Role of Toxicology in Criminal Investigations Lyndsey Knoy, D-ABFT-FT Forensic Scientist Washington State Toxicology Laboratory Lyndsey.Knoy@wsp.wa.gov

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?

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

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

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

Cases Handled Death Investigations Overdoses Suicides Poisonings Traffic Fatalities Homicides Undetermined

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

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! 2016- 14,617 cases Court Testimony for 2017 1,529 hours for 308 appearances LK: 139 hours for 26 cases

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

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

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

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-18-00001) Maintains chain of custody

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

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

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

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

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

GC Analysis of Volatile Mix

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

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

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

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

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

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

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

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

Extracted Sample Through GC/MS NPD

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

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

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

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

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

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

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- 0.01 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- 0.01 mg/L for diazepam, temazepam, nordiazepam, zopiclone, alprazolam, clonazepam, lorazepam, oxazepam, midazolam, chlordiazepoxide, triazolam, flurazepam, flunitrazepam; 0.02 mg/L for quetiapine

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

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

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

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

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

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

Reports TWO pages! or more Agency owns report

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

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

Questions/Concerns? Contact us! (206) 262-6100 toxlab@wsp.wa.gov Lyndsey.Knoy@wsp.wa.gov