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Bertholf, Ph.D. Associate Professor of Pathology Chief of Clinical Chemistry & Toxicology University of Florida College of Medicine Health Science Center/Jacksonville Why test babies for cocaine?Prevalence of maternal cocaine use Effect of in utero cocaine exposure on fetal and postnatal development Medical/social/legal interventions $-Y0X"Prevalence of maternal cocaine useo1998 Household Survey revealed that 1.8 million Americans use cocaine (0.8% of population >12 years old) 2% of 18-25 year-olds; 1.2% of 26-35 year-olds 0.5% of women are cocaine users Estimates of the prevalence of cocaine use during pregnancy range from less than 2% to more than 15% Reasons for variations Geographic/socioeconomic/racial Methods used to detect use tiZOZ|Z;ZZiO|;%Is prenatal cocaine exposure harmful?Extensive literature on clinical effects of prenatal cocaine exposure Confounding factors are almost always present Poor prenatal care Concomitant drug use (alcohol, nicotine) Reliable animal models do not exist Sheep Pig Rodent\tZ<Z$ZZt<$0Well documented physiological effects of cocaine\Vasoconstriction Decreases uterine blood flow in sheep Cerebral vasodilation and hypertension in newborn sheep and pigs Neurotransmitter deficiencies ACTH and corticosterone response in rats Dopamine transporter binding in mice Consistent observations in cocaine exposed neonates Intrauterine growth retardation (LBW) Prematurity Complications/LOS ZgZZNZ4ZDZZgN4E  >@ R   Does cocaine cross the placenta?Studies in pregnant sheep and rodents indicate that both cocaine and its metabolites cross the placenta Fetal levels approximately 15% of maternal There is also evidence that the placenta has AChE activity May have a protective effect Placental transfer of cocaine does not appear to be flow-limited Cocaine can be measured in the amniotic fluid, but there is disagreement over whether it originates solely from fetal urinethZ+Z;ZZZh+; !Other sources of cocaine exposure?Breast Milk Passive inhalation Intentional/accidental ingestion ?Detecting prenatal cocaine exposure: Analytical considerations=Cocaine metabolism Quantitative thresholds Choice of specimen   Cocaethylene Quantitative thresholdsFor immunochemical methods, quantitative thresholds are established by calibration Pure compound or mixture of isomers Semi-quantitative results for real specimens Purely quantitative results are only available using specific methods GC/MS HPLC GC\SZQZFZZSQF -Detecting prenatal cocaine exposure: MethodsYImmunochemical methods RIA, ELISA EMIT FPIA CEDIA POC Spectrometric methods GC/MS HPLC GC\ZZZZ Immunochemical methodsMost are calibrated to a threshold of 300 ng benzoylecgonine per mL of urine Corresponds to the SAMHSA specifications May be adapted to other specimens Most are designed to measure benzoylecgonine in urine Benzoylecgonine is not the principal metabolite in all specimensTMK6AMK6AP*q 1Chromatographic methodsQuantitative GC/MS analysis (by isotope dilution) is considered the reference method Sensitivity <10 ng/mL Requires extensive specimen preparation (extraction, derivatization, isotopically-labeled internal standards) HPLC has similar sensitivity to GC/MS applications, but detection is less specific Does not require preparation of volatile derivatives GC is robust and sensitive, but lacks detector specificity Nitrogen/Phosphorus detectors are very sensitive but expensiveUZZSZ5Z;Z?ZUS5;?  Pe5  .Detecting prenatal cocaine exposure: SpecimensYMaternal specimens Urine Amniotic fluid Hair Neonatal specimens Urine Meconium Hair Nails\ZZZZF   Cocaine in maternal specimensMaternal urine is easy to collect, and measurements are reliable Usually reflects exposure within the past 2-3 days Uncontaminated amniotic fluid is harder to obtain More direct measure of fetal exposure (may be substitute for screening fetal urine) Pharmacokinetics of cocaine and metabolites in amniotic fluid have not been established. Hair is the most difficult specimen to analyze, but shows promise as a clinical specimen Incorporation of cocaine in hair has been widely documentedAZ3Z2ZZYZ<ZA32Y<  Cocaine in neonatal specimensUrine Sometimes difficult to collect Can be extracted from diapers Meconium More difficult to collect uncontaminated specimens Can also be extracted from diapers Hair Shows promise, but methods are not widely available Nails Most data on postmortem specimensZ=Z ZVZZ4ZZ"Z= V4  "C Urine drug testing in neonatesMTypically uses immunoassays configured for workplace drug testing Positive threshold >300 ng/mL (benzoylecgonine) Most studies indicate that neonatal urine drug screening at conventional thresholds fails to detect up to half of cocaine-exposed infants Results of neonatal urine cocaine screening usually correlate with maternal urineBBZ0ZZB0>Z!Use of lower screening thresholdsUrine collection from diapersSeveral studies have documented the correlation between chemistry and microscopic laboratory results on bag-collected and diaper-extracted urine Cocaine, benzoylecgonine, and ecgonine methyl ester can be extracted from diapers with methanol.Z, :Meconium drug testing   &Is meconium testing better than urine? 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Hair testingOther specimens$Distribution of cocaine metabolites!Which specimens are most stable?Comparison of sensitivities Conclusions  Fonts UsedDesign TemplateEmbedded OLE Servers Slide Titles&_Roger BertholfRoger Bertholf  !"#$%&'()*+,-./0123456789:;<=>?@ABCDEFGHIJKLMNOPQRSTUVWXYZ[\]^_`abcdefghijklmnopqrstuvwxyz{|}~      !"#$%&'()*+,-./0123456789:;<=>?@ABCDEFGHIJKLMNOPQRSTUVWXYZ[\]^_`abcdefghijklmnopqrstuvwxyz{|}~      !"#$%&'()*+,-./0123456789:;<=>?@ABCDEFGHIJKLMNOPQRSTUVWXYZ[\]^_`abcdefghijklmnopqrstuvwxyz{|}~      !"#$%&'()*+,-./0123456789:;<=>?@ABCDEFGHIJKLMNOPQRSTUVWXYZ[\]^_`abcdefghijklmnoprstuvwxyz{|}~Root EntrydO)PictureshCurrent UserSummaryInformation(q\JPowerPoint Document(DocumentSummaryInformation8