DETERMINATION ON WHETHER A SUBSTANCE IS A HAZARDOUS SUBSTANCE PURSUANT TO SECTION 26 OF THE HSNO ACT 29 October 2009 Application Code ERMA200009 Application Type To determine whether a substance is hazardous under Section 26 of the Hazardous Substances and New Organisms Act 1996 (“the Act”).
Applicant Ministry of Health
Date Application Received 24 July 2009
Consideration Date 29 October 2009 Considered by A Committee of the Authority (“the Committee”) Purpose of the Application To determine whether preparations of Salvia Divinorum are hazardous.
1 Application process
1.1 The Agency received an application requesting a formal determination on whether preparations of Salvia Divinorum are hazardous substances under section 26 of the HSNO Act.
1.2 The application was formally received on 24 July 2009.
1.3 The evaluation of the application was undertaken by the ERMA New Zealand project team (“the Agency”) which comprised the following staff members: Matthew Allen Advisor (Hazardous Substances) Tonderai Kaitano Advisor (Hazardous Substances)
1.4 The report was reviewed and signed out by: Lynne Waterson Applications Manager (Hazardous Substances).
1.5 No external experts were used in the consideration of this application.
1.6 The following members of the Authority considered the application: Helen Atkins (Chair) and Shaun Ogilvie. 1.7 The information available to the Committee comprised: the application; and this report. 2 Background to the application Purpose of the application 2.1 To determine whether preparations of Salvia Divinorum (i.e. preparations of Salvia Divinorum that contain the substance salvinorin-A) are hazardous.
Identification of Salvinorin-A 2.2 Salvia Divinorum is a perennial herb of the mint family native to Mexico which is now also grown in many countries (Drug Enforcement Administration, 2008). It has gained popularity as a widely-available substance of abuse (Butelman et al., 2009). 2.3 Salvia Divinorum produces several closely related neoclerodane diterpenes, with salvinorin A being the psychoactive compound that can be extracted from Salvia Divinorum. 2.4 Salvinorin A is a potent naturally occurring hallucinogen and has been described as “the most potent naturally occurring hallucinogen known, rivalling the synthetic hallucinogen lysergic acid diethylamide in potency” (Sheffler and Roth, 2003). 2.5 Fresh leaves of this plant may be chewed, prepared as an aqueous infusion (possibly to be inhaled or injected), or dried and smoked (Valdes, 1994). 2.6 The structural formula of and general information on salvinorin-A can be found below in Table 2.1 and Figure 1. Table 2.1: Identification of Salvinorin-A. CAS number: 83729-01-5 IUPAC name 2H-Naphtho(2,1-c)pyran-7-carboxylic acid, 9- (acetyloxy)-2- (3-furanyl)dodecahydro-6a,10b- dimethyl-4,10-dioxo-, methyl ester, (2S- (2alpha,4aalpha,6abeta,7beta,9beta,10aalpha,10bbeta))- Common name Salvinorin-A Synonyms Divinorin A Salvinorin
Molecular formula C23H28O8 Molecular weight 432.47 Known uses Used as an illicit hallucinogen. Being investigated for use in treatment of certain psychiatric disorders such as schizophrenia and Alzheimer‟s disease, as well as various mood disorders (Vortherms and Roth, 2006). HSNO classification 6.9A
ERMA New Zealand Section 26 Determination - ERMA200009 Page 2 of 19
Figure 1: Structural formula of Salvinorin-A
3 Hazardous property assessment Hazard classification 3.1 The Agency has based its assessment of preparations of Salvia Divinorum on the ingredient salvinorin-A, and has not taken into consideration the toxicity of possible additional components e.g. solvents, co-formulants in preparations.
3.2 The Agency does not have information on amounts likely to be present in preparations and thus our classifications are based on whether there is sufficient evidence to suggest that salvinorin-A has the intrinsic potential to trigger any of the HSNO least degrees of hazard. The Agency notes that salvinorin-A is found in the leaves of Salvia Divinorum at concentrations ranging from 0.89 to 3.7 mg/g and up to about 0.15 mg/g in the stems (Siebert, 2004).
3.3 Using information contained in the referenced literature (Appendix 1), the Agency has classified Salvinorin-A (refer to Appendix 2). The summary of this classification is detailed in Table 3.1:
Table 3.1: Summary of the Agency’s classifications of Salvinorin-A
Hazardous Property Agency’s Classificati Component(s) Classification on Method driving classification Explosiveness No data# N/A No data Flammability No data# N/A No data Physical Oxidising No data# N/A No data Metal corrosive No data# N/A No data 6.1 oral No data# N/A No data 6.1 dermal No data# N/A No data 6.1 inhalation No data# N/A No data Toxicity 6.3/8.2 Skin irritation/corrosion No data# N/A No data 6.4/8.2 Eye irritation/corrosion No data# N/A No data 6.5 Respiratory sensitization No data# N/A No data
ERMA New Zealand Section 26 Determination - ERMA200009 Page 3 of 19 Hazardous Property Agency’s Classificati Component(s) Classification on Method driving classification 6.5 Contact sensitisation No data# N/A No data 6.6 Mutagenicity No data# N/A No data 6.7 Carcinogenicity No data# N/A No data 6.8 Reproductive developmental toxicity No data# N/A No data 6.9 Target organ systemic toxicity 6.9A Test data Salvinorin A 9.1 Aquatic ecotoxicity No data# N/A No data Aquatic Persistence No data# N/A No data Bioaccumulative No data# N/A No data Ecotoxicity 9.2 Soil ecotoxicity No data# N/A No data Soil Persistence No data# N/A No data 9.3 Terrestrial vertebrate ecotoxicity No data# N/A No data 9.4 Terrestrial invertebrate ecotoxicity No data# N/A No data
# No data represents insufficient data or lack of data for this endpoint.
3.4 A summary of the hazardous property assessment is provided below. Refer to Appendix 2 for further information.
Class 1: Substances with Explosive Properties 3.5 No information was located to determine whether salvinorin-A is explosive. It is therefore considered there is no information to classify salvinorin-A for explosivity.
Classes 2-4: Flammability 3.6 No information was located to determine whether salvinorin-A is flammable. It is therefore considered there is no information to classify salvinorin-A for flammability.
Class 5: Oxidisers/Organic Peroxides 3.7 No information was located to determine whether salvinorin-A has oxidising properties. It is therefore considered there is no information to classify salvinorin-A for oxidising properties.
Sub-class 8.1: Metal Corrosiveness 3.8 No information was located to determine whether salvinorin-A is corrosive to metals. It is therefore considered there is no information to classify salvinorin-A for corrosivity to metals.
ERMA New Zealand Section 26 Determination - ERMA200009 Page 4 of 19 Class 6: Toxicity Sub-class 6.1 – Acute toxicity 3.9 No information was located to determine whether salvinorin-A is an acute oral, dermal or inhalation toxicant (6.1). It is therefore considered there is no information to classify salvinorin-A for acute toxicity.
Sub-class 6.3 – Skin Irritation 3.10 No information was located to determine whether salvinorin-A is a skin irritant (6.3A). It is therefore considered there is no information to classify salvinorin-A for skin irritancy.
Sub-class 6.4 – Eye Irritation 3.11 No information was located to determine whether salvinorin-A is an eye irritant (6.4A). It is therefore considered there is no information to classify salvinorin-A for eye irritancy.
Sub-class 6.5 – Respiratory (6.5A) and Contact (6.5B) Sensitisation 3.12 No information was located to determine whether salvinorin-A is a respiratory sensitiser (6.5A). It is therefore considered there is no information to classify salvinorin-A for respiratory sensitisation. 3.13 No information was located to determine whether salvinorin-A is a contact sensitiser (6.5B). It is therefore considered there is no information to classify salvinorin-A for contact sensitisation.
Sub-class 6.6 – Mutagenicity 3.14 No information was located to determine whether salvinorin-A is a mutagenic toxicant (6.6). It is therefore considered there is no information to classify salvinorin-A for mutagenicity.
Sub-class 6.7 – Carcinogenicity 3.15 No information was located to determine whether salvinorin-A is a carcinogenic toxicant (6.7). It is therefore considered there is no information to classify salvinorin-A for carcinogenicity.
Sub-class 6.8 – Reproductive/Developmental Toxicity 3.16 No information was located to determine whether salvinorin-A is a reproductive or developmental toxicant (6.8). It is therefore considered there is no information to classify salvinorin-A for Reproductive/Developmental Toxicity.
Sub-class 6.9 – Target Organ Toxicity
ERMA New Zealand Section 26 Determination - ERMA200009 Page 5 of 19 3.17 A number of literature studies and reviews have been sourced investigating the effects of preparations of Salvia Divinorum and salvinorin-A: 3.17.1 Salvinorin-A targets the central nervous system in non-human primates, consistent with observations in humans; 3.17.2 Very low doses of salvinorin-A exert target organ effects in non-human primates, supporting a classification of 6.9A, rather than 6.9B; 3.17.3 A decrease in locomotor activity over a 30 minute time period was observed in rats for salvinorin-A administered by intraperitoneal injection. Although the exposure route is not relevant to humans, these findings are consistent with observations in humans and are supportive of a 6.9A classification;
3.17.4 Salvinorin-A targets specific receptors (κ1-opioidreceptor and κ2-opioidreceptor (weak interaction)), supporting classification of salvinorin-A as a target organ toxicant; 3.17.5 Information contained in a US Drug Enforcement Agency review indicates that absorption of salvinorin-A across the oral mucosa, or smoking pure salvinorin-A affects the central nervous system, causing psychic effects and adverse physical effects, supporting a target organ toxicant classification (6.9A), specifically affecting the central nervous system. 3.18 The Agency considers that the findings of all the studies reviewed support 6.9A classification of salvinorin-A, by the inhalation route and when leaves are chewed and retained in the mouth.
Summary of the Class 6 Toxicity classifications of salvinorin-A and preparations of Salvia Divinorum 3.19 In summary, based on the available information, the Agency considers that Salvinorin-A, should be classified as a target organ toxicant, 6.9A. Based on the potency of salvinorum-A, the Agency considers that preparations from Salvia Divinorum should also be classified as a target organ toxicant, 6.9A, provided they contain salvinorum-A.
Class 9: Ecotoxicity and environmental fate Sub-class 9.1 – Aquatic ecotoxicity, fate and degradation 3.20 No information was located to determine whether salvinorin-A is an aquatic ecotoxicant (9.1). It is therefore considered there is no information to classify salvinorin-A for aquatic ecotoxicity. 3.21 No information was located to determine whether salvinorin-A is persistent or bioaccumulative in the aquatic environment.
Sub-class 9.2 – Soil ecotoxicity and terrestrial fate 3.22 No information was located to determine whether salvinorin-A is a soil ecotoxicant (9.2). It is therefore considered there is no information to classify salvinorin-A for soil ecotoxicity.
ERMA New Zealand Section 26 Determination - ERMA200009 Page 6 of 19 3.23 No information was located to determine whether salvinorin-A is persistent in the soil environment.
Sub-class 9.3 – Terrestrial vertebrate ecotoxicity 3.24 No information was located to determine whether salvinorin-A is toxic to terrestrial vertebrates (9.3). It is therefore considered there is no information to classify salvinorin-A for terrestrial vertebrate ecotoxicity.
Sub-class 9.4 – Terrestrial invertebrate ecotoxicity 3.25 No information was located to determine whether salvinorin-A is toxic to terrestrial invertebrates (9.4). It is therefore considered there is no information to classify salvinorin-A for terrestrial invertebrate ecotoxicity.
Summary of the Class 9 Ecotoxicity classifications of salvinorin-A and preparations of Salvia Divinorum 3.26 The Agency considers that there is no information to classify salvinorin-A or preparations of Salvia Divinorum as ecotoxic substances.
ERMA New Zealand Section 26 Determination - ERMA200009 Page 7 of 19 Recommended Determination
If the Authority is of the view that “Preparations from Salvia Divinorum” and salvinorin-A are hazardous under the HSNO Act, then the Authority may: a) determine that “Preparations from Salvia Divinorum” and salvinorin-A as defined are hazardous pursuant to section 26 of the HSNO Act 1996; and b) direct the Chief Executive to arrange for notice of this determination to be placed in the Gazette.
Lynne Waterson, Applications Manager, Hazardous Substances Group Date:
Determination by the Authority
The recommended determination is approved.
Helen Atkins Chair Date: 29 October 2009
ERMA New Zealand Section 26 Determination - ERMA200009 Page 8 of 19 APPENDIX 1: REFERENCES
ERMA New Zealand (2008a) User Guide to HSNO Thresholds and Classifications. ERMA New Zealand, Wellington.
Butelman, E.R., Prisinzano, T.E., Deng, H., Rus, S., Kreek, M.J. (2009). Unconditioned Behavioural Effects of the Powerful -Opioid Hallucinogen Salvinorin A in Nonhuman Primates: Fast Onset and Entry into Cerebrospinal Fluid. Journal of Pharmacology and Experimental Therapeutics. 328: 588-597.
Drug Enforcement Administration (2008). Drugs and Chemicals of Concern: Salvia Divinorum and Salvinorin A. Office of Diversion Control. United States Department of Justice. http://www.deadiversion.usdoj.gov/drugs_concern/Salvia_d/Salvia_d.htm.
Hooker, J.M, Xu, Y., Schiffer, W., Shea, C., Carter, P. and Fowler, J.S. (2008). Pharmacokinetics of the potent hallucinogen, salvinorin A in primates parallels the rapid onset and short duration of effects in humans. Neuroimage. 41: 1044-50.
Butelman, E.R., Mandau, M., Tidgewell, K., Prisinzano, T.E., Yuferov, V. and Kreek, M.J. (2007). Effects of Salvinorin A, a -Opioid Hallucinogen, on a Neuroendocrine Biomarker Assay in Nonhuman Primates with High -Receptor Homology to Humans. The Journal of Pharmacology and Experimental Therapeutics. 320: 300–306.
Klimisch, HJ, Andreae, E, Tillman, U (1997). A systematic approach for evaluating the quality of experimental and ecotoxicological data. Regulatory Toxicology and Pharmacology 25: 1–5.
Ansonoff, M.A., Zhang, J., Czyzyk, T., Rothman, R.B., Stewart, J., Xu, H., Zjwiony, J., Siebert, D.J., Yang, F., Roth, B.L. and Pintar, J.E. (2006). Antinociceptive and Hypothermic Effects of Salvinorin A Are Abolished in a Novel Strain of -Opioid Receptor-1 Knockout Mice. Journal of Pharmacology and Experimental Therapeutics. 318: 641–648.
Carlezon, Jr., W.A., Beguin, C., DiNieri, J.A., Baumann, M.H., Richards, M.R., Todtenkopf, M.S., Rothman, R.B., Ma, Z., Lee, D.Y.-W. and Cohen B.M. (2006). Depressive-Like Effects of the -Opioid Receptor Agonist Salvinorin A on Behavior and Neurochemistry in Rats. The Journal Of Pharmacology and Experimental Therapeutics. 316: 440–447.
Siebert, D.J. (2004). Localization of Salvinorin A and Related Compounds in Glandular Trichomes of the Psychoactive Sage, Salvia Divinorum. Annals of Botany. 93: 763±771.
Sheffler, D.J. and Roth, B.L. (2003). Salvinorin A: the „magic mint‟ hallucinogen finds a molecular target in the kappa opioid receptor. Trends in Pharmacological Sciences. 24: 107- 109.
Valdés, L.J. (1994). Salvia Divinorum and the unique diterpene hallucinogen, Salvinorin (divinorin) A. Journal of Psychoactive Drugs. 26: 277-83.
Salvia Divinorum. Center for Substance Abuse Research, University of Maryland. http://www.cesar.umd.edu/cesar/drugs/Salvia.asp (accessed 25/06/09).
ERMA New Zealand Section 26 Determination - ERMA200009 Page 9 of 19
Vortherms, T.A. and Roth, B.L. (2006). Salvinorin A: From Natural Product to Human Therapeutics. Molecular Interventions. 6: 257-265.
Chavkin, C., Sud, S., Jin, W., Stewart, J., Zjawiony, J.K., Siebert, D.J., Toth, B.A., Hufeisen, S.J. and Roth, B.L. (2004). Salvinorin A, an Active Component of the Hallucinogenic Sage Salvia Divinorum is a Highly Efficacious -Opioid Receptor Agonist: Structural and Functional Considerations. The Journal of Pharmacology and Experimental Therapeutics. 308: 1197-1203.
ERMA New Zealand Section 26 Determination - ERMA200009 Page 10 of 19 APPENDIX 2: HAZARD CLASSIFICATION
Classification of preparations of Salvia Divinorum
The Agency does not have information on amounts likely to be present in preparations and thus our classifications are based on whether there is sufficient evidence to suggest that salvinorin A has the intrinsic potential to trigger any of the HSNO least degrees of hazard. Note that salvinorin A is found in the leaves at concentrations ranging from 0.89 to 3.7 mg/g and up to about 0.15 mg/g in the stems (Siebert, 2004).
Details of the components and the methods used to derive the classifications are presented in Table A2.1. The relevant sections of the User Guide to Thresholds and Classifications under the HSNO Act (ERMA 2008a) that describe the mixture rules are listed in Table A2.2.
The Agency has provided a summary of the toxicity data in Table A2.5.
Data quality – overall evaluation
The Agency has adopted the Klimisch et al (1997) data reliability scoring system for evaluating data used in the hazard classification and risk assessment of chemicals (section 1.2.4 in ERMA 2008a). The data used by The Agency to classify “Preparations of Salvia Divinorum” are predominantly from publications in peer-reviewed journals and also reviews by overseas regulatory agencies.
Table A2.0: Physical and chemical properties of “Preparations of Salvia Divinorum”. Test “Preparations of Salvia Divinorum” Method Reference Appearance The plant Salvia Divinorum belongs to the mint family. It No data Application has hollow square stems, large dark green leaves, and occasional white flowers with purple bracts. When sequentially extracted with hexane and acetone from ground leaves, salvinorin A appears as a white crystalline solid (Carlezon Jr., et al., 2006). Odour No data - - Density at 20°C No data - Surface tension No data - - pH No data - Dynamic viscosity No data - - Flash point No data - - Auto flammability No data - - Explosive properties No data - -
ERMA New Zealand Section 26 Determination - ERMA200009 Page 11 of 19
Table A2.1: Summary of the toxicity and ecotoxicity hazard classifications of “Preparations of Salvia Divinorum”. Hazardous Property Agency’s Classification Classification Component(s) driving Method classification 6.1 oral No data N/A No data 6.1 dermal No data N/A No data 6.1 inhalation No data N/A No data 6.3/8.2 Skin No data N/A No data irritation/corrosion 6.4/8.2 Eye No data N/A No data irritation/corrosion 6.5 Respiratory No data N/A No data sensitization 6.5 Contact No data N/A No data sensitisation 6.6 Mutagenicity No data N/A No data 6.7 Carcinogenicity No data N/A No data 6.8 Reproductive No data N/A No data developmental toxicity 6.9 Target organ 6.9A Test data Salvinorin A systemic toxicity 9.1 Aquatic ecotoxicity No data N/A No data Aquatic Persistence No data N/A No data Bioaccumulative No data N/A No data 9.2 Soil ecotoxicity No data N/A No data Soil Persistence No data N/A No data 9.3 Terrestrial No data N/A No data vertebrate ecotoxicity 9.4 Terrestrial No data N/A No data invertebrate ecotoxicity ND= no data, NA= not applicable
Table A2.2: Location of mixture rules within the HSNO Thresholds and Classifications User Guide (V2.0. March 2008). Hazard User Guide to HSNO Thresholds and Classifications Reference Subclass 6.1 Acute Toxicity Part V, Chapter 10, Page 12 Subclass 6.3/8.2 Skin Irritancy/Corrosivity Part V, Chapter 11, Page 7 Subclass 6.4/8.3 Eye Irritancy/Corrosivity Part V, Chapter 12, Page 9 Subclass 6.5 Contact and Respiratory Sensitisation Part V, Chapter 13, Page 8 Subclass 6.6 Mutagenicity Part V, Chapter 14, Page 5 Subclass 6.7 Carcinogenicity Part V, Chapter 15, Page 8 Subclass 6.8 Reproductive Developmental Toxicity Part V, Chapter 16, Page 11 Subclass 6.9 Target Organ Systemic Toxicity Part V, Chapter 17, Page 10 Subclass 9.1 Aquatic Ecotoxicity Part VI, Chapter 19, Page 18 Subclass 9.2 Soil Ecotoxicity Part VI, Chapter 20, Page 8 Subclass 9.3 Terrestrial Vertebrate Ecotoxicity Part VI, Chapter 21, Page 7 Subclass 9.4 Terrestrial Invertebrate Ecotoxicity Part VI, Chapter 22, Page 5
ERMA New Zealand Section 26 Determination - ERMA200009 Page 12 of 19 Identity of the Active Ingredient The active component in preparations of Salvia Divinorum is salvinorin A. Information available on this component is summarised in the following table.
Table A2.3: Identification of salvinorin A. CAS number: 83729-01-5 IUPAC name 2H-Naphtho(2,1-c)pyran-7-carboxylic acid, 9-(acetyloxy)-2- (3- furanyl)dodecahydro-6a,10b-dimethyl-4,10-dioxo-, methyl ester, (2S- (2alpha,4aalpha,6abeta,7beta,9beta,10aalpha,10bbeta))- Common name Salvinorin A Synonyms Divinorin A Salvinorin Molecular formula C23H28O8 Molecular weight 432.47 Structural formula and Significant impurities: S. Divinorum produces several closely related neoclerodane diterpenes as shown opposite (Siebert, 2004). However, salvinorin A is responsible for the psychoactive effects.
Known uses Used as an illicit hallucinogen. Being investigated for use in treatment of certain psychiatric disorders such as schizophrenia and Alzheimer‟s disease, as well as various mood disorders (Vortherms and Roth, 2006). HSNO classification 6.9A
ERMA New Zealand Section 26 Determination - ERMA200009 Page 13 of 19 Biological Hazards: Class 6 Toxicity
Table A2.5: Summary of toxicity data on Salvia Divinorum and Salvinorin A. Acute Toxicity Studies
Type of Study: Acute effects of salvinorin A
Species: unspecified non-human primates
Test Guidelines: None
Study Summary: The study focused on the characterization of overt effects of salvinorin A, such as sedation (operationally defined as unresponsiveness to environmental stimuli) and postural relaxation, previously observed with centrally penetrating -agonists in nonhuman primates. Salvinorin A was administered intravenously at doses of 0.01–0.1 mg/kg. At 0.032 mg/kg, salvinorin A could enter the central nervous system (as reflected in cisternal cerebrospinal fluid) within 1 minute, inducing effects such as facial relaxation and ptosis.
Reference Source: Butelman, E.R., Prisinzano, T.E., Deng, H., Rus, S., Jeanne, Kreek M.J. (2009). Unconditioned Behavioral Effects of the Powerful -Opioid Hallucinogen Salvinorin A in Nonhuman Primates: Fast Onset and Entry into Cerebrospinal Fluid. Journal of Pharmacology and Experimental Therapeutics. 328: 588-597.
Comments The findings from this study provide evidence that the test material triggers 6.9 target organ classification. Although no detail was available relating to the exact test species, the material used and the experimental procedure, the reported findings can be relied upon to support a 6.9 classification. The article is published in a recognised scientific journal (of the American Society for Pharmacology and Experimental Therapeutics). Furthermore, the findings are consistent with observations reported in individuals taking salvinorin A (Chavkin et al., 2004).
Target Organ Toxicity
Type of Study: Effects of salvinorin A on behaviour and neurochemistry in rats
Test Species: 129 male Sprague-Dawley rats.
Test Substance: Salvinorin A, extracted from dried S. Divinorum leaves using hexane and acetone then evaporated to give the crystalline solid. Determined by authors to be >99% pure.
Dose level: Test substance was dissolved in a mixture of dimethyl sulfoxide (DMSO) with distilled water and administered by intraperitoneal injection. For the Forced Swim Test, dose levels were 0.125–2.0 mg/kg body weight given 1, 19, and 23 h after the first exposure.
Guidelines: Study conducted in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals (National Institutes of Health Publication 85-23, 1996) as well as McLean Hospital and National Institute on Drug Abuse-Intramural Research Program policies.
Tests carried out by the Authors include: Forced Swim Test (FST); Locomotor Activity studies; and In vivo microdialysis studies.
Results 1. There was a statistically significant increase in immobility at 0.25 mg/kg, 0.5 mg/kg, 1.0 mg/kg and 2.0
ERMA New Zealand Section 26 Determination - ERMA200009 Page 14 of 19 mg/kg; 2. Swimming behaviour was significantly decreased at all doses from 0.25 to 2.0 mg/kg; 3. There were no effects on climbing or diving behaviours 4. There were no treatment-related differences in locomotor activity in an open field at any of the doses tested, 5. There was a significant decrease in locomotor activity levels in all groups over the course of the 30-min test period (the Agency has interpreted point 4 above as referring to differences in locomotor activity between different groups, and point 5 to mean that for each treatment group, locomotor activity decreased significantly during the 30-min test period). The increase in immobility behaviour was accompanied by a decrease in swimming behaviours. This specific effect is opposite to that seen with SSRIs, which decrease immobility and increase swimming behaviours; 6. In the in vivo microdialysis studies, injection of 1.0 mg/kg salvinorin A produced rapid decreases in dopamine that were detectable for the entire 2 hour test period, whereas 0.125 mg/kg salvinorin A had no effect at any time point.
Reference Source: Carlezon, Jr., W.A., Beguin, C., DiNieri, J.A., Baumann, M.H., Richards, M.R., Todtenkopf, M.S., Rothman, R.B., Ma, Z., Lee, D.Y.-W. and Cohen B.M. (2006). Depressive-Like Effects of the -Opioid Receptor Agonist Salvinorin A on Behavior and Neurochemistry in Rats. The Journal Of Pharmacology and Experimental Therapeutics. 316: 440–447.
Klimisch Score: 1.
Comments Although the study was not conducted in accordance with OECD guidelines, some national guidelines were followed and the experimental procedure is described in sufficient detail. Although the route of exposure used in this study is not relevant to humans, the results of this study support 6.9A classification of salvinorin A targeting the central nervous system. This is also consistent with observations in humans.
Type of Study: Efficacy and potency of salvinorin A in non-human primates
Test Species: Rhesus monkeys (Macaca mulatta)
Test Substance: Salvinorin A extracted from dried S. Divinorum leaves.
Dose levels: 0.0032–0.056 mg/kg bw
Comment on Method: Serum prolactin levels have been used in nonhuman primates to study the potency, receptor selectivity, and apparent efficacy of -agonists in vivo, and so the authors have used the same approach in this assay.
Results Before intravenous injection of the vehicle (1:1:8 ethanol/Tween 80/sterile water v/v; 0.16 ml/kg), prolactin levels in males were relatively stable, averaging 15.5 ± 4.2 ng/ml. Female subjects in follicular phase had similar preinjection baselines (mean 15.1 ± 3.0 ng/ml); Vehicle administration did not have a significant effect on the prolactin levels. In males: Injection of salvinorin A caused robust dose- and time-dependent increases in prolactin levels; Salvinorin A effects were observable within 5 minutes of injection, peaking at 15 minutes before subsequently declining gradually over a 120-minute interval. All treated animals except for the lowest dose exhibited statistically significant difference in prolactin levels at 5, 15, and 30 minutes compared with those given the vehicle. Only the highest dose group showed statistically significant difference at 60 min. By 90 and 120 min after salvinorin A administration, no significant differences were detected. Subcutaneous administration of salvinorin A at 0.032 mg/kg in male subjects resulted in much smaller prolactin release and a slower onset than that observed by the intravenous route. The peak effect only observed at 60 minutes post-injection Follicular phase females:
ERMA New Zealand Section 26 Determination - ERMA200009 Page 15 of 19 At 0.0032 mg/kg by intravenous injection salvinorin A produced larger prolactin elevation compared with males; At 0.032 mg/kg salvinorin A by intravenous injection there were even greater prolactin elevation compared with males; When administered by subcutaneous injection, 0.032 mg/kg salvinorin A produced robust prolactin release from 15 minutes after administration, lasting at least 120 minutes.
Authors concluded that salvinorin A is equipotent and equieffective to the synthetic high-efficacy -agonist U69,593. They further concluded that gonadally intact females are more sensitive to salvinorin A than males.
Reference Source: Butelman, E.R., Mandau, M., Tidgewell, K., Prisinzano, T.E., Yuferov, V. and Kreek, M.J. (2007). Effects of Salvinorin A, a -Opioid Hallucinogen, on a Neuroendocrine Biomarker Assay in Nonhuman Primates with High -Receptor Homology to Humans. The Journal of Pharmacology and Experimental Therapeutics. 320: 300–306.
Comments Although no study guidelines were cited by the authors, the experimental procedure was described in sufficient. The findings of this study demonstrate that salvinorin A exerts effects at doses as low as 0.032 mg/kg bw, the extent of the effect varies between sexes and route of administration. This supports the 6.9A classification assigned in the study summarised previous. 6.9A is assigned rather than 6.9B based on the effects being seen at very low doses.
Absorption, Distribution and Metabolism Studies
Type of Study: Pharmacokinetics
Species: 6 female baboons
Test Guidelines: None
Study Summary: Salvinorin A was labelled with C-11 by acylation of salvinorin B with [11C]-acetyl chloride to study its kinetic behaviour in the brain. This labelled salvinorin A was administered to 6 female baboons. Positron emission tomography (PET) studies indicated rapid brain uptake reaching a peak accounting for 3.3% of the total administered dose in 40 seconds and clearing with a half-life of 8 minutes. Authors reported distribution of labelled salvinorin A throughout the brain, with the highest concentrations in the cerebellum and to a lesser extent, the visual cortex. Regarding metabolism and excretion, authors suggested at least two modes of metabolism and excretion involving the renal and biliary systems. Authors concluded that the exceptionally rapid uptake and brief duration of salvinorin A in the brain match the time-course of visual hallucinations reported for S. Divinorum when smoked by humans. Furthermore, the effects of salvinorin A may occur at <10 μg in the human brain, emphasizing its remarkable potency.
Reference Source: Hooker, J.M., Xu, Y., Schiffer, W., Shea, C., Carter, P., Fowler J.S. (2008). Pharmacokinetics of the potent hallucinogen, salvinorin A in primates parallels the rapid onset and short duration of effects in humans. Neuroimage. 41: 1044-50.
Klimisch Score: 4 (not assignable – only the abstract available). However, the article is published in a recognised, peer-reviewed journal. Therefore, meaningful conclusions can still be drawn from the study regarding the absorption, distribution, metabolism and excretion of salvinorin A.
Comments Information on the effects of administering salvinorin A to the animals was not available. Therefore, this study cannot be used to determine toxicological classification of salvinorin A. However, the suggestion by the authors that effects of this substance may be seen at less than 10 μg in the human brain is consistent with the toxicological profile of xenobiotics that target specific receptors. This also demonstrates the potency of salvinorin A.
Receptor Binding Studies
ERMA New Zealand Section 26 Determination - ERMA200009 Page 16 of 19
Type of Study: Effects of salvinorin A in -Opioid Receptor-1 Knockout (KOR-1 KO) Mice
Test Species: KOR-1 KO mice derived from mating either KOR-1 heterozygous mutant mice maintained on a mixed C57BL6/JX129S6 background or wild-type and KOR-1 mutant mice maintained on an inbred 129S6 background. Subsequent behavioural experiments used wild-type and KOR-1 KO mice of both sexes maintained on the 129S6 background.
Test Guidelines: The study was conducted in accordance with guidelines of the Institutional Care and Use Committees of the University of Medicine and Dentistry of New Jersey, the Robert Wood Johnson Medical School and the National Institute on Drug Abuse, National Institutes of Health in facilities accredited by the American Association for the Accreditation of Laboratory Animal Care.
Test Material: Salvinorin A was isolated from dried leaves of S. Divinorum following the method of Valdes et al (1994).
Results: Salvinorin A was found to selectively bind to the 1-opioid receptor and to weakly interact with the 2- opioid receptor; Following intracerebroventricular injection over a dose range of 1 to 30 µg, salvinorin A produced antinociception (reduced sensitivity to painful stimuli) in wild-type mice but not in a novel strain of - opioid receptor knockout mice. Salvinorin A reduced rectal body temperature in wild type mice, similar to conventional -opioid receptor agonists.
Reference Source: Ansonoff, M.A., Zhang, J., Czyzyk, T., Rothman, R.B., Stewart, J., Xu, H., Zjwiony, J., Siebert, D.J., Yang, F., Roth, B.L. and Pintar, J.E. (2006). Antinociceptive and Hypothermic Effects of Salvinorin A Are Abolished in a Novel Strain of -Opioid Receptor-1 Knockout Mice. Journal of Pharmacology and Experimental Therapeutics. 318: 641–648.
Comments Although the route of exposure used in this study is not relevant to humans as it by-passes the blood-brain barrier, the findings of this study demonstrate that salvinorin A targets a specific receptor andthat CNS-related effects reported in other studies are not just random findings. Thus, this study supports classification of salvinorin A as a target organ toxicant. Review by other Agencies
Type of Study: Review by a US Drug Enforcement agency
Summary: Salvinorin A is found primarily in the leaves, and to a lesser extent in the stems of Salvia Divinorum. When chewed, the leaf mass and juice are maintained within the cheek area where absorption occurs across the lining of the oral mucosa. Effects first appear within 5 to 10 minutes. Smoking pure salvinorin A at a dose of 200-500 μg results in effects within 30 seconds which last for about 30 minutes. Some reported psychic effects include: Perceptions of bright lights, vivid colours and shapes; Body movements and body or object distortions; Dysphoria; Uncontrolled laughter; A sense of loss of body; Overlapping realities; Hallucinations. Adverse physical effects reported include: Lack of coordination; Dizziness; Slurred speech.
Regulatory Status
ERMA New Zealand Section 26 Determination - ERMA200009 Page 17 of 19 Salvinorin A and/or Salvia Divinorum have been placed under regulatory controls in several countries including Australia, Belgium, Denmark, Estonia, Finland, Italy, Japan, Spain, Sweden and several states in the US.
Reference Source: Drugs and Chemicals of Concern: Salvia Divinorum and Salvinorin A (2008). US Department of Justice. Drug Enforcement Administration, Office of Diversion Control. http://www.deadiversion.usdoj.gov/drugs_concern/Salvia_d/Salvia_d.htm.
Comments Although this review by the US Drug Enforcement Agency is a summary of information obtained from various sources, it provides fairly reliable information indicating that salvinorin A is a target organ toxicant, specifically affecting the central nervous system. This is consistent with the toxicological properties of salvinorin A inferred from other sources as well. Taken together, these reported findings provide evidence that preparations of Salvia Divinorum should be classified as target organ toxicants. The fact that these findings were reported in humans makes a case for 6.9A classification.
Conclusion on Classification Preparations of Salvia Divinorum trigger 6.9A classification, targeting the central nervous system.
Type of Study: Review by the Center for Substance Abuse Research, University of Maryland
The review identified the following as some of the effects of salvinorin A: Intense hallucinations, such as sensations of lightness of the body, twisting, spinning, floating, flying, traveling through time and space; Effects that occur only when the eyes are closed, including visual hallucinations of various patterns and shapes; Dizziness; Nausea; Lack of coordination; Slurred speech and awkward sentence patterns; Decreased heart rate; Chills.
Reference Source: Salvia Divinorum. Center for Substance Abuse Research, University of Maryland. http://www.cesar.umd.edu/cesar/drugs/Salvia.asp (accessed 25/06/09).
Comments: These effects identified above support classification of S. Divinorum as a 6.9 target organ toxicant.
No data was available on ecotoxicity and environmental fate.
ERMA New Zealand Section 26 Determination - ERMA200009 Page 18 of 19 Conclusion The findings of all the studies reviewed support 6.9A classification of preparations of Salvia Divinorum by the inhalation route and when leaves are chewed and retained in the mouth. However, there was no data to determine the systemic toxicity of salvinorin A apart from its effects on the central nervous system and the consequent psychological manifestation. Nevertheless, these psychological effects may impair an individual‟s judgment, making the user more susceptible to accidents and injury.
ERMA New Zealand Section 26 Determination - ERMA200009 Page 19 of 19