Involvement Of Alcohol And/Or Controlled Substances In Driving Related Offenses

Involvement Of Alcohol And/Or Controlled Substances In Driving Related Offenses Felix Adatsi, Ph.D. MICHIGAN DEPARTMENT OF STATE POLICE FORENSIC SCIENCE DIVISION

MSP Toxicology Unit Lansing Laboratory Analysis of blood and other biological specimens for alcohol and drugs of abuse. Analysis of beverages for alcohol Supervisors Alcohol Analysis: Dr. Felix Adatsi, Toxicologist Drug Analysis: Dr. Michele Glinn, Toxicologist Telephone: (517) 322-6600

Quick Facts MSP Toxicology Unit is an accredited Forensic Toxicology Laboratory routinely participates in external proficiency tests has established uniform guidelines for evidence collection, handling, and transport to the laboratory utilizes current state-of-the-art instruments and techniques in the analysis of submitted samples

Administrative Rules Department of State Police promulgates uniform rules for the administration of chemical tests for law enforcement (Sec. 625a(6)(g)) Alcohol and drug testing of biological and non-biological specimens Tests, expression of results, acceptable techniques, calibration, and collection of samples.

Collection of Evidence MSP sole distributor of evidence collection kits to law enforcement and other agencies Evidence collection kits meet generally accepted and current scientific standards Evidence collection kits contain Form FSD-93 for additional instructions

FSD-93 Instructions FOR BLOOD SAMPLES To Physician Or Other Qualified Medical Person 1.Do not use alcohol or alcoholic solution to sterilize skin surface,needle or syringe. 2.Draw two tubes of venous blood from subject in presence of law enforcement officer,and tell the subject IN THE OFFICER S PRESENCE that no alcohol was used in sterilizing skin surface,needle or syringe.lowly invert blood collection tube(s)several times to distribute the sodium fluoride/potassium oxalate preservative. 3.Complete blood specimen label(s)by entering name of subject,date and time of blood collection,and your name in ink. 4.In the presence of subject,hand tube(s)of blood and label(s)to law enforcement officer for signing,packaging and transfer to the laboratory. To Law Enforcement Officer 1.Review accompanying information sheet and be sure all information is supplied before sealing and mailing. Sign the Label(s)in the place provided and attach to the tubes. 2.Place the tube(s)into the cardboard holder from which they were taken,for mailing protection.seal the tube(s)and holder in the zip-lock plastic bag.complete the FSD- 93 and place it in the cardboard mailing box. Seal the box with the provided seals. Mail the sealed box using First Class U.S.mail.

FSD-93 Instructions FOR URINE SAMPLES To Law Enforcement Officer 1.THE URINE SPECIMEN SHALL BE COLLECTED IN THE PRESENCE OF AN OFFICER to be certain the subject does not contaminate the specimen and to insure that the subject EMPTIES THE BLADDER. 2.The urine test requires the subject to provide two samples collected at least 30 minutes apart.the urine samples shall be collected in separate bottles and identified as Specimen Bottle #1 and Specimen Bottle #2. 3.a. Specimen Bottle #1 :SUBJECT COMPLETELY EMPTIES THE BLADDER INTO ONE OF THE BOTTLES. b.wait FOR A LEAST 30 MINUTES. c. Specimen Bottle #2.SUBJECT COMPLETELY EMPTIES THE BLADDER A SECOND TIME INTO THE SECOND BOTTLE. 4.TIGHTEN CAPS FIRMLY.Complete urine specimen labels by entering name of subject, date and time of urine collection,and your name on the labels and attach to bottles.seal the bottles in plastic zip-lock bag,then place the bag with samples in the cardboard mailing box.complete the FSD-93 and place into the mailing box.seal the box with the provided seals.mail the sealed box using First Class U.S.mail. THE 2 BOTTLES MAY BE USED FOR LIQUID SAMPLES OTHER THAN URINE (i.e.a BEVERAGE FOR OPEN INTOXICANTS).

Chain of Custody All outgoing MSP evidence collection kits pre-sealed with integrity seal Evidence collected pursuant to administrative rules and instructions on Form FSD-93 Evidence labels provided in kit are used to document information on subject and establish chain-of-custody Completed evidence labels are attached to individual specimens

Chain of Custody Samples that are sent to the laboratory shall be sealed in a manner that ensures their integrity (R 325.2675(6)) Labeled individual specimens and completed FSD-93 form are secured in evidence collection kit Evidence collection kit and contents are sealed with kit shipping seals Sealed evidence collection kit and contents are delivered to the laboratory

Specimen Collection Alcohol blood urine vitreous humor other body fluids & tissues

Specimen Collection Blood: acquisition of sample is invasive useful for demonstrating under the influence

Specimen Collection Urine: end product of metabolism non-invasive sample collection Vitreous: useful in death cases less chance of contamination

Alcohol

Alcohols Carbon-containing compounds with an oxygen-hydrogen bond (-OH) For example: methanol (wood alcohol) ethanol (grain alcohol; beverage alcohol)» chemical structure: CH 3 CH 2 OH isopropanol (rubbing alcohol)

Chemistry of Alcoholic Beverages Alcoholic Beverage: Active ingredient in alcoholic beverage is ethanol Any potable liquid that contains 0.5% to 95% ethanol Proof Term used to describe alcoholic beverages Twice the percent concentration: 100 proof = 50% ethanol

Chemistry of Alcoholic Beverages Beverage %Ethanol Beer 3.2-5.0 Wines 7.0-14.0 Whiskies 40.0-75.0 Vodkas 40.0-50.0 Gins 40.0-49.0

Alcoholic Beverages In General: One beer equals one glass of wine One beer equals one mixed drink One beer equals one shot of spirits

Not a Standard Drink!

Expressing Blood Alcohol Concentration Name gram milligram liter milliliter deciliter Abbreviation g mg L ml dl 1 gram = 1000 mg 1 liter = 1000 ml 1 deciliter = 100 ml

Expressing Blood Alcohol Blood alcohol results expressed as grams alcohol per 100 milliliters of blood A blood alcohol of 248 mg/dl will be converted into grams per 100 milliliters: 1 gram = 1000 mg 1 deciliter = 100 ml Concentration Therefore, 248 mg/dl is equal to 0.248 g/100 ml

Blood Alcohol Testing Techniques Gas Chromatograph State of the Art Direct injection or Headspace analysis Whole blood Analyzers & Kits Abbot Diagnostics TDx ethanol kit CalBiochem-Behring Ethyl Alcohol kit Sigma Diagnostics Alcohol (ethanol) kit others Enzymatic methods

Basic Gas Chromatography Sample preparation step Sample inlet and sampling devices

Basic Gas Chromatography Column Long, coiled tube that is internally coated with chemical Substances passing through the column are separated in time

Basic Gas Chromatography Carrier Gas Introduced into column under pressure Carries samples through the column Purges column after analyte passes through Detector & Data System Flame Ionization Detector (FID) PC

Alcohol Analysis Known volume of blood plus internal standard is placed in a clean vial and sealed Vial is heated Ethanol and other volatiles evaporate into space above the blood in the sealed vial The vapor above the blood in the vial (headspace) is analyzed for ethanol content No direct physical contact is made between blood sample and any part of the GC

Alcohol Analysis Each sample is analyzed twice by two separate and independent gas chromatographs Two chromatograms are generated

RT: 0.00-4.99 SM: 7G 100 80 Relative Intensity 60 40 2.28 3.61 NL: 1.50E6 Right_FID Analog 101104CAL 04 20 0 0.24 0.56 0.84 1.17 1.46 1.87 2.54 3.07 3.36 3.90 4.35 4.86 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 Time (min) A chromatogram Qualitative: Quantitative: provides What How is much? it? Qualitative and Quantitative information Retention Area countime - Established Compared to by calibration standardscurve -prepared Column from specific known standards

Alcohol Analysis Results must agree within 0.01 The lower of the two results is reported

Sample ID: 0.118 g/100ml Data File: 101104CAL04 Curr Data Path: C:\Xcalibur\data\Cal Stn 1\\ Acquisition Date: 10/11/04 14:54:16 Inst Method: C:\Xcalibur\methods\Alcohol_HS.meth Proc Method: C:\Xcalibur\methods\Alcohol Barcode: Unreadable Vial: Tray1:4 RT: 0.00-4.99 10 0 80 Relative Intensity 60 40 SM: 7G 2.28 3.61 NL: 1.50 E6 Right_FID Analog 10 110 4 C AL 04 20 0 0.24 0.56 0.84 1.17 1.4 6 1.8 7 2.54 3.07 3.36 3.90 4.35 4.86 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 Time (min) Name Expected RT RT Component Type Component Found Area Calculated Amount Methanol N/A N/A Target Compound No N/A N/A Isopropanol N/A N/A Target Compound No N/A N/A Acetone N/A N/A Target Compound No N/A N/A Ethanol 2.26 2.28 Target Compound Yes 1020965 0.118 n-propanol 3.60 3.61 Internal Standard Yes 5088918 N/A Comments: Samples are reported in g/100ml 50 µl sample/800 µl n-propyl alcohol @ 0.02 g/100 ml Column = BAC-1 30m x 0.53mm ID Oven temp = 45 o C Syringe temp = 70 o C Inlet temp = 125 o C Sample incubation temp = 70 o C

Calibration Curve Ethanol Y = -0.00212407+1.72226*X R^2 = 0.9999 W: Equal 1.0 0.8 Area Ratio 0.6 0.4 0.2 0.0 0.0 0.1 0.2 0.3 0.4 0.5 0.6 g/100ml

Analytical Methods Calibration of instrument involves use of known standard ethanol solutions Multi-point calibration using 8 standard ethanol solutions to generate calibration curve Acceptability of calibration curve dependent on correlation coefficient (CC) Calibration curve VERIFIED using external ethanol control solutions External controls: aqueous control & blood control

Calibration Curve Ethanol Y = -0.00212407+1.72226*X R^2 = 0.9999 W: Equal 1.0 0.8 Area Ratio 0.6 0.4 0.2 0.0 0.0 0.1 0.2 0.3 0.4 0.5 0.6 g/100ml

Analytical Methods External alcohol controls do NOT set calibration curve and have no effect on CC Column does not set calibration curve

Analytical Methods Calibrators and Controls (both internal and external) Prepared from stock 100% ethanol by dilution with de-ionized water Stock 100% ethanol used once or twice only per batch of calibrators and controls Calibrators and Controls are dilute solutions of stock 100% ethanol

Analytical Methods Escape of ethanol from solution by evaporation minimized Calibrators and Controls not taken out directly from stock 100% ethanol daily

Pharmacology & Toxicology of Ethanol Central Nervous System (CNS) Depressant CNS is system most severely affected by alcohol. Intensity of effect on CNS is proportional to alcohol level Other Systems: Muscular, Eye, Cardiovascular, Hepatic, Renal, Skin, etc. Euphorigenic

Clinical Signs/Symptoms Subclinical. No apparent influence. Behavior nearly normal. Talkative; decrease inhibitions; decrease in judgement, attention and control; increased self confidence, slowing of reaction time. BAC (g/dl) 0.01-0.05 0.03-0.12 Loss of critical judgement; impairment of memory and comprehension; impaired balance; reduced visual acuity; ataxia and unsteady gait. 0.09-0.25 Confusion; disorientation; mental confusion; increased muscular incoordination; slurred speech Stupor; severe visual impairment; vomiting; inability to stand or walk; sleep. Coma; respiratory arrest; anethesia; incontinence; complete unconsciousness and possible death. 0.18-0.30 0.25-0.40 0.35-0.50 Adapted from: K.M. Dubowski. Stages of Acute Alcoholic Influence/Intoxication

Pharmacology & Toxicology of Ethanol Induction of Tolerance Effectiveness or potency of alcohol diminishes after continuous or chronic consumption May be Dispositional (metabolic), Functional (constitutional) Physical Dependence (Addiction) Need and continuous craving for alcohol

Toxicokinetics of Ethanol Absorption Occurs in stomach Most rapid and significant in small intestine Rate of absorption of ethanol into blood influenced by several factors Food has significant effect on rate of absorption of ethanol into blood Absorption occurs over a period of time

Toxicokinetics of Ethanol Distribution Uniformly distributed throughout all tissues and body fluids Equilibrium is rapidly established BAC can be estimated using Widmark Formula A = WrCT/0.8

Toxicokinetics of Ethanol Metabolism Liver is main organ responsible for metabolizing alcohol Enzyme dependent Alcoholics metabolize at faster rates than social drinkers

Toxicokinetics of Ethanol Elimination Decline in blood alcohol concentrations from metabolism Influence by several factors: race, gender, age, physiological status, etc. Elimination rates higher in chronic consumers or alcoholics Average rate for males = 0.015 g/dl/h Average rate for females = 0.018 g/dl/h

Concentration-Time Profile of Ethanol in Blood Absorptive phase Post-absorptive phase BAC (g/dl) fasting fed 90 180 270 Time (min)

Alcohol in Whole Blood vs. Serum Serum = (whole blood) - (blood cells/clotting factors) Plasma = (whole blood) - (blood cells) Serum alcohol and plasma alcohol concentrations are greater than whole blood alcohol concentration Serum and plasma alcohol concentrations can be converted to equivalent whole blood result

Whole Blood and Serum SPIN Serum Whole Blood Blood cells

Whole Blood and Serum Blood containing alcohol Whole Blood Lower alcohol level Serum Higher alcohol level

Alcohol in Whole Blood vs. Serum To convert serum alcohol to equivalent whole blood alcohol, divide serum alcohol by average conversion factor 1.16 Clinical or hospital laboratories usually test serum and or plasma for alcohol Forensic laboratories routinely test whole blood

Drugs

Analytical Methods Case history important in determining nature of poison Screening tests according to drug class (amphetamines, barbiturates, opiates, etc) Detailed analyses Confirmation using GC/MS Use of other instrumentation (e.g. carbon monoxide)

Analytical Methods First step in drug analysis involves screening of samples Technique involves Fluorescence Polarization Immunoassay (FPIA) Drug identification is by class (presumptive positive or negative outcomes)

Analytical Methods Second step involves extraction of drug(s) from biological specimens Technique involves solid-phase extraction (SPE) Sample passes through a column containing adsorbent material Drug(s) in sample bind to column matrix Drugs are washed off (eluted) from column with appropriate solvent

Analytical Method Final step involves analysis of eluted solvent by GC/MS Mass spectrum molecular fingerprint of identified drug is produced Confirmation is established by comparing with database Method is sensitive and specific

Quality Assurance/Quality Control Evidence Collection R325.2675 outlines acceptable techniques: Antiseptic should not contain alcohol or alcoholic solution Blood should be drawn with a sterile dry needle into specimen tube Specimen tube contains sodium fluoride alone or in combination with other preservatives or anticoagulants Urine should be collected into clean glass or plastic container with top

Quality Assurance/Quality Control Analysis Protocol driven, ensures uniformity and consistency Standards and controls for calibration and to check precision and accuracy Instrument maintenance logs Proficiency testing Uniform test reporting system Peer review of all out-going reports

Disposition of Evidence Evidence may be discarded after 180 days Request to retain or hold evidence honored but hold time not indefinite Request for independent analysis by opposing counsel must be approved by prosecutor Court orders for independent analysis Return of evidence may be denied due to questionable integrity outside laboratory

FOIA, Subpoenas, Court Orders Business record maintained on each case and filed Requests for information on cases channeled through office of FOIA Requests by subpoenas usually will seek approval from prosecutor s office Prompt compliance with Court Orders

If the law has made you a witness, remain a man of science. You have no victim to avenge, no guilty or innocent person to convict or save, you must bear your testimony within the limits of science. ~ Dr. P.C.H. Brouardel 19th Century French Medico-legalist