ORAL FLUID DRUG TESTING Dean F. Fritch, Ph.D., DABFT, DABCC-TC Manager of Technical & Analytical Services

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1 detection technologies for the new millennium ORAL FLUID DRUG TESTING Dean F. Fritch, Ph.D., DABFT, DABCC-TC Manager of Technical & Analytical Services TESTING FOR DRUGS OF ABUSE Tests for imairment/drug-induced effect 4Blood 4Saliva 4Performance Tests for recent drug exosure 4 4Saliva Tests for historical drug exosure 4Hair 4Sweat Why Test Saliva? Easily and raidly obtained Minimal invasion of rivacy Difficult to adulterate Short drug detection time rovides evidence of recent drug exosure Presence of active drug indicates otential for ongoing drug effect at time of testing PHYSIOLOGY OF ORAL FLUID PRODUCTION Three Glands: Parotid, Submandibular, and Sublingual Two Tyes of Secreting Cells: 4Serous cells secrete water fluid containing electrolytes and amylase 4Mucous cells secrete mucins (mucoroteins and mucoloysaccharides) PHYSIOLOGY OF ORAL FLUID PRODUCTION Saliva flow varies from.5 to.5 L/day ORAL FLUID COLLECTION Sitting/swabbing Flow stimulated collection Resting flow:. to ml/min Can be stimulated to eak flow of aroximately ml/min Saliva collection devices Pure arotid saliva collection (asiration)

2 DRUG TRANSPORT INTO SALIVA Passive diffusion (most drugs) Physiochemical (Ka, liosolubility molecular weight) lasma rotein binding H (increases with stimulation) Normal H (u to 7.8) Filtration through ores in membrane INFLUENCE OF H Weakly acidic drugs most influence when Ka is less than 8.5 and close to H of saliva Butabarbital Ka 7.9 Butalbital Ka 7.6 Phenobarbital Ka 7.2 THC Ka 9.5 INFLUENCE OF H Weakly basic drugs most influence when Ka is more than 5.5 and close to H of saliva Heroin Ka 7.6 Morhine Ka 8. Oxycodone Ka 8.5 Cocaine Ka 8.6 PCP Ka 8.5 Amhetamines Ka 9.9 Benzodiazeines ka 2-4 Relative concentrations of Drug and Metabolites in Various Matrices COCAINE Metabolic Profile HEROIN Metabolic Profile Blood Saliva Sweat Hair Blood Saliva Sweat Hair COC BE EME CE Heroin 6-MAM Morhine M-3-G, M-6-G 2

3 METHAMPHETAMINE Metabolic Profile CANNABINOIDS Metabolic Profile Blood Saliva Sweat Hair Blood Saliva Sweat Hair Methamhetamine Amhetamine THC -H THC THCA WINDOW OF DETECTION COCAINE Window of Detection Dose Blood Day Time between doses Saliva - 2 Days Route of administration Sweat 3 Days 4-3 Days Matrix Hair 25 Days., Goal of Study: Determine Effect of Reeated Cocaine Dosing on Detection Times Comare detection times for cocaine and benzoylecgonine (BZE) in oral fluid and urine in two grous of subjects Grou : Single dose study (crossover design) Six subjects received singe dose of cocaine 25 mg IV 32 mg IN 42 mg SM Grou 2: Reeated dosing study Six subjects received daily escalating doses of oral cocaine to maximum tolerable doses Reached daily maximum doses of 375 mg to 2 mg cocaine Secimens collected following last dose [Analysis by GC-MS (LOD = )] Reeated Cocaine Dosing A.) Cocaine B.) BZE Hours Accumulation and biexonential elimination of cocaine contribute to greater increase in oral fluid detection times comared to urine 3

4 Hours Single and Reeated Doses of Cocaine: Detection Times Error bars = SEM N=6 X4 SD OF COC 8 RD OF COC 8 SD OF BZE 8 X7 RD OF BZE 8 X2 SD UR RD UR BZE 5 BZE Conclusions Reeated dosing increased oral fluid detection times by factor X4-7 comared to X2 for urine Single dose studies underestimate detection times Oral fluid detection times will be significantly longer for detection of cocaine abuse in realistic settings where multile dosing occurs DETECTION OF MARIJUANA: HOW LONG CAN THC LAST? THC ELIMINATION (Baseline and 4 Hours) After single use u to 2 to 24 hours Baseline Elimination Profiles (t=4 hours) Data available showing that urine may not be much better No data yet on heavy users - Not Percentage of Secimens Percentage of Secimens exected to be as long as urine! < > THC Concentration Ranges by GC/MS () < > THC Concentration Ranges by GC/MS () THC ELIMINATION ( Hours and 24 Hours) Elimination Profiles (t= hours) Elimination Profiles (t=24 hours) Percentage of Secimens Percentage of Secimens SERUM VS ORAL FLUID < > < THC Concentration Ranges by GC/MS () > THC Concentration Ranges by GC/MS () 4

5 Saliva/Serum (S/P) Correlation Issues Saliva H due to method of collection Remains of orally ingested, smoked or internasally administered drugs Ethanol 3 minutes Smoked or Internasal 4-8 hours S/P Ratios Cocaine to.4 (BE is a zwitterion and not effected by changes in salivary H) THC. (high rotein binding revents transfer from serum) Oiates 3.8 to.2 Amhetamines 4 to 2.6 PCP 3 to.5 Barbiturates.2 to.5 Benzodiazeines.2 to.5 deending on Benzo Intercet Collection-- OraSure Technologies URINE VS ORAL FLUID Intercet Samle Volume Collector contains 8 µl reservative buffer Swab collects ~ 4 µl saliva Results in :3 dilution of saliva Positive Rates by Drug January - June 23 Drug Testing Index - Intercet Gen. Workforce Federal (n=527k) (n=2,8k) (n=6k) Total Positives 4.62% 5.% 2.5% Marijuana 3.8% 3.2%.39% Cocaine.32%.74%.58% Oiates.9%.34%.9% Amhetamines.47%.46%.29% PCP.3%.3%.4% Comarable results for all drug targets Drug Testing Index courtesy of Quest Diagnostics, Inc., Teterboro, NJ 5

6 Prevalence by Drug Prevalence by Reason % 3.% 2.5% 2.% General UA 25.% 2.% 5.% General UA.5%.%.%.5% 5.%.% Marijuana Cocaine Oiates Amhetamines PCP.% Pre-emloyment Random Post-accident Susicion Follow-u Return-to-duty / Correlation Diluted urines: Dr. Barry samle reorted before the Oversight and Investigations Committee that a drug ositive secimen is twice as likely to have a creatinine of less than 2 mg/dl. Other means of urine adulteration Chronic users will be detected longer eriods of time LABONE ONE S DATA From 2//99 to 4/3/99: 35,743 Secimens Tested 53 Nitrites (.7%) 5 Urinaids (.%) 29 Pyridine (.2%) 96 Substituted (.6%) Total 87 Secimens (.28%) and,857 (3.8%) Dilute Secimens Closed Room Study Design THC Passive Studies Subjects Active smokers: five healthy, male cannabis users Passive subjects: four healthy, drug-free males Staff members (secimen collections, monitored study) Conditions: sealed room, 36 m 3 (3 m X 4 m X 3 m) Active users: smoked single cannabis cigarette.75% THC mixed with tobacco Purchased in The Netherlands from a commercial source 2 minute smoking eriod 4 hour session Passive subjects: remained in sealed room for the 4 hour session Located aroximately.5 m from smokers 6

7 Closed Room Study Design (cont.) Secimen collection (timed from end of smoking) Oral fluid (baseline,, 5, 45, 75, 5, 35, 65, 95, and 225 min) (baseline,, and 225 min) Oral fluid collection with Intercet DOA Oral Secimen Collection Device THC oral fluid analysis (OraSure Technologies) Screened with the Cannabinoids Intercet MICRO-PLATE Enzyme Immunoassay; cutoff = 3 (adjusted for neat concentration) Confirmation by GC-MS-MS; LOD/LOQ =.75 THC analysis: GC-MS MS-MS (THCCOOH); LOD/LOQ = THC Intercet Passive Room Study (linear scale) Minutes Red = 2 DHHS cutoff 225 Log THC Intercet. -25 Passive Room Study (log scale) Minutes Red = 2 DHHS cutoff Room Study Summary Passive inhalation THC oral fluid concentrations ranged <.75 to 26.4 Present u to 3 min after exosure All urine secimens from assive subjects tested negative Active smokers THC in oral fluid was X to X2 fold higher than assive Generally ositive throughout 4 hour session secimens ositive at 4 hours Pattern of decline of THC was bi-hasic suggesting oral deot effect for THC The Van The Van Motor Vehicle Study Design Subjects Active smokers: four healthy, male cannabis users Passive subjects: four healthy, drug-free males Staff members (secimen collections, monitored study) Conditions: Unventilated van, 5.3 m 3 (.7m x.8m x 5.m) Passive subjects: Each assive subject sat next to a smoker in van Active users: each smoked single cannabis cigarette 7

8 Motor Vehicle Study Design (cont.) Study I 5.4% THC mixed with tobacco (39.5 mg) Purchased in The Netherlands from a commercial source 2 minute smoking eriod 8 hour session; remained in van for first hour Study II.4% THC, no tobacco (83.2 mg) Purchased in The Netherlands from a commercial source 2 minute smoking eriod 8 hour session; exited van immediately after smoking Motor Vehicle Study Design (cont.) Secimen collection (timed from end of smoking) Oral fluid (baseline,, 5, 3, 45, 6, 75, 9, 5, 2, 5, 8, 2, 24 minutes, 6 & 8 hours (baseline, 6, 2, 24 minutes, 6 & 8 hours) Oral fluid collection with Intercet DOA Oral Secimen Collection Device THC oral fluid analysis (OraSure Technologies) Screened with the Cannabinoids Intercet MICRO-PLATE Enzyme Immunoassay; cutoff = 3 (adjusted for neat concentration) Confirmation by GC-MS-MS; LOD/LOQ =.75 THC analysis: GC-MS MS-MS (THCCOOH); LOD/LOQ = 9 8 Passive Van Study (linear scale) Passive Van Study (log scale) THC Intercet Red = 2 DHHS Cutoff Orange = Exosed devices Log THC Intercet Hours Hours Red = 2 DHHS Cutoff Orange = Exosed devices THC in Intercet (corrected) Intercet Study Comarison Hours Red = 2 DHHS Cutoff Green = Van Study II Van Study Summary Passive inhalation THC oral fluid concentrations ranged <.75 to 82 when collected inside Van, none exceeded.75 when collected outside van Present u to 3 min after exosure when collected in the resence of smoke, none when subjects removed from smoke during collections All urine secimens from assive subjects tested negative Active smokers THC in oral fluid was X to X2 fold higher than assive Generally ositive throughout 8 hour session secimens not all ositive until 6 and 8 hours Pattern of decline of THC was bi-hasic suggesting oral deot effect for THC 8

9 Conclusions Conclusions Passive conditions were designed to reresent extreme exosure conditions Initial high levels of THC are raidly cleared (active) Second hase decline of THC after active use suggests formation of deot of THC in oral cavity Overall, these studies indicate that collection of secimens outside an area of ossible smoke contamination is necessary for accurate determination of assive exosure effects on oral fluid testing. Absent extreme conditions, assive exosure to marijuana smoke is not a credible exlanation for a ositive oral fluid test. 9

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