1. CHEMICAL CHARACTERISATION OF AMINOINDAN(E) PRODUCTS PURCHASED FROM THE INTERNET JACQUELINE L. STAIR SENIOR LECTURER IN ANALYTICAL CHEMISTRY CENTRE FOR CLINICAL PRACTICE, SAFE MEDICINES AND DRUG MISUSE RESEARCH

2. AMINOINDANES

3. Full chemical characterisation of the pure substances by Casale et al. (2011; 2012) Products often characterised as part of a diverse group 19 products were analysed by Baron et al. (2012) Primarily GC-MS analysis

4. Assi, S.; Fergus, S.; Halsey, S.; Stair, J.L. “On-spot identification of so-called ‘legal highs’ using handheld Raman spectroscopy”. The Ever-Changing World of Psychoactive Drugs, Budapest, March 2012.

5. OVERVIEW METHOD 1: Identification of volatile molecular constituents using Gas Chromatography Mass Spectrometry (GC-MS) METHOD 2: Determination of non-volatile molecular constituents and quantification using Liquid Chromatography Photo Diode Array (LC-PDA) METHOD 3: Identification and quantification of elemental constituents using Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES)

6. AMINOINDANE PRODUCTS

7. METHOD 1: VOLATILE MOLECULAR CONSTITUENTS 0.1 mg/ml solutions prepared in methanol Analysed using Varian 450 GC/240 MS Varian FactorFour 5% phenyl-methyl capillary column (30m × 0.25mm × 0.25µm) GC Temperature Programme

8. Findings: 6 products matched their label claim Only 1-2 substances were detected No 5-IAI was detected 5 samples contained 2-AI Findings: 6 products matched their label claim Only 1-2 substances were detected No 5-IAI was detected 5 samples contained 2-AI methylphenidate methiopropamine (MPA) Label:MDAI MPA Label:MDAI MPA Label:Aminoindan Caffeine Cola Vera Aminoacid Complex Label:Aminoindan Caffeine Cola Vera Aminoacid Complex

9. METHOD 2: NON-VOLATILE MOLECULAR CONSTITUENTS 0.5 mg/ml solutions prepared in methanol:water (50/50 v/v) Analysed using Perkin Elmer Flexar with PDA (λ=210 nm, FR=1.5 ml min -1 ) Phenomenex core Kinetex 5µ XB C18 column (150 × 4.6 mm) HPLC Gradient Method

10. LC METHOD DEVELOPMENT Chromatogram of 2-AI, 5-IAI, MDAI, & CAF (0.5 mg/ml) using Kinetex XB-C18. 2-AI 5-IAI MDAI CAF

11. ProductLabel GC-MS identification HPLC %w/w of Sample 12-AI Methyl phenidate X Caffeine25.8 ± 0.3 22-AI 66.9 ± 0.5 35-IAI MDAI22.4 ± 0.8 Caffeine23 ± 2 45-IAI2-AI50.5 ± 0.3 5MDAI 32.6 ± 0.5 6MDAI2-AI75.7 ± 0.7 7MDAI 84 ± 2 8MDAI-- 9Pink Panther MDAI18.9 ± 0.9 MPA72.0 ± 0.3 10 Pink Champagne 2-AI10.9 ± 0.4 Caffeine11.6 ± 0.6 11Blurrberry 2-AI 29 ± 2 Caffeine14 ± 2 Findings: HPLC work confirmed number of constituents found by GC-MS. Aminoindanes: 11-84 %w/w Caffeine: 12-25 %w/w Results indicate inorganic additives may be present Findings: HPLC work confirmed number of constituents found by GC-MS. Aminoindanes: 11-84 %w/w Caffeine: 12-25 %w/w Results indicate inorganic additives may be present

12. METHOD 3: ELEMENTAL CONSTITUENTS 0.1 g used for each analysis Samples digested using conc. HNO 3 and MW. Samples diluted and filtered Analysed using Varian ICP-OES with radial torch position. ICP Torch

13. WHICH ELEMENTS ARE PRESENT? Semi-Quant analysis of all samples H Hydrogen He Helium Li Lithium Be Beryllium B Boron C Carbon N Nitrogen O Oxygen F Fluorine Ne Neon Na Sodium Mg Magnesium Al Aluminium Si Silicon P Phosphorus S Sulfur Cl Chlorine Ar Argon K Potassium Ca Calcium Sc Scandium Ti Titanium V Vanadium Cr Chromium Mn Manganese Fe Iron Co Cobalt Ni Nickel Cu Copper Zn Zinc Ga Gallium Ge Germanium As Arsenic Se Selenium Br Bromine Kr Krypton Rb Rubidium Sr Strontium Y Yttrium Zr Zirconium Nb Niobium Mo Molybdenum Tc Technetium Ru Ruthenium Rh Rhodium Pd Palladium Ag Silver Cd Cadmium In Indium Sn Tin Sb Antimony Te Tellurium I Iodine Xe Xenon Cs Caesium Ba Barium lanthanide Series Hf Hafnium Ta Tantalum W Tungsten Re Rhenium Os Osmium Ir Iridium Pt Platinum Au Gold Hg Mercury Tl Thallium Pb Lead Bi Bismuth Po Polonium At Astatine Rn Radon Fr Francium Ra Radium Actinide Series Rf Rutherfordium Db Dubnium Sg Seaborgium Bh Bohrium Hs Hassium Mt Meitnerium UnmUuuUubUuq

14. WHICH ELEMENTS ARE PRESENT? Semi-Quant analysis of all samples H Hydrogen He Helium Li Lithium Be Beryllium B Boron C Carbon N Nitrogen O Oxygen F Fluorine Ne Neon Na Sodium Mg Magnesium Al Aluminium Si Silicon P Phosphorus S Sulfur Cl Chlorine Ar Argon K Potassium Ca Calcium Sc Scandium Ti Titanium V Vanadium Cr Chromium Mn Manganese Fe Iron Co Cobalt Ni Nickel Cu Copper Zn Zinc Ga Gallium Ge Germanium As Arsenic Se Selenium Br Bromine Kr Krypton Rb Rubidium Sr Strontium Y Yttrium Zr Zirconium Nb Niobium Mo Molybdenum Tc Technetium Ru Ruthenium Rh Rhodium Pd Palladium Ag Silver Cd Cadmium In Indium Sn Tin Sb Antimony Te Tellurium I Iodine Xe Xenon Cs Caesium Ba Barium lanthanide Series Hf Hafnium Ta Tantalum W Tungsten Re Rhenium Os Osmium Ir Iridium Pt Platinum Au Gold Hg Mercury Tl Thallium Pb Lead Bi Bismuth Po Polonium At Astatine Rn Radon Fr Francium Ra Radium Actinide Series Rf Rutherfordium Db Dubnium Sg Seaborgium Bh Bohrium Hs Hassium Mt Meitnerium UnmUuuUubUuq

15. CALCIUM AND MAGNESIUM Ca Mg

16. SELECTED METALS

17. LEAD ICH. Draft consensus guideline for elemental impurities Q3D Current Step 2b version. 26 July 2013.

18. CONCLUSIONS For the products analysed, 6 out of 11 matched their label claim and no 5-IAI was detected. Quantitative analysis showed concentration of aminoindanes ranged from 11-84 %w/w. Elemental analysis using ICP-OES detected a range of constituents, but further work is needed.

19. ACKNOWLEDGEMENTS Katy Kellett Dr. Jade D. Owen (Melbourne Scientific) Dr. Sulaf Assi (University of Bournemouth) Dr. Suzanne Fergus Dr. Mark Scott Prof. Fabrizio Schifano Sheelagh Halsey (Thermo Scientific) University of Hertfordshire