Lyndsey Knoy, D-ABFT-FT Forensic Scientist Washington State Toxicology Laboratory Lyndsey.Knoy@wsp.wa.gov
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?
WSP FLSB Crime Lab Toxicology Lab Breath Test Section DRE Program
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
Death Investigations Overdoses Suicides Poisonings Traffic Fatalities Homicides Undetermined
Police Investigations DUI/DRE Drug Investigations Drug Endangered Children Liquor and Cannabis Board Drug Facilitated Sexual Assaults
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
Sample receipt and log-in (Bob Smith, Former PEC) Data Analysis (Andrew Gingras) Create Report Lab analysis (Katie Harris) Expert Testimony Agency/Prosecutor
1. Sample receipt and accession 2. Alcohol analysis 3. Drug screen analysis 4. Confirmation analysis 5. Specialty drug analysis 6. Prepare report 7. Review by supervisors and manager
US Postal Service Courier (UPS / FedEx) Hand delivered Campus mail
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
Each case entered in LIMS database ST# is used to track samples in the laboratory Currently about 80 cases are received daily
1. Sample receipt and accession 2. Alcohol analysis 3. Drug screen analysis 4. Confirmation analysis 5. Specialty drug analysis 6. Prepare report 7. Review by supervisors and manager
Forensic collection of sample Gray top tube Preservative (NaF) & anticoagulant (C 2 K 2 O 4 ) FDA Regulated Certificates from the manufacturer Experts can testify to hearsay Samples are not clotted Vacuum Expiration Date: refers to vacuum
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
Alcohol is a volatile When heated, volatiles enter the vapor phase Vapor is injected onto a long-thin column Compounds on column separated by physicalchemical properties (size, shape, volatility, charge) Ethanol has a unique retention time Distinguishes between ethanol and other volatiles Isopropanol from swabs Acetone from diabetes
Calibration curve for both instruments (r 2 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
1. Sample receipt and accession 2. Alcohol analysis 3. Drug screen analysis 4. Confirmation analysis 5. Specialty drug analysis 6. Prepare report 7. 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
6 classes screened Cocaine metabolite Opiates Benzodiazepines Barbiturates Cannabinoids Amphetamines 4 classes screened for urine (amphetamines and cannabinoids omitted)
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 specimens require extraction procedure Calibrators, controls and blank extracted with each set Positive results must be confirmed
Alkaline (basic) drug screen Samples are extracted through a series of steps Calibrators, controls and blanks extracted Purifies and concentrates drugs
Analysis by Gas Chromatography/Mass Spectrometry (GC/MS and GC/NPD) Separates drugs based on unique chemical properties Provides unique chemical fingerprint for drugs
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
1. Sample receipt and accession 2. Alcohol analysis 3. Drug Screen analysis 4. Confirmation analysis 5. Specialty drug analysis 6. Prepare report 7. Review by supervisors and manager
1.Extraction of Sample 2.Analysis of sample using GC/MS; GC/FID; LCMS; LCMSMS
1. Sample receipt and accession 2. Alcohol analysis 3. Drug screen analysis 4. Confirmation analysis 5. Specialty drug analysis 6. Prepare report 7. 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
Analysts must be certified for analysis of these drugs Typically, solid phase extraction (SPE) Blank, calibrators, controls extracted with each set of cases
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
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
Refrigerated while not testing Samples removed from fridge only while preparing for tests Evidence maintained in locked personal fridges or in evidence vault
1. Sample receipt and accession 2. Alcohol analysis 3. Drug Screen analysis 4. Confirmation analysis 5. Specialty drug analysis 6. Preparation of report 7. Review by supervisors and manager
Review all data files from sequence Blanks are blank Negatives have ISTD Calibration curve is acceptable (R 2 0.99) Controls are within range Identify/quantifies drug(s) present All data is reviewed by other analysts and/or supervisors
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
Analyst, dates, and methodology Any test not performed by the primary analyst is indicated below the specific test result
TWO pages! or more Agency owns report
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
Testimony! 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
(206) 262-6100 toxlab@wsp.wa.gov Lyndsey.Knoy@wsp.wa.gov