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The Isolation, Identification, and Quantitation of Dimethyltryptamine Jack A. Fasanello* and Andrea D. Placke [Presented in Part at the 33rd Annual NEAFS Meeting, Bolton Landing, NY, ABSTRACT: Dimethyltryptamine (DMT) was extracted from the root bark of Mimosa hostilis via three methods, using methanol (direct or via Soxhlet) and acetic acid (direct only), respectively. The product from the direct methanol extraction was used in both qualitative and quantitative analysis, while the product from the acetic acid extraction (isolated in crystal form after workup) was used for qualitative analysis. FTIR/ATR, GC/MS, GC/IRD, 1H-NMR, and HPLC data are presented. Quantitative analysis by 1H-NMR and HPLC indicated 0.9 percent and 0.8 percent DMT, respectively, in the analyzed samples. KEYWORDS: Mimosa hostilis, Dimethyltryptamine, DMT, Extraction, Analysis, Forensic Chemistry Introduction Tryptamines are substituted indole compounds which are both naturally occurring and synthetically manufactured. Many tryptamines, including dimethyltryptamine (DMT, Figure 1), have hallucinogen properties, and are therefore listed as Schedule I drugs under the U.S. Controlled Substances Act (21 CFR 1308.11). DMT is present in many plants and their seeds, including in Mimosa hostilis and Psychotria viridis [1-3], and can be abused by smoking, injection, or ingestion of either these natural materials or their crude or purified extracts, either alone or in combination with other extracts (e.g., Ayahuasca [4].) Mimosa hostilis and similar natural plant materials are not formally controlled (by name) in the United States; however, they are controlled (Schedule I) if they are shown to contain DMT or other controlled hallucinogens. Despite their controlled status, a number of DMT-containing natural products, including Mimosa hostilis, are openly marketed on the Internet.
Figure 1. Structure of Dimethyltryptamine (DMT; C12H16N2, m.w. = 188.27). Clandestine DMT extraction laboratories are occasionally seized by law enforcement agencies [e.g., 5]. The basis of this report was the seizure of an unknown plant material (Photo 1) at a clandestine MDMA (Ecstasy) laboratory in rural Pennsylvania. GC/MS analysis of a methanolic extraction of the material identified DMT. Upon debriefing, the defendant in the case indicated that material was root bark from Mimosa hostilis. Similar seizures of this material have been made at other clandestine laboratory sites in the United States, and subsequent analyses of those exhibits confirmed that they also contained DMT.
Photo 1. Mimosa hostilis Root Bark Seized at Clandestine Lab in Pennsylvania. Experimental Methanol Extraction: The root bark was cut into small pieces then ground in a blender to produce a very fine powder. For direct extraction, methanol was added to the powder, heated to 60°C with stirring for 1 hour, and then filtered. This step was repeated three more times, except the re-extractions were carried out for only 5 - 10 minutes each. The combined extracts were evaporated to a residue over steam, then reconstituted as needed for analysis. For Soxhlet extraction, the powdered material was placed in an extraction thimble, placed in a Soxhlet, and extracted with 50 mL of methanol for approximately 50 volumes. The solvent was evaporated to a residue over steam, then reconstituted as needed for analysis. Acetic Acid Extraction: The root bark was cut into small pieces then ground in a blender to produce a very fine powder. A 3% acetic acid solution was added to the powder, and the resulting suspension was stirred for approximately two hours. The solution was filtered and transferred to a separatory funnel, made basic with sodium hydroxide, and then extracted with methylene chloride. The methylene chloride solution was isolated, and the aqueous later was re-extracted with a second volume of methylene chloride. The combined extracts were dried over magnesium sulfate, filtered, and evaporated to give a crystalline material. Fourier Transform Infrared with Attenuated Total Reflectance (FTIR/ATR) Gas Chromatograph/Mass Spectrometer (GC/MS) Gas Chromatograph/Infrared Detector (GC/IRD) Proton Nuclear Magnetic Resonance (1H-NMR) High Performance Liquid Chromatography (HPLC) Results and Discussion The extraction of DMT from Mimosa hostilis was completed using two different solvents, methanol (direct or via Soxhlet) and acetic acid (direct only). The methanol extraction gave the maximum recovery of DMT for qualitative and quantitative analysis; however, the extract included other soluble plant impurities. The extraction efficiency using methanol was identical whether done directly or via Soxhlet. The acetic acid extraction gave a very clean, pure product, but in lower yield versus the methanol extraction. FTIR/ATR: The crystals from the acetic acid extraction procedure produced a clean spectrum (Figure 2). GC/MS: DMT eluted at 6.06 minutes using the described method. The spectra showed a base peak at m/z = 58 GC/IRD: DMT eluted at 6.88 minutes using the described method (Figure 5). 1H-NMR (Qualitative): The singlet at 2.35 ppm is due to the two N-methyl groups, the two triplets at 2.70 ppm and 2.95 ppm correspond to the alpha and beta methylene groups. The multiplet at 7.00 ppm corresponds to protons 2, 5, and 6 on the indole. Finally, the two doublets at 7.25 ppm and 7.50 ppm correspond to protons 4 and 7 on the indole. A slight shift was observed in the extract versus a DMT standard; this was due to pH differences (the spectrum was obtained from DMT acquired using the acetic acid extraction procedure, which involved an acid base workup). (Figure 6). (Qualitative): Using the direct methanol extract, DMT was determined to be 0.9% weight/weight in Mimosa hostilis (Figure 7 and Table 1). Using the direct methanol extract, DMT was determined to be 0.9% weight/weight in Mimosa hostilis (Figure 8 and Table 2). HPLC: DMT eluted in under 3 minutes. Using the methanol extract, DMT was determined to be 0.8% weight/weight in Mimosa hostilis (Figure 9 and Table 3). Acknowledgments The authors would like to thank the Laboratory Director Thomas Blackwell, Supervisory Chemists Christopher Guglielmo and Ann Marie O’Neill, Senior Forensic Chemist Michelle Camilleri, and Forensic Chemists Christopher Benintendo and Ken Fuentecilla (all of this laboratory), and Senior Forensic Chemist Patrick Hays (DEA Special Testing and Research Laboratory, Dulles, VA). References 1. Duke JA, Vásquez, R. Amazonian Ethnobotanical Dictionary. CRC Press, Boca Raton, FL:1994. 2. Schultes RE, Hoffmann A. The Botany and Chemistry of Hallucinogens, 2nd ed., Charles C. Thomas, Springfield, IL:1980. 3. Blackledge RD, Taylor CM. Psychotria viridis - A botanical source of dimethyltryptamine. Microgram Journal 2003;1(1-2):18-22. 4. Casale JF, Koles JE. Analysis of ayahuasca (“Santo Daime”). Microgram 1995;28(9):296.* 5. Anonymous. Clandestine dimethyltryptamine (DMT) laboratory seized in Hollywood, California. Microgram Bulletin 2007;40(7):65-6. * Law Enforcement Restricted Publication.
Figure 2. FTIR/ATR of a DMT Standard (Top Trace) and DMT from the Acetic Acid Extraction - - - - - - - - - -
Figure 3. GC/MS Total Ion Chromatogram of DMT (Methanol Extract).
Figure 4. GC/MS Data of DMT (Methanol Extract). [Note: Molecular Ion at m/z = 188.]
Figure 5. GC/IRD Data of DMT (Methanol Extract). [Note: DMT eluted at 6.88 Minutes.] - - - - - - - - - -
Figure 6a. Full-Scale NMR Data of DMT (Acetic Acid Extract). - - - - - - - - - -
Figure 6b. Expanded Spectrum from 2 to 4 ppm. See Results and Discussion for Peak Assignments.
Figure 6c. Expanded Spectrum from 6.5 to 7.6 ppm. See Results and Discussion - - - - - - - - - -
Figure 7a. NMR Quantitation of DMT (Direct Methanol Extract); See Table 1.
Figure 7b. NMR Quantitation of DMT (Direct Methanol Extract); Expansion; See Table 1. - - - - - - - - - -
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Figure 8a. NMR Quantitation of DMT (Methanol - Soxhlet Extract); See Table 2.
Figure 8b. NMR Quantitation of DMT (Methanol - Soxhlet Extract); Expansion; See Table 2.
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Figure 9. HPLC Quantitation of DMT (Methanol Extract); See Table 2.
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