- Email: [email protected]
Underground pill testing, down under
Forensic Science International 151 (2005) 53–58 www.elsevier.com/locate/forsciint
Underground pill testing, down under Andrew M. Camilleria,*, David Caldicottb a
Forensic Science South Australia, 21 Divett Place, Adelaide, Australia Emergency Department, Royal Adelaide Hospital, Adelaide, Australia
b
Received 22 April 2004; received in revised form 7 July 2004; accepted 9 July 2004 Available online 1 September 2004
Abstract At a recent South Australian rave, results reported to users from on-site pill-testing, using pill-testing kits, were compared with GCMS analysis of a scraping from the same pill. The presence of an ecstasy-like substance or methylamphetamine was correctly reported to users in 100% of pills that contained those substances. However only 11% of pills with combinations of illicit substances had both substances correctly identified. Ketamine was particularly problematic with identification occurring in only 18% of pills and in some instances, the colorimetric response obtained from ketamine was confused with the response from methylamphetamine. This study also allowed a comparison between pill design and composition encountered at the rave with those submitted to the forensic laboratory over a 6-month period including the month the rave was held. MDMA was present in 68% of pills at the rave and 89% of pills submitted by the police. Ketamine was present in 27 and 26% of pills, respectively and was often combined with other substances. The combinations of illicit substances were identical apart from one police-pill seizure that contained MDMA combined with PMA. This combination has not been previously encountered in South Australia. The pill designs observed at the rave differed significantly from the designs on pills submitted for testing by police. These differences limit the use of pill comparison charts as an alternative identification tool to colorimetric pill testing in South Australia. # 2004 Elsevier Ireland Ltd. All rights reserved. Keywords: Ecstasy; Ketamine; Pills; Pill-testing kits; Spot-test reagents; Rave
1. Introduction During the late 1980s, 3,4-methylenedioxymethamphetamine (MDMA) sold in pills known as ‘Ecstasy’ became associated with enhancing the experience from rave and dance parties [1]. The types of illicit substances now commonly associated with this scene have become many and varied with several studies showing that pills often contain other substances including MDA, MDEA, ketamine and amphetamines [2,3]. These pills are sold using the ‘brand’ * Corresponding author. Tel.: +61 8 82267700; fax: +61 8 2267777. E-mail address: [email protected] (A.M. Camilleri).
name associated with the design of the pill despite the ‘brand’ providing no information about the drug content [2]. In South Australia, pills seized by South Australia Police (SAPOL) and analysed at Forensic Science South Australia (FSSA) also show this varied content with MDMA, MDA and methylamphetamine regularly detected. More recently, ketamine has emerged as the second most prevalent illicit drug in pills after MDMA and is frequently combined with methylamphetamine and/or caffeine [4,5]. The designer amphetamine PMA is an ongoing problem in South Australia with 11 reported fatalities from the drug since 1995 [6]. Significant seizures over the previous 12 months, of pills and powder containing PMA or PMA combined with methylamphetamine and ketamine or MDMA, show that PMA manufacture and distribution is continuing.
0379-0738/$ – see front matter # 2004 Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.forsciint.2004.07.004
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With the diverse range and the concerning trend towards combinations of illicit substances within pills in South Australia, users are faced with even greater uncertainty about the content of their pills and acknowledge this as a risk associated with their pill usage [5]. Pill-testing kits have been available for purchase for many years and are marketed to users through various internet sites such as EZ test [7] and DanceSafe [8]. The kits are supplied with at least one spottest reagent, which can provide an indication of the identity of the illicit substance contained within a pill. The spot-test reagents included in these kits are identical to those commonly used by law enforcement personnel and these reagents have been effective in presumptively identifying a range of illicit substances in the forensic environment [9,10]. Each reagent produces a visual colour change when a chemical reaction between the illicit substance and the reagent occurs. This result is then matched to a colour chart showing the expected colour change of the reagent with various illicit substances. The limitations associated with identifying drugs, using spot-test reagents, are well documented [3,10]. Factors contributing to inaccurate results include false positive color reactions and variations in the response produced by differing drug concentration or salt form. The colorimetric response produced is of a subjective nature and its interpretation can alter based on the experience of the analyst, whether the analyst is drug-affected at the time of interpretation and the type, or lack of lighting available when the test is performed. The effectiveness of these kits is further diminished by the increasing frequency of combinations of illicit substances within pills. The predominant color change masks or interferes with the color change from other substances resulting in these other substances remaining unidentified [3]. On-site pill-testing programs provided by trained analysts are endorsed at rave and dance parties in many countries in Europe. Funding is supplied to these programs based on their potential to reduce the harm associated with pill consumption. These programs rely on a range of identification techniques such as the accurate but time-consuming and expensive analytical instrumentation GCMS or HPLC through to pill-testing kits and pill comparison charts [11,12]. Raves in the USA and UK also have on-site pill testing however, as in Australia, pill testing contravenes legislation regarding the possession and supply of controlled substances. Testing in these countries is therefore limited and relies on volunteer harm reduction organizations using results obtained from pill-testing kits performed by analysts who may be inadequately trained. The non-profit organization Enlighten, a dance-scene advocacy group, provides pill testing (with colorimetric kits) free of charge at some Australian raves. Enlighten generally combines the results from two spots test reagents, the Marquis reagent and the Mandelin reagent to inform users of the illicit substances contained within their pill. For the first time in Australia, Enlighten was able to provide pill testing at a South Australian rave, with the understanding of
local authorities, on the premise of harm reduction to users. The rave chosen had an estimated attendance of 10,000 people and was held at a remote outdoor location. Enlighten reported the results of their pill testing to users and these results were later compared with GCMS analysis conducted at FSSA on a scraping from the same pill. This allowed for a direct evaluation of the accuracy of on-site pill testing in Australia and of how effective the kits are at identifying combinations of illicit substances within one pill. The study also provided an insight into the current South Australian rave drug market and allowed for comparison of this market with SAPOL pill seizures.
2. Materials and methods The promoters of the rave were approached by the Royal Adelaide Hospital Designer Drug Academic Research Collaboration (RADAR) and gave support for this collaboration to provide pill testing for those in attendance. Ravesafe SA, an organization delivering harm reduction information to people in the dance party scene, provided an information booth at the rave. This was identified as the most appropriate site to set up a pill-testing booth, as the organization is well known and respected by dance-party goers. The booth was lit using a fluorescent light placed directly above a desk upon which the colorimetric testing was to occur. Individuals attending the tent were asked if they would like to participate in the study by providing a small scraping from a pill they intended to consume. They were then informed that the scraping would be subjected to two separate colorimetric tests to indicate the content of that pill and a further laboratory test at a later time. The pill was handed to a member of Enlighten, and the characteristic design (front and back), shape and color of the pill were recorded prior to testing. Three separate scrapings were taken from the outer surface of each pill and the pill was then returned to the user. One scraping was stored in a sealed glass phial, for transportation and subsequent submission to FSSA for testing. The second and third scrapings were placed onto a clean white tile and one scraping was subjected to 1–2 drops of Marquis reagent while the other scraping was subjected to 1–2 drops of Mandelin reagent. The results of the two spot tests were interpreted by Enlighten, recorded and reported to the individual. Every result reported by Enlighten was provided with a disclaimer indicating that the result did not mean the pill was safe, it did not mean that the pill was pure and it did not indicate how much of that substance was present in the pill. If the result differed to what the individual had expected, they were asked if they would still consume their pill. An amnesty bin was also provided for users who wished to dispose of their pill. The scrapings submitted to FSSA were extracted into methanol and 1 ml of the extract was injected into an Agilent Technologies 6890 N gas chromatograph fitted with a 5973
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mass selective detector. This was equipped with a HP-1 capillary column (15 m 0.25 mm 0.25 mm) with helium carrier gas at 6 psi. The flow of the split valve was 50 ml/ min; injector temperature 300 8C; oven temperature 90 8C for 3 min with a 45 8C/min ramp to 300 8C with a hold of 1 min. This method of analysis is used by FSSA for the screening of unknown samples submitted by SAPOL for analysis. It is successful in identifying a wide variety of illicit substances, including substances commonly identified in pills, as part of routine drug identification. The retention time and mass spectrum of ions above 10% abundance detected in the methanol extract of the pill scrapings were matched to the retention time and mass spectrum of ions above 10% abundance of pure drug standards analyzed using the same temperature program. The pure drug standards were purchased locally or manufactured in-house and validated against a purchased pure drug standard. An illicit substance was not detected in five of the extracts (all of the same pill design). Some of these extracts were reanalyzed or re-extracted using an alternate technique and using the same GCMS with variations in the temperature programs to screen for other components in the scrapings. The above temperature program with an extended temperature hold of 11 min at 300 8C was used to analyze one of the methanol extracts and another methanol extract, which had been made alkaline using one drop of concentrated ammonium solution. This temperature program has been successful in identifying compounds of higher boiling points such as anabolic and androgenic steroids. Another of the methanol extracts was acidified using one drop of dilute hydrochloric acid. This extract was analyzed on a low-temperature program with the oven temperature at 40 8C for 2 min with a 20 8C/min ramp to 100 8C. The flow of the split valve was 13 ml/min. This temperature program has been validated as successful in detecting gamma-butyrolactone (GBL), an
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indicator of the presence of 4-hydroxybutanoic acid (GHB). This extract was also analyzed on a higher temperature program with the oven temperature isothermal at 280 8C for 5 min. This temperature program has also been validated as successful in detecting lysergide.
3. Results and discussion 3.1. GC/MS analysis of tablet scrapings A total of 84 pills with 32 characteristic logos were presented for testing. The results of the colorimetric testing and GCMS analysis of the pills are supplied in Table 1. Quantitative analysis of the components from each scraping was not possible due to the limited sample size provided for analysis. MDMA was detected in 55 pills (65%) with 15 individual logos and was the most prevalent illicit substance detected. Ketamine was the second most common illicit substance, present in a total of 22 pills (26%) with 15 different logos. Ketamine was almost always combined with other substances with only two pills found to contain only ketamine. Ketamine was most commonly combined with methylamphetamine, which was never present alone, and this combination of substances together with caffeine, in most cases, accounted for 14 pills (17%) with 10 different logos. A total of four pills (5%) contained MDA while PMA was not detected in any of the submitted scrapings. An illicit substance was not detected in five pills (6%) all bearing the same design. Some users reported hallucinogenic effects after consuming these pills and said that they were being marketed as ‘‘Tripstasy’’, a mixture of LSD and ecstasy. No controlled substances, including lysergide, were detected in these pills. Forensic Chemistry Queensland Health Scientific Services recently analyzed pills that resembled the color and logo of the five pills presented
Table 1 Comparison of GCMS testing of pills with the results reported by Enlighten Number of pills
Number of designs
Results reported by Enlighten
Results of GCMS testing
49 5 1 4 7 5 1 2 1 1 5 1 2
12 2 1 1 4 4 1 2 1 1 1 1 1
MDXa MDX MDX MDX Amphetamine/methylamphetamine Amphetamine/methylamphetamine Amphetamine/methylamphetamine Amphetamine/methylamphetamine, ketamine Amphetamine/methylamphetamine, Ketamine Ketamine No results No result Not tested
MDMA MDMA and ketamine MDMA and caffeine MDA Methylamphetamine, ketamine and caffeine Methylamphetamine and ketamine Ketamine and caffeine Methylamphetamine, ketamine and caffeine Ketamine Ketamine No drugs detected Ephedrine Pseudoephedrine sulfate and chlorpheniramine maleateb
a b
MDX = MDA, MDMA, MDEA. Tablet identified using E-MIMS, version 4.00.0500, MediMedia Australia Pty Limited.
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for testing at the rave. Initial screening indicated the presence of lysergide in these pills; however, this had not been confirmed at the time of writing. No surface of these pills had been discolored, as is often the case for LSD presented in this form, and the pills appeared to be homogenous (K. Blakey and M. Edirisinghe, personal communication). It is likely that the scrapings taken from the five pills presented at the rave did contain LSD; however, the concentration of LSD in the scraping was insufficient for detection using the extraction and analytical procedure performed. Two pills (2%) submitted for testing were identified as the pharmaceutical preparation ‘‘Demazin’’, which is a cold and flu preparation containing pseudoephedrine sulfate and chlorpheniramine maleate. The users who submitted these pills for analysis reported that they were being sold as ‘Ecstasy’. One tablet (1%) was identified as a pharmaceutical preparation containing ephedrine; however, the user was aware of the content of the pill prior to purchase. Five plastic bags containing paste or powdered material were also presented to Enlighten for testing at the rave. Methylamphetamine was detected in the material in four of these bags while the material in the fifth bag contained MDMA. The results from these five bags are not included in Table 1 and are not discussed further. 3.2. Evaluation of the results from spot-test reagents A comparison between the results reported by Enlighten and the GCMS analysis of the scrapings taken from pills is shown in Table 1. Enlighten was correctly able to report the presence of an ecstasy type drug (MDX) or amphetamine/ methylamphetamine in the 73 different pills that contained those drugs. The identification of ketamine was not as successful however, as only four of the 22 pills (18%) that contained ketamine were correctly reported. Further problems associated with the identification of ketamine were encountered when Enlighten reported the presence of amphetamine/methylamphetamine in two separate pills in which GCMS analysis only identified ketamine. It is possible that the scraping supplied was insufficient to detect the low level of amphetamine/methylamphetamine in the pill; however, it is more likely that the colorimetric response was misinterpreted. Enlighten was only able to correctly identify a combination of illicit substances in two of the 19 pills (11%) containing multiple illicit substances (caffeine was not considered an illicit substance). A small amount of the Mandelin reagent used by Enlighten on the night was retained in a sealed glass phial to test its accuracy against pure drug standards. A drop of the Mandelin reagent was added to two small separate samples of pure methylamphetamine hydrochloride and two small separate samples of pure ketamine hydrochloride on a white porcelain tile. Initially a burst of pink color was observed within 5–10 s when added to methylamphetamine hydrochloride with the color slowly changing to aqua over 1–2 min. An orange color was observed when the Mandelin
reagent was added to ketamine hydrochloride remaining this color over time. The two initial colors are easily distinguishable when compared under adequate lighting conditions and can be more easily distinguished if the colorimetric response is reported 2 min after the addition of the Mandelin reagent. However, as discussed earlier, a number of variables including drug concentration, interfering color reactions from combinations of substances and the salt form of the drug can produce different results to those reported from pure drug standards. Combine these variables with the poor lighting conditions commonly encountered in a rave setting and analysts who are unlikely to have actually seen the color response obtained from a pure drug sample and the accuracy of results reported using spot-test reagents is significantly diminished. For on-site testing using pill-testing kits to be a useful tool in identification of substances within pills, analysts who are reporting the results must be adequately trained to identify differences between the colorimetric results of illicit substances. The authors are currently investigating the interaction between colorimetric results obtained when combinations of illicit substances are present in an attempt to determine whether pill-testing kits can provide accurate substance identification in these instances. Due to limited access to funding, organizations such as Enlighten can only afford to interpret results obtained from pill-testing kits. The accuracy of the results reported needs to be increased without significantly increasing costs or delaying reporting results if pill testing is to provide users with more definitive information. Interpretation of more spot-test reagent results could help to identify more substances within a pill. Obviously, the use of more reagents is limited by the amount of sample the user is willing to supply but this method is much less expensive and time consuming compared to upgrading to on-site GCMS, HPLC or equivalent analysis. Another advantage of incorporating more spot-test reagents is that they are more likely to detect the presence of low-dose drugs such as LSD. No drugs were detected in the GCMS analysis of five of the scrapings supplied for testing at the rave even when further extraction and reanalysis was performed. Adding a spot-test reagent such as the Van Urk reagent to these scrapings would have been more successful in identifying the presence of the LSD and if used on-site, the results could have been immediately conveyed to the user rather than giving the user no result. 3.3. Differences between SAPOL seizures and pills tested at the rave The types of illicit substances and combinations of these substances in pills presented for testing at the rave are consistent with SAPOL pill seizures analysed at FSSA over a 6-month period including the month the rave was held (Table 2). MDMA was the most common drug detected at the rave (68%, if one does not include the three pharmaceutical preparations) with a higher percentage seized by SAPOL (89%). In both cases, the majority of pills containing
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Table 2 Contents of pills submitted to Forensic Science South Australia (FSSA) for testing by South Australia Police (SAPOL) over a 6-month period including the month the rave was held Number of pills
Number of designs
Substances detected
3862 859 153 22 14 533 37 12 3 7
35 3 1 4 5 6 1 3 1 3
MDMA MDMA and ketamine MDMA and PMA MDA Methylamphetamine, ketamine and caffeine Methylamphetamine and ketamine Methylamphetamine and paracetamol Ketamine and caffeine Ketamine Caffeine
MDMA contained only this drug; however, some pills also contained ketamine or caffeine. On one occasion, a SAPOL seizure uncovered MDMA combined with PMA, which is the first time this combination of substances has been identified in South Australia. Ketamine was the second most common drug present at the rave (27%) with a comparable distribution in SAPOL seizures (26%). Ketamine was almost always combined with another active ingredient such as methylamphetamine and or caffeine at both the rave and in the SAPOL seizures. A total of 62 different pill designs were submitted to FSSA by SAPOL for testing over this period. A number of the designs appeared to be attempts to replicate an already established logo but the color, score lines or the actual design differed slightly. Pill manufacturers are replicating common designs attempting to capitalize on the reputation of that logo in the market. The type, combination or concentration of illicit substances in these pills can vary significantly. A user may find it difficult to identify these design differences and therefore consider a pill to be ‘safe’ based on their own or others previous use or a pill-testing kit result. If the concentration or content of these pills varies unexpectedly, there is an increased possibility of adverse effects or overdose for the user. Despite the limitations discussed for onsite pill-testing programs using pill-testing kits, it still may be useful in providing users with information about differences in constitution of pills that visually appear to be identical. The majority of pill designs encountered at the rave were not represented as part of SAPOL seizures during this 6month period. It is possible that a large proportion of pills presented for testing at the rave are sourced from local producers who have limited pill-manufacturing operations. Within this 6-month period, three well-equipped clandestine laboratories with the capability of producing significant quantities of MDMA were uncovered by SAPOL despite no MDMA manufacturing laboratories being uncovered in the previous 4 years. Tablet producing equipment was not present at two of these laboratories while tablet production in the third laboratory was not efficient in generating large numbers of pills. These smaller batches of locally produced
pills are likely to be consumed rapidly and are less likely to be detected as part of SAPOL seizure data than pills imported into Australia in large quantities. It is apparent that the seizure data complied by FSSA does not truly reflect the pill market especially in the rave community in South Australia. Pill comparison is therefore not an alternative option to pill testing using colorimetric spot-test reagents in South Australia.
4. Conclusions The limitations of using pill-testing kits as a harmreduction tool, highlighted by the results reported at the rave, have been discussed. The kits however can still provide users with some valuable information about the content of their pills by identifying the presence or absence of ecstasylike substances. This can be especially useful for users in identifying variations in the composition of pills that are purchased based on the ‘brand’ associated with the pill logo. In the current market, where it is common to encounter combinations of substances within pills, the kits regularly fail to identify all of the illicit substances present. This is especially dangerous if a user considers their pill ‘safe’ because one substance is accurately identified while high concentrations of substances such as PMA or ketamine are not. Another significant concern undermining the validity of on-site pill testing is the misinterpretation of the colorimetric spot-test result even when performed by an ‘experienced’ analyst. Interpretation of the ketamine colorimetric response was particularly problematic often being masked by other substances and on occasions, was confused with the colorimetric response of methylamphetamine. The success of this form of on-site pill testing can be improved by providing training to analysts, researching the accuracy of the kits when combinations of substances are present and increasing the number of spot-test reagents used especially when unusual results are obtained. There is limited scope in South Australia for using pill comparisons rather than pill-testing kits as an alternative method of identification. This was confirmed through
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limited overlap between the design types seized by SAPOL and those encountered at the rave. The process of on-site pill testing, even if the harm-reduction potential is questionable, still has value as it provides information about the rave scene in South Australia, which could not be determined based on SAPOL pill seizures. It also shows that the manufacture and distribution of locally produced pills that are rapidly consumed is occurring on a regular basis.
References [1] T. Randall, Ecstasy-fueled ‘rave’ parties become dances of death for English youths, J. Am. Med. Assoc. 268 (1992) 1505–1506. [2] J.C. Cole, M. Bailey, H.R. Sumnall, G.F. Wagstaff, L.A. King, The content of ecstasy tablets: implications for the study of their long-term effects, Addiction 97 (2002) 1531– 1536. [3] A.R. Winstock, K. Wolff, J. Ramsey, Ecstasy pill testing: harm minimization gone too far? Addiction 96 (2001) 1139–1148.
[4] Australian Crime Commission, Australian Illicit Drug Report 2001–2002, Commonwealth of Australia, 2003. [5] K. Quinn, C. Breen, B. White, Illicit tablet market in Victoria. Party Drug Trends Bulletin June 2004 (http://ndarc.med. unsw.edu.au/ndarc.nsf/website/IDRS.bulletins. 2004). [6] D.G.E. Caldicott, N.A. Edwards, A. Kruys, K.P. Kirkbride, R.W. Byard, M. Prior, R.J. Irvine, Dancing with ‘‘death’’: p-methoxyamphetamine overdose and its acute management, Clin. Tox. 41 (2003) 143–154. [7] http://www.eztest.com/mambo. [8] http://www.dancesafe.org. [9] E.G.C. Clarke, Clarke’s Isolation and Identification of Drugs in Pharmaceuticals, Body Fluids and Post-Mortem Material, 2nd ed. The Pharmaceutical Press, London, 1986, pp. 128– 147. [10] C.L. O’Neal, D.J. Crouch, A.A. Fatah, Validation of 12 chemical spot tests for the detection of drugs of abuse, Forensic. Sci. Int. 109 (2000) 189–201. [11] G. Burkhart, On-site pill-testing interventions in the European Union, European Monitoring Centre for Drugs and Drug Addiction, 2001. [12] A. Benschop, M. Rabes, D.J. Korf, Pill Testing, Ecstasy And Prevention. A Scientific Evaluation in Three European Cities, Rozenberg Publisher, Amsterdam, 2002.
Underground pill testing, down under Andrew M. Camilleria,*, David Caldicottb a
Forensic Science South Australia, 21 Divett Place, Adelaide, Australia Emergency Department, Royal Adelaide Hospital, Adelaide, Australia
b
Received 22 April 2004; received in revised form 7 July 2004; accepted 9 July 2004 Available online 1 September 2004
Abstract At a recent South Australian rave, results reported to users from on-site pill-testing, using pill-testing kits, were compared with GCMS analysis of a scraping from the same pill. The presence of an ecstasy-like substance or methylamphetamine was correctly reported to users in 100% of pills that contained those substances. However only 11% of pills with combinations of illicit substances had both substances correctly identified. Ketamine was particularly problematic with identification occurring in only 18% of pills and in some instances, the colorimetric response obtained from ketamine was confused with the response from methylamphetamine. This study also allowed a comparison between pill design and composition encountered at the rave with those submitted to the forensic laboratory over a 6-month period including the month the rave was held. MDMA was present in 68% of pills at the rave and 89% of pills submitted by the police. Ketamine was present in 27 and 26% of pills, respectively and was often combined with other substances. The combinations of illicit substances were identical apart from one police-pill seizure that contained MDMA combined with PMA. This combination has not been previously encountered in South Australia. The pill designs observed at the rave differed significantly from the designs on pills submitted for testing by police. These differences limit the use of pill comparison charts as an alternative identification tool to colorimetric pill testing in South Australia. # 2004 Elsevier Ireland Ltd. All rights reserved. Keywords: Ecstasy; Ketamine; Pills; Pill-testing kits; Spot-test reagents; Rave
1. Introduction During the late 1980s, 3,4-methylenedioxymethamphetamine (MDMA) sold in pills known as ‘Ecstasy’ became associated with enhancing the experience from rave and dance parties [1]. The types of illicit substances now commonly associated with this scene have become many and varied with several studies showing that pills often contain other substances including MDA, MDEA, ketamine and amphetamines [2,3]. These pills are sold using the ‘brand’ * Corresponding author. Tel.: +61 8 82267700; fax: +61 8 2267777. E-mail address: [email protected] (A.M. Camilleri).
name associated with the design of the pill despite the ‘brand’ providing no information about the drug content [2]. In South Australia, pills seized by South Australia Police (SAPOL) and analysed at Forensic Science South Australia (FSSA) also show this varied content with MDMA, MDA and methylamphetamine regularly detected. More recently, ketamine has emerged as the second most prevalent illicit drug in pills after MDMA and is frequently combined with methylamphetamine and/or caffeine [4,5]. The designer amphetamine PMA is an ongoing problem in South Australia with 11 reported fatalities from the drug since 1995 [6]. Significant seizures over the previous 12 months, of pills and powder containing PMA or PMA combined with methylamphetamine and ketamine or MDMA, show that PMA manufacture and distribution is continuing.
0379-0738/$ – see front matter # 2004 Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.forsciint.2004.07.004
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With the diverse range and the concerning trend towards combinations of illicit substances within pills in South Australia, users are faced with even greater uncertainty about the content of their pills and acknowledge this as a risk associated with their pill usage [5]. Pill-testing kits have been available for purchase for many years and are marketed to users through various internet sites such as EZ test [7] and DanceSafe [8]. The kits are supplied with at least one spottest reagent, which can provide an indication of the identity of the illicit substance contained within a pill. The spot-test reagents included in these kits are identical to those commonly used by law enforcement personnel and these reagents have been effective in presumptively identifying a range of illicit substances in the forensic environment [9,10]. Each reagent produces a visual colour change when a chemical reaction between the illicit substance and the reagent occurs. This result is then matched to a colour chart showing the expected colour change of the reagent with various illicit substances. The limitations associated with identifying drugs, using spot-test reagents, are well documented [3,10]. Factors contributing to inaccurate results include false positive color reactions and variations in the response produced by differing drug concentration or salt form. The colorimetric response produced is of a subjective nature and its interpretation can alter based on the experience of the analyst, whether the analyst is drug-affected at the time of interpretation and the type, or lack of lighting available when the test is performed. The effectiveness of these kits is further diminished by the increasing frequency of combinations of illicit substances within pills. The predominant color change masks or interferes with the color change from other substances resulting in these other substances remaining unidentified [3]. On-site pill-testing programs provided by trained analysts are endorsed at rave and dance parties in many countries in Europe. Funding is supplied to these programs based on their potential to reduce the harm associated with pill consumption. These programs rely on a range of identification techniques such as the accurate but time-consuming and expensive analytical instrumentation GCMS or HPLC through to pill-testing kits and pill comparison charts [11,12]. Raves in the USA and UK also have on-site pill testing however, as in Australia, pill testing contravenes legislation regarding the possession and supply of controlled substances. Testing in these countries is therefore limited and relies on volunteer harm reduction organizations using results obtained from pill-testing kits performed by analysts who may be inadequately trained. The non-profit organization Enlighten, a dance-scene advocacy group, provides pill testing (with colorimetric kits) free of charge at some Australian raves. Enlighten generally combines the results from two spots test reagents, the Marquis reagent and the Mandelin reagent to inform users of the illicit substances contained within their pill. For the first time in Australia, Enlighten was able to provide pill testing at a South Australian rave, with the understanding of
local authorities, on the premise of harm reduction to users. The rave chosen had an estimated attendance of 10,000 people and was held at a remote outdoor location. Enlighten reported the results of their pill testing to users and these results were later compared with GCMS analysis conducted at FSSA on a scraping from the same pill. This allowed for a direct evaluation of the accuracy of on-site pill testing in Australia and of how effective the kits are at identifying combinations of illicit substances within one pill. The study also provided an insight into the current South Australian rave drug market and allowed for comparison of this market with SAPOL pill seizures.
2. Materials and methods The promoters of the rave were approached by the Royal Adelaide Hospital Designer Drug Academic Research Collaboration (RADAR) and gave support for this collaboration to provide pill testing for those in attendance. Ravesafe SA, an organization delivering harm reduction information to people in the dance party scene, provided an information booth at the rave. This was identified as the most appropriate site to set up a pill-testing booth, as the organization is well known and respected by dance-party goers. The booth was lit using a fluorescent light placed directly above a desk upon which the colorimetric testing was to occur. Individuals attending the tent were asked if they would like to participate in the study by providing a small scraping from a pill they intended to consume. They were then informed that the scraping would be subjected to two separate colorimetric tests to indicate the content of that pill and a further laboratory test at a later time. The pill was handed to a member of Enlighten, and the characteristic design (front and back), shape and color of the pill were recorded prior to testing. Three separate scrapings were taken from the outer surface of each pill and the pill was then returned to the user. One scraping was stored in a sealed glass phial, for transportation and subsequent submission to FSSA for testing. The second and third scrapings were placed onto a clean white tile and one scraping was subjected to 1–2 drops of Marquis reagent while the other scraping was subjected to 1–2 drops of Mandelin reagent. The results of the two spot tests were interpreted by Enlighten, recorded and reported to the individual. Every result reported by Enlighten was provided with a disclaimer indicating that the result did not mean the pill was safe, it did not mean that the pill was pure and it did not indicate how much of that substance was present in the pill. If the result differed to what the individual had expected, they were asked if they would still consume their pill. An amnesty bin was also provided for users who wished to dispose of their pill. The scrapings submitted to FSSA were extracted into methanol and 1 ml of the extract was injected into an Agilent Technologies 6890 N gas chromatograph fitted with a 5973
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mass selective detector. This was equipped with a HP-1 capillary column (15 m 0.25 mm 0.25 mm) with helium carrier gas at 6 psi. The flow of the split valve was 50 ml/ min; injector temperature 300 8C; oven temperature 90 8C for 3 min with a 45 8C/min ramp to 300 8C with a hold of 1 min. This method of analysis is used by FSSA for the screening of unknown samples submitted by SAPOL for analysis. It is successful in identifying a wide variety of illicit substances, including substances commonly identified in pills, as part of routine drug identification. The retention time and mass spectrum of ions above 10% abundance detected in the methanol extract of the pill scrapings were matched to the retention time and mass spectrum of ions above 10% abundance of pure drug standards analyzed using the same temperature program. The pure drug standards were purchased locally or manufactured in-house and validated against a purchased pure drug standard. An illicit substance was not detected in five of the extracts (all of the same pill design). Some of these extracts were reanalyzed or re-extracted using an alternate technique and using the same GCMS with variations in the temperature programs to screen for other components in the scrapings. The above temperature program with an extended temperature hold of 11 min at 300 8C was used to analyze one of the methanol extracts and another methanol extract, which had been made alkaline using one drop of concentrated ammonium solution. This temperature program has been successful in identifying compounds of higher boiling points such as anabolic and androgenic steroids. Another of the methanol extracts was acidified using one drop of dilute hydrochloric acid. This extract was analyzed on a low-temperature program with the oven temperature at 40 8C for 2 min with a 20 8C/min ramp to 100 8C. The flow of the split valve was 13 ml/min. This temperature program has been validated as successful in detecting gamma-butyrolactone (GBL), an
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indicator of the presence of 4-hydroxybutanoic acid (GHB). This extract was also analyzed on a higher temperature program with the oven temperature isothermal at 280 8C for 5 min. This temperature program has also been validated as successful in detecting lysergide.
3. Results and discussion 3.1. GC/MS analysis of tablet scrapings A total of 84 pills with 32 characteristic logos were presented for testing. The results of the colorimetric testing and GCMS analysis of the pills are supplied in Table 1. Quantitative analysis of the components from each scraping was not possible due to the limited sample size provided for analysis. MDMA was detected in 55 pills (65%) with 15 individual logos and was the most prevalent illicit substance detected. Ketamine was the second most common illicit substance, present in a total of 22 pills (26%) with 15 different logos. Ketamine was almost always combined with other substances with only two pills found to contain only ketamine. Ketamine was most commonly combined with methylamphetamine, which was never present alone, and this combination of substances together with caffeine, in most cases, accounted for 14 pills (17%) with 10 different logos. A total of four pills (5%) contained MDA while PMA was not detected in any of the submitted scrapings. An illicit substance was not detected in five pills (6%) all bearing the same design. Some users reported hallucinogenic effects after consuming these pills and said that they were being marketed as ‘‘Tripstasy’’, a mixture of LSD and ecstasy. No controlled substances, including lysergide, were detected in these pills. Forensic Chemistry Queensland Health Scientific Services recently analyzed pills that resembled the color and logo of the five pills presented
Table 1 Comparison of GCMS testing of pills with the results reported by Enlighten Number of pills
Number of designs
Results reported by Enlighten
Results of GCMS testing
49 5 1 4 7 5 1 2 1 1 5 1 2
12 2 1 1 4 4 1 2 1 1 1 1 1
MDXa MDX MDX MDX Amphetamine/methylamphetamine Amphetamine/methylamphetamine Amphetamine/methylamphetamine Amphetamine/methylamphetamine, ketamine Amphetamine/methylamphetamine, Ketamine Ketamine No results No result Not tested
MDMA MDMA and ketamine MDMA and caffeine MDA Methylamphetamine, ketamine and caffeine Methylamphetamine and ketamine Ketamine and caffeine Methylamphetamine, ketamine and caffeine Ketamine Ketamine No drugs detected Ephedrine Pseudoephedrine sulfate and chlorpheniramine maleateb
a b
MDX = MDA, MDMA, MDEA. Tablet identified using E-MIMS, version 4.00.0500, MediMedia Australia Pty Limited.
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for testing at the rave. Initial screening indicated the presence of lysergide in these pills; however, this had not been confirmed at the time of writing. No surface of these pills had been discolored, as is often the case for LSD presented in this form, and the pills appeared to be homogenous (K. Blakey and M. Edirisinghe, personal communication). It is likely that the scrapings taken from the five pills presented at the rave did contain LSD; however, the concentration of LSD in the scraping was insufficient for detection using the extraction and analytical procedure performed. Two pills (2%) submitted for testing were identified as the pharmaceutical preparation ‘‘Demazin’’, which is a cold and flu preparation containing pseudoephedrine sulfate and chlorpheniramine maleate. The users who submitted these pills for analysis reported that they were being sold as ‘Ecstasy’. One tablet (1%) was identified as a pharmaceutical preparation containing ephedrine; however, the user was aware of the content of the pill prior to purchase. Five plastic bags containing paste or powdered material were also presented to Enlighten for testing at the rave. Methylamphetamine was detected in the material in four of these bags while the material in the fifth bag contained MDMA. The results from these five bags are not included in Table 1 and are not discussed further. 3.2. Evaluation of the results from spot-test reagents A comparison between the results reported by Enlighten and the GCMS analysis of the scrapings taken from pills is shown in Table 1. Enlighten was correctly able to report the presence of an ecstasy type drug (MDX) or amphetamine/ methylamphetamine in the 73 different pills that contained those drugs. The identification of ketamine was not as successful however, as only four of the 22 pills (18%) that contained ketamine were correctly reported. Further problems associated with the identification of ketamine were encountered when Enlighten reported the presence of amphetamine/methylamphetamine in two separate pills in which GCMS analysis only identified ketamine. It is possible that the scraping supplied was insufficient to detect the low level of amphetamine/methylamphetamine in the pill; however, it is more likely that the colorimetric response was misinterpreted. Enlighten was only able to correctly identify a combination of illicit substances in two of the 19 pills (11%) containing multiple illicit substances (caffeine was not considered an illicit substance). A small amount of the Mandelin reagent used by Enlighten on the night was retained in a sealed glass phial to test its accuracy against pure drug standards. A drop of the Mandelin reagent was added to two small separate samples of pure methylamphetamine hydrochloride and two small separate samples of pure ketamine hydrochloride on a white porcelain tile. Initially a burst of pink color was observed within 5–10 s when added to methylamphetamine hydrochloride with the color slowly changing to aqua over 1–2 min. An orange color was observed when the Mandelin
reagent was added to ketamine hydrochloride remaining this color over time. The two initial colors are easily distinguishable when compared under adequate lighting conditions and can be more easily distinguished if the colorimetric response is reported 2 min after the addition of the Mandelin reagent. However, as discussed earlier, a number of variables including drug concentration, interfering color reactions from combinations of substances and the salt form of the drug can produce different results to those reported from pure drug standards. Combine these variables with the poor lighting conditions commonly encountered in a rave setting and analysts who are unlikely to have actually seen the color response obtained from a pure drug sample and the accuracy of results reported using spot-test reagents is significantly diminished. For on-site testing using pill-testing kits to be a useful tool in identification of substances within pills, analysts who are reporting the results must be adequately trained to identify differences between the colorimetric results of illicit substances. The authors are currently investigating the interaction between colorimetric results obtained when combinations of illicit substances are present in an attempt to determine whether pill-testing kits can provide accurate substance identification in these instances. Due to limited access to funding, organizations such as Enlighten can only afford to interpret results obtained from pill-testing kits. The accuracy of the results reported needs to be increased without significantly increasing costs or delaying reporting results if pill testing is to provide users with more definitive information. Interpretation of more spot-test reagent results could help to identify more substances within a pill. Obviously, the use of more reagents is limited by the amount of sample the user is willing to supply but this method is much less expensive and time consuming compared to upgrading to on-site GCMS, HPLC or equivalent analysis. Another advantage of incorporating more spot-test reagents is that they are more likely to detect the presence of low-dose drugs such as LSD. No drugs were detected in the GCMS analysis of five of the scrapings supplied for testing at the rave even when further extraction and reanalysis was performed. Adding a spot-test reagent such as the Van Urk reagent to these scrapings would have been more successful in identifying the presence of the LSD and if used on-site, the results could have been immediately conveyed to the user rather than giving the user no result. 3.3. Differences between SAPOL seizures and pills tested at the rave The types of illicit substances and combinations of these substances in pills presented for testing at the rave are consistent with SAPOL pill seizures analysed at FSSA over a 6-month period including the month the rave was held (Table 2). MDMA was the most common drug detected at the rave (68%, if one does not include the three pharmaceutical preparations) with a higher percentage seized by SAPOL (89%). In both cases, the majority of pills containing
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Table 2 Contents of pills submitted to Forensic Science South Australia (FSSA) for testing by South Australia Police (SAPOL) over a 6-month period including the month the rave was held Number of pills
Number of designs
Substances detected
3862 859 153 22 14 533 37 12 3 7
35 3 1 4 5 6 1 3 1 3
MDMA MDMA and ketamine MDMA and PMA MDA Methylamphetamine, ketamine and caffeine Methylamphetamine and ketamine Methylamphetamine and paracetamol Ketamine and caffeine Ketamine Caffeine
MDMA contained only this drug; however, some pills also contained ketamine or caffeine. On one occasion, a SAPOL seizure uncovered MDMA combined with PMA, which is the first time this combination of substances has been identified in South Australia. Ketamine was the second most common drug present at the rave (27%) with a comparable distribution in SAPOL seizures (26%). Ketamine was almost always combined with another active ingredient such as methylamphetamine and or caffeine at both the rave and in the SAPOL seizures. A total of 62 different pill designs were submitted to FSSA by SAPOL for testing over this period. A number of the designs appeared to be attempts to replicate an already established logo but the color, score lines or the actual design differed slightly. Pill manufacturers are replicating common designs attempting to capitalize on the reputation of that logo in the market. The type, combination or concentration of illicit substances in these pills can vary significantly. A user may find it difficult to identify these design differences and therefore consider a pill to be ‘safe’ based on their own or others previous use or a pill-testing kit result. If the concentration or content of these pills varies unexpectedly, there is an increased possibility of adverse effects or overdose for the user. Despite the limitations discussed for onsite pill-testing programs using pill-testing kits, it still may be useful in providing users with information about differences in constitution of pills that visually appear to be identical. The majority of pill designs encountered at the rave were not represented as part of SAPOL seizures during this 6month period. It is possible that a large proportion of pills presented for testing at the rave are sourced from local producers who have limited pill-manufacturing operations. Within this 6-month period, three well-equipped clandestine laboratories with the capability of producing significant quantities of MDMA were uncovered by SAPOL despite no MDMA manufacturing laboratories being uncovered in the previous 4 years. Tablet producing equipment was not present at two of these laboratories while tablet production in the third laboratory was not efficient in generating large numbers of pills. These smaller batches of locally produced
pills are likely to be consumed rapidly and are less likely to be detected as part of SAPOL seizure data than pills imported into Australia in large quantities. It is apparent that the seizure data complied by FSSA does not truly reflect the pill market especially in the rave community in South Australia. Pill comparison is therefore not an alternative option to pill testing using colorimetric spot-test reagents in South Australia.
4. Conclusions The limitations of using pill-testing kits as a harmreduction tool, highlighted by the results reported at the rave, have been discussed. The kits however can still provide users with some valuable information about the content of their pills by identifying the presence or absence of ecstasylike substances. This can be especially useful for users in identifying variations in the composition of pills that are purchased based on the ‘brand’ associated with the pill logo. In the current market, where it is common to encounter combinations of substances within pills, the kits regularly fail to identify all of the illicit substances present. This is especially dangerous if a user considers their pill ‘safe’ because one substance is accurately identified while high concentrations of substances such as PMA or ketamine are not. Another significant concern undermining the validity of on-site pill testing is the misinterpretation of the colorimetric spot-test result even when performed by an ‘experienced’ analyst. Interpretation of the ketamine colorimetric response was particularly problematic often being masked by other substances and on occasions, was confused with the colorimetric response of methylamphetamine. The success of this form of on-site pill testing can be improved by providing training to analysts, researching the accuracy of the kits when combinations of substances are present and increasing the number of spot-test reagents used especially when unusual results are obtained. There is limited scope in South Australia for using pill comparisons rather than pill-testing kits as an alternative method of identification. This was confirmed through
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limited overlap between the design types seized by SAPOL and those encountered at the rave. The process of on-site pill testing, even if the harm-reduction potential is questionable, still has value as it provides information about the rave scene in South Australia, which could not be determined based on SAPOL pill seizures. It also shows that the manufacture and distribution of locally produced pills that are rapidly consumed is occurring on a regular basis.
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[4] Australian Crime Commission, Australian Illicit Drug Report 2001–2002, Commonwealth of Australia, 2003. [5] K. Quinn, C. Breen, B. White, Illicit tablet market in Victoria. Party Drug Trends Bulletin June 2004 (http://ndarc.med. unsw.edu.au/ndarc.nsf/website/IDRS.bulletins. 2004). [6] D.G.E. Caldicott, N.A. Edwards, A. Kruys, K.P. Kirkbride, R.W. Byard, M. Prior, R.J. Irvine, Dancing with ‘‘death’’: p-methoxyamphetamine overdose and its acute management, Clin. Tox. 41 (2003) 143–154. [7] http://www.eztest.com/mambo. [8] http://www.dancesafe.org. [9] E.G.C. Clarke, Clarke’s Isolation and Identification of Drugs in Pharmaceuticals, Body Fluids and Post-Mortem Material, 2nd ed. The Pharmaceutical Press, London, 1986, pp. 128– 147. [10] C.L. O’Neal, D.J. Crouch, A.A. Fatah, Validation of 12 chemical spot tests for the detection of drugs of abuse, Forensic. Sci. Int. 109 (2000) 189–201. [11] G. Burkhart, On-site pill-testing interventions in the European Union, European Monitoring Centre for Drugs and Drug Addiction, 2001. [12] A. Benschop, M. Rabes, D.J. Korf, Pill Testing, Ecstasy And Prevention. A Scientific Evaluation in Three European Cities, Rozenberg Publisher, Amsterdam, 2002.