Scott K.S., Nakahara Y.
National Institute of Health Sciences 1-18-1, Kamiyoga, Setagaya-ku, Tokyo(158), Japan
The hair analysis of eight benzodiazepines (chlordiazepoxide, diazepam, estazolam, flunitrazepam, flura-zepam, medaflura-zepam, oxazepam and triazolam) using rat hair was investigated. Each of the benzodiazepines was injected daily into three Dark Agouti rats for 10 days at 10 mg/kg. The back hair of the rats was sha-ved prior to the first injection and again on the 28th day after the initial administration. To investigate the optimum extraction conditions, 10 mg hair samples incorporated with diazepam, flurazepam or medaze-pam were extracted by 7 different methods (Proteinase K, methanol-ammonia, methanol-trifluoroacetic acid, Soerensens buffer, 1M NaOH, ^-glucuronidase/ arylsulfatase, Biopurase). The method which yiel-ded the highest recoveries for the three drugs investigated was an acidic methanol extraction. Using this extraction procedure, the incorporation rates (ICR: the ratio of the hair concentration and the plasma AUC) of the eight benzodiazepines into rat hair were investigated. The ICRs ranged were found to range from 0.002 for flunitrazepam to 0.049 for flurazepam.
Incorporation of toluene and xylene metabolites into rat hair
Saito T.. Kusakabe T., Takeichi S.Department of Forensic Medicine, Tokai University School of Medicine, Bohseidai, Isehara, Kanagawa 259-1193,Japan
Although methamphetamine is mainly used as drug of abuse, thinner abuse by the young generation is also a social problem in Japan. Hair analysis of narcotic drugs including methamphetamine greatly advanced for recent decade. However, hair analysis of organic solvent abuse such as thinner have not been studied.
Thinner is mainly composed from toluene and xylenes, and we studied the incorporation of those meta-bolite hippuric acid (HA) and o-, m-, and p-methyl hippuric acids (o-, m-, p-MUA) into hair.
In the present study, we studied in vivo binding of HA and o-, m-, and p-MHA to dark agouti rats hair. The back hair of rat was shaved with animal electric shaver before exposure. Studies performed in vivo expo-sed of 30 min/day at 3 kinds of different concentration toluene and xylenes for 10 times/2 weeks. The newly grown hair was tweezed out from the root with tweezers at 7th of the last exposure. Hair samples were then washed, extracted, derivatized, and analyzed by GC-MS. HA and o-, m-, and p-MRA were not detected in the unexporsed rats hair. However, the each metabolite concentration in the hair also changed with the dependence on the exposure concentration. Mean concentrations ranged from ND (not detected) to 7.6 ng/mg, from ND to 13.8 ng/mg, from ND to 10.1 ng/mg, and from ND to 9.2 ng/ml hair for HA, o-, m-, and p-MHA, respectively.
These results indicated that the each metabolite concentration in hair is an effective index of thinner expo-sure.
Annales de Toxicologie Analytique, vol. XTV, n° 3, 2002
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Tandem mass spectrometry for the analysis of drugs of abuse in human hair
Sims D.N.. Stockham P.C.Forensic Science SA, 21 Divett Place, Adelaide SA 5000, Australia
As part of a study to monitor the drug use of patients undergoing a heroin withdrawal trial we investiga-ted methods which would allow drug screening to be conducinvestiga-ted on hair segments. Patients were under-going maintenance treatment with methadone, 1-alpha-acetylmethadol (LAAM) or naltrexone. In order to assess patients self reported drug use hair samples were collected and examined to reveal drug use over the preceding month. Drugs of particular interest were heroin, monoacetylmorphine, morphine, ampheta-mine, methylamphetaampheta-mine, methylenedioxy-methylamphetamine and diazepam. We sought a simple extraction step followed by identification of multiple drugs in a single chromatographic run.
Approximately 20 mg of each hair sample was incubated overnight at 45° C in methanol. The solvent was evaporated to dryness and the residue derivatized with pentafluoropropionic anhydride. The concentration of heroin and its metabolites in hair are typically in the range 0.1-5 ng/mg. Thus for 20 mg hair samples the total amount of drug present is typically 2-100 ng. Accurate determination of these drug concentra-tions requires sensitive assays. Trial analyses were conducted by gas chromatography/mass spectrometry in both selected ion monitoring mode (SIM) using a benchtop quadrupole and tandem MS mode using an ion trap. Minimal sample preparation of the hair produced some difficult matrices. Tandem MS provided improved signal to noise ratios for low drug concentrations compared with SIM. In addition, a product ion spectrum has the potential to provide a higher quality identification than the 3 ion ratios commonly used in SIM. Approximately 300 hair samples have been successfully analysed using tandem MS.
Determination of opiates and amphetamine in hair of detoxification and methadone treat-ment patients addicted to home made "polish heroin"
Stanaszek R.(!>, Piekoszewski W.( l 2 ), Karakiewicz J3,(3), Kozielec T.(3) (1) Institute of Forensic Research, Cracow, Poland
(2) Department of Clinical and Industrial Toxicology, School of Medicine, Jagiellonian University, Cracow, Poland
(3) Deparment of Family Medicine, Pomeranian Medical Academy, Szczecin, Poland
One of the most commonly abused form of opiates in Poland is a liquid home made product, called pot" or "Polish heroin", produced from poppy straw or poppy head juice. The main constituents of "kom-pot" are morphine (range 1.2-50 mg/ml), acetylcodeine (0-12.5 ng/ml), codeine (0,2-^1 mg/ml), 6-MAM (0-2.5 mg/ml), and heroin (0.2 - 2 mg/ml). Although the new trend among the Polish opiates addicts is to use the solid heroin for instance "brown sugar" 71 % of the examined patients appeared to be mixed depen-dent on opiates, amphetamines, cannabis, solvents and other drugs.
The main aim of the study was to evaluate and validate the analytical procedures of determination of main opiates (morphine, codeine, 6-MAM), methadone and amphetamines amphetamine (AMP), ephedrine (EP), methamphetamine (MA), methcathinone (MTC), PMA, MDA, MDMA, MDEA) in hair. The elabo-rated methods were applied to analyse hair samples of 107 subjects from the detoxification and methado-ne treatment programme form West Pomeranian Region (Szczecin) and 6 children living with addicted parents.
For determination of opioids hair stands after decontamination with isopropanol, phosphate buffer and dichloromethane were cut into 2 cm segments and pulverised in a ball mill. The samples were than hydro-lysed in 0,1 M HCI (50° C, overnight), extracted on Bond Elute Certify columns and derivatised with the mixture of PFPA and PFPOH. Extracts were analysed by ion trap GC/MS in a full scan EI mode. In case of amphetamines determination hair samples were decontaminated. The target substances were extracted with n-bytyl chloride after alkaline (IM NaOH) digestion and analysed with LC-APCI-MS in SIM mode.
The ranges for measured concentrations (ng/mg) were 0.3-32.4 for morphine, 12,5 for codeine, 0.5-6.1 for 6-MAM, 2.2-80.0 for methadone, 0.15-62.52 for AMP, 0.17-17.28 for EP, 0.10-16.52 for MA, 0.52-2.08 for MTC, 0.10-0.88 MDMA and 0.10-0.16 for MDEA. There were no cases which were positi-ve for PMA and MDA. The results of sectional analysis agreed with the self reported drug histories. The results show that morphine is the predominant opiate metabolite found in hair of home made "Polish heroin" abusers. In addition the good correlation was found between medical examination and review and the results of hair analysis for drugs of abuse. The application of the developed methods in monitoring of the abstinence in the methadon treatment programs revealed 40 % of these who did not comply. They usually took amphetamines (37 %) and opiates less frequently (19 %) during the substitution therapy. High concentration of methadone in hair may be due to high doses of this drug applied in the therapy.
The analysis of hair samples taken from children who were living with parents addicted to "Polish heroin"
were negative both for opiates and amphetamines.
Annales de Toxicologie Analytique, vol. XIV, n° 3, 2002