BUFOTENINE Latest Revision: August 16, 25 H N HO N CH 3 CH 3 1. SYNONYMS CFR: Bufotenine CAS #: Base: 487-93-4 Other Names: N,N-dimethylserotonin, 5-hydroxy-N,N-dimethyltryptamine, mappine 2. CHEMICAL AND PHYSICAL DATA 2.1. CHEMICAL DATA Form Chemical Formula Molecular Weight Melting Point ( o C) Base C 12 H 16 N 2 O 24.3 146-147 Oxalate Hydrate C 12 H 16 N 2 O C 2 H 4 O 2 H 2 O 312.32 97-99 2.2. SOLUBILITY Form A C E H M W Base *** *** SS *** FS I Oxalate Hydrate VSS SS I I FS VS A = acetone, C = chloroform, E = ether, H = hexane, M = methanol and W = water, VS = very soluble, FS = freely soluble, S = soluble, PS = sparingly soluble, SS = slightly soluble, VSS = very slightly soluble and I = insoluble Note: Bufotenine is soluble in dilute acids and alkalis. 3. SCREENING TECHNIQUES 3.1. COLOR TESTS REAGENT Marquis COLOR PRODUCED Green-brown
Van Urks Violet to dark purple 3.2. THIN LAYER CHROMATOGRAPHY Visualization Van Urk's reagent COMPOUND RELATIVE R F SYSTEM TLC 4 bufotenine 1. dimethyltryptamine 1.3 3.3. GAS CHROMATOGRAPHY Method BUF-GCS1 Instrument: Column: Carrier gas: Gas chromatograph operated in split mode with FID 5% phenyl/95% methyl silicone 12 m x.2 mm x.33µm film thickness Hydrogen at 1. ml/min Temperatures: Injector: 27 C Detector: 28 C Oven program: 1) 165 C initial temperature for 1. min 2) Ramp to 28 C at 3 C/min 3) Hold final temperature for 3.8 min Injection Parameters: Split Ratio = 6:1, 1 µl injected Samples are to be dissolved in 4:1 chloroform: methanol and filtered. COMPOUND RRT COMPOUND RRT dimethylsulfone.15 caffeine.78 methamphetamine.24 lidocaine.83 ephedrine.35 procaine.95
3,4-MDA.45 psilocin.95 3,4-MDMA.5 bufotenine 1. (3.85 min) benzocaine.52 cocaine 1.1 acetaminophen.63 tetracosane 1.2 phenacetin.64 heroin 2.84 dimethyltryptamine.73 quinine 1.67 3.5. HIGH PERFORMANCE LIQUID CHROMATOGRAPHY Method BUF-LCS1 Instrument: Column: Detector: Flow: High performance liquid chromatograph equipped with diode array Waters Xterra RP18 (4.5 x 15 mm, 3.5 µm) UV, 21 nm 1. ml/min Injection Volume: 5µL Buffer: Mobile Phase:.1% (v/v) Trifluoroacetic acid in water 8% Buffer : 2% Acetonitrile Samples are to be dissolved in methanol and filtered with a.45 micron filter. COMPOUND RRT COMPOUND RRT psilocybin.71 psilocin 1.31 bufotenine 1. (2.32 min) 3.4. CAPILLARY ELECTROPHORESIS 4 Method BUF-CES1 Injection Solvent: 3.75 mm sodium phosphate monobasic ph = 3.2. Standard Solution Preparation: Accurately weigh and prepare a standard solution of bufotenine oxalate hydrate at.5 mg/ml in methanol.
Sonicate the methanol solution for 5 min. Take 1 ml of the methanol solution and add it to 11 ml of injection solvent. Filter with a.45 µm or smaller filter. Sample Preparation: Weight out an appropriate amount of sample into an Erlenmeyer flask. Add enough methanol to cover the sample material and sonicate the sample solution for 1 hour. Filter 2-3 ml of the methanol solution with a.45 µm or smaller filter. Take 1 ml of the filtered methanol solution and add it to 11 ml of injection solvent. Filter with a.45 µm or smaller filter. Target the sample concentration to be approximately the sample concentration of the sample if possible. Mode: Column: Dynamically Coated Capillary Electrophoresis 6.2 cm x 5 µm fused silica capillary (5 cm to detector) Run Buffer: Microsolv DEA custom chiral run buffer ph to 1.8 Detector: Voltage: Temperature: Injection: Run Time: Rinse Time: UV, 222 nm 1 kv 15 C aired cooled 6.2 s hydrodynamic injection of sample at.5 psi 5. s hydrodynamic injection of water at.2 psi 13 min Flush with.1 N NaOH for 1 min Flush with water for 1 min Flush with Microsolv CElixer A for 1 min Flush MicroSolv DEA custom chiral ph to 1.8 for 2 min
COMPOUND RMT COMPOUND RMT psilocin.93 psilocybin 1.94 bufotenine 1. (6.64 min) 4. SEPARATION TECHNIQUES Bufotenine is an indole alkaloid obtained from the seeds and leaves of Piptadenia peregrine and P. macrocarpa. It has also been isolated from the skin glands of toads (Bufo species). The base is typically seen as white crystalline powder and the oxalate hydrate as a lavender to brown powder. The oxalate hydrate is slightly soluble in chloroform and can be separated from chloroform insoluble material for analysis. 5. QUANTITATIVE DATA N/A 6. QUALITATIVE DATA 6.1. ULTRAVIOLET SPECTROPHOTOMETRY SOLVENT MAXIMUM ABSORBANCE (NM) Aqueous Acid 278 See spectra on the following pages for FT-IR, Mass Spectrometry, Nuclear Magnetic Resonance, and Vapor Phase IR. 7. REFERENCES 1) Moffat, A., Osselton, M.D., Widdop, B., Clarke s Analysis of Drugs and Poisons, 3rd Edition, The Pharmaceutical Press, 24. 2) Mill, T., Robertson, J.C., Instrumental Data for Drug Analysis., 2 nd Edition, Elsevier Science Publishing Co, 1987. 3) Budavari, S., The Merck Index, 12 th Edition, Merck and Co., Inc., 1996. 4) Lurie, I.S., et al., Electrophoresis 24, 25, 158-1591. *** = data not available 8. ADDITIONAL RESOURCES Forendex Wikipedia
1587.58 3656.8 3513.37 933.72 154.19 783.77 2947.34 2776.57 148.96 1461.3 Absorbance 117.7 415.76 432.11 474.14 1199.16 794.27 639.96 73.8 1459.21 1584.7 934.82 961.25 761.83 %T 171.95 1373.22 156.98 828.8 86.98 FTIR (ATR, single bounce): Bufotenine Oxalate Hydrate 32 scans, 4 cm -1 resolution 95 9 85 8 75 7 65 6 55 5 45 4 35 3 4 35 3 25 2 15 1 5 Wavenumbers (cm-1) Vapor Phase FTIR: Bufotenine 4 scans, 8 cm -1 resolution, Flow Cell Temperature 26 C.3.25.2.15.1.5 4 37 34 31 28 25 22 19 16 13 1 7 Wavenumber (cm-1)
Relative Abundance Abundance MS: Bufotenine 115 11 15 1 95 9 85 8 75 7 65 6 55 5 45 4 35 3 25 2 15 58 Scan 271 (1.969 min): DKR1.D 1 5 m/ z--> 24 42 146 13 77 13 91 117 16 5 66 171 187 4 5 6 7 8 9 1111213141516171819221 Liquid Chromatography/Mass Spectrometry (ESI): Bufotenine bufotenine2_581773439 #145 RT: 1.98 AV: 1 NL: 1.21E7 F: + c ESI Full ms [ 8.-225.] 1 25.5 9 8 7 6 5 4 3 2 1 14.8 13.45 11. 26.5 122.56 143.1 151.9 11.88 16.22 132.59 196.91 82.83 94.84 12.11 171.38 181.1 29.2 221.82 8 9 1 11 12 13 14 15 16 17 18 19 2 21 22 m/z
Relative Abundance Relative Abundance Liquid Chromatography/Mass Spectrometry (ESI): Bufotenine Source Fragmentation on at 3V bufotenine2_581773439 #144 RT: 1.97 AV: 1 NL: 4.67E6 F: + c ESI sid=3. Full ms [ 8.-225.] 1 16.2 9 8 7 6 5 4 3 2 1 81.8 196.77 18.77 161.23 122.44 142.94 154.89 24.92 11.1 13.4 195.85 99.8 151.93 17.6 25.88 8 9 1 11 12 13 14 15 16 17 18 19 2 21 22 m/z 221.64 Liquid Chromatography/Mass Spectrometry (ESI): Bufotenine MS/MS 25 m/z @ 35% Normalized Collision Energy bufotenine2_581773439 #142 RT: 1.95 AV: 1 NL: 2.37E6 F: + c ESI Full ms2 25.@35. [ 8.-225.] 1 16.13 9 8 7 6 5 4 3 2 1 161.26 134.5 174.71 181.55 197.67 24.66 8 9 1 11 12 13 14 15 16 17 18 19 2 21 22 m/z
6.68 6.71 6.97 7.11 7.18 7.2 3.1 3.31 3.31 3.34 3.36 4.96. 2.88 Relative Abundance Liquid Chromatography/Mass Spectrometry (ESI): Bufotenine MS 3 25 m/z @ 35% Normalized Collision Energy 16 m/z @ 35% Normalized Collision Energy bufotenine2_581773439 #143 RT: 1.96 AV: 1 NL: 2.72E4 F: + c ESI Full ms3 25.@35. 16.@35. [ 5.-225.] 1 16.8 9 8 132.7 7 6 5 4 3 2 1 133.37 6 8 1 12 14 16 18 2 22 m/z NMR (Proton): Bufotenine Oxalate Hydrate 2 mg/ml in CD 3 OD with TMS, 4 MHz 14 13 12 11 1 9 8 7 6 5 4 3 2 1-1 -2 Chemical Shift (ppm)
166.78 151.56 125.1 128.8 133.11 13.17 18.95 112.86 113.8 59.1 21.93 43.49 6.71 6.71 6.69 6.68 3.8 3.36 7.2 7.18 6.97 6.97 3.12 3.1 3.34 7.11 3.32 3.31 3.31 Nuclear Magnetic Resonance (Proton): Bufotenine Oxalate Hydrate 7.6 7.5 7.4 7.3 7.2 7.1 7. 6.9 6.8 6.7 6.6 6.5 6.4 6.3 Chemical Shift (ppm) 3.6 3.55 3.5 3.45 3.4 3.35 3.3 3.25 3.2 3.15 3.1 3.5 3. Chemical Shift (ppm) NMR (Carbon): Bufotenine Oxalate Hydrate 2 mg/ml in CD 3 OD with TMS, 4 MHz 24 23 22 21 2 19 18 17 16 15 14 13 12 11 1 9 8 7 6 5 4 3 2 1-1 -2 Chemical Shift (ppm)