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Planta Med 2019; 85(05): 431-438
DOI: 10.1055/a-0828-8387
Natural Product Chemistry and Analytical Studies
Original Papers
Georg Thieme Verlag KG Stuttgart · New York

Analysis of Terpenes in Cannabis sativa L. Using GC/MS: Method Development, Validation, and Application

Elsayed A. Ibrahim
1   National Center for Natural Products Research, University of Mississippi, University, MS, USA
2   Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy, Suez Canal University, Ismailia, Egypt
,
Mei Wang
1   National Center for Natural Products Research, University of Mississippi, University, MS, USA
,
Mohamed M. Radwan
1   National Center for Natural Products Research, University of Mississippi, University, MS, USA
3   Pharmacognosy Department, Faculty of Pharmacy, Alexandria University, Alexandria, Egypt
,
Amira S. Wanas
1   National Center for Natural Products Research, University of Mississippi, University, MS, USA
4   Pharmacognosy Department, Faculty of Pharmacy, Minia University, Minia, Egypt
,
Chandrani G. Majumdar
1   National Center for Natural Products Research, University of Mississippi, University, MS, USA
,
Baharthi Avula
1   National Center for Natural Products Research, University of Mississippi, University, MS, USA
,
Yan-Hong Wang
1   National Center for Natural Products Research, University of Mississippi, University, MS, USA
,
Ikhlas A. Khan
1   National Center for Natural Products Research, University of Mississippi, University, MS, USA
5   Department of Biomolecular Sciences, School of Pharmacy, University of Mississippi, University, MS, USA
,
Suman Chandra
1   National Center for Natural Products Research, University of Mississippi, University, MS, USA
,
Hemant Lata
1   National Center for Natural Products Research, University of Mississippi, University, MS, USA
,
Ghada M. Hadad
2   Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy, Suez Canal University, Ismailia, Egypt
,
Randa A. Abdel Salam
2   Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy, Suez Canal University, Ismailia, Egypt
,
Amany K. Ibrahim
6   Pharmacognosy Department, Faculty of Pharmacy, Suez Canal University, Ismailia, Egypt
,
Safwat A. Ahmed
6   Pharmacognosy Department, Faculty of Pharmacy, Suez Canal University, Ismailia, Egypt
,
Mahmoud A. ElSohly
1   National Center for Natural Products Research, University of Mississippi, University, MS, USA
7   Department of Pharmaceutics and Drug Delivery, School of Pharmacy, University of Mississippi, University, MS, USA
› Author Affiliations
Further Information

Publication History

received 15 May 2018
revised 04 December 2018

accepted 18 December 2018

Publication Date:
15 January 2019 (online)

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Abstract

Terpenes are the major components of the essential oils present in various Cannabis sativa L. varieties. These compounds are responsible for the distinctive aromas and flavors. Besides the quantification of the cannabinoids, determination of the terpenes in C. sativa strains could be of importance for the plant selection process. At the University of Mississippi, a GC-MS method has been developed and validated for the quantification of terpenes in cannabis plant material, viz., α-pinene, β-pinene, β-myrcene, limonene, terpinolene, linalool, α-terpineol, β-caryophyllene, α-humulene, and caryophyllene oxide. The method was optimized and fully validated according to AOAC (Association of Official Analytical Chemists) guidelines against reference standards of selected terpenes. Samples were prepared by extraction of the plant material with ethyl acetate containing n-tridecane solution (100 µg/mL) as the internal standard. The concentration-response relationship for all analyzed terpenes using the developed method was linear with r2 values > 0.99. The average recoveries for all terpenes in spiked indoor cultivated samples were between 95.0 – 105.7%, with the exception of terpinolene (67 – 70%). The measured repeatability and intermediate precisions (% relative standard deviation) in all varieties ranged from 0.32 to 8.47%. The limit of detection and limit of quantitation for all targeted terpenes were determined to be 0.25 and 0.75 µg/mL, respectively. The proposed method is highly selective, reliable, and accurate and has been applied to the simultaneous determination of these major terpenes in the C. sativa biomass produced by our facility at the University of Mississippi as well as in confiscated marijuana samples.

Key words

Cannabis sativa - Cannabaceae - terpenes - GC-MS - method development - method validation

Supporting Information

    LOD, LOQ and retention times (Table 1S), regression data (Table 2S), inter- and intraday precision and accuracy parameters (Tables 5S–10S), and calibration curves for targeted terpenes with the regression equation and correlation coefficient (r 2) for each curve (Fig. 1S) are available as Supporting Information.

  • Supporting Information
 

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