BUFOTENINE Latest Revision: August 16, 2005

Home , Psilocin, Psilocybin

H N

CH3 HO N

CH3

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 (oC)

Base C12H16N2O 204.3 146-147

Oxalate Hydrate C12H16N2O·C2H4O2·H2O 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 COLOR PRODUCED Marquis Green-brown

Van Urks Violet to dark purple

3.2. THIN LAYER CHROMATOGRAPHY

Visualization

Van Urk's reagent

RELATIVE RF COMPOUND SYSTEM TLC 4

bufotenine 1.0

dimethyltryptamine 1.3

3.3. GAS CHROMATOGRAPHY

Method BUF-GCS1

Instrument: Gas chromatograph operated in split mode with FID

Column: 5% phenyl/95% methyl silicone 12 m x 0.2 mm x 0.33µm film thickness

Carrier gas: Hydrogen at 1.0 mL/min

Temperatures: Injector: 270°C Detector: 280°C Oven program: 1) 165°C initial temperature for 1.0 min 2) Ramp to 280°C at 30°C/min 3) Hold final temperature for 3.8 min

Injection Parameters: Split Ratio = 60:1, 1 µL injected

Samples are to be dissolved in 4:1 chloroform: methanol and filtered.

COMPOUND RRT COMPOUND RRT dimethylsulfone 0.15 caffeine 0.78 methamphetamine 0.24 lidocaine 0.83 ephedrine 0.35 procaine 0.95

3,4-MDA 0.45 psilocin 0.95

3,4-MDMA 0.50 bufotenine 1.00 (3.85 min) benzocaine 0.52 cocaine 1.10 acetaminophen 0.63 tetracosane 1.20 phenacetin 0.64 heroin 2.84 dimethyltryptamine 0.73 quinine 1.67

3.5. HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

Method BUF-LCS1

Instrument: High performance liquid chromatograph equipped with diode array

Column: Waters Xterra RP18 (4.5 x 150 mm, 3.5 µm)

Detector: UV, 210 nm

Flow: 1.0 mL/min

Injection Volume: 5µL

Buffer: 0.1% (v/v) Trifluoroacetic acid in water

Mobile Phase: 80% Buffer : 20% Acetonitrile

Samples are to be dissolved in methanol and filtered with a 0.45 micron filter.

COMPOUND RRT COMPOUND RRT psilocybin 0.71 psilocin 1.31 bufotenine 1.00 (2.32 min)

3.4. CAPILLARY ELECTROPHORESIS4

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 0.5 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 0.45 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 0.45 µm or smaller filter. Take 1 mL of the filtered methanol solution and add it to 11 mL of injection solvent. Filter with a 0.45 µm or smaller filter. Target the sample concentration to be approximately the sample concentration of the sample if possible.

Mode: Dynamically Coated Capillary Electrophoresis

Column: 60.2 cm x 50 µm fused silica capillary (50 cm to detector)

Run Buffer: Microsolv DEA custom chiral run buffer pH to 1.8

Detector: UV, 222 nm

Voltage: 10 kV

Temperature: 15°C aired cooled

Injection: 6.2 s hydrodynamic injection of sample at 0.5 psi 5.0 s hydrodynamic injection of water at 0.2 psi

Run Time: 13 min

Rinse Time: Flush with 0.1 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 0.93 psilocybin 1.94 bufotenine 1.00 (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, 2004. 2) Mill, T., Robertson, J.C., Instrumental Data for Drug Analysis., 2nd Edition, Elsevier Science Publishing Co, 1987. 3) Budavari, S., The Merck Index, 12th Edition, Merck and Co., Inc., 1996. 4) Lurie, I.S., et al., Electrophoresis 2004, 25, 1580-1591. *** = data not available

8. ADDITIONAL RESOURCES

Forendex

Wikipedia

FTIR (ATR, single bounce): Bufotenine Oxalate Hydrate 32 scans, 4 cm-1 resolution

828.08

%T 1056.98

60 860.98

1701.95 1373.22

934.82

1584.07

961.25 761.83

50 1459.21

794.27 703.80

40 639.96

35 1199.16

415.76 474.14

4000 3500 3000 2500 2000 1500 1000 500 432.11 Wavenumbers (cm-1)

Vapor Phase FTIR: Bufotenine 4 scans, 8 cm-1 resolution, Flow Cell Temperature 260°C

0.03

1170.07 0.025

0.02

0.015

Absorbance

2776.57

2947.34

1480.96 1461.30

0.01

783.77

3656.8

1054.19

3513.37 933.72

0.005 1587.58

0 4000 3700 3400 3100 2800 2500 2200 1900 1600 1300 1000 700 Wavenumber (cm-1)

MS: Bufotenine Abundance

Scan 271 (1.969 min): DKR001.D 1150000 58

1100000 1050000

1000000

950000 900000

850000 800000

750000

700000 650000

600000

550000 500000

450000 400000 350000

300000 250000 200000

150000 100000

50000 42 146 204 91 50 66 77 103 117 130 160 171 187 0 40 50 60 70 80 90 100110120130140150160170180190200210 m/ z-->

Liquid Chromatography/Mass Spectrometry (ESI): Bufotenine bufotenine2_050817073439 #145 RT: 1.98 AV: 1 NL: 1.21E7 F: + c ESI Full ms [ 80.00-225.00] 205.05 100

40 RelativeAbundance

30 130.45 104.80 20 110.00 206.05 122.56 143.01 10 151.90 110.88 160.22 132.59 196.91 82.83 94.84 102.11 171.38 181.01 209.02 221.82 0 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 m/z

Liquid Chromatography/Mass Spectrometry (ESI): Bufotenine Source Fragmentation on at 30V bufotenine2_050817073439 #144 RT: 1.97 AV: 1 NL: 4.67E6 F: + c ESI sid=30.00 Full ms [ 80.00-225.00] 160.20 100

40 RelativeAbundance

20 196.77 221.64 180.77 10 161.23 122.44 142.94 154.89 204.92 130.40 195.85 99.08 110.01 81.08 151.93 170.06 205.88 0 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 m/z

Liquid Chromatography/Mass Spectrometry (ESI): Bufotenine MS/MS 205 m/z @ 35% Normalized Collision Energy bufotenine2_050817073439 #142 RT: 1.95 AV: 1 NL: 2.37E6 F: + c ESI Full ms2 [email protected] [ 80.00-225.00] 160.13 100

40 RelativeAbundance

10 161.26 134.05 174.71 181.55 197.67 204.66 0 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 m/z

Liquid Chromatography/Mass Spectrometry (ESI): Bufotenine MS3 205 m/z @ 35% Normalized Collision Energy 160 m/z @ 35% Normalized Collision Energy bufotenine2_050817073439 #143 RT: 1.96 AV: 1 NL: 2.72E4 F: + c ESI Full ms3 [email protected] [email protected] [ 50.00-225.00] 160.08 100

132.07 80

40 RelativeAbundance

133.37 10

0 60 80 100 120 140 160 180 200 220 m/z

NMR (Proton): Bufotenine Oxalate Hydrate

20 mg/mL in CD3OD with TMS, 400 MHz

2.88

0.00

4.96

7.11

3.31

6.97

7.18

7.20

3.34

3.10

3.36

6.71 6.68

14 13 12 11 10 9 8 7 6 5 4 3 2 1 0 -1 -2 Chemical Shift (ppm)

Nuclear Magnetic Resonance (Proton): Bufotenine Oxalate Hydrate

3.31

3.32

7.11

3.34

3.10

6.97

7.18

3.12

6.97

7.20

3.36

3.08

6.71

6.68 6.69

7.6 7.5 7.4 7.3 7.2 7.1 7.0 6.9 6.8 6.7 6.6 6.5 6.4 6.3 3.60 3.55 3.50 3.45 3.40 3.35 3.30 3.25 3.20 3.15 3.10 3.05 3.00 Chemical Shift (ppm) Chemical Shift (ppm)

NMR (Carbon): Bufotenine Oxalate Hydrate

20 mg/mL in CD3OD with TMS, 400 MHz

43.49

103.17

113.08

21.93

112.86

59.10

125.01

151.56

166.78

108.95

128.80 133.11

240 230 220 210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 -10 -20 Chemical Shift (ppm)