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A Weekly Science Blog & Podcast focused on utilizing publicly available research to expand our understanding of entheogens.

  • Ian Bollinger

Salvia divinorum & the unique entheogen: Salvinorin A

https://www.sciencedirect.com/science/article/pii/S1021949814000180?via%3Dihub

Scientists (Po-Xiang Lin et al) investigated a number of Salvia species native to Taiwan to see if they contained the main entheogenic compound salvinorin A typically found in the leaves of S. divinorum. They imported S. divinorum dried leaf, extracts, and a standard of salvinorin A and were unable to detect with High-Performance Liquid Chromatography (HPLC) salvinorin A in endemic Taiwan species.

#HPLC #salvia #salvinorin #terpene




Project Proposal and Author Note

The Scientists recognized a large importing of Salvia divinorum dried leaves and concentrated extracts into the country, and were concerned about whether endemic species of Salvia might contain the main entheogen found in S. divinorum leaves, the terpene salvinorin A. Their proposal can be inferred as: endemic species to Taiwan might become sources for recreational use of the entheogenic terpene. Or as they put it: "The aim of this study was therefore to employ high performance liquid chromatography (HPLC) to detect the presence of salvinorin A in substantial amounts in S. divinorum and endemic species of Salvia in Taiwan."

S. divinorum is more readily accessible than any other entheogen discussed up until now; speaking personally, S. divinorum practice can be a potent experience. Again, the Scientists in Taiwan specifically pursued this project due to increases in recreational use of the entheogen. Traditional use of this entheogen has been in place for generations, and there are case-study experts in its use. When it comes to entheogens, it should be "know...not no", says vocal psychonaut Lawrence "Lorenzo" Hagerty curator of the Psychedelic Salon podcast. Treating entheogen use with care, respect, and trust can lead to very positive experiences; the opposite is true when they are avoided.



'Classically Psychoactive' Entheogens

Almost every compound that is documented as producing what is, subjectively, described as 'psychedelic' typically comes from the stimulation of the serotonin 2A (5-HT2A) receptors. These receptors are found largely in the central nervous system (CNS), typically where large amounts of serotonin are found like in areas known to modulate cognitive processes, working memory, and attention. [3,4,5]

The activity of these 'classical psychedelics' is in their similarity in structure to serotonin or dopamine. One structural similarity lies in the ring structures, the six-membered benzene ring of dopamine and the paired five-and-six-membered indole ring of serotonin; these two ring structures are mirrored in mescaline and DMT respectively (Tbl 1). Another notable structural similarity is the amine arm depicted to the right of the ring structure (Tbl 1). These structures play a major role in the lock-and-key mechanism of 5-HT2A receptor binding and subjective psychoactive effects; from normal functioning of the default mode network to the synesthetic experience of tasting sounds.



Unique Properties of Salvia divinorum

Found endemically to to a specific region of the Sierra Mazateca mountains in Oaxaca, Mexico (Fig 1); S. divinorum is still used by the Mazatec, primarily through shamanic ceremonial practice intended for curing or divination [6]. The use of S. divinorum is an experience of strong disassociation while still semi-present and capable of acting; while subjectively experienced in different ways it is an almost inescapable experience when ingested. Traditionally ceremonial practice see leaves consumed by rolling and placing in the cheeks, the leaves would breakdown through saliva and chewing and the terpenes would slowly dissolve through swallowing and tissue absorption. Recreational consumption is typically seen in the form of smoking dried leaves or concentrates, as seen in the samples the researchers chose to utilize in this research (Fig 2).

As previously stated, 'classical' psychedelics, like those discussed last week from the seminal Shulgin Texts: PiHKAL and TiHKAL, due to their similar structures, act on the serotonin receptors in the central nervous system, altering how sensory signals are processed. The main entheogenic compound in S. divinorum is salvinorin A, a terpene (Fig 1) which is quite different from those 'classical psychedelics' in structure due to it is a large interwoven chain of carbon, hydrogen, and oxygen; but lacking the 'classical psychedelic' hallmark nitrogen (Tbl 1). This large terpene complex binds to, also known as an agonist of, kappa-opioid receptors (κ-opioid receptors, KOR). These receptors are widely distributed throughout the (CNS) with high concentrations being found in the prefrontal cortex, the hippocampus and areas centralized around the brains stem and play a major role in sensory perception, consciousness, pain modulation, motor control, and mood [7, 8].




High-Performance Liquid Chromatography (HPLC) Analysis

While previously mentioned in blog posts as a method of chemical analysis, HPLC shares a lot of similarities with Thin-Layer Chromatography (TLC) in principle. For TLC there is the Mobile Phase, the Stationary Phase, and the Post-Run Treatments that define the major steps of the analysis process; with HPLC there is overlap in the use of a Mobile Phase and a Stationary phase, but instead of Post-Run Treatments the sample is then analyzed with a spectrophotometer for activity when exposed to specific light frequencies, sometimes even ranges of frequencies.

The mobile phase is not placed into a chamber, instead it is pumped through a small filament under pressure, the process used to be called High-Pressure Liquid Chromatography due to this distinction. Instead of there being a plate, the Stationary Phase is packed into a column, run in line with the Mobile Phase, which allows some compounds to adhere to the inner pores of the packed column more so than others and thus regulating the speeds at which compounds through the column. As compounds are selectively separated and moving through the Stationary and Mobile phases the compounds are sent through a spetcrophotometer for various forms of spectral analysis from absorbance, reflectance, to fluorescence as a signal.

The whole of HPLC instrumentation is typically vertically stacked as there is a system of Mobile Phase Reservoirs, Pumps, Auto-Sampling Robot, Stationary Phase Chambers, Detector, and sometimes add-ons as well (Fig 3). This signal is then out put as a graph depicting that light reactivity over time (Fig 4). Through the utilization of pure compounds it is possible to identify those same compounds in natural extracts; when those pure target compounds are used in parallel with non-natural ones it is possible to then quantify the target compound (Fig 4). Notice how the use of 4-bromonitrobenzene came out noticeably earlier, ~37min mark, than the larger terpene salvinorin a, ~47min (Fig 4); this is an example of size preventing quick 'movement' of salvinorin A through the mobile phase and stationary phase. The added bonus of HPLC is the small sample size, typically in the 0.1-10uL range, allow for smaller preparations and lower solvent waste relative to TLC. All of these factors come together in the HPLC methodology to create a highly useful and extremely versatile form of chemical analysis that has largely become the staple of industrial and production chemical testing.




Conclusion and Caveats

The final results presented depict interesting insights into the concentrations of salvinorin A in numerous different Salvia species (Tbl 3). Most notable is the fact that those products sold and advertised as 'concentrates' do not have the potency depicted on their labels. This is an important fact to note for anything purchased online, or from a source that is unverified for quality; again "Know...".

Right out of the gate the group notes that the storage conditions of the samples might have impacted observable data; there were no other concerns of note they voice. They do use a limited number of samples (5) in the process of potency determination, which is small for more than a preliminary review; as always deeper data sets help provide better insights. The statistical analysis of potency concentrations provided (mean plus and minus standard deviation) is very applicable, but to be robust requires larger data sets.

While the Scientists did not achieve their proposed goal of identifying native species that might become recreationally abused, they still voice concern over import of S. divinorum dried leaves and extracts. For the record, I disagree with some of the conclusions the Scientists draw between depression and S. divinorum recreational use; I feel the structure of the paper implies that 'abuse' of the entheogen leads to depression. They say "Some reports have indicated that patients using S. divinorum can experience psychotic symptoms and schizophrenia, and one study reported that S. divinorum users had a higher incidence rate of depression than that of nonusers [9]." As a person with depression might seek to alleviate that depression through escape or modulation of mood (through KOR stimulation from salvinorin A) might be a form of personal treatment for, more than a symptom of, S. divinorum use.



Proposal Success or Failure?

There are no Salvia species native to Taiwan that show detectable concentrations of salvinorin A to be utilized as recreational drugs.



Reviewing the data and the conclusions put forth, the hypothesis proposed by the Scientists was supported. The reason for acceptance are quoted below (emphasis and parentheticals mine):


"The results of this study demonstrated that salvinorin A could be detected in dry leaves of S. divinorum and its related concentrated extract products, but not in the dry leaves of endemic Salvia species of Taiwan, including S. arisanensis, S. coccinea, S. hayatana, S. japonica, S. nipponica, S. scapiformis, S. tashiroi, and S. keitaoensis."


"The results indicated that, under the collection time and conditions, Salvia species endemic to Taiwan may not become recreational drugs in Taiwan."


"The results indicated that, under the collection time and conditions, Salvia species endemic to Taiwan may not become recreational drugs in Taiwan."



References

1) Casselman I, Nock CJ, Wohlmuth H, Weatherby RP, Heinrich M. From local to global - Fifty years of research on Salvia divinorum. Journal of Ethnopharmacology February 2015, 151(2): 768-783.

2) Otis B. Pictures of cultivated S. divinorum for spiritual practice. Sacred Garden Community Church December 2020, personal request.

3) Aghajanian GK, Marek GJ. Serotonin, via 5-HT2A receptors, increases EPSCs in layer V pyramidal cells of prefrontal cortex by an asynchronous mode of glutamate release. Journal of Brain Research April 1999, 825(1–2): 161–71.

4) Marek GJ, Wright RA, Gewirtz JC, Schoepp DD. A major role for thalamocortical afferents in serotonergic hallucinogen receptor function in the rat neocortex. Journal of Neuroscience 2001, 105(2): 379–92.

5) Bortolozzi A, Díaz-Mataix L, Scorza MC, Celada P, Artigas F. The activation of 5-HT receptors in prefrontal cortex enhances dopaminergic activity. Journal of Neurochemistry December 2005, 95(6): 1597–607.

6) Valdés, LJ. III; Díaz, JL; Paul, AG. Ethnopharmacology of ska Maria Pastora (Salvia divinorum, Epling and Jativa-M). Journal of Ethnopharmacology May 1983, 7(3): 287–312.

7) Wang YH, Sun JF, Tao YM, Chi ZQ, Liu JG. The role of kappa-opioid receptor activation in mediating antinociception and addiction. Acta Pharmacologica Sinica September 2010, 31(9): 1065–70.

8) Mansour A, Fox CA, Akil H, Watson SJ. Opioid-receptor mRNA expression in the rat CNS: anatomical and functional implications. Trends in Neurosciences January 1995, 18(1): 22–9.

9) L.T. Wu, G.E. Woody, C. Yang, et al. Recent national trends in Salvia divinorum use and substance-use disorders among recent and former Salvia divinorum users compared with nonusers. Substance Abuse Rehabilitation 2011, 53-68.


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