Analyte Targeting for NPS Identification in Seized Drugs and Toxicology

Transcription

1 Analyte Targeting for NPS Identification in Seized Drugs and Toxicology Alex J. Krotulski, MS Research Scientist Center for Forensic Science Research & Education

2 Background Explosion and expansion of NPS nationally and internationally

3 Chemical Variations Historically common among a variety of chemical applications and drug discovery fields Emerging NPS often share similar backbones or structural features with other NPS or more common drugs of abuse vs. vs.

4 Chemical Variations Challenges: Seemingly endless variations Rate of turnover Incorporation in analytical method/testing procedures Unknown pharmacological effects

5 Chemical Variations Advantages: Similarities in chemical classification or characterization Expected retention time range Characteristic fragmentation patterns Ease of chemical synthesis for vendors*

6 Analyte Targeting

7 Analyte Targeting Systematic approach to identifying compounds of interest Drugs of abuse NPS Steps in the process: Sample preparation/extraction Analytical method and instrumentation Data processing

8 Analyte Targeting Approach is going to differ for NPS Instrumental capabilities / platform available Knowledge about the drug market Justification dependent Availability of standard reference material Accreditation guidelines / operating procedures And so on...

9 Instrumentation

10 Instrumentation Key to NPS discovery Type of instrumentation or access to instrumentation is dependent on agency, jurisdiction, country, etc. Importance of diverse data sharing Importance of collaboration and joint efforts

11 Instrumentation (USA) Typical Crime Laboratory Infrared spectroscopy (IR) Gas chromatography mass spectrometry (GC-MS) Typical Toxicology Laboratory Gas chromatography mass spectrometry (GC-MS) Liquid chromatograph mass spectrometry (LC-MS) Liquid chromatograph tandem mass spectrometry (LC-MS/MS) Advanced Laboratories High resolution accurate mass spectrometry (HRAMS) Nuclear magnetic resonance (NMR)

12 Gas Chromatography Mass Spectrometry Non-targeted analytical technique for the detection of volatile compounds Drugs of abuse, NPS, therapeutic drugs, etc. Robust and reproducible Highly reproducible fragmentation patterns Library searching Inter-laboratory information sharing Structural elucidation*

13 High Resolution Accurate Mass Spectrometry Emerging analytical technique for the detection of drugs of abuse and NPS Time-of-flight (TOF) and OrbiTrap mass analyzers Allows for formula elucidation Analyte Formula Nominal Mass Exact Mass Difference Methoxyacetyl Fentanyl C 22 H 28 N 2 O AB-005 C 23 H 32 N 2 O ~100 ppm Non-targeted screening applications

14 Nuclear Magnetic Resonance Analytical technique for chemical characterization Ability to perform a variety of experiments 1 H, 13 C, 14 N, 19 F, etc. 2D: COSY, NOESY, etc. 60 MHz, 300 MHz, 900 MHz, etc. Allows for formula structural elucidation Typically highest utility in combination with GC-MS or LC-QTOF

15 Combined Workflow GC-MS Good screening technique/library search capability LC-QTOF Provides additional level of specificity/formula elucidation NMR Structural confirmation Positional isomer differentiation

16 Data Processing

17 Data Processing Strategies for identifying NPS Detection is only the first step 1. Targeted data processing Mass list, retention time, library/database 2. Non-targeted data processing Peak finding strategies, software algorithms 3. Manual data processing Analyst-specific approach

18 Analyte Targeting Targeted data processing Can be most useful approach for NPS identification and/or discovery in biological samples Typically the simplest processing approach Samples are often a soup of NPS, drugs, metabolites, endogenous species pick out what you want to see

19 Analyte Targeting Manual data processing Most useful approach for NPS identification and/or discovery in seized drug exhibits Manufacturer: pure powders typically 1 substance Distribution: combinations upwards of 20 components

20 Casework Examples

21 Example Seized Drug Collaboration with USA ports of entry Unknown or suspicious powders sent for identification Workflow (Manual Data Processing): Acid/base extraction Initial analysis by GC-MS For suspected NPS, analysis by LC-QTOF For structural confirmation, analysis by NMR

22 Example Seized Drug Initial analysis by GC-MS

23 Example Seized Drug Initial analysis by GC-MS Unknown Analyte Internal Standards

24 Example Seized Drug Initial analysis by GC-MS

25 Example Seized Drug For suspected NPS, analysis by LC-QTOF

26 Example Seized Drug For suspected NPS, analysis by LC-QTOF Unknown Analyte Internal Standards

27 Example Seized Drug TOF MS Data

28 Example Seized Drug MS/MS Data (MS/MS Acquisition: High Sensitivity)

29 Example Seized Drug Retention Time: min Synthetic cannabinoid? Suspected Formula: C 25 H 33 N 3 O Consistent with drug of abuse or NPS Prominent Fragment Ion: C 10 H 15 + Is this fragment in our library database?

30 Example Seized Drug Excel Spreadsheet used to populate XIC List

31 Example Seized Drug AKB-48 (APINACA) 5F-JWH-018 Adamantyl Analogue

32 Example Seized Drug Piecing the puzzle back together C8H5N2O+ C15H17N2O+ +C7H12

33 Example Seized Drug Piecing the puzzle back together Formula: C 25 H 33 N 3 O Adamantyl: C 10 H 15 Indazole: C 8 H 4 N 2 O Carbon Chain: C 7 H 13 What s missing? Nitrogen (+H) Linker

34 Example Seized Drug Adamantyl-CHMINACA Structure confirmed by NMR

35 Example Toxicology Biological sample extracts received from NMS Labs Testing procedures for confirmation of NPS Workflow (Targeted Data Processing): Sample preparation/extraction) At NMS Labs Re-analysis by LC-QTOF only Complex, tiered data processing strategies Mass list and library database Continual updating as NPS standards become available

36 Example Toxicology Library/Database Name Formula Exact mass Fragment accurate masses Retention time Library spectra >700 compounds NPS: >500 DoA: 66 Other: 175

37 Example Toxicology

38 Example Toxicology Sample 1

39 Example Toxicology Sample 2

40 Discussion/Conclusions

41 Discussion/Conclusions You can only identify the NPS you are detecting or processing for Analyte targeting is dependent on and customizable to the platform, analyst, discipline, etc. Intra-laboratory data sharing is extremely beneficial for identifying emerging NPS SWGDRUG, Cayman Chemical, European Project Response, NPS Discovery, etc. Small number of laboratories reporting NPS for the first time

42 Discussion/Conclusions Throwing a wrench into NPS identification: Are you identifying the true NPS contained within the sample? What s in a sample/exhibit vs. what is detected Synthetic cannabinoid metabolite in blood 25X-NBOH by GC-MS

43 Questions? Contact Information Alex J. Krotulski, MS