Hair analyses. Hair analysis beyond workplace testing. Ia: Fatal steroid abuse. Identification of stanozolol in hair

Transcription

1 Hair analyses Hair analysis beyond workplace testing Detlef Thieme IDAS Kreischa IRM + FTC Munich. anabolic (steroids, clenbuterol). kinetic (segmentation and profiling) 3. genetic (amitriptyline vs.nortriptyline Ia: Fatal steroid abuse Identification of stanozolol in hair Bodybuilder 996: Administration of.5g/month (endogenous prod.-5mg/day) Cardiac infarction Heart weight ~ 55 g Exact administration data available mg Apr-8 androlone Decanoate Testosterone (4 Esters) Metandienone Metenolone Enantate Stanozolol Stanozolol (8 pg/mg hair) 6.8E6 5: CH3 3.4E6.E : 3: 4: 5: 6: 7: 8: '--Stanozolol (6 pg/mg hair).9e5 5:49 3' CH3 9.7E4.E : 3: 4: 5: 6: 7: 8: date Mai-. Steroid-precursor C R Identification of testosterone esters H Testosterone esters. Active Anabolic Steroids Androstendiol CH 3 Std. Mixture (ng) Prop () Iso () Dec (4) PhP () 5.4E5 4.3E5 3.3E5.E5.E5 Testosterone Stanozolol.E : : 3: 4: 5: 6: 7: 3. Inaktive Metabolites C Suspicious Hair Sample () (.4) (5) (.8).7E5.4E5.E5 6.8E4 H H Phase : Androsterone H Phase : Androsterone glucuronid : : 3: 4: 5: 6: 7: 3.4E4.E

2 Ib: Stallion licensing and hair testing Clenbuterol Profiles in Hair (after 5 therapeutic doses) H Cl Cl C CH H C(CH3)3 5 d d 35 d 4 d 9 d d 5 d 36 d Mane Hair Segment [cm] androlone in horse hair Stallion Licensing androlone hair concentration in untreated stallions >5 Concentration Concentration [pg/mg] Age [a] androlone concentration in mane hair after transdermal treatment day 84: expected maximum Physiology of Skin, Hair and Sweat Eccrine Sweat Glands: High Amount Water, Electrolytes Skin Surface Thermoregulation Apocrine Glands: Small Amount ily Liquid Hair Follicle Bacterial Degradation dorants, Pheromones

3 Bioavailability I Transdermal Application of 9-orandrost-4-ene-diol Bioavailability II ral Application of 9-orandrost-4-ene-diol Blood conc [ng/ml] MB: orandrosterone AS: androlone PC: orandrostendiol Blood conc [ng/ml] Phase-MB: orandrosterone-glucuronide. 3 time [h] 3 time [h] androlone in horse hair Stallion Licensing Relevance in doping control? > Doping according to WADA: the detection of a prohibited substance or metabolites or markers of that substance in the athlete s specimen, the use of prohibited substances or methods, possession or trafficking prohibited substances or Concentration Conclusions Anabolics may be identified in hair Parent compound > metabolite Information about applied compound (e.g. differentiation of steroid esters) refusing without compelling justification to submit a sample. 3

4 . Illegal administration of clozapine Hair profile of clozapine 3 cm segments of a hair strand at least 5 victims duration > year date of termination was relevant analyses started 6 weeks later Clozapin [pg/mg] 5 5 Cl H Single hair analysis, reproducibility Segments of.5 mm, taken -ov-3 Second hair collection. ov.3 vs. 3. Mai 4 Individual hair growth rate of.66 cm / month ± 7% 6 6 7d+X 4 conc [pg/mg] Segment [o] Segment [o] Miniaturization Segment length -5 cm -5 mm weight 5- mg -5 µg Solvent 5 ml 3 µl HPLC analytical micro LC.5e4 Intensity, cps Fast Chromatography.5 mm segment ~ 6 µg hair ~7 fg clozapine on column.64 clozapin ~3 pg/mg desmethyl-mb

5 Hair Blank Desmethyl-metabolite 9 LD ~ 5 fg on column 7 6 Clozapin orclozapin 4 Intensity, cps Conc Clocapin [pg/mg] Conc. orclozapin [a.u.] Hair Segment [count] Maximum (?) resolution mm (~ days per data point) 3 5 Conclusions Accurate long term profiles of hair concentrations may be obtained from analyses of single hairs Conc. [pg/mg] Haar Segment [r] 3. Amitriptyline in hair of >3 children Illegal administration by a nurse Side effects: dizziness, somnolence, 8 unconsciousness, speech disorders, 6 diarrhoea, constipation 4 Large variations between children Investigation started 3-4 weeks after last administration [pg/cm] 6 4 Critical Timeframe Ami-T or-t 5 5 5

6 ortriptyline Profile (reproducibility) Ami-T or-t hair conc. [pg/cm] [pg/cm] Critical Timeframe Amitriptyline Biotransformation Amitriptyline - (E/Z)- Hydroxy-Amitriptyline CYP D6 78./7. Max..e4 cps. 5.9.e4 Amitriptyline 94./5. Max.. cps. -- Amitriptyline E Z /33. Max..e5 cps Max. cps. 8./ CYP C9 CYP D6 CYP C9.8e5 ortriptyline ortriptyline E Z H ortriptyline - (E/Z)- Hydroxy-ortriptyline -T/T Variation of biotransformation (9 children) AT T E--T Z--T E--AT Z--AT T/AT 5 5 6

7 Conclusions 3 Quantitation ratios in hair are reliable and highly reproducible Hair concentrations reflect bioavailability better than urinary levels Metabolite ratios examined in hair are consistent markers for individual biotransformation Limitation (uncertain incorporation via sweat) Citalopram in hairof a complient patient Hair Concentration Theory Segment [cm] C F 7