Light Spectrum Effects on Metabolite Profile of Cannabis

As the FDA progresses toward a regulatory framework for hemp based cannabinoid products, I’ll touch on some unique research from the production side, focusing on lighting and hydration strategies in greenhouse settings and provide examples of how formulary might help drive the selection. Look for discussions on extraction methods or strain/chemovar choice at another time.

At a 2019 Cannabis Science Conference East talk, typical production methods for cannabis production were described as follows (Kern, 2019):

  • High light intensity and CO2 concentration
  •  Photoperiod
    • Vegetative growth at 18 hours
    • Flowering < 12 hours
    • Use of Blue light during flowering

The blue light treatment is used to optimize several factors:

  • Increase terpenes with all blue light last three days of flower development
  • Blue light at end of day can increase leaf expansion increasing photosynthesis and plant mass yield
  • Blue light at night can slightly increase ∆9-tetrahydrocannabinol (THC)

Research by Mosaleeyanon et al. (2005) showed that light impacts secondary metabolite production, with increased photosynthetic rates leading to increased total hypericin (hypericin + pseudohypericin) concentration in St. Johnswort.

Hawley et al. (2018) experimented with the use of subcanopy lighting (SCL) of red/blue (“Red-Blue”) or red-green-blue (“RGB”) light versus no SCL in a greenhouse production cannabis. They used a plant layout design to limit the amount of additional radiation from the subcanopy and to prevent the treatment light from influencing other plants. They ran the experiment without gyping (removal of the bottom 20 cm of stems). The SCL treatment created differences in the lower canopy metabolite concentrations:

Neon Cannabis Leaf
  • 9-THC and ∆9-THCA significantly increased using both RGB and RB.
  • CBD, CBG did not increased by either treatment.
  • Terpenes alpha-pinene and borneol significantly increased in RGB.
  • cis-Nerolidol significantly increased in both RGB and RB.

The SCL treatment created differences in upper canopy concentrations:

  • cis-Nerolidol significantly increased in both RGB and RB.
  • Alpha-pinene, limonene, myrcene and linalool significantly increased in RGB.

From a QA perspective, the RB SCL treatment also provided the most consistent levels of cannabinoids and terpenes in both upper and lower canopy.

Hawely et al. (2018) suggested previous research (Miller et al., 1995: Zur et al., 2000) had demonstrated light spectra rich in green light was largely absorbed by terpenes, and that the plant had to up regulate the biosynthetic precursors in response to increased green light stimulus. The precursor molecules turn out to be  part of both terpene and cannabinoid metabolic pathways, leading to an enriched biosynthetic stream. Greater precursors available led to increased production of both class of metabolites

Two examples of how formulary might take advantage of ecological influences

CBC and CBG concentrations have been shown to be present in leaves at equal or greater levels than in the flowers (Bernstein, Gorelick and Koch, 2019). This suggests that lighting and harvesting strategies may include adding leaf material to an extract and/or optimizing the impact of different light spectra on leaf metabolite profiles.Taking it a step further, cold water extraction of immature leaf of selectively bred cannabis chemotypes yields enriched CBC fraction (Potter, 2009)

Since CBC is the second most prevalent cannabinoid compound found in cannabis (Russo 2011), and research has shown topical anti-inflammatory activity (De Petrocellis et al., 2012: Cascio & Pertwee 2014; Oláh et al., 2016), and sebum reduction in acne, then leaf material might be combined with flowers in an extract. And the impact of under canopy lighting should also be investigated more extensively to determine if the leaf yield of cannabinoids and terpenes can be improved. Applying the RBC-SCL strategy to increase terpenes, the following enriched fractions might improve the results as an anti-acne topical: alpha-pinene as an effective antibacterial (Appendino et al., 2008); limonene, pinene and linalool decreasing sebum/sebocytes (Biro et al., 2009).

CBG displays moderate 5-HT1A antagonist suggesting antidepressant properties (Formukong et al., 1988), and linalool and limonene also show anti-anxiety via 5-HT1A (Russo et al., 2005). Additionally, research on CBD supports the use of CBD as an anxiolytic (Bergamaschi et al., 2011; Shannon et al., 2019; Siloti et al., 2019).

In a second approach to using ecological factors to alter metabolite profiles in cannabis, Caplan et al. (2019) applied drought stress  to container-grown cannabis plants, gradually drying substrate under controlled environment for 11 days. The treatment resulted in greater metabolite yield per unit growing area:

  • THCA 43% higher than the control,
  • CBDA 47% higher
  • ∆9-tetrahydrocannabinol (THC) 50% higher
  • cannabidiol (CBD) 67% higher

Experimenting with the yield in various chemovars based on multiple ecological inputs will make for a challenging project. The use of ecological inputs as one strategy for creating treatment specific strains might eventually be combined with chemovar selection and extraction protocols to create a highly specific therapeutic product.

References:

  1. Appendino G, Gibbons S, Giana A, Pagani A, Grassi G, Stavri M, et al. Antibacterial cannabinoids from Cannabis sativa: a structure-activity study. J Nat Prod. 2008;71:1427–1430.
  2. Bergamaschi, M. M., Queiroz, R. H., Chagas, M. H., de Oliveira, D. C., De Martinis, B. S., Kapczinski, F., Quevedo J, Roesler R, Schröder N, Nardi AE, Martín-Santos R, Hallak JE, Zuardi AW, Crippa, J. A. (2011). Cannabidiol reduces the anxiety induced by simulated public speaking in treatment-naïve social phobia patients. Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology36(6), 1219–1226. doi:10.1038/npp.2011.6.
  3. Bernstein, N, Gorelick, J, and Koch, S. (2019) Interplay between chemistry and morphology in medical cannabis (Cannabis sativa L.). Industrial Crops and Products. 129: 185-194. https://doi.org/10.1016/j.indcrop.2018.11.039
  4. Biro T, Olah A, Toth BI, Czifra G, Zouboulis CC, Paus R. Proceedings 19th Annual Conference on the Cannabinoids. Pheasant Run, St. Charles, IL: International Cannabinoid Research Society; 2009. Cannabidiol as a novel anti-acne agent? Cannabidiol inhibits lipid synthesis and induces cell death in human sebaceous gland-derived sebocytes; p. 28.
  5. Caplan, D., Dixon, M. and Zheng, Y. (2019) Increasing Inflorescence Dry Weight and Cannabinoid Content in Medical Cannabis Using Controlled Drought Stress. HortScience. 54(5): 964–969. DOI: https://doi.org/10.21273/HORTSCI13510-18
  6. Cascio, M. G., & Pertwee, R. G. (2014). Known pharmacological actions of nine nonpsychotropic phytocannabinoids. In R. G. Pertwee (Ed.), Handbook of cannabis (pp. 137–156). Oxford, UK: Oxford University Press. http://doi.org/10.1093/acprof: oso/9780199662685.003.0007.
  7. De Petrocellis, L., Orlando, P., Moriello, A. S., Aviello, G., Stott, C., Izzo, A. A., et al. (2012). Cannabinoid actions at TRPV channels: Effects on TRPV3 and TRPV4 and their potential relevance to gastrointestinal inflammation. Acta Physiologica, 204(2), 255–266. http://doi.org/10.1111/j.1748-1716.2011.02338.x.
  8. Formukong, E. A., Evans, A. T., & Evans, F. J. (1988). Analgesic and antiinflammatory activity of constituents of Cannabis sativa L. Inflammation, 12(4), 361–371.
  9. Hawley, D., Graham, T., Stasiak, M., & Dixon, M. (2018). Improving Cannabis Bud Quality and Yield with Subcanopy Lighting, HortScience horts, 53(11), 1593-1599. Retrieved Oct 11, 2019, from https://journals.ashs.org/hortsci/view/journals/hortsci/53/11/article-p1593.xml.
  10. Kern, R. (2019) Moving Towards Sustainable Cultivation Practices, Agate Biosciences. 2019 Cannabis Science Conference East. Baltimore, MD.
  11. Marcu, JP. (2016) Chapter 62 – An Overview of Major and Minor Phytocannabinoids. Ed. Victor R. Preedy. Neuropathology of Drug Addictions and Substance Misuse. Volume 1: Foundations of Understanding, Tobacco, Alcohol, Cannabinoids and Opioids (pp. 672-678). King’s College London, London, UK. Academic Press. https://doi.org/10.1016/B978-0-12-800213-1.00062-6
  12. Miller, E.S., G. Mackinney, and F.P. Zscheile. 1935. Absorption spectra of alpha and beta carotenes and lycopene1. Plant Physiol. 10:375–381.
  13. Mosaleeyanon, K., S.M.A. Zobayed, F. Afreen,and T. Kozai. 2005. Relationships between net photosynthetic rate and secondary metabolite contents in St. John’s wort. Plant Sci. 169:523–531.
  14. Oláh A, Markovics A, Szabó-Papp J, Szabó PT, Stott C, Zouboulis CC, Bíró T. (2016) Differential effectiveness of selected non-psychotropic phytocannabinoids on human sebocyte functions implicates their introduction in dry/seborrhoeic skin and acne treatment. Exp Dermatol. 25(9):701-7. doi: 10.1111/exd.13042. https://www.ncbi.nlm.nih.gov/pubmed/27094344.
  15. Potter DJ (2009). The propagation, characterisation and optimisation of Cannabis sativa as a phytopharmaceutical. PhD, King’s College, London, 2009.
  16. Russo EB, Burnett A, Hall B, Parker KK. Agonistic properties of cannabidiol at 5-HT-1a receptors. Neurochem Res. 2005;30:1037–1043.
  17. Russo, EB. (2011) Taming THC: potential cannabis synergy and phytocannabinoid-terpenoid entourage effects. British J Pharmacology. 163: 1344-1364.
  18. Shannon, S., Lewis, N., Lee, H., & Hughes, S. (2019). Cannabidiol in Anxiety and Sleep: A Large Case Series. The Permanente journal23, 18–041. doi:10.7812/TPP/18-041.
  19. Silote GP, Sartim A, Sales A, Eskelund A, Guimarães FS, Wegener G, Joca S. (2019) Emerging evidence for the antidepressant effect of cannabidiol and the underlying molecular mechanisms. J Chem Neuroanat. 98:104-116. doi: 10.1016/j.jchemneu.2019.04.006. Epub 2019 Apr 27.
  20. Zur, Y., A.A. Gitelson, O.B. Chivkunova, and M.N. Merzlyak. 2000. The spectral contribution of carotenoids to light absorption and reflectance in green leaves. Pap. Natl. Resources 2:272.

The Humanity of Movement

The joy of moment has always been a huge part of my life. I was a hiding go seek champion at a young age, frustrating my elder siblings and cousins; I started out boxing at 11 years old. I added soccer and martial arts in middle and high school respectively, continuing to explore my love of movement. I found expression for my own darkness with combat martial arts in the ring and on the street. And somewhere during that time I fell in love with modern dance. I studied for several years at George Mason University, but in the end moved into a more predictable career path.

This recent NY Times review about Mikhail Baryshnikov’s coaching of dancers in a role of The Dreamer in “Opus 19” written for him by Jerome Robbins, had me re-thinking my own journey thru movement:

The quality Robbins was after — here and in other ballets — connects with a dancer’s way of marking movement, or executing the steps halfway so that a performance is not presentational, but human.

[youtube https://www.youtube.com/watch?v=NP37HntlhTA]

Watching Taylor Stanley dance, the beauty of his movement and the emotional depth of the choreography covers up an amazing athleticism. What he is able to do with his body, the control, the strength…

It had always seemed to me that play was at the heart of movement. Now I would expand that understanding to include healing.

We move thru the world carrying dark and light in varying ratios, depending on life circumstances and choices we make. Of late I’ve found ways to immerse myself in my own darkness as a creative act as opposed to an embrace of violence – dancing the blues, or surprisingly, lifting weights. The challenge of pushing beyond my preconceived limits requires some letting go, and the companionship of that darkness is welcome at the edge.

So I’ll leave you with a blues number that has been my partner in varying guises of late and hope it finds you smiling and wanting to move!

[youtube https://www.youtube.com/watch?v=FjqmcW929to]