement for value’s relationship to the mean of a group of values.

Results

In the first round, eight out of fourteen laboratories employed an LC-MS while the remaining six used GC-MS. Differently, in the second round, six out of eleven laboratories employed a GC-MS while the remaining five used LC-MS. In the first round, only 28.6 % laboratories achieved an acceptable performance (Z-score±2), and all of them used LC-MS as analytical method. In the second round, none of the laboratories achieved an acceptable performance. Satisfactory results, based on Z-scores, were generally low (0.0–75.0 %), with only one exception of 100 % for THCA-A determination in sample 22-1B. In the second round, three false negatives (two THC and one CBD by GC-MS determination) were reported while no false positives were described in the blank sample. The two rounds yielded a mean ERR% of 42 % approximately and a mean CV% around 70 % in GC-MS determination. When applying LC-MS determination, the two rounds yielded a mean ERR% of 36 % approximately and a mean CV% around 33 %.

Conclusions

The obtained results underline the need for a clear and consistent protocol to be adopted by all laboratories intending to include the titration of oily cannabis-based products into their routinely analytical techniques. This emphasis on methodology standardization and participation to quality control schemes is essential for ensuring reliable and accurate measurements, ultimately enhancing the overall effectiveness and reliability of medical cannabis treatments.

Keywords: cannabinoids; external quality assessment; cannabis oil

Corresponding author: Simona Pichini, National Centre on Addiction and Doping, Italian National Institute of Health, V.Le Regina Elena 299, 00161 Rome, Italy, E-mail: simona.pichini@iss.it

Acknowledgments

We thank Antonella Bacosi, Simonetta Di Carlo, Chiara Fraioli, Laura Martucci, and Michele Sciotti for their technical assistance.

Research ethics: Not applicable.
Informed consent: Not applicable.
Author contributions: The authors have accepted responsibility for the entire content of this manuscript and approved its submission.
Competing interests: The authors state no conflict of interest.
Research funding: None declared.
Data availability: Not applicable.

References

1. Dei, CM, Casagni, E, Casiraghi, A, Minghetti, P, Fornasari, DMM, Ferri, F, et al.. Phytocannabinoids profile in medicinal cannabis oils: the impact of plant varieties and preparation methods. Front Pharmacol 2020;11:570616. https://doi.org/10.3389/fphar.2020.570616.Search in Google Scholar PubMed PubMed Central

2. Gouvêa-Silva, JG, Costa-Oliveira, CD, Ramos, YJ, Mantovanelli, DF, Cardoso, MS, Viana-Oliveira, LD, et al.. Is there enough knowledge to standardize a Cannabis sativa L. medicinal oil preparation with a high content of cannabinoids? Cannabis Cannabinoid Res 2023;8:476–86. https://doi.org/10.1089/can.2022.0076.Search in Google Scholar PubMed

3. Pacifici, R, Marchei, E, Salvatore, F, Guandalini, L, Busardò, FP, Pichini, S. Evaluation of cannabinoids concentration and stability in standardized preparations of cannabis tea and cannabis oil by ultra-high performance liquid chromatography tandem mass spectrometry. Clin Chem Lab Med 2017;55:1555–63. https://doi.org/10.1515/cclm-2016-1060.Search in Google Scholar PubMed

4. Pacifici, R, Marchei, E, Salvatore, F, Guandalini, L, Busardò, FP, Pichini, S. Evaluation of long-term stability of cannabinoids in standardized preparations of cannabis flowering tops and cannabis oil by ultra-high-performance liquid chromatography tandem mass spectrometry. Clin Chem Lab Med 2018;56:94–6. https://doi.org/10.1515/cclm-2017-0758.Search in Google Scholar PubMed

5. Pacifici, R, Marchei, E, Salvatore, F, Guandalini, L, Busardò, FP, Pichini, S. Stability of cannabinoids in cannabis FM1 flowering tops and oil preparation evaluated by ultra-high performance liquid chromatography tandem mass spectrometry. Clin Chem Lab Med 2019;57:e165–8. https://doi.org/10.1515/cclm-2018-1145.Search in Google Scholar PubMed

6. Elsohly, MA, Slade, D. Chemical constituents of marijuana: the complex mixture of natural cannabinoids. Life Sci 2005;78:539–48. https://doi.org/10.1016/j.lfs.2005.09.011.Search in Google Scholar PubMed

7. Radwan, MM, Chandra, S, Gul, S, ElSohly, MA. Cannabinoids, phenolics, terpenes and alkaloids of cannabis. Molecules 2021;26:2774. https://doi.org/10.3390/molecules26092774.Search in Google Scholar PubMed PubMed Central

8. Russo, EB. Cannabinoids in the management of difficult to treat pain. Therapeut Clin Risk Manag 2008;4:245–59. https://doi.org/10.2147/tcrm.s1928.Search in Google Scholar PubMed PubMed Central

9. Goodman, S, Wadsworth, E, Schauer, G, Hammond, D. Use and perceptions of cannabidiol products in Canada and in the United States. Cannabis Cannabinoid Res 2022;7:355–64. https://doi.org/10.1089/can.2020.0093.Search in Google Scholar PubMed PubMed Central

10. Veit, M. Quality requirements for medicinal cannabis and respective products in the European Union – Status Quo. Planta Med 2023;89:808–23. https://doi.org/10.1055/a-1808-9708.Search in Google Scholar PubMed

11. DECRETO 9 novembre 2015 Funzioni di Organismo statale per la cannabis previsto dagli articoli 23 e 28 della convenzione unica sugli stupefacenti del 1961, come modificata nel 1972. (15A08888) (GU Serie Generale n.279 del 30-11-2015). https://www.gazzettaufficiale.it/eli/id/2015/11/30/15A08888/sg;jses [Accessed 07 March 2024].Search in Google Scholar

12. Romano, LL, Hazekamp, A. Cannabis oil: chemical evaluation of an upcoming cannabis-based medicine. Cannabinoids 2013;1:1–11.Search in Google Scholar

13. Citti, C, Ciccarella, G, Braghiroli, D, Parenti, C, Vandelli, MA, Cannazza, G. Medicinal Cannabis: principal cannabinoids concentration and their stability evaluated by a high performance liquid chromatography coupled to diode array and quadrupole time of flight mass spectrometry method. J Pharm Biomed Anal 2016;128:201–9. https://doi.org/10.1016/j.jpba.2016.05.033.Search in Google Scholar PubMed

14. Casiraghi, A, Roda, G, Casagni, E, Cristina, C, Musazzi, UM, Franzè, S, et al.. Extraction method and analysis of cannabinoids in cannabis olive oil preparations. Planta Med 2018;84:242–9. https://doi.org/10.1055/s-0043-123074.Search in Google Scholar PubMed

15. Calvi, L, Pentimalli, D, Panseri, S, Giupponi, L, Gelmini, F, Beretta, G, et al.. Comprehensive quality evaluation of medical Cannabis sativa L. inflorescence and macerated oils based on HS-SPME coupled to GC-MS and LC-HRMS (q-exactive orbitrap®) approach. J Pharm Biomed Anal 2018;150:208–19. https://doi.org/10.1016/j.jpba.2017.11.073.Search in Google Scholar PubMed

16. Baratta, F, Simiele, M, Pignata, I, Ravetto Enri, L, D’Avolio, A, Torta, R, et al.. Cannabis-based oral formulations for medical purposes: preparation, quality and stability. Pharmaceuticals 2021;14:171. https://doi.org/10.3390/ph14020171.Search in Google Scholar PubMed PubMed Central

17. Silva Sofrás, FM, Alonso, R, Retta, DS, Di Leo Lira, P, Desimone, MF, van Baren, CM. Development and validation of a simple, fast, and accessible HPLC-UV method for cannabinoids determination in Cannabis sativa L. extracts and medicinal oils. Curr Pharmaceut Des 2023;29:1918–28. https://doi.org/10.2174/1381612829666230809094304.Search in Google Scholar PubMed

18. Cangemi, G, Bucchioni, P, Tripodi, G, Barco, S, Franceschini, P, Sbarbaro, IM. Analysis of cannabinoids concentration in cannabis oil galenic preparations in the Liguria Region: experimental project of the regional reference laboratories. Biochim Clin 2020;44:367–73.Search in Google Scholar

19. Carcieri, C, Tomasello, C, Simiele, M, De Nicolò, A, Avataneo, V, Canzoneri, L, et al.. Concentration variability in cannabis olive oil galenic preparations. J Pharm Pharmacol 2018;70:143–9. https://doi.org/10.1111/jphp.12845.Search in Google Scholar PubMed

20. Palermiti, A, Cafaro, A, Barco, S, Bucchioni, P, Franceschini, P, Cusato, J, et al.. Analysis of cannabinoids concentration in cannabis oil galenic preparations: harmonization between three laboratories in Northern Italy. Pharmaceuticals 2021;14:462. https://doi.org/10.3390/ph14050462.Search in Google Scholar PubMed PubMed Central

21. Pichini, S, Ventura, M, Pujadas, M, Ventura, R, Pellegrini, M, Zuccaro, P, et al.. HAIRVEQ: an external quality control scheme for drugs of abuse analysis in hair. Forensic Sci Int 2004;145:109–15. https://doi.org/10.1016/j.forsciint.2004.04.025.Search in Google Scholar PubMed

22. Ventura, M, Pichini, S, Pujadas, M, Ventura, R, Di Giovannandrea, R, Zuccaro, P, et al.. Four years’ experience in external proficiency testing programs for hair testing of drugs of abuse in Italy (HAIRVEQ) and comparison with the Society of Hair Testing program in 2005. Ther Drug Monit 2007;29:11–9. https://doi.org/10.1097/ftd.0b013e31802bb100.Search in Google Scholar

23. Ventura, M, Stramesi, C, Pichini, S, Ventura, R, Pujadas, M, Di Giovannandrea, R, et al.. Hairveq 2006: evolution of laboratories’ performance after different educational actions. Forensic Sci Int 2008;176:2–8. https://doi.org/10.1016/j.forsciint.2007.06.022.Search in Google Scholar PubMed

24. Ventura, M, Ventura, R, Pichini, S, Leal, S, Zuccaro, P, Pacifici, R, et al.. ORALVEQ: external quality assessment scheme of drugs of abuse in oral fluid: results obtained in the first round performed in 2007. Forensic Sci Int 2008;182:35–40. https://doi.org/10.1016/j.forsciint.2008.09.016.Search in Google Scholar PubMed

25. A.Li.Sa. Alisa - Sistema Sanitario Regione Liguria. Delibera Regionale n.78/2018 dell’11/04/2018: Aggiornamento indirizzi per la prescrizione, la preparazione e la somministrazione della cannabis ad uso medico a carico del SSR. https://www.sifoweb.it/images/pdf/attivita/sezioni-regionali/liguria/normativa_2018/Dlb_78-2018.pdf [Accessed 07 march 2024].Search in Google Scholar

26. Manca, A, Palermiti, A, Mula, J, De, VED, Zeaiter, S, Simiele, M, et al.. A description of Cannabinoid levels in Cannabis oil by high-performance liquid chromatography-mass spectrometry in a reference laboratory of North-Italy. Phytomedicine 2022;102:154218. https://doi.org/10.1016/j.phymed.2022.154218.Search in Google Scholar PubMed

27. Dussy, FE, Hamberg, C, Luginbühl, M, Schwerzmann, T, Briellmann, TA. Isolation of Delta9-THCA-A from hemp and analytical aspects concerning the determination of Delta9-THC in cannabis products. Forensic Sci Int 2005;149:3–10. https://doi.org/10.1016/j.forsciint.2004.05.015.Search in Google Scholar PubMed

28. Vernich, F, Stefani, L, Fiorelli, D, Mineo, F, Pallocci, M, Treglia, M, et al.. Trends in illicit cannabis potency based on the analysis of law enforcement seizures in the Southern Area of Rome. Toxics 2023;11:648. https://doi.org/10.3390/toxics11080648.Search in Google Scholar PubMed PubMed Central

29. Marchei, E, Tittarelli, R, Pellegrini, M, Rotolo, MC, Pacifici, R, Pichini, S. Is “light cannabis” really light? Determination of cannabinoids content in commercial products. Clin Chem Lab Med 2020;58:e175–7. https://doi.org/10.1515/cclm-2020-0040.Search in Google Scholar PubMed

30. Baudone, I, Corsini, C, Petricciani, G, Sbarbaro, IM, Franceschini, P, Bucchioni, P. Titration of galenic preparations of cannabis oil: the experience of the Levante Ligure reference laboratory. Biochim Clin 2024. Ahead of print. https://doi.org/10.19186/BC.2024.18Search in Google Scholar

Received: 2024-03-07

Accepted: 2024-04-22

Published Online: 2024-05-13

Published in Print: 2024-10-28

You are currently not able to access this content.

Articles in the same Issue

https://doi.org/10.1515/cclm-2024-0311

Keywords for this article

cannabinoids; external quality assessment; cannabis oil