Summary Report

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.

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.

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

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

Sample 1 Analysis

Identified substances for Test Sample 1

Code Identified Substances QAIJNY Amphetamine QEQYJH Amphetamine QSFQ61 amphetamine QWVW6W Amphetamine R4L0AU - RTKSN8 Amfetamine SDXS6Q amfetamine SSWJUS amphetamine TINTMK amphetamine UHHHHH amfetamine VXDMPD Amfetamine VXE8ED Amphetamine WEKKGR amfetamine YKNUPK Amphetamine YXHCCC Amphetamine ZFOMDD Amfetamine

Statement of findings for Test Sample 1

Code Amfetamine 1Y8ESP +

2HHVHH ANP Examination of the sample yielded none. 343Y74 + 1- Screnning test (EIA) positive for AMFETAMINE. 2- GC/MSD retention time and relation mass (m/z) the reference material AMFETAMINE are equivalent to those the sample 2012/2/BS-1. 3MDQMM + Screening and Confirmation by GC/MS technique : Positive 6DD7OZ + The concentration of Caffeine is 2278.5ng/mL 882968 ANP

8881Z8 + Positive Amphetamine. 88TQG8 + illegal substance on the base of law in this country 8TTL6I + Speed consumption 8UBRN7 + amphetamine 9999CX - Sample contain methamphetamine. 9S1TDC -

A0W14T ANP Only by screeing- AII8CE + Hubo consumo de anfetamina BIWZEX + POSITIVE FOR AMPHETAMINE BJWNAC +

2012/2-BS (7) Copyright (c) 2013 UNODC INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISE (ICE) Statement of findings for Test Sample 1 Code Amfetamine C2GUMY + We found amfetamine in BS-1 as a controlled substance CJMESS + Amfetamine, MDEA CSYRRB + take of amfetamine DWOKZI -

EUV89W + The sample contains amfetamine. EWEPEK +

EXJ929 +

FNEFK2 + Anphetamine consumption FROTTQ +

GGMB9O + metanfetamina found below cut-off GP9YSG + CONFIRMATION BY GC-MS H37NYU +

HSEQDA + In the urine sample marked: UNODC International Collaborative Exercises (ICE) Round IC 2012/2Sample BS-1 Urine lyophilised. 122 - 2013, detected the presence of an amphetamine HVT14V + IN THE URINE SAMPLE ANALYZED WAS DETECTED AMFETAMINE I8ATWC + Amfetamine was detected by thin layer chromatography and confirmed by GC/MS. I8G88M ANP use of amfetamines IQK1D5 + The coincidence between the retentions times and mass fragmentograms from a reference material analysed in the same conditions that the biological sample indicates trhe persence of Amphetamine.

2012/2-BS (8) Copyright (c) 2013 UNODC INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISE (ICE) Statement of findings for Test Sample 1 Code Amfetamine IYHZ7L + NON STANDARD PER QUANTITATIVA JCIMSY + Findings of amfetamine in urine are consistent with previous use of the drug. KAMTKU + 1.Results of screening test (EIA) were positive for amphetamines. 2. Retention time and fragmentation pattern obtained with the biological sample are equivalent to those obtained with "amphetamine" (Refererence Material)

KJKKSB + Results indicate that the person consumed amfetamine. KOKZEC + Test sample 2012/2/BS-1 was found to contain controlled drugs therefore positive for amphetamine for confirmatory test using Agilent GC-MS Quadrupole with concentration value of 3810.84 ng/ml at a cut-off value of 250 ng/ml. LRRNF7 + Sample is positive for Amfetamine LTNHUP +

M0KY06 +

M9MMMM + Amfetamine was detected, no other drugs as methamfetamine or drugs metabolizing to amfetamine were identified, this indicate that amfetamine was consumed MZB6AZ -

NYNLTC + positive QAIJNY +

QEQYJH + Positive for amphetamine; cut-off 200 ng/ml QSFQ61 +

QWVW6W +

QZUT7S +

2012/2-BS (9) Copyright (c) 2013 UNODC INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISE (ICE) Statement of findings for Test Sample 1 Code Amfetamine R4L0AU - - RTKSN8 + The amfetamine could come from the intake of the drug itself of from the metabolism of other drugs such a metamfetamine. SDXS6Q +

SSWJUS + positive for amphetamine TINTMK +

UHHHHH + a person was consumed amfetamine, benzfetamin, metamfetamin or selegiline VR925K +

VXDMPD +

VXE8ED + BS-1 was positive for Amphetamine WEKKGR + The results indicate either the use of amphetamine or not recent use of methamphetamine.

YKNUPK + Amphetamine YXHCCC + Amphetamine in 2012/2/BS-1 sample was identified by GCMS. ZFOMDD + Amfetamine is used for recreationnal purposes It's a controlled narcotic

Identification methods for Test Sample 1 Legend: S ... used for Screening I ... used for Identification SI ... used for both

Analytical Technique Codes Code 100 110 120 150 161 170 171 180 230 231 210 1Y8ESP S I I 2HHVHH S I 343Y74 S I 3MDQMM S I 6DD7OZ S I 882968 S 8881Z8 S I 88TQG8 S I S immunochemical test 8TTL6I S I I 8UBRN7 SI I I 9999CX S I I 9S1TDC S I A0W14T S I AII8CE SI I S Randox BIWZEX S SI BJWNAC SI S immunochromatography C2GUMY S I I CJMESS I S Dipro Druglab screen CSYRRB S I I DWOKZI S I I I EUV89W SI S Multi-Drug Screen Test Panel EWEPEK I EXJ929 SI FNEFK2 S I FROTTQ I S I GGMB9O SI I GP9YSG S I H37NYU S I I I HSEQDA S I HVT14V SI I I I8ATWC SI I I8G88M S I IQK1D5 S I IYHZ7L S I

2012/2-BS (11) Copyright (c) 2013 UNODC INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISE (ICE) Identification methods for Test Sample 1 Code 100 110 120 150 161 170 171 180 230 231 210 JCIMSY S I I KAMTKU S I KJKKSB S I I KOKZEC I I S Drug Test Kit LRRNF7 S S I I LTNHUP S S I M0KY06 S I I I M9MMM S I S Lateral Flow Immunoassay M MZB6AZ S I NYNLTC SI QAIJNY S I I QEQYJH S I QSFQ61 S I QWVW6 SI I I W QZUT7S S I R4L0AU SI RTKSN8 SI I I I SDXS6Q S S I I SSWJUS S SI TINTMK S I UHHHHH I VR925K S I VXDMPD I I VXE8ED S I WEKKGR S I SI I YKNUPK S S I I YXHCCC SI I ZFOMDD S I I

2012/2-BS (12) Copyright (c) 2013 UNODC INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISE (ICE) Summary for BS-1 The response summary of all 62 participating laboratories for the identification of BS-1, frequency of use of screening and identification techniques and false positives and negatives: Identification of BS-1:

H Response Summary Participants: 62

Amfetamine: 53

False positives for Test Sample 1 (controlled substances and/or metabolites)

Code Name Technique CJMESS MDEA 150 9999CX Metamfetamine 150/180

False negatives for BS-1 (with technique(s)) Code - 9S1TDC (180), DWOKZI (171/180), MZB6AZ (180), R4L0AU (180), 9999CX (150/180), 2HHVHH (ANP), I8G88M (180)

Frequency of use of screening and identification techniques:

Response Summary Participants: 62 Section 100 110 120 150 161 170 171 180 230 231 210 Screening 2 38 9 5 0 1 0 7 1 1 6 3.2% 61.3% 14.5% 8.1% 0.0% 1.6% 0.0% 11.3% 1.6% 1.6% 9.7% Identification 0 4 2 6 9 3 2 57 2 12 0 0.0% 6.5% 3.2% 9.7% 14.5% 4.8% 3.2% 91.9% 3.2% 19.4% 0.0% # labs using 2 38 9 10 9 3 2 57 3 12 6 this technique 3.2% 61.3% 14.5% 16.1% 14.5% 4.8% 3.2% 91.9% 4.8% 19.4% 9.7%

Z-Scores for Test Sample 1

Amfetamine CODE MAD: 374.6 Conc. * AbsDev ZScore 1Y8ESP 3927.0 552.4 1.0 6DD7OZ 3384.5 9.9 0.0 88TQG8 1985.2 1389.4 2.5 8TTL6I 3078.0 296.6 0.5 AII8CE 2274.0 1100.6 2.0 CJMESS 6262.0 2887.4 5.2 CSYRRB 3545.1 170.5 0.3 Analytical techniques in BS quantification 603.0 2771.6 5.0 100% EXJ929 9.7 6.9 7.7 6.7 15.4 6.9 6.7 FNEFK2 2360.0 1014.6 1.8 6.5 11.5 80% 7.7 GGMB9O 3630.0 255.4 0.5

GP9YSG 3052.0 322.6 0.6 60% H37NYU 8000.0 4625.4 8.3 83.9 86.2 86.7 JCIMSY 3143.0 231.6 0.4 40% 76.9 80.8 KJKKSB 3000.0 374.6 0.7 20% KOKZEC 3810.8 436.2 0.8

LRRNF7 3167.0 207.6 0.4 0% M0KY06 2992.0 382.6 0.7 BS-1 BS-2 BS-3 BS-4 Benzoyl BS-4 Methyl GC/MS LC/MS Others NYNLTC 2328.0 1046.6 1.9 QAIJNY 3374.6 0.0 0.0 QEQYJH 3400.0 25.4 0.0 QWVW6W 3398.5 23.9 0.0 QZUT7S 3320.0 54.6 0.1 SDXS6Q 3511.0 136.4 0.2 TINTMK 3800.0 425.4 0.8 UHHHHH 1840.0 1534.6 2.8 VXE8ED 3480.0 105.4 0.2 YKNUPK 3157.0 217.6 0.4 ZFOMDD 3495.0 120.4 0.2

*) Concentration: ng/ml

Z-Score has been calculated as modified Z-Score as it is a robust method and therefore relatively insensitive to outliers. Modified Z-Score = 0.6745 * AbsDev/MAD AbsDev = Absolute value of the difference between a laboratory value and the median of all the laboratory values for the given drug; MAD = median absolute deviation about the median. Modified Z-scores with an absolute value of greater than 3.5 should be considered as potential outliers. Reference: Iglewicz, B. and Hoaglin, D. C.: 1993 How to Detect and Handle Outliers, American Society for Quality Control, Milwaukee, WI

Round: 2012/2 Substance: BS-1/Amfetamine Median = 3374.6 MAD = 374.6

Sample 2 Analysis

Identified substances for Test Sample 2

Code Identified Substances 1Y8ESP THCCOOH 2HHVHH Carboxy-Tetrahydrocannabinol 343Y74 QRU¨7+&FDUER[\OLFDFLG 3MDQMM 11-nor-delta9-THC-9-carboxylic acid 6DD7OZ QRU¨7+&FDUER[\OLFDFLGDQG&DIIHLQH 8881Z8 THCA 88TQG8 THC-9-carboxylic acid 8TTL6I Carboxy-THC 8UBRN7 11-nor-ƩTHC-9-carboxylic acid 9999CX tetrahidrokanabinolic acid A0W14T 11-nor-9THC carboxylic a AII8CE 11 nor delta 9 THC-carboxylic acid BIWZEX QRU¨7+&FDUER[\OLFDFLG BJWNAC Cannabinoides C2GUMY QRU¨7+&FDUER[\OLFDFLG CJMESS Cannabinoids CSYRRB 11-nor-delta9-THC-9-carboxylic acid DWOKZI 11-nor-delta9-THC-9-carboxylic acid EUV89W QRU¨7+&FDUER[\OLFDFLG EXJ929 THC-COOH 586 ng/ml FNEFK2 THCCOOH (THC metabolite) FROTTQ 11-NOR-ƩTETRAHYDROCANNABINOL-9-CARBOXYLICACID GGMB9O THCCOOH GP9YSG CANNABIS H37NYU THC METABOLITE HSEQDA QRU¨7+&FDUER[\OLFDFLG HVT14V QRU¨7+&FDUER[\OLFDFLG I8ATWC QRU¨7+&FDUER[\OLFDFLG I8G88M cannabinoids IYHZ7L THC-COOH JCIMSY 11-nor-delta9-THC-9-carboxylic acid KAMTKU 11-Nor-Delta-9-THC-Carboxylic acid KJKKSB QRU¨7+&FDUER[\OLFDFLG KOKZEC 11-nor-delta9-THC-9-carboxylic acid LRRNF7 QRU¨7+&FDUER[\OLFDFLG LTNHUP cannabinoids M9MMMM delta 9-THC-COOH QAIJNY c-THC QEQYJH 11-nor-delta9-THC-carboxylic acid QSFQ61 THC-COOH

Identified substances for Test Sample 2

Code Identified Substances QWVW6W 11nor delta 9tetrahidrocannabinol carboxylic acid R4L0AU THC RTKSN8 Carboxy-Delta-9-THC SDXS6Q QRU¨7+&FDUER[\OLFDFLG SSWJUS QRU¨7+&FDUER[\OLFDFLG TINTMK cannabis UHHHHH THC-COOH VXDMPD QRU¨7+&FDUER[\OLFDFLG VXE8ED THC-COOH WEKKGR 11-nor-ƩTHC-9-carboxylic acid YKNUPK 11-nor-delta-9-THC-9-carboxylic acid YXHCCC 11-nor-delta9-THC-9-carboxylic acid ZFOMDD 11-nor-delta 9-THC carboxylic acid (THCCOOH)

2012/2-BS (17) Copyright (c) 2013 UNODC INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISE (ICE) Statement of findings for Test Sample 2 Code QRU¨7+& carboxylic acid C2GUMY + :HIRXQGQRU¨7+&FDUER[\OLFDFLGLQ%6DVDFRQWUROOHGVXEVWDQFH CJMESS + 11-nor delta 9THC-9 COOH acid CSYRRB + take of cannabis DWOKZI +

EUV89W + 7KHVDPSOHFRQWDLQVQRU¨7+&FDUER[\OLFDFLG EWEPEK -

EXJ929 +

FNEFK2 + Cannabis consumption FROTTQ +

GGMB9O +

GP9YSG + CONFIRMATION BY GC-MS OF THC-METABOLITE H37NYU +

HSEQDA + Exercises (ICE) Round IC 2012/2Sample BS-2 Urine lyophilised. 122 - 2013, detected the presence RIQRU¨7+&FDUER[\OLFDFLG HVT14V + ,17+(85,1(6$03/($1$/<=(':$6'(7(&7('QRU¨7+&FDUER[\OLFDFLG I8ATWC + The drug was detected by Thin Layer Chromatography and confirmed by GC/MS. I8G88M ANP use of cannabinoids IQK1D5 ANP In inmunoassay a minor concentration to the cut off means that the analytical system did not detect the presence of the investigated substances.

Statement of findings for Test Sample 2

Code QRU¨7+& carboxylic acid 1Y8ESP + metabolite of THC 2HHVHH + Examination of the sample revealed the presence of Carboxy-Tetrahydrocannabinol, a metabolite of cannabis. 343Y74 + 1- Screnning test (EIA) positive for Cannabinoids . 2- GC/MSD retention time and relation mass (m/z) the reference material 11QRU¨9-THC-9- carboxylic acid are equivalent to those the sample 2012/2/BS-2. 3MDQMM + Screening by Immunoassay technique : Positive. Confirmation by GC/MS technique : Positive 6DD7OZ + The concentration of Caffeine is 3945ng/mL 882968 ANP

8881Z8 + Positive Tetrahydrocannabinol Acid (THCA). 88TQG8 + it is not a controlled drugs in this country 8TTL6I + Cannabis use 8UBRN7 + cannabinoids 9999CX + Sample contain tetrahidrokannnabinolic acid. 9S1TDC ANP

A0W14T +

AII8CE + Hubo consumo de marihuana BIWZEX + 326,7,9()25QRU¨7+&FDUER[\OLFDFLG BJWNAC +

INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISE (ICE) Statement of findings for Test Sample 2 Code QRU¨7+& carboxylic acid C2GUMY + :HIRXQGQRU¨7+&FDUER[\OLFDFLGLQ%6DVDFRQWUROOHGVXEVWDQFH CJMESS + 11-nor delta 9THC-9 COOH acid CSYRRB + take of cannabis DWOKZI +

EUV89W + 7KHVDPSOHFRQWDLQVQRU¨7+&FDUER[\OLFDFLG EWEPEK -

EXJ929 +

FNEFK2 + Cannabis consumption FROTTQ +

GGMB9O +

GP9YSG + CONFIRMATION BY GC-MS OF THC-METABOLITE H37NYU +

2012/2-BS (20) Copyright (c) 2013 UNODC INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISE (ICE) Statement of findings for Test Sample 2 Code QRU¨7+& carboxylic acid IYHZ7L +

JCIMSY + Findings of 11-Nor-delta-9-tetrahydrocannabinol-9-carboxylic acid (a pharmacologically inactive metabolite of tetrahydrocannabinol, the active drug present in cannabis) are consistent with previous use of cannabis. KAMTKU + 1.Results of screening test (EIA) were positive for Cannabinoids. 2. Retention time and fragmentation pattern obtained with the biological sample are equivalent to those obtained with "11-Nor-Delta-9-Carboxylic acid" (Refererence Material)

KJKKSB + Results indicate that the person consumed marijuana and/or hashish. KOKZEC + Test sample 2012/2/BS-2 was found to contain controlled drugs therefore positive for 11-nor-delta9- THC-9-carboxylic acid both from the screening at a cut-off value of 50 ng/ml using screen test device an immunoassay competitive binding technique and confirmatory test using Agilent GC-MS Quadrupole with a concentration value of 658.46 ng/ml at a cut-off value of 15 ng/ml. LRRNF7 + 6DPSOHLVSRVLWLYHIRUQRU¨7+&FDUER[\OLFDFLG LTNHUP +

M0KY06 +

M9MMMM + delta 9-THC-COOH indicates that THC was consumed. MZB6AZ ANP

NYNLTC - negative QAIJNY +

QEQYJH + Positive for THC-COOH; cut-off 15 ng/ml QSFQ61 +

QWVW6W +

2012/2-BS (21) Copyright (c) 2013 UNODC INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISE (ICE) Statement of findings for Test Sample 2 Code QRU¨7+& carboxylic acid QZUT7S +

R4L0AU + - RTKSN8 + The detection of Carboxy-Delta-9-THC in urine indicates consumption of cannabinoids. SDXS6Q +

SSWJUS + SRVLWLYHIRUQRU¨7+&FDUER[\OLFDFLG TINTMK +

UHHHHH + a person was consumed marijuana or hashish VR925K +

VXDMPD +

VXE8ED + BS-2 was positive for THC-COOH (THCA) WEKKGR +

The results indicate the use of cannabis.

YKNUPK + 11-nor-delta-9-THC-9-carboxylic acid YXHCCC + 11-nor-delta9-THC-9-carboxylic acid was identified by GCMS ZFOMDD + THCCOOH is metabolite of delta 9 THC. Results indicates a cannabis consumption. THC is a mind altering drug ans often used for recreationnal purposes. Delta 9 THC is a controlled narcotic in this country

Identification methods for Test Sample 2

Legend: S ... used for Screening I ... used for Identification SI ... used for both

2012/2-BS (23) Copyright (c) 2013 UNODC INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISE (ICE) Identification methods for Test Sample 2 Code 100 110 120 150 161 170 171 180 230 231 210 JCIMSY S I I KAMTKU S I KJKKSB S I I KOKZEC I I S Drug Test Kit LRRNF7 S S I I LTNHUP S S I M0KY06 S I I I M9MMM S I S Lateral Flow Immunoassay M MZB6AZ S I NYNLTC SI QAIJNY S I I QEQYJH S I QSFQ61 S I QWVW6 SI I I W QZUT7S S I R4L0AU SI RTKSN8 SI I I I SDXS6Q S S I I SSWJUS S SI TINTMK S I UHHHHH I VR925K S I VXDMPD I I VXE8ED S I WEKKGR S I SI I YKNUPK S S I I YXHCCC SI I ZFOMDD S I I

Summary for BS-2 The response summary of all 62 participating laboratories for the identification of BS-2, frequency of use of screening and identification techniques and false positives and negatives: Identification of BS-2:

H Response Summary Participants: 62

QRU¨7+&FDUER[\OLFDFLG: 55

No False positives for Test Sample 2

False negatives for BS-2 (with technique(s)) Code - EWEPEK (180), NYNLTC (231)

Response Summary Participants: 62

Frequency of use of screening and identification techniques:

Section 100 110 120 150 161 170 171 180 230 231 210 Screening 2 38 9 5 0 1 0 7 1 1 6 3.2% 61.3% 14.5% 8.1% 0.0% 1.6% 0.0% 11.3% 1.6% 1.6% 9.7% Identification 0 4 2 6 9 3 2 57 2 12 0 0.0% 6.5% 3.2% 9.7% 14.5% 4.8% 3.2% 91.9% 3.2% 19.4% 0.0% # labs using 2 38 9 10 9 3 2 57 3 12 6 this technique 3.2% 61.3% 14.5% 16.1% 14.5% 4.8% 3.2% 91.9% 4.8% 19.4% 9.7%

Z-Scores for Test Sample 2

QRU¨7+&FDUER[\OLF acid CODE MAD: 141.3 Conc. * AbsDev ZScore 1Y8ESP 772.0 136.8 0.7 6DD7OZ 59.8 575.5 2.7 8881Z8 773.0 137.8 0.7 88TQG8 451.5 183.7 0.9 8TTL6I 480.0 155.2 0.7 AII8CE 705.0 69.8 0.3 CJMESS 135.0 500.2 2.4 Analytical techniques in BS quantification CSYRRB 1337.0 701.8 3.4 100% 720.0 84.8 0.4 9.7 6.9 7.7 6.7 EUV89W 15.4 6.9 6.7 586.0 49.2 0.2 6.5 11.5 EXJ929 80% 7.7 FNEFK2 1091.0 455.8 2.2 GGMB9O 739.0 103.8 0.5 60% GP9YSG 612.0 23.2 0.1 86.2 86.7 40% 83.9 80.8 H37NYU 135.0 500.2 2.4 76.9

I8ATWC 707.0 71.8 0.3 20% IYHZ7L 107.0 528.2 2.5 JCIMSY 502.0 133.2 0.6 0% BS-1 BS-2 BS-3 BS-4 Benzoyl BS-4 Methyl 660.0 24.8 0.1 KJKKSB GC/MS LC/MS Others KOKZEC 658.5 23.2 0.1 LRRNF7 568.0 67.2 0.3 M0KY06 293.0 342.2 1.6 QAIJNY 674.0 38.8 0.2 QEQYJH 380.0 255.2 1.2 QWVW6W 662.5 27.2 0.1 QZUT7S 769.0 133.8 0.6 R4L0AU 865.0 229.8 1.1 SDXS6Q 780.0 144.8 0.7 TINTMK 590.0 45.2 0.2 UHHHHH 420.0 215.2 1.0 YKNUPK 663.0 27.8 0.1 ZFOMDD 1037.0 401.8 1.9

*) Concentration: ng/ml

Round: 2012/2 Substance: BS-2/QRU¨7+&FDUER[\OLFDFLG Median = 635.2 MAD = 141.3

Sample 3 Analysis

Identified substances for Test Sample 3

Code Identified Substances 1Y8ESP morphine 2HHVHH Morphine 343Y74 Morphine 3MDQMM Morphine 6DD7OZ Morphine-3-O-glucuronide and Caffeine 8881Z8 Morphine 88TQG8 morphine 8TTL6I Morphine 8UBRN7 morphine 9999CX morphine 9S1TDC none A0W14T Morphine and/or metabolites AII8CE Morphine BIWZEX morphine BJWNAC Morphine C2GUMY Morphine and/or Metabolites CJMESS Opiates CSYRRB morphine EUV89W Morphine EWEPEK morfine EXJ929 Morphine 1019 ng/ml FNEFK2 Morphine, 6-monoacetilmorphine FROTTQ MORPHINE TOTAL GGMB9O MORFINE GP9YSG MORPHINE H37NYU MORPHINE - 6 glocrunide HSEQDA NO FOUND DRUGS HVT14V MORPHINE I8ATWC Morphine (free) I8G88M morphine, paracetamol IQK1D5 Morphine IYHZ7L MORFINA JCIMSY Morphine KAMTKU Morphine and Codeine KJKKSB Morphine KOKZEC Morphine LRRNF7 Morfine LTNHUP morphine M9MMMM Morphine MZB6AZ morphine

2012/2-BS (28) Copyright (c) 2013 UNODC INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISE (ICE) Identified substances for Test Sample 3 Code Identified Substances NYNLTC free morphine QAIJNY Morphine QEQYJH Morphine QSFQ61 morphine, codeine QWVW6W Free morphine R4L0AU Morphine RTKSN8 Morphine SDXS6Q morphine SSWJUS morphine TINTMK morphine UHHHHH morphine VXDMPD QRU¨7+&FDUER[\OLFDFLG0RUSKLQH VXE8ED Morphine WEKKGR morphine YKNUPK Morphine YXHCCC Morphine ZFOMDD Morphine

Statement of findings for Test Sample 3

Code Morphine 1Y8ESP +

2HHVHH + Examination of the sample revealed the presence of Morphine. 343Y74 + 1- Screnning test (EIA) positive for Opiates . 2- GC/MSD retention time and relation mass (m/z) the reference material Morphine are equivalent to those the sample 2012/2/BS-3. 3MDQMM + Screening by Immunoassay technique : Positive. Confirmation by GC/MS technique : Positive 6DD7OZ ANP The concentration of Caffeine is 9830ng/mL 882968 ANP

8881Z8 + Positive morphine. 88TQG8 + illegal substance on the base of DPR 309/90 8TTL6I + Heroin use can not be excluded, however morphine or opium consumption is also possible 8UBRN7 + morphine

9999CX + Sample contain morphine. 9S1TDC ANP Found amphetamine type stimulants on screening but unable to detect any drugs in confirmatory analysis. A0W14T ANP only screening AII8CE + Hubo consumo de morfina. BIWZEX + POSITIVE FOR morphine

BJWNAC +

2012/2-BS (30) Copyright (c) 2013 UNODC INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISE (ICE) Statement of findings for Test Sample 3 Code Morphine C2GUMY + We found Morphine and/or Metabolites in BS-3 as a controlled substance CJMESS + Codeine, Morphine

CSYRRB + take of morphine itself or a metabolic precursor DWOKZI -

EUV89W + The sample contains morphine and paracetamol.

EWEPEK +

EXJ929 + Only free morphine was measured FNEFK2 + Heroine consumption FROTTQ +

GGMB9O + SOUGHT ONLY TOTAL MORFINE GP9YSG + CONFIRMATION BY GC-MS H37NYU -

HSEQDA ANP Exercises (ICE) Round ICE 2012/2 Sample BS-3 Urine lyophilised. 122 - 2013, no detected the presence of metabolites of opioids. HVT14V + IN THE URINE SAMPLE ANALYZED WAS DETECTED MORPHINE I8ATWC + Morphine (free) was detected by Thin Layer Chromatography and confirmed by GC/MS. I8G88M + use of morphine, paracetamol IQK1D5 + The coincidence between the retentions times and mass fragmentograms from a reference material analysed in the same conditions that the biological sample indicates trhe persence of Morphine.

JCIMSY + The presence of morphine is consistent with use of pure morphine or heroin. KAMTKU + 1.Results of screening test (EIA) were positive for opiates. 2. Retention time and fragmentation pattern obtained with the biological sample are equivalent to those obtained with "morphine and codeine" (Refererence Material)

KJKKSB + Results indicate that the person consumed heroin and/or morphine. KOKZEC + Test Sample 2012/2/BS-3 was found to contain controlled drugs for Morphine, positive from the screening at a cut-off value of 300 ng/ml using screen test device an immunoassay competitive binding technique and negative for the confirmatory test using Thermo Scientific LC-MS/MS with a concentration value of 1062.61 ng/ml (Free Morphine) and 1264.39 ng/ml (Total Morphine) at a cut- off value of 2000 ng/ml. LRRNF7 + Sample is positive for Morphine LTNHUP +

M0KY06 +

M9MMMM + Morphine was detected, no other metabolite were identified. MZB6AZ +

NYNLTC + positive QAIJNY + There is almost no difference between hydrolyzed and not hydrolyzed samples. Thus, morphine- glucuronides are not included in the BS-3. QEQYJH + Positive for morphine; cut-off 200 ng/ml QSFQ61 +

QWVW6W +

2012/2-BS (32) Copyright (c) 2013 UNODC INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISE (ICE) Statement of findings for Test Sample 3 Code Morphine (Total) QZUT7S +

R4L0AU + - RTKSN8 + The morphine detectio could come from the intake of the drug itsefl or from the metabolism of other drugs such as codeine and/or heroine. SDXS6Q +

SSWJUS + positive for morphine TINTMK +

UHHHHH + a person was consumed morphine, diamorphine or codeine VR925K +

VXDMPD +

VXE8ED + BS-3 was positive for Morphine WEKKGR + The results indicate either the use of morphine or not recent use of heroin.

YKNUPK + Morphine YXHCCC + Morphine was identified by GCMS ZFOMDD + Morphine is a powerful analgesic agent often used in first emergency care, but it's also an abused drug. It's the metabolite of many opiates (Heroin, codeine). Morphine is controlled narcotic in France

Identification methods for Test Sample 3

Legend: S ... used for Screening I ... used for Identification SI ... used for both

2012/2-BS (34) Copyright (c) 2013 UNODC INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISE (ICE) Identification methods for Test Sample 3 Code 100 110 120 150 161 170 171 180 230 231 210 JCIMSY S I I KAMTKU S I KJKKSB S I I KOKZEC I I S Drug Test Kit LRRNF7 S S I I LTNHUP S S I M0KY06 S I I I M9MMM S I S Lateral Flow Immunoassay M MZB6AZ S I NYNLTC SI QAIJNY S I I QEQYJH S I QSFQ61 S I QWVW6 SI I I W QZUT7S S I R4L0AU SI RTKSN8 SI I I I SDXS6Q S S I I SSWJUS S SI TINTMK S I UHHHHH I VR925K S I VXDMPD I I VXE8ED S I WEKKGR S I SI I YKNUPK S S I I YXHCCC SI I ZFOMDD S I I

Summary for BS-3 The response summary of all 62 participating laboratories for the identification of BS-3, frequency of use of screening and identification techniques and false positives and negatives: Identification of BS-3:

H Response Summary Participants: 62

Morphine: 55

False positives for Test Sample 3 (controlled substances and/or metabolites)

Code Name Technique 6DD7OZ Morphine-3-glucuronide 231 FNEFKZ 6-Monoacetyl-morphine 180 H37NYU Morphine-6-glucoronide 171/180 KAMTKU Codeine 180 QSFQ61 Codeine 180 VXDMPD 11-nor-delta-9-THC-carboxylic acid 180/150 VXE8ED Morphine-3-glucoronide 231 VXE8ED Morphine-6-glucoronide 231 CJMESS Codeine 150

False negatives for BS-3 (with technique(s)) Code - DWOKZI (171/180) , H37NYU (171/180)

Frequency of use of screening and identification techniques:

Response Summary

Participants: 62 Section 100 110 120 150 161 170 171 180 230 231 210 Screening 2 38 9 5 0 1 0 7 1 1 6 3.2% 61.3% 14.5% 8.1% 0.0% 1.6% 0.0% 11.3% 1.6% 1.6% 9.7% Identification 0 4 2 6 9 3 2 57 2 12 0 0.0% 6.5% 3.2% 9.7% 14.5% 4.8% 3.2% 91.9% 3.2% 19.4% 0.0% # labs using 2 38 9 10 9 3 2 57 3 12 6 this technique 3.2% 61.3% 14.5% 16.1% 14.5% 4.8% 3.2% 91.9% 4.8% 19.4% 9.7%

Z-Scores for Test Sample 3

Morphine (Total) Morphine CODE MAD: 195.9 CODE MAD: 195.9 Conc. * AbsDev ZScore Conc. * AbsDev ZScore 1Y8ESP 1100.0 96.4 0.3 CJMESS 1000.0 196.4 0.7 88TQG8 1182.7 13.7 0.0 CSYRRB 1390.4 194 0.7 8TTL6I 821.0 375.4 1.3 EXJ929 1019 177.4 0.6 AII8CE 1233.0 36.6 0.1 GP9YSG 1441 244.6 0.8 EUV89W 1530.0 333.6 1.1 I8ATWC 1434 237.6 0.8 FNEFK2 3280.0 2083.6 7.2 M0KY06 1001 195.2 0.7 GGMB9O 1637.0 440.6 1.5 QWVW6W 1002.2 194.2 0.7 GP9YSG 1441.0 244.6 0.8 ZFOMDD 1389 192.6 0.7 IYHZ7L 203.0 993.4 3.4

JCIMSY 1026.0 170.4 0.6 Analytical techniques in BS quantification 1264.4 67.9 0.2 KOKZEC 100% 9.7 6.9 7.7 6.7 15.4 LRRNF7 943.0 253.4 0.9 6.9 6.7 6.5 11.5 QAIJNY 1036.9 159.5 0.5 80% 7.7 QEQYJH 1640.0 443.6 1.5 60% QZUT7S 1210.0 13.6 0.0

R4L0AU 885.0 311.4 1.1 83.9 86.2 86.7 40% 76.9 80.8 SDXS6Q 1175.0 21.4 0.1 TINTMK 1000.0 196.4 0.7 20% UHHHHH 1700.0 503.6 1.7 0% YKNUPK 1293.0 96.6 0.3 BS-1 BS-2 BS-3 BS-4 Benzoyl BS-4 Methyl

GC/MS LC/MS Others *) Concentration: ng/ml

Round: 2012/2 Substance: BS-3/Morphine Median = 1196.4 MAD = 195.9 8

7.2

6 (value) 4 3.4 Z score

Modified 2 1.7 1.5 1.5 1.3 1.1 1.1 0.8 0.9 0.8 0.8 0.7 0.7 0.7 0.6 0.5 0.7 0.6 0.7 0.7 0.3 0.2 0.3 0.0 0.1 0.1 0.0 0

AII8CE JCIMSY EXJ929 QAIJNY 8TTL6I CJMESS IYHZ7L QZUT7S 1Y8ESP KOKZEC GP9YSG GP9YSG QEQYJH FNEFK2 SDXS6Q CSYRRB M0KY06 LRRNF7 YKNUPK R4L0AU 88TQG8 I8ATWC TINTMK EUV89W ZFOMDD GGMB9O Laboratories UHHHHH QWVW6W

Sample 4 Analysis

Code Identified Substances 1Y8ESP benzoylecgonine 2HHVHH Benzoylecgonine 343Y74 Benzoylecgonine 3MDQMM Benzoylecgonine, Methylecgonine 6DD7OZ Benzoylecgonine and Caffeine 8881Z8 BE 88TQG8 benzoylecgonine, methyacgonine 8TTL6I Benzoyl-ecgonine 8UBRN7 benzoylecgonine 9999CX benzoilecgonine,methylecgonine benzoylecgonine,methylecgonine

9S1TDC none A0W14T Benzoylecgonine-Methylecgonine AII8CE Benzoylecgonine and methylecgonine BIWZEX Benzoylecgonine, Methylecgonine BJWNAC cocaine metabolites C2GUMY Benzoylecgonine CJMESS Cocaine and metabolites CSYRRB benzoylecgonine and methylecgonine DWOKZI Methylecgonine EUV89W Benzoylecgonine and Methylecgonine EWEPEK cocaine, benzoilecgonine EXJ929 Benzoyl-ecgonine 1844 ng/ml FNEFK2 Cocaine FROTTQ BENZOYLECGONINE, METHYLECGONINE GGMB9O BENZOILECGININE GP9YSG COCAINE HSEQDA BENZOYLECGONINE HVT14V BENZOYLECGONINE I8ATWC Benzoylecgonine and Methylecgonine I8G88M methylecgonine, benzoylecgonine, paracetamol IQK1D5 Benzoylecgonine IYHZ7L BENZOILECGONINA JCIMSY Benzoylecgonine and Methylecgonine KAMTKU Benzoilecgonine KJKKSB Benzoylecgonine, methylecgonine KOKZEC Benzoylecgonine LRRNF7 Benzoylecgonine, Methylecgonine LTNHUP benzoylecgonine, methylecgonine M9MMMM Benzoylecgonine, Methylecgonine MZB6AZ Anhidroecgonine methyl ester

2012/2-BS (39) Copyright (c) 2013 UNODC INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISE (ICE) Identified substances for Test Sample 4 Code Identified Substances NYNLTC benzoylecgonine and methylecgonine QAIJNY Benzoylecgonine, Methylecgonine QEQYJH Benzoylecgonine QSFQ61 cocaine, methylecgonine QWVW6W Benzoylecgonine and Methylecgonine R4L0AU Benzoylecgonine and Methylecgonine RTKSN8 Benzoylgonine/metilecgonine SDXS6Q benzoylecgonine, methylecgonine SSWJUS Benzoylecgonine, Methylecgonine and Paracetamol (Acetaminophen) TINTMK cocaine UHHHHH benzoylecgonine, paracetamol VXDMPD QRU¨7+&FDUER[\OLFDFLG VXE8ED Benzoylecgonine WEKKGR benzoylecgonine, methylecgonine YKNUPK Benzoylecgonine YXHCCC Methylecgonine ZFOMDD Benzoylecgonine ans methylecgonine

Statement of findings for Test Sample 4

Code Benzoylecgonine Methylecgonine 1Y8ESP + + metabolite of cocaine 2HHVHH + - Examination of the sample revealed the presence of benzoylecgonine, a metabolite of cocaine. 343Y74 + - 1- Screnning test (EIA) positive for Cocaine and/or metabolites . 2- GC/MSD retention time and relation mass (m/z) the reference material Benzoylecgonine are equivalent to those the sample 2012/2/BS-4. 3MDQMM + + Screening by Immunoassay technique : Positive. Confirmation by GC/MS technique : Positive 6DD7OZ + - The concentration of Caffeine is 9275ng/mL 882968 ANP ANP

8881Z8 + ANP Positive Benzoylecognine (BE). 88TQG8 + + it is not a controlled drugs in Italy 8TTL6I + ANP Cocain use 8UBRN7 + ANP benzoylecgonine and paracetamol 9999CX + + Sample contain cocaine metabolites-benzoylecgonine, methylecgonine. 9S1TDC ANP ANP Found cannabiniods on screening but unable to detect any drugs in confirmatory analysis. A0W14T + +

AII8CE + + Hubo consumo de Cocaina. BIWZEX + + POSITIVE FOR Benzoylecgonine, Methylecgonine BJWNAC + +

2012/2-BS (41) Copyright (c) 2013 UNODC INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISE (ICE) Statement of findings for Test Sample 4 Code Benzoylecgonine Methylecgonine C2GUMY + ANP We found Benzoylecgonine in BS-4 as a controlled substance CJMESS + + Benzoylecgonine, Methylecgonine CSYRRB + + take of cocaïne DWOKZI - +

EUV89W + + The sample contains benzoylecgonine, methylecgonine and paracetamol. EWEPEK + ANP

EXJ929 + ANP As the concentration was over the highest point of the calibration curvethe the sample was diluted 1:1 by negative urine, processed, and analysed FNEFK2 + + Detection of cocaine and its metabolites benzoilecgonine and ecgonine metilester. Cocaine consumption FROTTQ + +

GGMB9O + ANP FOUND COCAINE GP9YSG + ANP CONFIRMATION BY GC-MS OF BENZOYLECGONINE H37NYU - -

HSEQDA + ANP Exercises (ICE) Round ICE 2012/2 Sample BS-4 Urine lyophilised. 122 - 2013, detected the presence of an metabolite of cocaine (Benzoylecgonine). HVT14V + ANP IN THE URINE SAMPLE ANALYZED WAS DETECTED BENZOYLECGONINE I8ATWC + + Benzoylecgonine and Methylecgonine were detected by Thin Layer Chromatography and confirmed by GC/MS. I8G88M + + use of cocaine metabolites, paracetamol

2012/2-BS (42) Copyright (c) 2013 UNODC INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISE (ICE) Statement of findings for Test Sample 4 Code Benzoylecgonine Methylecgonine IQK1D5 + - The coincidence between the retentions times and mass fragmentograms from a reference material analysed in the same conditions that the biological sample indicates trhe persence of Benzoylecgonine. IYHZ7L + + METHYLECGONINA JCIMSY + + Findings of metabolites, benzoylecgonine and ecgonine methyl ester (inactive breakdown products of cocaine), in urine are consistent with previous use of cocaine. KAMTKU + - 1.Results of screening test (EIA) were positive for cocaine and metabolites. 2. Retention time and fragmentation pattern obtained with the biological sample are equivalent to those obtained with "Benzoylecgonine" (Refererence Material)

KJKKSB + + Results indicate that the person consumed cocaine. KOKZEC + ANP Test sample 2012/2/BS-4 was found to contain controlled drugs therefore positive for Benzoylecgonine both from the screening at a cut-off value of 300 ng/ml using screen test device an immunoassay competitive binding technique and confirmatory test using Thermo Scientific LC- MS/MS with a concentration value of 2482.39 ng/ml at a cut-off value of 100 ng/ml. LRRNF7 + + Sample is positive for Benzoylecgonine and Methylecgonine LTNHUP + + metabolites of cocaine M0KY06 + +

M9MMMM + + Benzoylecgonine and Methylecgonine indicate that cocaine was consumed. MZB6AZ - -

NYNLTC + + positive QAIJNY + +

QEQYJH + - Positive for benzoylecgonine; cut-off 200 ng/ml QSFQ61 - +

2012/2-BS (43) Copyright (c) 2013 UNODC INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISE (ICE) Statement of findings for Test Sample 4 Code Benzoylecgonine Methylecgonine QWVW6W + +

QZUT7S + +

R4L0AU + + - RTKSN8 + + The detection of benzoylecgonine and mehylecgonine in a urine sample indicates consumption of the drug cocaine. SDXS6Q + +

SSWJUS + + positive for Benzoylecgonine, Methylecgonine and Paracetamol (Acetaminophen) TINTMK + +

UHHHHH + - a person was consumed cocaine and paracetamol VR925K ANP ANP

VXDMPD - -

VXE8ED + + BS-4 was positive for Cocaine metabolites WEKKGR + + The presence of benzoylecgonine and methylecgonine indicates the use of cocaine.

YKNUPK + ANP Benzoylecgonine YXHCCC - + Methylecgonine was identified by GCMS. ZFOMDD + + Results indicate cocaine consumption. Cocaine is an illegal recreationnal drug. It's controlled narcotic in this country

Identification methods for Test Sample 4

Legend: S ... used for Screening I ... used for Identification SI ... used for both

2012/2-BS (45) Copyright (c) 2013 UNODC INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISE (ICE) Identification methods for Test Sample 4 Code 100 110 120 150 161 170 171 180 230 231 210 JCIMSY S I I KAMTKU S I KJKKSB S I I KOKZEC I I S Drug Test Kit LRRNF7 S S I I LTNHUP S S I M0KY06 S I I I M9MMM S I S Lateral Flow Immunoassay M MZB6AZ S I NYNLTC SI QAIJNY S I I QEQYJH S I QSFQ61 S I QWVW6 SI I I W QZUT7S S I R4L0AU SI RTKSN8 SI I I I SDXS6Q S S I I SSWJUS S SI TINTMK S I UHHHHH I VR925K S I VXDMPD I I VXE8ED S I WEKKGR S I SI I YKNUPK S S I I YXHCCC SI I ZFOMDD S I I

2012/2-BS (46) Copyright (c) 2013 UNODC INTERNATIONAL QUALITY ASSURANCE PROGRAMME (IQAP) INTERNATIONAL COLLABORATIVE EXERCISE (ICE) Summary for BS-4 The response summary of all 62 participating laboratories for the identification of BS-4, frequency of use of screening and identification techniques and false positives and negatives:

Identification of BS-4:

Response Summary Participants: 62

Benzoylecgonine: 53 Methylecgonine: 37

FFalse positives for Test Sample 4 (controlled substances and/or metabolites)

Code Name Technique VXDMPD 11-nor-delta-9-THC-carboxylic acid 180/150

False negatives for BS-4 (with technique(s)). Benzoylecgonine Code - DWOKZI (171/180), H37NYU (171/180), MZB6AZ (180), QSFQ61 (180), , VXDMPD (180/150), YXHCCC (180) Methylecgonine Code - H37NYU (171/180), IQK1D5 (180), 2HHVHH (180), 343Y74 (180), 6DD7OZ (231), 8881Z8 (180), KAMTKU (180), MZB6AZ (180), QEQYJH (180), UHHHHH (180), VXDMPD(180/150)

Frequency of use of screening and identification techniques:

Z-Scores for Test Sample 4

Benzoylecgonine Methylecgonine CODE MAD: 420.0 MAD: 84.1 Conc. * AbsDev ZScore Conc. * AbsDev ZScore 1Y8ESP 2785.0 505.0 0.8 1123.0 0.0 0.0 6DD7OZ 2785.0 505.0 0.8 88TQG8 2199.5 80.5 0.1 8TTL6I 2000.0 280.0 0.5 AII8CE 3925.0 1645.0 2.6 1252.0 129.0 1.0 CSYRRB 2202.6 77.4 0.1 1207.1 84.1 0.7 EUV89W 2280.0 0.0 0.0 290.0 833.0 6.7 EXJ929 1844.0 436.0 0.7 FNEFK2 1750.0 530.0 0.9 GGMB9O 400.0 1880.0 3.0 GP9YSG 1749.0 531.0 0.9 I8ATWC 3360.0 1080.0 1.7 3680.0 2557.0 20.5 IYHZ7L 1230.0 1050.0 1.7 JCIMSY 2289.0 9.0 0.0 1236.0 113.0 0.9 KOKZEC 2482.4 202.4 0.3 LRRNF7 956.0 1324.0 2.1 668.0 455.0 3.7 M0KY06 2297.0 17.0 0.0 1114.0 9.0 0.1 NYNLTC 1143.0 1137.0 1.8 1100.0 23.0 0.2 QAIJNY 1693.2 586.8 0.9 939.9 183.1 1.5 QEQYJH 2700.0 420.0 0.7 QWVW6W 2413.3 133.3 0.2 1178.1 55.1 0.4 QZUT7S 2180.0 100.0 0.2 1193.0 70.0 0.6 R4L0AU 1980.0 300.0 0.5 1050.0 73.0 0.6 SDXS6Q 2280.0 0.0 1160.0 37.0 0.3 TINTMK 2000.0 280.0 0.5 600.0 523.0 4.2 VXE8ED 3140.0 860.0 1.4 YKNUPK 2220.0 60.0 0.1 ZFOMDD 2695.0 415.0 0.7 1182.0 59.0 0.5

*) Concentration: ng/ml

Analytical techniques in BS quantification

100% 9.7 6.9 7.7 6.7 15.4 6.9 6.7 6.5 11.5 80% 7.7

60%

83.9 86.2 86.7 40% 76.9 80.8

20%

0% BS-1 BS-2 BS-3 BS-4 Benzoyl BS-4 Methyl

Round: 2012/2 Substance: BS-4/Benzoylecgonine Median = 2280 MAD = 420.0

Round: 2012/2 Substance: BS-4/Methylecgonine Median = 1123 MAD = 84.1

The Laboratory and Scientific Section UNODC, International Centre Wagramer Strasse 5, P.O. Box 500 A-1400, Vienna, AUSTRIA Email: [email protected]