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Mobility has long been recognised as a key feature of later prehistoric communities in eastern Eurasia. Isotope analysis of human hair offers new potential for studying individual mobility patterns within these communities. Hair samples from individuals of the Tashtyk culture buried in the Oglakhty cemetery in southern Siberia (third to fourth centuries AD) reveal variations in diet during the last months of their lives. Millet and fish were important in summer and autumn, [C.sub.3] plants and meat and dairy products at other times of year. The results indicate strong seasonal shifts in diet, and seasonal movement between different areas. Keywords: Siberia, third-fourth centuries AD, isotope analysis, hair, diet, mobility, Introduction Southern Siberia is a mosaic of forest and steppes, with mountain ranges, vast plains, depressions, rolling hills and large river systems. From the earliest times, this diversity in the [...]

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2009.12.027 Byline: Marie Katrine Klose Nielsen, Sys Stybe Johansen, Petur Weihe Dalsgaard, Kristian Linnet Keywords: UPLC-TOF-MS; Hair analysis; Drugs of abuse; Pharmaceuticals Abstract: An UPLC-TOF-MS method for simultaneous screening and quantification of 52 drugs in hair was developed and validated. The selected drugs represent the most common classes of pharmaceuticals and drugs of abuse such as amphetamines, analgesics, antidepressants, antipsychotics, benzodiazepines, cocaine, ketamine and opioids. Hair samples were extracted with methanol:acetonitrile:ammonium formate (2mM, 8% acetonitrile, pH 5.3) overnight at 37[degrees]C. The target drugs were separated and quantified using a Waters ACQUITY UPLC coupled to a Waters Micromass LCT Premier XE Time-of-Flight mass spectrometer. Total chromatographic run time was 17min. The data were treated with the MassLynx software ChromaLynx XS and QuanLynx for automated identification and quantification, respectively. The limits of detection ranged from 0.01 to 0.10ng/mg using a 10-mg hair sample and the limit of quantification was 0.05ng/mg for 87% of the analytes. A good linear behaviour was achieved for most of the analytes in the range from LOQ to 10 or 25ng/mg except for the amphetamines. The method showed an acceptable precision and trueness, since the obtained CV and BIAS values were [less than or equal to]25% for 81% of the analytes. The extraction recoveries for 92% of the analytes ranged between 84 and 106% and the extraction recoveries for all analytes were better than 60%. The method was applied to 15 autopsy hair samples from forensic investigations showing a wide abuse pattern of many pharmaceuticals and drugs of abuse within a period of less than three months. The present study demonstrated that the combination of accurate mass and retention time can provide good selectivity, which demonstrates that the TOF instrument is adequate for both screening and quantification purposes. Furthermore, it was shown that screening with the ChromaLynx XS software is less sensitive and selective for some analytes than the QuanLynx software, especially in low concentrations. Author Affiliation: Section of Forensic Chemistry, Department of Forensic Medicine, Faculty of Health Sciences, University of Copenhagen, Frederik VIs vej 11, DK-2100, Denmark Article History: Received 11 June 2009; Accepted 10 August 2009

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2009.12.041 Byline: M. Pellegrini (a), M.C. Rotolo (a), S. La Grutta (b), F. Cibella (b), O. Garcia-Algar (c), A. Bacosi (a), G. Cuttitta (b), R. Pacifici (a), S. Pichini (a) Keywords: Environmental tobacco smoke; Hair nicotine; Urinary cotinine Abstract: We investigated acute and chronic exposure to environmental tobacco smoke (ETS) in a cohort of young adolescents using urinary cotinine and hair nicotine testing after recent implementation of Italian smoke free legislation. Study subjects were 372 Italian young adolescents, between 10 and 16 years of age from the principal city of Sicily, Palermo. Urine and hair samples were collected between November 2005 and May 2006, when the legislation to ban smoking in all the enclosed places of employment (including bars, restaurants, pubs) was completely enforced. An exhaustive questionnaire including sociodemographic characteristics and active and passive exposure to cigarette smoking was completed. Urinary cotinine was analyzed by radioimmunoassay and hair nicotine by a validated GC/MS method. Based on urinary cotinine results, 2.1% and 89% of the study participants, respectively, showed non-exposure and low acute exposure to ETS, whereas only 1.6% presented very high exposure or a hidden active smoking habit in the recent past. Hair nicotine disclosed non-exposure and low exposure to ETS in 11.8% and 65.6% of the young adolescents, respectively, taking into consideration a larger time-window. High repeated exposure, suggesting active smoking in some cases was observed in 8.6% of the study subjects. Hair nicotine was inversely related to educational level of the adolescents' parents. Overall, due to the implementation of smoke-free legislation and information campaign against smoking, a significant trend toward low exposure to ETS was observed in this study cohort with no association between exposure to ETS and respiratory illnesses. Author Affiliation: (a) Department of Therapeutic Research and Medicines Evaluation, Istituto Superiore di Sanita, V.le Regina Elena 299, 00161 Rome, Italy (b) Consiglio Nazionale delle Ricerche, Istituto di Biomedicina e Immunologia Molecolare, Palermo, Italy (c) Unitat de Recerca Infancia i Entorn (URIE), Paediatric Service, IMIM-Hospital del Mar, Barcelona, Spain Article History: Received 17 June 2009; Accepted 5 August 2009

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2009.12.026 Byline: Thomas Nadulski (a), Simona Bleeck (a), Johannes Schrader (b), Wolf-Rainer Bork (c), Fritz Pragst (a) Keywords: 11-Nor-delta-9-tetrahydrocannabinol-9-carboxylic acid ethyl ester; Alcohol marker; Cannabinoids; Hair analysis Abstract: 11-Nor-[DELTA].sup.9-tetrahydrocannabinol-9-carboxylic acid ethyl ester (THC-COOEt) can be presumed to be a mixed metabolite formed during combined consumption of cannabinoids and alcohol. In order to examine this hypothesis, THC-COOEt and its deuterated analogue D.sub.3-THC-COOEt were synthesized as reference substance and internal standard from the corresponding carboxylic acids and diazoethane and methods were developed for the sensitive detection of THC-COOEt in plasma and hair based on gas chromatography-electron impact mass spectrometry after silylation with N-methyl-N-tert-butyldimethylsilyl-trifluoroacetamide and gas chromatography-negative chemical ionization mass spectrometry (GC-NCI-MS) as well as tandem mass spectrometry (GC-NCI-MS-MS) after derivatization with pentafluoropropionyl anhydride. The methods were applied for THC-COOEt determination to plasma samples from 22 drunk driving cases which contained both ethanol (0.30-2.16mg/g) and THC-COOH (15-252ng/mL) as well as to 12 hair samples from drug fatalities which were both positive for THC (0.09-2.04ng/mg) and fatty acid ethyl esters as markers of chronic alcohol abuse (0.70-6.3ng/mg). In none of these samples THC-COOEt could be found with limits of detection of 0.3ng/mL in plasma and 2pg/mg in hair in 11 samples using GC-NCI-MS and 0.2pg/mg in one sample using GC-NCI-MS. Therefore, the use of this compound as a marker for combined cannabis and alcohol consumption could not be achieved. Author Affiliation: (a) Institute of Legal Medicine, University Hospital Charite, Hittorfstr. 18, 14195 Berlin, Germany (b) Central Institute of Medical Services of German Armed Forces, Scharnhorststr. 14, 10115 Berlin, Germany (c) Forensic Science Institute Berlin, LKA KT 41, Tempelhofer Damm 12, 12101 Berlin, Germany Article History: Received 13 July 2009; Accepted 20 July 2009

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2009.12.034 Byline: Lisa O'Brien (a)(b), Carina Baumer (c), Detlef Thieme (c)(d), Hans Sachs (c)(d), Gideon Koren (a)(b)(e) Keywords: Pregnancy; Antidepressant metabolism; Hair; Segmental analysis Abstract: Depression and other psychiatric illnesses are common during pregnancy and are often treated with antidepressants. Physiological changes of pregnancy may alter the pharmacokinetics of medications and ultimately affect the dose required to maintain effective therapy. Human hair offers a safe, non-invasive way to monitor long term systemic exposures to medications. Author Affiliation: (a) Division of Clinical Pharmacology and Toxicology, Hospital for Sick Children, Toronto, Canada (b) Institute of Medical Science, University of Toronto, Canada (c) Forensic Toxicological Centre, Munich, Germany (d) Institute of Forensic Medicine, Munich, Germany (e) Department of Pediatrics, Hospital for Sick Children, Toronto, Canada Article History: Received 5 June 2009; Accepted 12 August 2009

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2009.12.035 Byline: L. Morini (a), E. Marchei (b), F. Vagnarelli (c), O. Garcia Algar (d), A. Groppi (a), L. Mastrobattista (b), S. Pichini (b) Keywords: Fatty acid ethyl esters; Ethyl glucuronide; Ethyl sulfate; Fetal alcohol spectrum disorders Abstract: This study investigated ethyl glucuronide (EtG) and ethyl sulfate (EtS) concentration in meconium and in maternal and neonatal hair (HEtG and HFAEEs, respectively) as potential markers of intrauterine exposure to ethanol together with meconium fatty acid ethyl esters (FAEEs) in a cohort of 99 mother-infant dyads, 49 coming from the Arcispedale of Reggio Emilia (Italy) and 50 from the Hospital del Mar of Barcelona (Spain). FAEEs, EtG and EtS were measured in meconium samples using liquid chromatography-tandem mass spectrometry. A head space-solid phase microextraction-gas chromatography-mass spectrometry was used to test HEtG and HFAEEs in hair samples from mothers and their newborns. Eighty-two meconium samples (82.8%) tested positive for EtG, 19 (19.2%) for EtS while 22 (22.2%) showed FAEEs levels higher than 2nmol/g, the cut-off used to differentiate daily maternal ethanol consumption during pregnancy from occasional or no use. Although EtG and EtS in meconium did not correlate with total FAEEs concentration, a good correlation between EtG, EtS and ethyl stearate was observed. Moreover, EtG correlated well with ethyl palmitoleate, while EtS with ethyl laurate, myristate and linolenate. Neither maternal nor neonatal hair appears as good predictors of gestational ethanol consumption and subsequent fetal exposure in these mother-infant dyads. In conclusion, these data show that meconium is so far the best matrix in evaluating intrauterine exposure to ethanol, with EtG and EtS being potentially good alternative biomarkers to FAEEs. Author Affiliation: (a) Department of Legal Medicine and Public Health, University of Pavia, Pavia, Italy (b) Department of Therapeutic Research and Medicine Evaluation, Istituto Superiore di Sanita, Roma, Italy (c) Arcispedale Santa Maria Nuova, Reggio Emilia, Italy (d) Unitat de Recerca Infancia i Entorn (URIE), Paediatric Service, IMIM-Hospital del Mar, Barcelona, Spain Article History: Received 17 June 2009; Accepted 17 August 2009

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2009.12.025 Byline: Cynthia Coulter, James Tuyay, Margaux Taruc, Christine Moore Keywords: Hair; Drugs of abuse; Immunoassay Abstract: A semi-quantitative analytical screening procedure for the determination of cocaine, amphetamines, opiates, and delta-9-tetrahydrocannabinol in hair has been developed. The procedure employs an aqueous extraction buffer, uses only 10mg of hair, requires 2h of incubation for the extraction to occur, and multiple drug classes can be screened using enzyme linked immunosorbent assays. Hair calibration standards were prepared around the recommended cut-off concentrations of the Society of Hair Testing. All drug classes showed excellent linearity over the concentration range tested, indicating that immunochemical screening can be used in a semi-quantitative mode for hair analysis using an aqueous buffer, rapid extraction and a small amount of hair. Author Affiliation: Immunalysis Corporation, 829 Towne Center Drive, Pomona, CA 91767, USA Article History: Received 9 July 2009; Accepted 27 July 2009

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2009.12.046 Byline: M. Moller (a)(b), K. Aleksa (a), P. Walasek (a), T. Karaskov (a), G. Koren (a)(b) Keywords: Solid-phase microextraction; Hair analysis; Opiates; Gas chromatography-mass spectrometry Abstract: Opiate hair analysis continues to prove difficult due to the scarcity of hair sample and low drug concentrations. For this reason, we developed a sensitive method utilizing headspace solid-phase microextraction (HS-SPME) coupled with gas chromatography-mass spectrometry (GC-MS) for the detection of three principle opiates; codeine, morphine, and 6-acetylmorphine. Author Affiliation: (a) Motherisk Program, Division of Clinical Pharmacology & Toxicology, Hospital for Sick Children, , Toronto, Ontario Canada, M5G1X8 (b) Department of Pharmacology & Toxicology, University of Toronto, Toronto, Ontario, Canada Article History: Received 9 June 2009; Accepted 21 July 2009

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2009.12.036 Byline: Robert Kronstrand (a), Ingrid Nystrom (a), Malin Forsman (a), Kerstin Kall (b) Keywords: Hair analysis; Driver's licence regranting; LC-MS-MS Abstract: When being convicted for petty drug offence or driving under the influence of drugs in Sweden, the driving license may be suspended. To regain the license, the person has to prove that he or she has been drug free during an observation period. This is controlled by urine samples taken at several occasions. However, the risk of manipulation and the risk of false negative urine samples are high. In addition, many people find it difficult or embarrassing to urinate when observed. Hair sampling might therefore be a welcome option to this procedure, with its easy sampling and minimal risk of manipulation. The longer detection window may also provide better information to the physician. The aim of this work was to evaluate if clients preferred hair samples to urine and to investigate practical and interpretive problems or advantages with hair samples. Ninety-nine hair samples and 198 urine samples were collected from 84 clients during the 12 month study period. Hair samples were divided into either one segment (0-3cm) or two segments (0-3 and 3-6cm) depending on the length. The hair samples were screened with LC-MS-MS for 20 drugs and confirmation of positive results were performed with GC-MS or LC-MS-MS. The results were compared to urine samples taken at two occasions during the observation period. To cover the timeframe of the urine samples hair was collected 2 weeks after the second sample. The urine samples were analysed with immunochemical screening and positive results confirmed with GC-MS or LC-MS-MS. Seventy-four clients presented with negative results in both urine and hair. Hair analysis identified illegal drugs at seven different occasions whereas urine failed to identify any illegal drugs. However the thresholds used may still be too high to find sporadic use as clients that admitted to use drugs sporadically presented with drug concentrations lower than the agreed thresholds but above the limit of detection. This implicates that the physician must have an understanding and knowledge of the limitations of the screening methods used. Another important outcome was that the clients approved of hair sampling considering it a better means to prove their drug abstinence. In addition, both the clients and the clinicians thought hair sampling easier than urine sampling. We believe that hair analysis can offer several advantages compared to urine analysis for clinicians working with driving license regranting. Author Affiliation: (a) National Board of Forensic Medicine, Dep. of Forensic Toxicology, Linkoping, Sweden (b) Clinic for Dependency Disorders, University Hospital, Linkoping, Sweden Article History: Received 7 July 2009; Accepted 5 August 2009

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2009.12.033 Byline: Pascal Kintz (a), Julie Evans (b), Marion Villain (a), Vincent Cirimele (a) Keywords: Hair; Contamination; Decontamination; Methadone; Child; Administration Abstract: Methadone is not licensed for use in children though it can be employed for the management of neonatal opiate withdrawal syndrome. During the last 2 years, our laboratory has been asked to test for methadone and EDDP, its major metabolite, in hair from children that were admitted to hospital unconscious and where methadone had already been identified in a body fluid (4 cases) or where the children were deceased and evidence of methadone overdosage having already been established (2 cases). In all of these cases, segmental analysis revealed approximately the same amount of drug along the hair lock. As a consequence, contamination was considered as an issue and interpretation of the results was a challenge that deserves particular attention. After decontamination with dichloromethane and segmentation the hair was cut into small pieces, incubated overnight at 40[degrees]C, liquid-liquid extracted and analysed with LC-MS/MS, using 2 transitions per compound. The LOQ for both methadone and EDDP was 10pg/mg. In the first series involving children admitted to hospital, the following results were obtained: acents case 1: 4x1cm section, methadone at 0.05-0.08ng/mg, no EDDP detected, acents case 2: 4x1cm section, methadone at 0.13-0.15ng/mg, EDDP at 0.02ng/mg, acents case 3: 3x1.5cm section, methadone at 0.07-0.09ng/mg, EDDP at 0.01-0.03ng/mg, acents case 4: 6x2cm section, methadone at 0.06-0.13ng/mg, EDDP at 0.02-0.03ng/mg. The following concentrations were obtained from the children who had died following a methadone overdose: acents case 5: 2x2cm section, methadone at 0.53-0.58ng/mg, no EDDP detected, acents case 6: 4x1cm section, methadone at 0.44-0.77ng/mg, EDDP at 0.04-0.06ng/mg. The first observation is that all these concentrations are low by comparison with those observed in adults on methadone maintenance therapy. However, the more surprising observation is the relative homogenous concentrations along the hair locks in each specific case. This raises concerns around the possibility that contamination could have occurred prior to sampling and makes it hard to reach a conclusion regarding the possibility of repeated methadone exposure in the months prior the incidents. In these cases it was impossible to conclude that the children were deliberately administered methadone. The results of the analysis of hair could indicate that they were in an environment where methadone was being used and where the drug was not being handled and stored with appropriate care. The homogenous concentrations found on segmental analyses could be indicative of external contamination that may have arisen not only from direct contamination with the drug but also via contamination with body fluids at the post mortem or from sweat produced close to the time of the incident. In view of these results we concluded that a single determination should not be used firmly to discriminate long-term exposure to a drug. Author Affiliation: (a) Laboratoire ChemTox, 3 rue Gruninger, 67400 Illkirch, France (b) ROAR Forensics Ltd, Malvern, UK Article History: Received 10 June 2009; Accepted 24 August 2009

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2009.07.023 Byline: Ronald Agius (a), Thomas Nadulski (a), Hans-Gerhard Kahl (a), Johannes Schrader (b), Bertin Dufaux (a), Michel Yegles (c), Fritz Pragst (d) Keywords: Ethyl glucuronide; Hair analysis; Forensic validation; HS-SPME-GC-MS/MS; Alcohol marker; Quality control; Error estimation Abstract: The analysis of ethyl glucuronide (EtG) in hair is a powerful tool for chronic alcohol abuse control because of the typical wide detection window of the hair matrix and due to the possibility of segmentation, allowing evaluation of alcohol consumption in different periods. Additionally, EtG in hair is often the only diagnostic parameter of choice for alcohol abuse when other clinical parameters such as ALT, AST, [gamma]GT and CDT (asialotransferrin and disialotransferrin) are in the normal range and EtG in urine negative. In this paper, we describe the development, optimization and validation of a new method based on hair extraction with water, clean-up by solid phase extraction (SPE), derivatization with heptafluorobutyric anhydride and headspace solid-phase microextraction (HS-SPME) in combination with GC-MS/MS according to forensic guidelines. The assay linearity of EtG was confirmed over the range from 2.8 to 1000pg/mg hair, with a coefficient of determination (r.sup.2) above 0.999. The LLOQ was 2.8pg/mg and the LLOD was 0.6pg/mg. An error profile calculated according to the 'Guide to the Expression of Uncertainty in Measurement' (GUM) at 99% confidence intervals for the range 5-750pg/mg hair did not exceed 10%. This range corresponds to more than 98% of the positive samples analysed. Author Affiliation: (a) Labor Krone, Siemensstr. 40, 32105, Bad Salzuflen, Germany (b) Sanitatsdienst der Bundeswehr Kiel, AuAenstelle Berlin, Germany (c) Laboratoire National de Sante, Toxicologie, Luxembourg (d) Institut fur Rechtsmedizin, Charite Berlin, Germany Article History: Received 12 June 2009; Accepted 28 July 2009

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2009.12.045 Byline: Jin Young Kim, Soon Ho Shin, Jae Il Lee, Moon Kyo In Keywords: Phenylalkylamine; Micro-pulverized extraction; Hair analysis; GC-MS Abstract: A gas chromatography-mass spectrometric (GC-MS) method was developed and validated for the determination of five psychotropic phenylalkylamine derivatives (amphetamine, AP; methamphetamine, MA; 3,4-methylenedioxyamphetamine, MDA; 3,4-methylenedioxymethamphetamine, MDMA; norketamine, NKT) in human hair. Hair samples (10mg) were washed with distilled water and acetone, mechanically pulverized for 1.5min with a bead mill, and then incubated in 1mL of methanol under ultrasonication at 50[degrees]C for 1h. The resulting solutions were evaporated to dryness, derivatized using heptafluorobutyric anhydride (HFBA) at 50[degrees]C for 30min, and analyzed by GC-MS. The linear ranges were 0.1-20.0ng/mg for AP and MA and 0.05-20.0ng/mg for MDA, MDMA, and NKT, with the coefficients of determination (r.sup.2 0.9982). The intra-day and inter-day precisions were within 11.5% and 12.8%, respectively. The intra-day and inter-day accuracies were -4.1% to 5.8% and -6.6% to 4.2%, respectively. The limits of detections (LODs) for each compound were lower than 0.028ng/mg. The recoveries were in the range of 78.9-101.2%. Based on these results, the method proved to be effective for the rapid and simple determination of phenylalkylamine derivatives in hair specimens. Author Affiliation: Drug Analysis Laboratory, Forensic Science Division, Supreme Prosecutors' Office, 706 Banporo, Seocho-gu, Seoul 137-730, Republic of Korea Article History: Received 5 June 2009; Accepted 25 July 2009

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2009.12.040 Byline: R. Gow (a), S. Thomson (b), M. Rieder (b)(c), S. Van Uum (b), G. Koren (a)(b)(d)(e) Keywords: Hair; Cortisol; Chronic stress; Hypothalamic-pituitary-adrenal axis Abstract: Hair analyses for exogenous compounds, specifically drugs of abuse, have been a useful tool in detecting long-term drug exposure. More recently, studies have delved into the exposure of endogenous compounds in hair. Cortisol is synthesized in the adrenal cortex in response to stress-induced activation of the hypothalamic-pituitary-adrenal (HPA) axis. While catecholamines generally indicate acute stress, cortisol can be used as an indicator for sub-acute and chronic stress. Studies on the effects of chronic stress are most often subjective in nature, relying on questionnaires asking the participant to recall on past stressors. This can lead to the issue of recall and reporting bias. A new objective measure of chronic stress is needed for a more accurate understanding of the effects of chronic stress on the body. This review uses emerging evidence to describe the usefulness of hair analysis for cortisol and discusses the current methods used. Author Affiliation: (a) Department of Physiology and Pharmacology, University of Western Ontario, Canada (b) Department of Medicine, Schulich School of Medicine and Dentistry, University of Western Ontario, Canada (c) CIHR-GSK Chair in Pediatric Clinical Pharmcology, Children's Health Research Institute, London, Ontario, Canada (d) Ivey Chair in Molecular Toxicology, University of Western Ontario, Canada (e) Department of Clinical Pharmacology/Toxicology, Hospital for Sick Children, Toronto, Ontario, Canada Article History: Received 12 June 2009; Accepted 2 September 2009

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2009.12.037 Byline: Marjorie Cheze, Aurelie Muckensturm, Guillaume Hoizey, Gilbert Pepin, Marc Deveaux Keywords: Re-offending; Sedatives; Drug-facilitated crime; Hair; LC-MS/MS Abstract: The authors present 3 cases that demonstrate a return to DFC following periods of inactivity. The offences occurred in Paris and its suburbs and in each of the cases there were two distinct periods of activity by the offenders with 2, 8 and 22 victims attributed to each of the perpetrators. To 20mg of decontaminated and cut hair, 100pg/mg of clonazepam-d4 was added as internal standard. Hair specimens were extracted with CH.sub.2Cl.sub.2/ether after incubation overnight at 56[degrees]C in pH 7.6 buffer. Extractions were performed on blood and urine using Toxi-tube A.sup.[R] with 5ng/mL of clonazepam-d4. The residues were analyzed by LC-ESI-MS/MS. Calibration curves in blood and urine (0.5-500ng/mL) were prepared by spiking aliquots of blank fluids (r.sup.2 0.9816 for all drugs). LOD in body fluids ranged 0.5-10ng/mL. Calibration curves in hair (0.5-100pg/mg) were prepared by spiking aliquots of blank hair (r.sup.2 0.9877 for all drugs). LOD in hair ranged 0.5-5pg/mg. Case #1: Two young women were raped with an interval of approximately 1 year between the incidents. Lorazepam (present, Segmental hair analysis permits retrospective information on drug exposure and should be considered in the investigation of drug-facilitated crimes not only to prove single exposure but also when there has been any appreciable delay in samples being obtained for analysis. Indeed, in 56% cases reported in this paper, due to the long time that elapsed between offences and the opportunity to obtain samples for analysis hair analysis was considered the only viable matrix to investigate the possibility of drug involvement in the crimes. Our experience demonstrates that the incidence of re-offending in DFC after a period of inactivity (often due to imprisonment) may be of concern, notably in big cities. Author Affiliation: Laboratoire TOXLAB, 7 rue Jacques Cartier, F-75018 Paris, France Article History: Received 3 July 2009; Accepted 9 September 2009 Article Note: (footnote) [star] Paper presented at the 14th meeting of the SoHT, Roma, June 2009.

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2009.12.042 Byline: M. Falcon (a), F. Valero (a), M. Pellegrini (b), M.C. Rotolo (b), G. Scaravelli (b), J. Joya (c), O. Vall (c), O. Garcia Algar (c), A. Luna (a), S. Pichini (b) Keywords: Drug abuse; Pregnancy; Induced abortion Abstract: Drug abuse is a worldwide phenomenon with significant health and socioeconomic impact and it is of particular concern in women of reproductive age and in pregnant women. We aimed to investigate the prevalence of drug use by serum and hair testing in a cohort of pregnant women at 12th week gestation who decided voluntarily to interrupt their pregnancy and to investigate the relationship between drug exposure and induced abortions (IA), repeated IA and contraception. The study was conducted in an obstetrics clinic authorised to perform IA in Murcia, Spain during an 18 months period (2007-2009). Apart from serum and/or hair testing, the 142 women enrolled in the study completed a detailed questionnaire regarding drug, alcohol and tobacco use in the previous 3 months. Serum and hair samples were analyzed by gas chromatography mass spectrometry assays. Hair and serum samples showed a 30% overall positivity to drugs of abuse. Of these samples, 20.4, 14.1, 4.2 and 1.4% were positive for cannabinoids, cocaine, opiates, and MDMA, respectively, with polydrug use in 5.6% cases. In this cohort, a positive association was found between drug use and repeated IA. The results highlight the need for promoting pregnancy planning for young women in general, especially when consuming psychoactive substances as well as promote safe and accessible contraception in women of reproductive age. In women requesting IA, specific drug abuse counselling should be implemented. Author Affiliation: (a) Legal Medicine, University of Murcia, Spain (b) Istituto Superiore di Sanita, Rome, Italy (c) Paediatric Service, URIE, Hospital del Mar, and Universitat AutA[sup.2]noma, Barcelona, Spain Article History: Received 17 June 2009; Accepted 25 July 2009

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2009.12.024 Byline: Joey Gareri, Gideon Koren Keywords: Hair; Fetal; Neonatal; Toxicology; Drugs Abstract: Neonatal hair is a clinically important toxicological matrix, as it allows determination of in utero drug exposure. This paper serves to review the physiological development of the hair follicle and hair production during fetal life. An understanding of the mechanisms and timing of hair development in the prenatal period is critical to effectively assessing the time window of exposure determination associated with toxicological analysis of neonatal hair. Author Affiliation: Division of Clinical Pharmacology & Toxicology, Hospital for Sick Children, 555 University Avenue, Toronto, Ontario, M5G 1X8, Canada Article History: Received 12 June 2009; Accepted 21 July 2009

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2009.12.039 Byline: R. de la Torre (a)(b), E. Civit (a), F. Svaizer (c), A. Lotti (c), M. Gottardi (c), M. Miozzo (c) Keywords: Hair testing; Immunological methods; Opiates; Cannabinoids; Ecstasy; Cocaine; Methadone; Amphetamines; Sample preparation Abbreviations: MDMA, 3,4-methylenedioxymethamphetamine; MDA, 3,4-methylenedioxyhamphetamine; MDEA, 3,4-methylenedioxyethylamphetamine; THC, delta-9-tetrahydrocannabinol; THCCOOH, 11-nor-delta-9-carboxy-tetrahydrocannabinol; CBN, cannabinol; CBD, cannabidiol; LOQ, limit of quantification; 6-MAM, 6-monoacetylmorphine; EDDP, 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine; SPE, solid phase extraction; BSTFA, bis(trimethylsilyl)-trifluoroacetamide; TMCS, trimethyl-chlorosilane; TFAA, trifluoroacetic acid anhydride; ELISA, enzyme linked immunosorbent assay; EIA, enzyme immunoassay Abstract: Drug testing in hair usually requires a rather complex sample treatment before drugs are amenable to analysis by either immunological and/or chromatographic coupled to mass spectrometry methods. Immunological methods applied are usually dedicated to hair analysis as analytes present in this matrix are not always the same present in urine. Comedical s.a.s. laboratories recently commercialized reagents (VMA-T) purposely designed for hair sample treatment which are compatible with current immunometric methods used for urine drug testing. This is possible as some analytes (6-MAM and cocaine) present in hair after sample treatment are converted to those detected in urine (morphine and benzoylecgonine). A correlation study for several drug classes performed in two laboratories with 32 clinical and 12 spiked drug free (controls) hair samples shows that implementation of the method on clinical chemistry analyzers is easy and that results obtained by different operators and instruments are comparable and reproducible. The main advantage of VMA-T method is the possibility to simultaneously extract from hair main drug classes, in a period of time lower than 2h and its compatibility with immunological methods applied in urine drug testing. Author Affiliation: (a) Institut Municipal d'InvestigatiA[sup.2] Medica (IMIM-Hospital del Mar), Human Pharmacology and Clinical Neurosciences Research Group, Neuropsychopharmacology Program, Barcelona, Spain (b) Universitat Pompeu Fabra (CEXS-UPF), Barcelona, Spain (c) Laboratorio di Sanita Pubblica (LSP) - Azienda Provinciale Servizi Sanitari, Trento, Italy Article History: Received 23 July 2009; Accepted 24 September 2009

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2009.12.043 Byline: Isabela Tarcomnicu (a), Alexander L.N. van Nuijs (a), Katrien Aerts (a), Mireille De Doncker (b), Adrian Covaci (a)(c), Hugo Neels (a)(b) Keywords: Ethyl glucuronide; Meconium; Hair; Alcohol marker; HILIC-MS/MS; Solid-phase extraction Abstract: Ethyl glucuronide (EtG) detection in non-conventional matrices, such as hair and meconium, can provide useful information on alcohol abuse over a long time frame, for example during pregnancy or after a withdrawal treatment. This study reports on the development, validation and application of a new hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS) method for the analysis of EtG in meconium and hair. For each matrix, the sample preparation and the chromatographic separation were thoroughly optimised. Additionally, experiments with reversed-phase liquid chromatography were also performed in the development stages. Analyses were carried out using a Phenomenex Luna HILIC column (150mmx3mm, 5[mu]m) and a mobile phase composed by ammonium acetate 2mM and acetonitrile, in gradient. Different SPE cartridges (Oasis MAX, Oasis WAX, aminopropyl silica) and solvents were tested in order to obtain the highest recoveries and cleanest extracts. Optimal results were obtained for meconium with aminopropyl cartridges, while for hair an incubation of 16h with 2mL of water and acetonitrile (50/50, v/v) provided good results. The analytical method was validated for both matrices (meconium and hair) by assessing linearity, precision, accuracy, recovery and limit of quantification. The calibration curve concentrations ranged from 50 to 1200pg/mg for meconium and from 20 to 1000pg/mg for hair. Real meconium and hair samples were analyzed and results were consistent with literature. Author Affiliation: (a) Toxicological Centre, University of Antwerp, Universiteitsplein 1, 2610 Antwerp, Belgium (b) Toxicology Laboratory, ZNA Stuivenberg, Lange Beeldekenstraat 267, 2060 Antwerp, Belgium (c) Laboratory for Ecophysiology, Biochemistry and Toxicology, Department of Biology, University of Antwerp, Groenenborgerlaan 171, 2020 Antwerp, Belgium Article History: Received 29 July 2009; Accepted 12 September 2009

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2009.12.029 Byline: Silke SuAe (a), Carl M. Selavka (b), Tom Mieczkowski (c), Fritz Pragst (d) Keywords: Alcohol markers; Carbohydrate-deficient transferrin (CDT); Fatty acid ethyl esters; Gamma glutamyltransferase (GGT); Hair analysis Abstract: For diagnosis of chronic alcohol abuse, fatty acid ethyl esters (FAEE) were determined in hair samples from 644 individuals, mainly parents from child protection cases. The analysis for ethyl myristate, ethyl palmitate, ethyl oleate and ethyl stearate was performed according to a validated procedure consisting of external degreasing by two times washing with n-heptane, extraction with a mixture of dimethylsulfoxide and n-heptane, separation and evaporation of the n-heptane layer, headspace solid phase microextraction of the residue after addition of phosphate buffer pH 7.6 and gas chromatography-mass spectrometry using deuterated internal standards. For interpretation, the sum of the concentrations of the four esters C.sub.FAEE was used with the cut-off's 0.5ng/mg for the proximal scalp hair segment 0-3cm or less and 1.0ng/mg for scalp hair samples with a length between 3 and 6cm and for body hair. C.sub.FAEE ranged from 0.11 to 31ng/mg (mean 1.77ng/mg, median 0.82ng/mg). The mean concentration ratio between the 4 esters was 8:45:38:9. 298 cases had C.sub.FAEE above the cut-off's. Self-reported drinking data were obtained in 553 of the cases in the categories abstinent (156 cases), moderate drinking (252 cases) and excessive drinking (145 cases). Median and box-plot data clearly demonstrate differentiation of these ingestor sub-populations by C.sub.FAEE. However, in the abstinent and moderate groups the consumption was frequently underreported (37 and 110 cases positive) whereas in the group self-reported excessive drinking 32 cases were negative. Comparison of C.sub.FAEE with carbohydrate-deficient transferrin (CDT) in 139 cases and gamma-glutamyltransferase (GGT) in 136 cases showed a good agreement in CDT- and GGT positive cases (27/28 and 32/41) but a large portion of the negative CDT- and GGT-results with positive hair test (44/100 and 48/95) which is explained mainly by the much shorter time window of CDT and GGT. No significant correlation was found between persons weight and C.sub.FAEE showing that the test is not biased against physical fitness or obesity. Furthermore, there was no statistically significant difference between scalp hair (541 samples) and hair from other body sites (84 samples). In conclusion, FAEE in hair appeared to be suitable markers for the detection of excessive drinking. However, as there is no proportionality between drinking amount and C.sub.FAEE, the additional use of other markers can increase the reliability of the interpretation. Author Affiliation: (a) Trimega Drogencheck, Sedanstr. 14, 89077 Ulm, Germany (b) 9 Hollywood Drive, Charlton, MA 01507, USA (c) Department of Criminology, University of South Florida, USA (d) University Hospital Charite, Institute of Legal Medicine, Hittorfstr. 18, 14195 Berlin, Germany Article History: Received 12 June 2009; Accepted 31 August 2009

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2009.12.028 Byline: F. Pragst (a), M. Rothe (b), B. Moench (b), M. Hastedt (a), S. Herre (a), D. Simmert (a) Keywords: Alcohol marker; Ethyl glucuronide; Fatty acid ethyl ester; Hair analysis Abstract: In this study the combined use of fatty acid ethyl esters (FAEE) and ethyl glucuronide (EtG) for diagnoses of chronically excessive alcohol abuse is investigated at 174 hair samples from driving ability examination, workplace testing and child custody cases for family courts and evaluated with respect to the basics of interpretation. Using the cut-off values of 0.50ng/mg for FAEE and 25pg/mg for EtG, both markers were in agreement in 75% of the cases with 103 negative and 28 positive results and there were 30 cases with FAEE positive and EtG negative and 13 cases with FAEE negative and EtG positive. As the theoretical basis of interpretation, the pharmacokinetics of FAEE and EtG is reviewed for all steps between drinking of ethanol to incorporation in hair with particular attention to relationships between alcohol dose and concentrations in hair. It is shown that the concentrations of both markers are essentially determined by the area under the ethanol concentration in blood vs. time curve AUC.sub.EtOH, despite large inter-individual variations. It is demonstrated by calculation of AUC.sub.EtOH on monthly basis for moderate, risky and heavy drinking that AUC.sub.EtOH increases very strongly in the range between 60 and 120g ethanol per day. This specific feature which is caused by the zero-order elimination of ethanol is a favorable prerequisite for a high discrimination power of the hair testing for alcohol abuse. From the consideration of the different profiles of FAEE and EtG along the hair and in agreement with the literature survey, a standardized hair segment 0-3cm is proposed with cut-off values of 0.5ng/mg for FAEE and 30pg/mg for EtG. This improves also the agreement between FAEE and EtG results in the cases of the present study. A scheme for combined interpretation of FAEE and EtG is proposed which uses the levels of abstinence and the double of the cut-off values as criteria in addition to the cut-off's. Considering the large variations in the relationship between ethanol dose and FAEE and EtG concentrations in hair, the combined use of both parameters strongly increases the accuracy of the diagnosis by mutual confirmation and identification of false positive or false negative results due to biological variations or analytical errors. Author Affiliation: (a) Institute of Legal Medicine, University Hospital Charite, Hittorfstr. 18, 14195 Berlin, Germany (b) Lipidomix GmbH, Berliner Allee 261-269, 13088 Berlin, Germany Article History: Received 12 June 2009; Accepted 22 July 2009

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2009.12.023 Byline: Volker Auwarter (a), Ariane Wohlfarth (a), Jessica Traber (a), Detlef Thieme (b), Wolfgang Weinmann (a) Keywords: Tetrahydrocannabinolic acid A; Tetrahydrocannabinol; Hair analysis, GC-MS; Contamination Abstract: Differentiation between external contamination and incorporation of drugs or their metabolites from inside the body via blood, sweat or sebum is a general issue in hair analysis and of high concern when interpreting analytical results. In hair analysis for cannabinoids the most common target is [DELTA]9-tetrahydrocannabinol (THC), sometimes cannabidiol (CBD) and cannabinol (CBN) are determined additionally. After repeated external contamination by cannabis smoke these analytes are known to be found in hair even after performing multiple washing steps. A widely accepted strategy to unequivocally prove active cannabis consumption is the analysis of hair extracts for the oxidative metabolite 11-nor-9-carboxy-THC (THC-COOH). Although the acidic nature of this metabolite suggests a lower rate of incorporation into the hair matrix compared to THC, it is not fully understood up to now why hair concentrations of THC-COOH are generally found to be much lower (mostly [DELTA]9-Tetrahydrocannabinolic acid A (THCA A) is the preliminary end product of the THC biosynthesis in the cannabis plant. Unlike THC it is non-psychoactive and can be regarded as a 'precursor' of THC being largely decarboxylated when heated or smoked. The presented work shows for the first time that THCA A is not only detectable in blood and urine of cannabis consumers but also in THC positive hair samples. A pilot experiment performed within this study showed that after oral intake of THCA A on a regular basis no relevant incorporation into hair occurred. It can be concluded that THCA A in hair almost exclusively derives from external contamination e.g. by side stream smoke. Elevated temperatures during the analytical procedure, particularly under alkaline conditions, can lead to decarboxylation of THCA A and accordingly increase THC concentrations in hair. Additionally, it has to be kept in mind that in hair samples tested positive for THCA A at least a part of the 'non-artefact' THC probably derives from external contamination as well, because in condensate of cannabis smoke both THC and THCA A are present in relevant amounts. External contamination by side stream smoke could therefore explain the great differences in THC and THC-COOH hair concentrations commonly found in cannabis users. Author Affiliation: (a) Institute of Forensic Medicine, University Medical Centre Freiburg, Albertstr. 9, 79104 Freiburg, Germany (b) Institute of Doping Analysis and Sports Biochemistry, Dresdner Str. 12, 01731 Kreischa, Germany Article History: Received 30 July 2009; Accepted 31 August 2009

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2009.09.016 Byline: Sabina Strano-Rossi (a), Francesco Botre (a)(b), Ana Maria Bermejo (c), Maria Jesus Tabernero (c) Keywords: Amphetamines; Designer drugs; Hair analysis; Chiral separation; Forensic toxicology Abstract: This paper presents a rapid and sensitive method for the determination and chiral separation of amphetamines and related designer drugs in hair samples. The substances are extracted from hair matrix by a 30min treatment with a saturated carbonate buffer at pH 10 under ultrasonication. A commercial chiral derivatizing agent, trifluoroacetyl-prolyl chloride, is then added to the solution that is directly extracted with hexane and subsequently analyzed by GC/MS in SIM mode. R and S isomers of amphetamine, methamphetamine, MDA, MDMA and MDEA can be separated and detected with a limit of detection of 0.1ng/mg for amphetamine, methamphetamine and MDA, and of 0.2ng/mg for MDMA and MDEA. The method was then applied to 12 samples from suspected amphetamines abusers, showing the presence of both isomers of amphetamine and MDMA in one sample (27 and 1.5ng/mg, respectively) and of MDMA in further eight samples, in concentrations ranging from traces to 2.7ng/mg. No differences were observed in the disposition of different isomers in hair. Author Affiliation: (a) Laboratorio Antidoping, Federazione Medico Sportiva Italiana, Largo Giulio Onesti 1, 00197 Rome, RM, Italy (b) Dipartimento per le Tecnologie, le Risorse e lo Sviluppo, Universita 'La Sapienza', Via del Castro Laurenziano 9, 00161 Rome, RM, Italy (c) Instituto de Medicina Legal, University of Santiago de Compostela, C/San Francisco s/n, 15782 Santiago de Compostela, Spain Article History: Received 20 April 2009; Revised 14 September 2009; Accepted 21 September 2009

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2009.08.003 Byline: Bruno Lemos Batista, Jairo Lisboa Rodrigues, Vanessa Cristina de Oliveira Souza, Fernando Barbosa Keywords: ICP-MS; Hair samples; Trace elements; Rapid treatment; Forensic; Poisoning Abstract: An ultrasound-assisted extraction method is proposed for the determination of trace elements in hair samples by inductively coupled plasma-mass spectrometry (ICP-MS) for forensic investigation. Prior to analysis, 25mg of hair samples were accurately weighed into (15mL) conical tubes. Then, 2mL of 20% HNO.sub.3 is added to the samples, sonicated at 2min (50W, 100% amplitude), and then further diluted to 10mL with Milli-Q water. Resulted diluted slurries are centrifuged and the analytes are directly determined in the supernatant. Calibrations against aqueous solutions were carried out with rhodium as internal standard. The method was successfully applied for the extraction of Al, As, Ba, Be, Cd, Co, Cr, Cu, Mn, Pb, Tl, U, V and Zn with a method detection limit (3s, n =20) of 0.1, 0.4, 0.2, 0.09, 0.08, 0.04, 0.1, 2.9, 1.0, 0.9, 0.04, 0.05, 0.1 and 4.2ng/g, respectively. Method accuracy is traceable to Certified Reference Materials (CRMs) 85 and 86 human hair from the International Atomic Energy Agency (IAEA). Additional validation data are provided based on the analysis of hair samples from the trace elements intercomparison program operated by the Institut National de Sante' Publique du Quebec, Canada. The proposed method is very simple and can be applied for forensic purposes with the elimination of sample digestion step prior to analysis. Then, a considerable improvement in the sample throughput is archived with the use of the proposed method. Author Affiliation: Laboratorio de Toxicologia e Essencialidade de Metais, Depto. de Analises Clinicas, Toxicologicas e Bromatologicas, Faculdade de CiA*ncias FarmacA*uticas de Ribeirao Preto-USP, Avenida do Cafe s/n, Monte Alegre, 14040-903, Ribeirao Preto-SP, Brazil Article History: Received 26 April 2009; Revised 21 July 2009; Accepted 1 August 2009

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2009.04.004 Byline: F. Garcia-Bournissen (a), M. Moller (a), M. Nesterenko (a), T. Karaskov (a), G. Koren (a) Keywords: Hair analysis; Cocaine; Half-life Abstract: Methods that employ detection of drugs of abuse in hair are important for monitoring compliance with drug abstinence. Understanding the mechanisms and timeline of drug disappearance from hair is critical for clinical and forensic application of hair testing. We aimed to evaluate the kinetics of disappearance of cocaine and its metabolite, benzoylecgonine (BE), from hair after discontinuation of drug use. Author Affiliation: (a) Motherisk Program, Division of Clinical Pharmacology & Toxicology, Department of Pediatrics, Hospital for Sick Children, University of Toronto, Canada Article History: Received 14 November 2008; Revised 24 March 2009; Accepted 1 April 2009

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2009.04.008 Byline: Vivek Sahajpal (a), S.P. Goyal (a), M.K. Thakar (c), R. Jayapal (b) Keywords: Wildlife forensics; Microscopy; Bovid; Hair Abstract: Dorsal guard hairs of 10 bovid species of India, listed under Schedule-I of Wildlife (Protection) Act 1972 of India and some of them quite frequently encountered in illegal trade, were studied using light microscopy. We discuss characteristics including colour, hair thickness, cuticular pattern, medulla pattern, medulla index, cross-section and scale count index for species characterisation/identification to deal wildlife offence cases. Although some species could be identified very easily based on one or few microscopic hair characteristics, however there were some overlaps of few hair characteristics among some species. Species like Pantholops hodgsonii could be characterised most easily, just based on cuticular pattern and similarly Capricornis sumatraensis could be characterised simply by medulla pattern. For species showing overlaps in some of the microscopic hair characteristics, a combination of all the characteristics was most useful. We suggest the use of maximum number of parameters for distinguishing sympatric and closely related species. In addition to wild species, hair characteristics of three domestic species have been examined and compared with the selected wild species. Author Affiliation: (a) Wildlife Forensic Lab, Wildlife Institute of India, Dehradoon 248001, India (b) Wildlife Institute of India, Dehradoon 248001, India (c) Forensic Science Dept. Punjabi University, Patiala 147002, India Article History: Received 6 May 2008; Revised 2 March 2009; Accepted 6 April 2009

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2009.04.003 Byline: Luca Morini (a), Lucia Politi (b), Silvia Acito (a), Angelo Groppi (a), Aldo Polettini (c) Keywords: Alcohol markers; Hair ethyl glucuronide; Carbohydrate-deficient transferring; Sensitivity and specificity Abstract: This study was designed with the aim to compare sensitivity and specificity of ethyl glucuronide in hair (HEtG) and carbohydrate-deficient transferrin (CDT) in serum as markers of heavy drinking. Eighty-six volunteers, including teetotalers, social, and heavy drinkers, were interviewed to evaluate their ethanol daily intake (EDI) during the last 2-week and 3-month periods. HEtG determination was performed by a fully validated LC-MS-MS procedure and ranged from Author Affiliation: (a) Department of Legal Medicine and Public Health, University of Pavia, Via Forlanini 12, I-27100 Pavia, Italy (b) Department of Anatomy, Histology, and Legal Medicine, University of Florence, Viale Morgagni 85, I-50134 Florence, Italy (c) Department of Medicine and Public Health, University of Verona, Piazza L.A. Scuro 10, 37134 Verona, Italy Article History: Received 5 November 2008; Revised 28 March 2009; Accepted 1 April 2009

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2008.12.016 Byline: F. Musshoff (a), W. Rosendahl (b), B. Madea (a) Keywords: Nicotine; Hair analysis; Mummy; Gas chromatography-mass spectrometry; Cocaine; Drugs Abstract: In the literature, there are reports about the presence of cocaine, nicotine and cannabinoids in the hair of ancient mummies from South America or Egypt. Most of the results have been criticised because of the use of improper analytical techniques or contamination of the sample material. Recently an exhibition of 70 mummies from around the globe was organized at the Reiss-Engelhorn-Museen in Mannheim, Germany. It shows exhibits assembled by one of the worldfamous mummy projects, involving various specialists for anthropology, pathology, radiology, molecular biology and toxicology. Within this project, hair samples of eight pre-Columbian mummies were analyzed for drugs using modern routine gas chromatographic-mass spectrometric techniques. The tests revealed negative results, except for nicotine in the hair of three mummies. Nicotine was measured in concentrations of 57.5ng/mg in the hair of a woman, 14.1ng/mg in the hair of a child and 11.4ng/mg in the hair of a further female mummy, but all cases revealed negative results for cotinine. The target analysis was performed with limits of detection of 0.04ng/mg for nicotine and 0.033ng/mg for cotinine. The washing solutions yielded negative results for both analytes, nicotine as well as cotinine. In our opinion, even with respect to negative results in the washing solutions, the present results cannot definitely confirm an active consumption with body passage in the life time of the analyzed mummies: An external contamination cannot be excluded, e.g. by transfer from smoking visitors or employees during the early collection history of the objects in the 19th century, as well as in their respective lifetime. Author Affiliation: (a) Institute of Forensic Medicine, Stiftsplatz 12, 53111 Bonn, Germany (b) Reiss-Engelhorn-Museum, 68159 Mannheim, Germany Article History: Received 9 July 2008; Revised 13 December 2008; Accepted 18 December 2008

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2008.12.012 Byline: Sooyeun Lee, Yonghoon Park, Wonkyung Yang, Eunyoung Han, Sanggil Choe, Miae Lim, Heesun Chung Keywords: Measurement uncertainty; Hair analysis; Methamphetamine; Amphetamine; Method precision Abstract: The measurement uncertainties (MUs) were estimated for the determination of methamphetamine (MA) and its main metabolite, amphetamine (AP) at the low concentrations (around the cut-off value of MA) in human hair according to the recommendations of the EURACHEM/CITAC Guide and 'Guide to the expression of uncertainty in measurement (GUM)'. MA and AP were extracted by agitating hair with 1% HCl in methanol, followed by derivatization and quantification using GC-MS. The major components contributing to their uncertainties were the amount of MA or AP in the test sample, the weight of the test sample and the method precision, based on the equation to calculate the mesurand from intermediate values. Consequently, the concentrations of MA and AP in the hair sample with their expanded uncertainties were 0.66[+ or -]0.05 and 1.01[+ or -]0.06ng/mg, respectively, which were acceptable to support the successful application of the analytical method. The method precision and the weight of the hair sample gave the largest contribution to the overall combined uncertainties of MA and AP, for each. Author Affiliation: National Institute of Scientific Investigation, 331-1 Sinwol-7-dong, Yangcheon-gu, Seoul 158-707 Republic of Korea Article History: Received 27 January 2008; Revised 15 December 2008; Accepted 18 December 2008

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2008.11.002 Byline: Michelle R. Vaughn (a), Roland A.H. van Oorschot (b), Swati Baindur-Hudson (c) Keywords: Hair colour; Digital image analysis; Reflective spectrophotometry; Forensic science; Genetics; Macroscopic; Investigative tool Abstract: While reflective spectrophotometry is an established method for measuring macroscopic hair colour, it can be cumbersome to use on a large number of individuals and not all reflective spectrophotometry instruments are easily portable. This study investigates the use of digital photographs to measure hair colour and compares its use to reflective spectrophotometry. An understanding of the accuracy of colour determination by these methods is of relevance when undertaking specific investigations, such as those on the genetics of hair colour. Measurements of hair colour may also be of assistance in cases where a photograph is the only evidence of hair colour available (e.g. surveillance). Using the CIE L.sup.* a.sup.* b.sup.* colour space, the hair colour of 134 individuals of European ancestry was measured by both reflective spectrophotometry and by digital image analysis (in V++). A moderate correlation was found along all three colour axes, with Pearson correlation coefficients of 0.625, 0.593 and 0.513 for L.sup.* , a.sup.* and b.sup.* respectively (p-values=0.000), with means being significantly overestimated by digital image analysis for all three colour components (by an average of 33.42, 3.38 and 8.00 for L.sup.* , a.sup.* and b.sup.* respectively). When using digital image data to group individuals into clusters previously determined by reflective spectrophotometric analysis using a discriminant analysis, individuals were classified into the correct clusters 85.8% of the time when there were two clusters. The percentage of cases correctly classified decreases as the number of clusters increases. It is concluded that, although more convenient, hair colour measurement from digital images has limited use in situations requiring accurate and consistent measurements. Author Affiliation: (a) Victoria University, School of Molecular Sciences, Hoppers Lane, Melbourne, Werribee, VIC 3030, Australia (b) Victorian Police Forensic Services Department, Macleod, VIC, Australia (c) Victoria University, School of Biomedical and Health Sciences, Melbourne, VIC, Australia Article History: Received 16 May 2008; Revised 5 November 2008; Accepted 7 November 2008

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2008.10.015 Byline: Sooyeun Lee, Rosa Cordero, Sue Paterson Keywords: 6-Monoacethylmorphine; Morphine; Heroin; Hair analysis; Metabolite-to-parent drug ratio Abstract: There is limited published data to aid interpretation of analytical findings from hair analysis. The aim of the study was to collate 6-monoacetylmorphine (6-AM) and morphine concentrations in head and pubic hair from heroin users and to propose reference ranges and relate these to the amount of heroin used. The ratio of morphine-to-6-AM was also investigated. A total of 82 head hair samples divided into 173 segments of various lengths and 15 pubic hair samples were collected at postmortem from heroin users. The hair was analysed using a previously published method. A statistical evaluation demonstrated that in head hair, the lower, middle and upper concentration ranges of 6-AM were 0.1-0.9, 0.9-12.5 and 12.5-154.1ng/mg and those of morphine were 0.1-0.8, 0.8-6.0 and 6.0-36.3ng/mg. In pubic hair, the lower, middle and upper concentration ranges of 6-AM were 0.2-0.5, 0.5-2.3 and 2.3-18.2ng/mg and those of morphine were 0.2-0.4, 0.4-2.4 and 2.4-13.3ng/mg. The morphine-to-6-AM ratio showed a large variation. The ratio tended to decrease from proximal to distal segments. The statistical results suggest low, middle and high concentration ranges which we propose can be used for estimating the amount of heroin consumed into corresponding low or occasional, regular or habitual and heavy or excessive drug use. The ratio of morphine-to-6-AM showed great variation and did not support the proposal that a ratio less than 0.77 is needed to prove ingestion of heroin. Author Affiliation: Toxicology Unit, Imperial College London, St Dunstan's Road, London W6 8RP, United Kingdom Article History: Received 20 August 2008; Revised 20 October 2008; Accepted 22 October 2008

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2007.03.002 Byline: Robert A. Anderson (a), Marinah M. Ariffin (a), Peter A.G. Cormack (b), Eleanor I. Miller (a) Keywords: Benzodiazepines; Hair; SPE; MISPE; LC-MS-MS Abstract: This preliminary study compares the benzodiazepine results for 10 post-mortem scalp hair samples using a classical solid-phase extraction (SPE) and a molecularly imprinted solid-phase extraction (MISPE) system. The hair samples selected for testing were from drug-related deaths where a positive benzodiazepine blood result was obtained. Samples were decontaminated with 0.1% sodium dodecyl sulfate, distilled water and dichloromethane, incubated overnight in methanol/25% aqueous ammonium hydroxide (20:1), extracted by SPE or MISPE and subsequently analysed by liquid chromatography-tandem mass spectrometry (LC-MS-MS). Both extraction methods detected diazepam, nordiazepam, oxazepam, temazepam and nitrazepam in the samples. Diazepam was detected in a greater number of samples using MISPE due to both its lower limit of detection (LOD) and higher extraction recovery as a result of excellent molecular recognition of the template (diazepam) imparted by the imprinting process. The selective recognition of two diazepam analogues, nordiazepam and oxazepam, was demonstrated using MISPE since they were also detected in a greater number of samples. In contrast, another diazepam analogue, temazepam, was detected in a greater number of samples using SPE since the LOD using this extraction was lower than with MISPE. Nitrazepam was detected in one sample using both extraction methods. Overall the MISPE and SPE hair results were in good qualitative agreement. For the samples, where both extraction methods detected nordiazepam, temazepam and oxazepam, the concentrations were always higher for SPE. This is probably due to the MIP procedure producing extracts with fewer matrix interferences than the extracts produced using the classical SPE method. MISPE could be used as a complementary method to classical SPE for the analysis of benzodiazepine positive hair samples collected from chronic users. Author Affiliation: (a) Forensic Medicine and Science, University of Glasgow, Joseph Black Building, University Place, Glasgow G12 8QQ, Scotland (b) WestCHEM, Department of Pure and Applied Chemistry, University of Strathclyde, Thomas Graham Building, 295 Cathedral Street, Glasgow G1 1XL, Scotland Article History: Received 4 October 2006; Revised 7 March 2007; Accepted 7 March 2007

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2006.09.003 Byline: Gisela Skopp (a), Peter Strohbeck-Kuehner (a), Karl Mann (b), Derik Hermann (b) Keywords: Cannabinoids; Self-report; Hair analysis; Correlation Abstract: Hair analysis has shown great potential in the detection and control of drug use. Whether an assay is of quantitative value roughly corresponding to the amount of drug consumed, is still a matter of debate. The present investigation was aimed at a possible relationship between the cannabinoid concentration in hair and the cumulative dose in regular users of cannabis. Hair samples from the vertex region of the scalp were obtained from 12 male regular users of cannabis, and 10 male subjects with no experience of cannabis served as controls. None of the subjects had his hair permed, bleached or colored. Cannabis users provided information on drug use such as the current cannabis dose per day, the cumulative cannabis dose of the last 3 months, as well as the frequency of cannabis use during the last year. The concentration of delta-9-tetrahydrocannabinol (THC), cannabinol (CBN) and cannabidiol (CBD) in hair was determined using gas chromatography-mass spectrometry. Cannabinoids were present in any hair sample of cannabis users, but were not detectable in control specimens. An increase in the amount of cannabinoids in hair with increasing dose was evident. The concentration of major cannabinoids (sum of THC, CBD and CBN) was significantly correlated to either the reported cumulative cannabis dose during the last 3 months or to the cannabis use during the last 3 months estimated from the daily dose and the frequency per year (r =0.68 or 0.71, p =0.023 or 0.014). A significant relationship between THC and the amount of cannabis used could not be established. As a conclusion, the sum of major cannabinoids in hair of regular users may provide a better measure of drug use than THC. Author Affiliation: (a) Institute of Legal Medicine and Traffic Medicine, Ruprecht-Karls University, Voss-Str. 2, 69115 Heidelberg, Germany (b) Central Institute of Mental Health, Department for Addictive Behavior and Addiction Medicine, P.O. Box 122120, Mannheim 68072, Germany Article History: Received 11 April 2006; Revised 26 August 2006; Accepted 11 September 2006

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2006.05.042 Byline: S. Amory (a)(b), C. Keyser (a)(b), E. Crubezy (b), B. Ludes (a) Keywords: Ancient DNA; STR; Y chromosome; Hair shaft; Siberia Abstract: The aim of this study was to determine if ancient hair shafts could be suitable for nuclear DNA analysis and to develop an efficient and straightforward protocol for DNA extraction and STR typing of ancient specimens. The developed method was validated on modern and forensic samples and then successfully applied on ancient hairs collected from Siberian mummies dating from the 16th to the early 19th centuries. In parallel extractions including or excluding a washing step were performed at least two times for each sample in order to evaluate the influence on the quantity of nuclear DNA yielded and on the typing efficiency. Twelve ancient individuals were analyzed through our approach and full and reliable profiles were obtained for four of them. These profiles were validated by comparison with those obtained from bone and teeth DNA extracted from the same ancient specimens. The present study demonstrates that the washing step cannot be considered as deleterious for DNA retrieval since the same results were obtained by the two approaches. This finding challenges the hypothesis that recoverable nuclear DNA is only found on the outer surface of hair shafts and provides evidence that nuclear DNA can be successfully extracted from ancient hair shafts. The method described here constitutes a promising way for non-invasive investigations in ancient DNA analysis for precious or historical samples as well as forensic casework analyses. Author Affiliation: (a) Laboratory of Molecular Anthropology, Institute of Legal Medicine of Strasbourg, 11 rue Humann, 67085 Strasbourg Cedex, France (b) Laboratory of Anthropo-biology, UMR 8555, CNRS, 39 allees Jules Guesde, 31400 Toulouse, France Article History: Received 5 May 2006; Revised 22 May 2006; Accepted 24 May 2006

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2006.04.015 Byline: Detlef Thieme (a)(b), Hans Sachs (a)(b) Keywords: Liquid chromatography-tandem mass spectrometry; Drug monitoring; Hair analysis; Segmentation; Antipsychotic drugs; Clozapine Abstract: The long-term administration of clozapine could be verified by fine segmentation and analysis of single hairs of one person to examine the history of a multiple poisoning case. Segments of 1-2.5mm length were extracted by ultrasonification in 30[mu]l of the mobile phase (mixture of methanol+water, 50+50). By application of isocratic liquid chromatography and using narrow bore columns (Synergy Polar-RP, Phenomenex), an acceleration and miniaturization of the HPLC-MS-MS assay could be achieved. Total amounts of clozapine down to 30fg (on column) and its desmethyl metabolite could be analysed in multiple reaction monitoring mode. According to typical sample amounts of [approximately equal to]16[mu]g, relevant hair concentrations higher than 1pg/mg were detected. Significant and reproducible concentration profiles along the hair fibres revealed characteristic administration cycles. The administration time course -in particular the time of its termination -- could be verified with a precision of a few days. The accuracy and reproducibility of the concentration profile was proven based on multiple investigations of single hairs. An individual hair growth rate of 0.55mm/day was determined with a relative standard deviation of 8% by comparison of concentration profiles in hairs collected after a time span of 165 days. Author Affiliation: (a) Institute of Forensic Medicine, Frauenlobstr. 7a, 80337 Munich, Germany (b) Forensic Toxicological Centre, Bayerstr. 53, 80335 Munich, Germany Article History: Received 3 December 2005; Revised 17 April 2006; Accepted 21 April 2006

We describe a fast method for the identification of the origin of native and chemically processed feathers, down, and hair and for distinguishing closely related species using enzyme digestion and MALDI-TOF mass spectrometry. Additionally we present two methods for the quantification of different identified bird and mammalian samples, respectively, in binary species mixtures. Without any prior cleaning or isolation of single proteins, enzymatical digests of feathers, down, and hair are performed. Fragments generated are analyzed by MALDI-TOF mass spectrometry, and peak groups of different selectivity are established for every animal species. For the identification of individual animal species, only unique species-specific peaks are accepted while for species classification of tinted or bleached hair, the group of semispecific peaks (SEMPs) is used. Samples from native animal species show a higher coincidence of digest peak masses of SEMPs than far-related species, indicating a phylogenetical relationship of the investigated structure proteins. Quantification of mixed binary avian samples is performed by identification of preweighed single feathers and down, followed by calculation of the gravimetric mass fractions. The composition of mixed binary mammalian samples is calculated from correlation of the quotient of the relative peak intensities or areas of these species and the quotient of corresponding gravimetric mass fractions. High accuracy is achieved by both quantification methods. The fast methods are well suited for industrial quality control for example.

This article evaluates the potential impact that the use of hair analysis may have on the drug prevalence estimations derived from survey research that has relied upon urinalysis as an indicator of the accuracy of self-reported drug use. The paper reviews the history and nature of the DUF and ADAM programs, the relationship between self-report drug use and urinalysis results for arrestee populations, and the outcome of a pilot study employing hair assays in lieu of urinalysis. The author concludes that hair analysis may have a significant impact on the estimations of drug use for cocaine, heroin, and amphetamines and is less likely to have an effect on estimations of marijuana use. The author recommends consideration of periodic use of hair analysis within the ADAM system to more accurately and effectively monitor drug use.

A noninvasive 5-hour hair-root-analysis test has been developed for fragile X syndrome in males. The test depends on lack of the FMR1 (ital) gene product (FMRP) in cells. In female patients with full mutation, FMRP is expressed in some hair roots, less than 55%. There is no overlap with normal female controls.