Recommended methods for the Identification and Analysis of Piperazines in Seized Materials MANUAL FOR USE BY NATIONAL DRUG ANALYSIS LABORATORIES Photo credits: UNODC Photo Library; UNODC/Ioulia Kondratovitch; Alessandro Scotti. Laboratory and Scientific Section UNITED NATIONS OFFICE ON DRUGS AND CRIME Vienna Recommended Methods for the Identification and Analysis of Piperazines in Seized Materials MANUAL FOR USE BY NATIONAL DRUG ANALYSIS LABORATORIES UNITED NATIONS New York, 2013 Note Operating and experimental conditions are reproduced from the original reference materials, including unpublished methods, validated and used in selected national laboratories as per the list of references. A number of alternative conditions and substitution of named commercial products may provide comparable results in many cases, but any modification has to be validated before it is integrated into laboratory routines. ST/NAR/47 Original language: English © United Nations, January 2013. All rights reserved, worldwide. The designations employed and the presentation of material in this publication do not imply the expression of any opinion whatsoever on the part of the Secretariat of the United Nations concerning the legal status of any country, territory, city or area, or of its authorities, or concerning the delimitation of its frontiers or boundaries. Mention of names of firms and commercial products does not imply the endorse- ment of the United Nations. This publication has not been formally edited. Publishing production: English, Publishing and Library Section, United Nations Office at Vienna. Acknowledgements UNODC’s Laboratory and Scientific Section (LSS), headed by Dr. Justice Tettey, wishes to express its appreciation and thanks to Ms. Pamela Smith for the prepara- tion of the first draft of this manual. LSS would also like to thank Mr. Jeffery Comparin of the United States Drug Enforcement Administration, Mr. Paul Loo, Mr. Chad Mehaux and Ms. Valérie Bélisle of the Canadian Border Services Agency and Dr. Mark Baron of the University of Lincoln, United Kingdom for their expert reviews and valuable contribution. The preparation of this manual was coordinated by Dr. Conor Crean and the con- tributions of UNODC staff and interns, Ms. Bridgette Webb and Mr. Diego Pazos are gratefully acknowledged. iii Contents Page 1. Introduction ..................................................... 1 1.1 Background .................................................. 1 1.2 Purpose and use of the Manual ................................ 2 2. General aspects ..................................................... 5 2.1 Description of the pure compounds. ............................ 5 2.2 Licit uses ..................................................... 8 2.3 Control status ................................................ 8 2.4 Illicit products/use ............................................ 8 2.5 Pharmacology ............................................ 9 3. Illicit manufacture of piperazines ..................................... 10 3.1 Illicit manufacture ......................................... 10 4. Qualitative and quantitative analysis of materials containing piperazines .. 12 4.1 Sampling .................................................... 12 4.2 Solubility ..................................................... 12 4.3 Screening tests ............................................... 13 4.4 Microcrystalline tests .......................................... 17 4.5 Thin-layer chromatography (TLC) .............................. 19 4.6 Gas chromatography (GC) with flame ionization detection (GC-FID) 23 4.7 Gas chromatography-mass spectrometry (GC-MS) ................ 25 4.8 Gas chromatography-infrared detection (GC-IRD) ................ 28 4.9 High pressure liquid chromatography (HPLC) .................... 29 4.10 Capillary electrophoresis (CE) .................................. 32 4.11 Fourier transform infrared (FTIR) spectroscopy .................. 33 5. Library information ................................................. 35 5.1 Ultraviolet (UV) spectrophotometry ............................. 35 5.2 GC-MS data for selected piperazines ............................ 35 Additional reading ..................................................... 37 References ............................................................ 43 v 1. Introduction 1.1 Background Piperazine, a heterocyclic six-membered ring compound which contains two nitro- gens in the 1 and 4 positions, is a cyclic member of the ethylenediamine group of molecules [1, 2]. The abuse of substituted derivatives of piperazine was first reported in the United States in 1996 and has, since then, spread to a number of countries worldwide [3]. The large scale use of synthetic derivatives of piperazine as substi- tutes or mimics of “ecstasy” started in New Zealand in the early 2000s and became common in Europe after 2004 [4]. The first piperazine derivative encountered was 1-benzylpiperazine (BZP), one of a group of phenyl and benzyl substituted piperazines that have become prevalent worldwide, especially in traditional 3,4-methylenedioxymethamphetamine (MDMA) markets. Other widely used piperazines include 1-(3-chlorophenyl)piperazine, (mCPP) and 1-(3-trifluoromethylphenyl)piperazine (TFMPP), the latter of which is commonly found in combination with BZP. BZP itself is a central nervous system stimulant with a potency of 10% that of d-amfetamine [4]. It has been reported to stimulate the release of dopamine, noradrenaline, and serotonin, and also inhibit their reuptake. The substances are thus amfetamine mimics and predominantly found in tablet form either alone, in com- bination with other piperazines or with amfetamine, cocaine, ketamine or MDMA. Neither BZP nor any other substituted piperazine are listed in the Schedules of the United Nations 1971 Convention on Psychotropic Substances. However, in 2007, the International Narcotics Control Board (INCB) requested the World Health Organization (WHO) to consider reviewing piperazine derived compounds for pos- sible scheduling under the 1971 Convention. Independently, many countries have introduced legislation controlling the use of BZP. This includes the United States and the countries of the European Union (EU), which submitted BZP for EU control in 2008 following a risk assessment using the early warning system of the European Monitoring Centre for Drugs and Drug Addiction (EMCDDA) [5]. 1 2 Recommended methods for the identification and analysis of piperazines in seized materials 1.2 Purpose and use of the Manual The present Manual is one in a series of similar publications dealing with the iden- tification and analysis of various classes of drugs under international control. These manuals are the outcome of a programme pursued by UNODC since the early 1980s, aimed at the harmonization and establishment of recommended methods of analysis for national drug analysis laboratories. This Manual was prepared taking into account the Commission on Narcotic Drugs 2012 resolution: 55/1 “Promoting international cooperation in responding to the challenges posed by new psychoactive substances”, which encourages the United Nations Office on Drugs and Crime and other relevant international organizations, upon request, to provide Member States with technical assistance, including by supporting forensic and toxicological capability, to respond to the challenges posed by new psychoactive substances. In accordance with the overall objective of the series, the present manual suggests approaches that may assist drug analysts in the selection of methods appropriate for the sample under examination and provide data suitable for the purpose at hand, leaving room also for adaptation to the level of sophistication of different laboratories and various legal requirements. The majority of methods included in this manual are validated, methods which have been used for a number of years in reputable laboratories and as part of inter-laboratory studies, collaborative exercises and pro- ficiency tests. The reader should be aware, however, that there are a number of other methods, including those published in the forensic science literature, which may also produce acceptable results. Any new method that is to be used in the reader’s laboratory must be validated and/or verified prior to routine use. In addition, there are a number of more sophisticated approaches, but they may not be necessary for routine operational applications. Therefore, the methods described here should be understood as guidance, that is, minor modifications to suit local circumstances should not affect the validity of the results. The choice of the meth- odology and approach to analysis, as well as the decision whether or not additional methods are required, remain with the analyst and may also be dependent on the availability of appropriate instrumentation and the level of legally acceptable proof in the jurisdiction within which the analyst works. Attention is also drawn to the vital importance of the availability to drug analysts of reference materials and books on drugs of abuse and the analytical techniques used for their identification. Moreover, the analyst must of necessity keep abreast of current trends in drug analysis, consistently following current analytical and foren- sic science literature. UNODC’s Laboratory and Scientific Section would welcome observations on the contents and usefulness of the present Manual. Comments and suggestions may be addressed to: Recommended methods for the identification and analysis