INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISES (ICE) Summary Report BIOLOGICAL SPECIMENS 2013/1 INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISES (ICE) Table of contents Introduction Page 3 Comments from the International Panel of Forensic Experts Page 3 Codes and Abbreviations Page 4 Sample 1 Analysis Page 5 Identified substances Page 5 Statement of findings Page 7 Identification methods Page 11 Summary Page 13 Z-Scores Page 14 Sample 2 Analysis Page 18 Identified substances Page 18 Statement of findings Page 20 Identification methods Page 24 Summary Page 26 Z-Scores Page 27 Sample 3 Analysis Page 33 Identified substances Page 33 Statement of findings Page 34 Identification methods Page 38 Summary Page 40 Z-Scores Page 41 Sample 4 Analysis Page 43 Identified substances Page 43 Statement of findings Page 44 Identification methods Page 48 Summary Page 50 Test Samples Information Samples Comments on samples Sample 1 To prepare BS-1, urine was spiked with Morphine sulphate (3057ng/ml, 2300ng base/ml), Codeine (1150ng base/ml) and 6-Monoacetylmorphine (576 ng base/ml). The spiked urine was dispensed in 50ml aliquots and lyophilised. Sample 2 To prepare BS-2, urine was spiked with Amfetamine sulfate (1562ng/ml, 1143ng base/ml), Nordazepam (1727ng/ml), Oxazepam (457ng/ml) and Temazepam (464ng/ml). The spiked urine was dispensed in 50ml aliquots and lyophilised. Sample 3 To prepare BS-3, urine was spiked with Sodium gammahydroxybutyrate (14427ng GHB/ml, ). The spiked urine was dispensed in 50ml aliquots and lyophilised Sample 4 BS-4 was a blank test sample prepared from urine containing no substance in the ICE menu. The sample contained urine spiked with Ephedrine hydrochloride (2700ng/ml, 2308ng base/ml). The spiked urine was dispensed in 50ml aliquots and lyophilised. Samples Substances Concentrations Comments on substances Sample 1 Morphine (Total) 2300 ng/ml 6-Monoacetylmorphine (6-MAM) 576 ng/ml Codeine 1150 ng/ml Sample 2 Nordazepam 1727 ng/ml Oxazepam 457 ng/ml Amfetamine 1143 ng/ml Temazepam 464 ng/ml Sample 3 gamma-Hydroxybutyric acid (GHB) 14427 ng/ml Sample 4 [blank sample] 2013/1-BS (2) Copyright (c) 2013 UNODC Introduction An important element of the UNODC International Quality Assurance Programme (IQAP) is the implementation of the International Collaborative Exercises (ICE). The exercises allow laboratories, from both developing and developed countries, to continuously monitor their performance in drug testing on a truly global scale. Two rounds are offered per year with each round presenting participants with four different test samples for analysis. This report provides information on the analytical results of laboratories participating in the Biological Specimens (BS) group. In order to maintain confidentiality, the participating laboratories have been assigned random “Web Codes”, which change every round. The analytical results returned by laboratories participating in ICE are evaluated by UNODC and a confidential report is provided to each laboratory on its own performance. The overall analytical results are reviewed by the UNODC’s International Panel of Forensic Experts which oversees the implementation of these exercises, and offers guidance and support in addressing relevant quality issues. The exercises provide an overview of the performance and capacity of participating laboratories and enable UNODC to tailor technical support in the laboratory sector for greatest impact. Comments from the International Panel of Forensic Experts Participation of laboratories Samples within both the Seized materials (SM) and Biological Specimens (BS) test group were sent to 167 laboratories in 57 countries. 68 laboratories participated in the BS test group and 58 (85%) laboratories submitted the results of their analysis in time for inclusion in the summary report, with 96% using the ICE portal for return of results. The almost universal usage of the ICE portal continues to be a high endorsement of its utility. Qualitative and Quantitative Analysis of the biological specimens test samples Overall, the level of performance in identification of the test samples was good given the inherently higher level of difficulty in the analysis of biological specimens compared to seized materials. The complexity of the test samples in ICE 2013/1 was increased in order to challenge the participants. BS-1 contained morphine, 6-monoacetylmorphine and codeine, which were identified by 79%, 79% and 91% of participants respectively. It is encouraging to note that 71% of participants identified all three components in BS-1 and only five false positive results were reported. BS-2 contained amphetamine and a mixture of three benzodiazepines (nordazepam, oxazepam and temazepam). While only 42% of participants identified all four components, 86% identified at least one of the benzodiazepines and 65% identified the amphetamine present. There were eight false positive results including four for diazepam. While the benzodiazepines present are metabolites of diazepam, their presence alone does not represent use of diazepam. BS-3 contained gammahydroxybutyric acid (GHB), a new substance on the ICE menu for the BS test group and 22% of participants identified its presence, while 64% of participants did not carry out analysis for identification of GHB. It is recognized that while GHB may not be commonly analysed by all participants, it has been included in the ICE menu as it represents an example of a more recently abused substance and at the Commission on Narcotic Drugs in March 2013, GHB was transferred from schedule IV to schedule II of the Convention of Psychotropic Substances of 1971. BS-4 was a blank test sample containing no substances from the ICE menu. Although it did contain ephedrine, a precursor to methamphetamine. 19% of participants identified the ephedrine and among five false positives, there was one for methamphetamine. The results for quantitative analysis within the BS test group were excellent. For BS-1 24 out of 25 (96%) laboratories quantified the morphine, 21 out of 22 (95%) quantified 6-MAM and 22 out of 23 (96%) quantified codeine with acceptable Z-scores. Within BS-2, the quantitation results were equally as good with 20 of 21 (95%) laboratories quantifying the amfetamine, 20 out of 22 (91%) the nordazepam, 13 of 16 (81%) the oxazepam and 14 out of 16 (88%) the temazepam with acceptable Z-scores. Within BS-3, 7 out of 9 (87%) participants quantified the GHB present with acceptable Z-scores. It is encouraging to note that of those laboratories who did carry out quantification, 52% performed quantification on at least seven of the eight substances in all test samples. The predominant analytical techniques used by participants for quantification within the BS test group were GC/MS (79%), followed by LC/MS (38%) and others at (17%). Laboratories are encouraged to carry out quantification as it can improve the quality of laboratory analyses and can be helpful in assessing the significance of the results. Also, laboratories reporting false positive or false negative results should investigate the reasons for this and corrective actions should be taken in order to continuously improve performance. Participation in ICE also helps in monitoring the effect of corrective actions. (3) INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISES (ICE) Codes and Abbreviations (+) “positive”: Indicates that the analyte is identified; for presumptive tests (e.g. colour reactions), indicates that a positive reaction was obtained. (-) “negative”: Indicates that the analyte is not identified. (ANP): Analysis not performed. Tech Code Name 100 Agglutination Techniques 110 Enzyme Immunoassay Techniques 120 Fluorescence Polarization Immunoassay 130 Radioimmunoassay 140 Colorimetric reactions 141 Marquis reagent (sulphuric acid, formaldehyde) 142 Cobalt thiocyanate 150 Thin Layer Chromatography 160 High/Ultra High Performance Liquid Chromatography 161 High Performance Liquid Chromatography with diode array detection 170 Gas Chromatography NPD 171 Gas Chromatography FID 172 Gas Chromatography ECD 180 Gas Chromatography/Mass Spectrometry 190 Fourier Transform Infrared Spectrometry 200 Spectrophotometry (visible, UV) 210 Others (specify) 211 NMR 220 Microcrystal test 230 Liquid chromatography/mass spectrometry 231 Liquid chromatography/tandem mass spectrometry 2013/1-BS (4) Copyright (c) 2013 UNODC INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISES (ICE) Sample 1 Analysis Identified substances for Test Sample 1 Code Identified Substances 0709WO 6-MAM, Morphine, Codeine 1C2WN2 opiates 1M39MI codeine, morphine, 6-monoacetylmorphine 2I1J7W codeine, 6 MAM and morphine 3ESBDT CODEINE, ACETYLMORPHINE 5A0LTW morphine, 6-MAM, codeine 5D8EIT Morphine, Heroin and Codeine 6H33U3 Codeine and 6-MAM 6ZZZCD morphine,codeine, 6-monoacethylmorphine 76JL37 morphine, 6-MAM, codeine 7HEM7O Codeine 98KDM8 Morphine, codeine and monoacetly-morphine ABBR1D Morphine,O6-monoacetylmorphine,codeine AKQ21O morphine, codeine and 6-acetyl-morphine ANXWGY Codeine, morphine and 6-Monoacethylmorphine BBRB0E Morphine BJG4OO 6-MAM, codeine, morphine BPL9ZY morphine, 6-MAM, codeine BVABMB 6-MAM, codeine, morphine CUH0QQ CODEINE, MORPHINE AND MORPHINE-6-ACETYL. CWOOM9 codeine, morphine, 6-mam CZJD27 morphine, codeine D7TTWV OPIÁCEOS DONO2Y 6-MAM, codeine, morphine ERWOWW Codeine FNSUKR 6-monoacetilmorfina, morfina, codeina ILS9UP Codeine, Morphine, 6-Monoacetylmorphine