1. Role of Toxicology in Criminal Investigations
Lyndsey Knoy, D-ABFT-FT
Forensic Scientist
Washington State Toxicology Laboratory

2. What Is Forensic Toxicology?
Forensic Toxicology is an interdisciplinary science that analyzes blood, fluid and/or tissues for the presence of alcohol, drugs, or poisons and applies the results of that analysis to answer the question:
Did drugs and/or alcohol contribute to a person’s impairment and/or their death?

3. Washington State Toxicology Laboratory
WSP
FLSB
Crime Lab
Toxicology Lab
Breath Test Section
DRE Program

4. Toxicology Laboratory
Seattle laboratory services WA, AK, OR
State Toxicologist – Fiona Couper
Lab Manager – Brianna Peterson
QA Manager –
Amanda Black
Technical Lead-
Brianne O’Reilly
2 Supervisors – Brian Capron, Brittany Thomas
14 Toxicologists
Support staff

5. State Toxicology Lab Services
Medical Examiners and Coroners
-Death Investigations
Law Enforcement Agencies
-DUI and DRE
Drug Facilitated Sexual Assaults
Liquor and Cannabis Control Board
Labor and Industry

6. Cases Handled Death Investigations Overdoses Suicides Poisonings
Traffic Fatalities
Homicides
Undetermined

7. Cases Handled Police Investigations DUI/DRE Drug Investigations
Drug Endangered Children
Liquor and Cannabis Board
Drug Facilitated Sexual Assaults

8. Caseload 2017– 15,947 cases! We receive ~1300 cases/month!
675 from AK, 226 from OR
DUI/DRE: 10,272
Top Drugs of 2017:
Ethanol
THC/COOH-THC
Methamphetamine/amphetamine
Morphine
LK: 1,496 cases!
,617 cases
Court Testimony for 2017
1,529 hours for 308 appearances
LK: 139 hours for 26 cases

9. The Toxicology Lab Data Analysis Sample receipt and log-in
(Andrew Gingras)
Sample receipt and log-in
(Bob Smith, Former PEC)
Create Report
Lab analysis
(Katie Harris)
Agency/Prosecutor
Expert Testimony

10. Sample Handling and Analysis
Sample receipt and accession
Alcohol analysis
Drug screen analysis
Confirmation analysis
Specialty drug analysis
Prepare report
Review by supervisors and manager

11. Receiving Samples US Postal Service Courier (UPS / FedEx)
Hand delivered
Campus mail

12. Sample Receipt and Log-In
Logged in as they arrive by a PEC
Photos are taken of mail packaging
and evidence seals
Each sample checked for name,
agency case#, volume and tampering
Each case receives a
Unique identifier number
(eg. ST )
Maintains chain of custody

13. Sample Receipt and Log-In
Each case entered in LIMS database
ST# is used to track samples in the laboratory
Currently about 80 cases are received daily

14. Sample Handling and Analysis
Sample receipt and accession
Alcohol analysis
Drug screen analysis
Confirmation analysis
Specialty drug analysis
Prepare report
Review by supervisors and manager

15. Blood Alcohol Forensic collection of sample Gray top tube Vacuum
Preservative (NaF) & anticoagulant (C2K2O4)
FDA Regulated
Certificates from the manufacturer
Experts can testify to hearsay
Samples are not clotted
Vacuum
Expiration Date: refers to vacuum

16. Ethanol Analysis All samples are tested for alcohol (ethanol)
Head-Space Gas Chromatography (HS/GC)
Run in duplicate
Two aliquots
Two instruments
Different columns
Different temperature programs
Calibrators for each run
Control and negative every ten samples

17. Ethanol Analysis Distinguishes between ethanol and other volatiles
Alcohol is a volatile
When heated, volatiles enter the vapor phase
Vapor is injected onto a long-thin column
Compounds on column separated by physical- chemical properties (size, shape, volatility, charge)
Ethanol has a unique retention time
Distinguishes between ethanol and other volatiles
Isopropanol from swabs
Acetone from diabetes

18. GC Analysis of Volatile Mix

19. Review of Results Calibration curve for both instruments (r2≥0.99)
Controls must be within 10% of the target
Blank cannot contain any peaks
Results from each case must agree within 10% of the mean from both results
Check for other volatiles in sample

20. Sample Handling and Analysis
Sample receipt and accession
Alcohol analysis
Drug screen analysis
Confirmation analysis
Specialty drug analysis
Prepare report
Review by supervisors and manager

21. Drug Screening – Enzyme Multiplied Immunoassay Technique (EMIT)
Chemical “lock and key” test
Qualitative
Class, not compound specific
Rule in/rule out for major CNS drug categories
Each drug has cut-off level: Blood cut off < urine cut-off
Drugs may be present below cut-off

22. EMIT Screening 6 classes screened
Cocaine metabolite
Opiates
Benzodiazepines
Barbiturates
Cannabinoids
Amphetamines
4 classes screened for urine (amphetamines and cannabinoids omitted)

23. Limits Zolpidem Diphenhydramine Dextromethorphan
Certain benzodiazepines: Lorazepam, Clonazepam
Fentanyl
Acidic/Neutral drugs: Anti-seizures, Soma
Most synthetics: bath salts, synthetic cannabinoids
SSRI drugs: Citalopram, Fluoxetine, Paroxetine, Venlafaxine, Sertraline
Most antipsychotics

24. Blood Drug Screen Blood specimens require extraction procedure
Calibrators, controls and blank extracted with each set
Positive results must be confirmed

25. BDS Drug Screen Alkaline (basic) drug screen
Samples are extracted through a series of steps
Calibrators, controls and blanks extracted
Purifies and concentrates drugs

26. BDS Drug Screen
Analysis by Gas Chromatography/Mass Spectrometry (GC/MS and GC/NPD)
Separates drugs based on unique chemical properties
Provides unique chemical fingerprint for drugs

27. Extracted Sample Through GC/MS
NPD

28. BDS Drug Screen Not absolutely comprehensive
Also has a cut-off, more sensitive than immunoassay
-distinguish specific member of drug class
Drugs may be present below cut-off
Drugs not confirmed reported as not detected

29. Sample Handling and Analysis
Sample receipt and accession
Alcohol analysis
Drug Screen analysis
Confirmation analysis
Specialty drug analysis
Prepare report
Review by supervisors and manager

30. Drug Confirmation Extraction of Sample
Analysis of sample using GC/MS; GC/FID; LCMS; LCMSMS

31. Sample Handling and Analysis
Sample receipt and accession
Alcohol analysis
Drug screen analysis
Confirmation analysis
Specialty drug analysis
Prepare report
Review by supervisors and manager

32. Specialty Tests for Drugs
Some drugs require special extractions for detection
THC
Cocaine metabolites
Some Opiates (morphine, hydromorphone)
Some Benzodiazepines (clonazepam)
Confirmation by GC/MS, LC/MS, or LC/MS/MS
Method of extraction and analysis allows greater sensitivity

33. Special Assays Analysts must be certified for analysis of these drugs
Typically, solid phase extraction (SPE)
Blank, calibrators, controls extracted with each set of cases

34. LOQs for Special Analytes
THC – 1 ng/mL
Zolpidem – 0.01 mg/L
Cocaine, BZE, cocaethylene – 0.01 mg/L
Methadone – 0.01 mg/L
PCP – 0.01 mg/L
Amines mg/L includes amphetamine, methamphetamine, MDA, MDMA
Barbiturates- 0.5 mg/L includes amobarbital, phenobarbital, pentobarbital, secobarbital, butalbital
Opiates: morphine, oxymorphone, oxycodone, codeine, hydrocodone= 0.01 mg/L; hydromorphone, 6-AM= 2 ng/mL
Benzodiazepines mg/L for diazepam, temazepam, nordiazepam, zopiclone, alprazolam, clonazepam, lorazepam, oxazepam, midazolam, chlordiazepoxide, triazolam, flurazepam, flunitrazepam; 0.02 mg/L for quetiapine

35. Testing for Designer Drugs
Currently, WSTL does not have testing protocols for designer drugs
Cases sent to two contract laboratories:
NMS Labs (Pennsylvania)
AIT Laboratories (Indiana)
Cases will be sent to contract lab if indicated on request form and no other drug(s) is detected

36. Storage of Samples Refrigerated while not testing
Samples removed from fridge only while preparing for tests
Evidence maintained in locked personal fridges or in evidence vault

37. Sample Handling and Analysis
Sample receipt and accession
Alcohol analysis
Drug Screen analysis
Confirmation analysis
Specialty drug analysis
Preparation of report
Review by supervisors and manager

38. Data Analysis Review all data files from sequence
Blanks are blank
Negatives have ISTD
Calibration curve is acceptable (R2≥0.99)
Controls are within range
Identify/quantifies drug(s) present
All data is reviewed by
other analysts and/or
supervisors

39. Preparation of Report
Report prepared after all testing is completed for each specific case
All batches of data– peer and supervisor reviewed prior to release
All compounds present above level of quantitation are reported
2 tests- screened and confirmed
Retention time-matched and fingerprint ID
Quantitative vs. Qualitative
Quantitative – gives value of drug present in blood
(0.04 mg/L or 3 ng/mL)
Qualitative – Positive without level
Parent drug and metabolites can be reported
Metabolites may or may not be active

40. Reports Analyst, dates, and methodology
Any test not performed by the primary analyst is indicated below the specific test result

41. Reports
TWO pages! or more
Agency owns report

42. Quality Control/Quality Assurance
Record keeping, compliance with SOPs, corrective actions
Quality Control
Proficiency testing (CAP and WSLH), accuracy of test
QA/QC system assures results are accurate, precise, and meaningful
Calibration Lab accredited by ASCLD/LAB
Tox Lab accredited by ABFT and ASCLD/LAB

43. Other Duties Testimony! Instrument maintenance
Entire state- Breath and blood
Alaska- Blood
Oregon- Blood
Instrument maintenance
Continuing Education  workshops, seminars, conferences
Educating the public  high schools, attorney conferences, continuing ed for police agencies, legislature

44. Questions/Concerns? Contact us!
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