INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISES (ICE) Summary Report BIOLOGICAL SPECIMENS 2012/2 INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISE (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 16 Identified substances Page 16 Statement of findings Page 18 Identification methods Page 23 Summary Page 25 Z-Scores Page 27 Sample 3 Analysis Page 28 Identified substances Page 28 Statement of findings Page 30 Identification methods Page 34 Summary Page 36 Z-Scores Page 37 Sample 4 Analysis Page 39 Identified substances Page 39 Statement of findings Page 41 Identification methods Page 45 Summary Page 47 Z-Scores Page 48 Test Samples Information Samples Comments on samples Sample 1 To prepare BS1, urine was spiked with amfetamine sulphate (4720 ng/ml, 3454 ng base/ml). The amfetamine sulphate was dissolved in water before adding to the urine and the spiked urine was dispensed in 50ml aliquots and lyophilised. Sample 2 7RSUHSDUH%6XULQHZDVVSLNHGZLWKQRU¨7+&FDUER[\OLFDFLG QJPO 7KHQRU¨ THC-9-carboxylic acid was dissolved in methanol before adding to the urine and the spiked urine was dispensed in 50ml aliquots and lyophilised. Sample 3 To prepare BS3, urine was spiked with morphine sulphate (1440ng/ml, 1150ng base/ml). The morphine sulphate was dissolved in water before adding to the urine and the spiked urine was dispensed in 50 ml aliquots and lyophilised. Sample 4 To prepare BS4, urine was spiked with benzoylecgonine hydrate (2370 ng/ml, 2230 ng base/ml) and methylecgonine (1150ng /ml). The benzoylecgonine hydrate was dissolved in water and the methylecgonine was dissolved in methanol before adding to the urine and the spiked urine was dispensed in 50 ml aliquots and lyophilised. Samples Substances Concentrations Comments on substances Sample 1 Amfetamine 3454 ng/ml Sample 2 QRU¨7+&FDUER[\OLFDFLG 800 ng/ml Sample 3 Morphine (Total) 1150 ng/ml Sample 4 Methylecgonine 1150 ng/ml Benzoylecgonine 2230 ng/ml This report contains the data received from laboratories participating in the current exercise. The results compiled in this report are not intended to be an overview of the quality of work and cannot be interpreted as such. These comments do not reflect the general state of the art within the profession. Participant results are reported using a randomly assigned "WebCode". This code maintains participant's anonymity, provides linking of the various report sections, and will change with every report. 2012/2-BS (2) Copyright (c) 2013 UNODC INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISE (ICE) 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. 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 55 countries. 68 laboratories in participated in the BS test group and 62 (91%) laboratories submitted the results of their analysis in time for inclusion in the summary report, with 97% 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. BS-1 contained amfetamine and 85% of participants identified the substance while there were two false positives for controlled substances (MDEA and metamfetamine). BS-2 contained 11-nor-∆9-THC-9-carboxylic acid, which was identified by 89% of participants and there were no false positive results reported for this test sample. BS-3 contained Morphine and this was identified by 89% of participants. There were a number of false positive results for this test sample, including four for morphine glucuronides, three for codeine and one each for 6-monoacetylmorphine and 11-nor-∆9-THC-9-carboxylic acid. BS-4 contained a mixture of two cocaine metabolites, benzoylecgonine and methylecgonine. The former was identified by 85% of participants, while 60% identifed the latter. Within BS-4, there was one false positive result for 11-nor-∆9-THC-9-carboxylic acid. With regard to false negative results, there were seven for BS-1, two for BS-2 and BS-3, six for benzoylecgonine in BS-4 and 11 for methylecgonine in BS-4. The results for quantitative analysis within the BS test group were excellent with 23 of 25 (92%, BS-1), 28 of 28 (100%, BS-2), 25 of 26 (96%, BS-3,) 25 of 25 (100%, BS-4, benzoylecgonine) and 11 of 15 (73%, BS-4, methylecgonine) participants performing quantification with acceptable Z-scores less than 3.5 (A modified Z-score was used for the statistical examination of the quantitative results, as it is a robust method based on absolute deviations from the median of all participant values and is relatively insensitive to outliers). It is encouraging to note that of those laboratories who did carry out quantification, 67% (22 of 31) performed quantification of at least one substance in all test samples. The predominant analytical techniques used by participants for quantifaction within the BS test group were GC/MS (83%) followed by LC/MS (8%) and others at (9%). 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. 2012/2-BS (3) Copyright (c) 2013 UNODC INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISE (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 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 2012/2-BS (4) Copyright (c) 2013 UNODC INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISE (ICE) Identified substances for Test Sample 1 Code Identified Substances 1Y8ESP amfetamine 343Y74 AMFETAMINE 3MDQMM Amphetamine 6DD7OZ Amphetamine and Caffeine 8881Z8 Amphetamine 88TQG8 amphetamine 8TTL6I Amphetamine 8UBRN7 amphetamine 9999CX methamphetamine 9S1TDC none A0W14T Amfetamine AII8CE Amfetamine BIWZEX AMPHETAMINE BJWNAC Amfetamine C2GUMY AMFETAMINE CJMESS Amphetamine type stimulans CSYRRB amfetamine EUV89W Amfetamine EWEPEK amfetamine EXJ929 Amphetamine 603 ng/ml FNEFK2 Anphetamine FROTTQ AMPHETAMINE GGMB9O AMFETAMINA GP9YSG AMPHETAMINE H37NYU Amphetamine HSEQDA Amfetamine HVT14V AMFETAMINE I8ATWC Amfetamine I8G88M amfetamines (amfetamine and/or tenamfetamine) IQK1D5 Amphetamine IYHZ7L AMFETAMINA JCIMSY Amfetamine KAMTKU Amphetamine KJKKSB amfetamine KOKZEC Amphetamine LRRNF7 Amfetamine LTNHUP amphetamine M9MMMM Amfetamine MZB6AZ amphetamine NYNLTC amphetamine 2012/2-BS (5) Copyright (c) 2013 UNODC INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE