the challenges of analytical method validation for hallucinogens and designer stimulants in...
DESCRIPTION
The Challenges of Analytical Method Validation for Hallucinogens and Designer Stimulants in Biological Samples Using LC-TOF Hosted by Agilent Technologies on October 8, 2012 Presented by Barry K Logan, Ph.D., NMS Labs National Director, Forensic ServicesTRANSCRIPT
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The Challenges of Analytical Method Validation for Hallucinogens and Designer Stimulants in Biological Samples Using LC-TOF
Barry K Logan, Ph.D., NMS Labs National Director, Forensic Services
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Goals
•The Need:
•Accurate, sensitive, versatile tools.
•Identification
•Detection
•Routine analysis
•Novel, complex, esoteric and designer drugs and other analytes in a variety of matrices and sample types.
•The solution:
•LCMS/MS
•LCTOF, LCQTOF
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NMS Labs Center for InnovationCrim. Blood Urine
JWH-018JWH-073
~35 Cpds.
Tim
e
CP47,497
HU210
JWH-018JWH-073JWH-019JWH-250
JWH-081JWH-122JWH-200JWH-210AM-2201AM-694RCS-4RCS-8
DEA scheduling
Professional activitiesClient Enquiries
Crim. Casework
Academic collaboration
International contacts
Vendor activity
JWH-018 metsJWH-073 mets
JWH-019 metJWH-250 metAM-2201 met
JWH-018 metsJWH-073 mets
CP47,497 (C8)
~17 Cpds.
CP47,497 (C7)HU-210
Internet purchases
Web Intelligence
Customer Feedback
~42 Cpds.
JWH-203JWH-022AM-2233AM-1248XLR-11UR-144AKB-48
A720,296
RCS-4 metJWH-122 metJWH-210 metJWH-081 metJWH-200 metJWH-250 metJWH-210 metJWH-081 metJWH-122 metJWH-398 met
Synthetic C
annabinoids
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NMS Labs Center for InnovationCrim. Blood/urine
MDMAMDAMDEA
TFMPPBZPDBZP5-MeO-DIPT…
~30 Cpds.
Tim
e
atropine benzylpiperazinedextromethorphandimethyltryptamineketamineMDAMDMAmescalinephencyclidinescopolamine2C-B 2C-B-FLY2C-T-75-MeO-DIPTAMTBDB MBDBMephedroneLSDbufoteninpsilocinsalvinorin Asalvinorin BTFMPP
DEA scheduling
Professional activitiesClient Enquiries
Crim. Casework
Academic collaboration
International contacts
Vendor activity
~10 Cpds.
Internet purchases
Web Intelligence
Customer Feedback
~70 Cpds.
Blood/urine Expanded
…MDPVMethyloneAmphetaminem-CPPO-desmethyltramadolDMAA3,4-DMMC4-MECPentylone2C-C2C-E2C-I2C-P2C-T-2Bromo-Dragon Fly5-MeO-DALTBuphedroneEthylonePetedronePhenazepam….
Stim
ulants and Hallucinogens
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The Scope
Bath Salts and Stimulant Designer Drugs – Expanded 43 Compounds
• Classical StimulantsAmphetamine, Methamphetamine, MDMA, MDEA, DOM,
DOB, PMA, Cocaine, Benzoylecgonine
• CathinonesCathinone, Mephedrone, Buphedrone, Methylone, Ethylone,
Butylone, Pentylone, Mmethcathinone, Methedrone, Naphyrone, 3-FMC, 4-FMC, 4-MEC, 3,4-DMMC,
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The Scope
Bath Salts and Stimulant Designer Drugs – Expanded 43 Compounds (Continued)
• 2C Series – Stimulants with Hallucinogenic Properties2C-B, 2C-C, 2C-E, 2C-H, 2C-I, 2C-N, 2C-P, 2C-T-2, 2C-T-7
• BenzylpiperazinesBZP, TFMPP, DBZP, MBZP, m-CPP
• OthersPhenazepam, Mitragynine, 7-OH Mitragynine,
O-desmethyltramadol, DMAA
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The Technology
Performance Checklist:
High Sensitivity High Performance High Resolution Mass Accuracy (>5ppm) User Friendly and Adaptable Data Reduction Tools Versatility Stable Performance Robust Design Technical Support Value/ROI
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The Technology
Liquid Chromatography-Time of Flight Mass Spectrometry (LCTOF/LCQTOF)
• Agilent 1200 HPLC system• Agilent 6230 with Jet Stream Technology
• Mass Hunter Software
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The Technology
Liquid Chromatography-Time of Flight Mass Spectrometry (LCTOF/LCQTOF)
• Agilent 1200 HPLC system• Agilent 6230 with Jet Stream Technology• Agilent 6530 QTOF• Mass Hunter Software
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Time of Flight (TOF)
6200/6500 Series Time of Flight Mass Spectrometry (LCTOF/LCQTOF)
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Analysis
Liquid Chromatography-Time of Flight Mass Spectrometry (LCTOF/LCQTOF) - Control
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•A sample is considered positive if the following are true:
The area is greater than or equal to 25% of the average area of the responses for the two decision point cut off calibrators (10ng/mL).
The retention times are within +/- 0.02 min of the average retention time of the decision point cut off calibrators.
The Mass Accuracy is within +/- 30ppm
Establishing Acceptance Criteria
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Analysis
Liquid Chromatography-Time of Flight Mass Spectrometry (LCTOF/LCQTOF) - Control
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Analysis
Liquid Chromatography-Time of Flight Mass Spectrometry (LCTOF/LCQTOF)
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Analysis
Liquid Chromatography-Time of Flight Mass Spectrometry (LCTOF/LCQTOF) - Control
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Analysis
Liquid Chromatography-Time of Flight Mass Spectrometry (LCTOF/LCQTOF) – Case 1
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Analysis
Liquid Chromatography-Time of Flight Mass Spectrometry (LCTOF/LCQTOF) – Case 1
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Analysis
Liquid Chromatography-Time of Flight Mass Spectrometry (LCTOF/LCQTOF) – Case 1
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Analysis
Liquid Chromatography-Time of Flight Mass Spectrometry (LCTOF/LCQTOF) – Case 1
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Analysis
Liquid Chromatography-Time of Flight Mass Spectrometry (LCTOF/LCQTOF) – Case 1
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Analysis
Liquid Chromatography-Time of Flight Mass Spectrometry (LCTOF/LCQTOF) – Case 1
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Analysis
Liquid Chromatography-Time of Flight Mass Spectrometry (LCTOF/LCQTOF) – Case 1
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Analysis
Liquid Chromatography-Time of Flight Mass Spectrometry (LCTOF/LCQTOF) – Case 1
Criterion Value
Mass Defect 161.46ppm
RT Diff 0.016min
Apparent Conc.
36ng/mL
Not Confirmed X
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Analysis
Liquid Chromatography-Time of Flight Mass Spectrometry (LCTOF/LCQTOF) – Case 2
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Analysis
Liquid Chromatography-Time of Flight Mass Spectrometry (LCTOF/LCQTOF) – Case 2
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Analysis
Liquid Chromatography-Time of Flight Mass Spectrometry (LCTOF/LCQTOF) – Case 2
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Analysis
Liquid Chromatography-Time of Flight Mass Spectrometry (LCTOF/LCQTOF) – Case 2
MDMA C11H15NO2
193.2423
MethedroneC11H15NO2
193.2423
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Analysis
Liquid Chromatography-Time of Flight Mass Spectrometry (LCTOF/LCQTOF) – Case 2
MDMA C11H15NO2
193.2423
MethedroneC11H15NO2
193.2423
Confirmed
by GCMS
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Qualitative Validation
Assay Validation Cut-off Verification Sensitivity and Specificity Interfering Substances Carryover Matrix Matching Stability
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Time of Flight
8756B/U Bath Salts and Stimulants Designer Drugs – Expanded
8085B/U Drug Impaired Driving/DRE Toxicology Bath Salts Add-On
• Test design based on:• 43 compounds• Regular updates based on trends• Compounds found in “bath salts” casework• Federally Scheduled Drugs• Covers additional drugs scheduled in many
states• Drugs emerging in Europe
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Designer Drugs Trends
NMS Labs Designer Drugs Trends ReportAugust 2012
http://www.nmslabs.com/services-forensic-designer-drug-trends
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Compare and Contrast
LCMSMS LCTOF LCQTOFTarget Compound Analysis
Comprehensive Screening
Target Compound analysis
Must set up scanning groups
Acquire all data and query database later
Acquires more limited data in QTOF mode
WYSIWYG Post hoc identification Post hoc identification
Compound specific transitions
Molecular formula information
Molecular formula information for fragments
Limited structural confirmation
No structural information
Limited Structural information
May differentiate isobars
TOF does not differentiate isobars
May differentiate isobars
Lower Cost Higher cost Even higher cost
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•NMS Labs• Alex Maggitti• Sherri Kacinko• Matthew McMullin• Fran Diamond
•Agilent• Tom Gluodenis• Jay Levine• Mary Cuddrye• John Hughes
Acknowledgements