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Analysis of pubic hair as an alternative specimen to scalp hair: A contamination issue

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Abstract

Pubic hair is often analyzed as an alternative to scalp hair to prove previous drug use. However, urine is a potential source of external contamination. In the present study, the concentrations of methamphetamine (MA) and amphetamine (AP) in both scalp and pubic hair from illegal MA users were compared. Furthermore, in order to investigate the external contamination of pubic hair by urine, MA and AP absorbed into pubic hair that had been contaminated with authentic urine from a MA user were measured using a previously validated method. The effect of shampoo-wash on the contaminated pubic hair was also examined. However, no correlation was found in the MA and AP concentrations between scalp and pubic hair from illegal MA users. As the number of contamination events by authentic urine increased, the concentrations of MA and AP in pubic hair increased. Both MA and AP were detected in the first methanol washes of the contaminated hair samples but were not detected in the second methanol washes. As the number of shampoo-washes of the contaminated pubic hair increased, the concentrations of MA and AP gradually decreased. Even though pubic hair can be used as an alternative to scalp hair to prove previous drug use, it should be avoided when estimating drug use history. It should be also noted that higher quantitative results in pubic hair do not necessarily represent heavier drug use.

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... The interpretation of an analytical hair result can be complicated by the presence of external drug contamination. This contamination can result from hair exposure to biological fluids such as sweat, blood or urine, indirect contamination after handling illicit drug powders or cannabis plant material, and exposure to crack cocaine (COC) or cannabis smoke [9][10][11][12][13][14]. Children living in a drug abusing environment are a prime example of the potential for the external contamination of hair [15]. ...
... The researchers found that the washes had no detectable level of drug, indicating that the shampoo had removed all of the surface drug contamination [10,55]. In another study, hair was briefly exposed to urine that was positive for amphetamines, resulting in hair concentrations above cut-off levels [11]. This hair was then decontaminated using a sequential wash with MeOH, water and MeOH. ...
... This hair was then decontaminated using a sequential wash with MeOH, water and MeOH. MA and amphetamine were only detected in the first MeOH wash, but were not detected in any of the washes when contamination was followed by treatment with shampoo [11]. Seemingly, washing hair with shampoo removes surface contamination available to solvents used in decontamination procedures [56]. ...
Article
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Hair is a mainstream specimen used in forensic toxicology to determine drug use and exposure. However, the interpretation of an analytical hair result can be complicated by the presence of external drug contamination. Decontamination procedures are included in hair analysis methods to remove external contamination, but the capacity of these washes to completely remove contamination for all drugs is controversial. It is evident that there is no consensus on the most effective decontamination procedure, nor can decontamination procedures consistently remove external drug contamination to less than reportable cut-offs for all analytes. ∆9-tetrahydrocannabinol deposited from cannabis smoke is mostly removed by organic solvents, whereas ionizable drugs are more effectively removed by an aqueous wash. Organizations such as the Society of Hair Testing recommend a hair decontamination procedure should include both an organic and aqueous washing step, which is in accordance with the reviewed literature. Studies involving a systematic evaluation of various solvents have shown that the most effective organic solvent was methanol and the most effective aqueous solvent contained sodium dodecyl sulfate detergent. If future systematic studies can demonstrate similar findings, a consensus on the most effective decontamination procedure for forensic hair analysis may be established.
... However, several studies involving both drugs and EtG determinations showed that different keratinic matrices may contain uneven concentrations of these analytes, since there are substantial inter-site differences in hair growth rate and growth cycle, and in the nature and activity of sweat and sebum glands [15]. Further variations could be ascribed to environmental exposure, washing habits and the use of special cosmetic products [16,17]. It was also shown that the contact with positive biological matrices, such as urine, can raise the EtG incorporation into hair samples by a factor of up to 400. ...
... For most social and heavy drinkers (cases 8,11,14,16,17,21,23,24,27,29,30 and 32), the measured EtG concentration in axillary hair was lower than in head hair and even below the LOD in five cases, while in a few other cases (9, 10 and 15), all relative to homeless people with poor hygiene, the opposite trend was observed. ...
Article
Ethyl glucuronide (EtG) is a direct metabolite of ethanol, frequently used as a biomarker of alcohol abuse. To this purpose, EtG is preferentially determined in hair samples, using a cut-off value of 30pg/mg to discriminate between social and heavy drinkers, as recently fixed by an international consensus conference. Although this cut-off value is assumed for head hair, alternative matrices, such as pubic, axillary and chest hair, are often analyzed when head hair is not available. Previous studies suggested that determination of EtG in various keratin matrices may lead to different results; growth cycle and rate, urine contamination, distribution of sebum glands and other environmental factors are likely to contribute to these differences. We analyzed more than 2700 samples (head, pubic, chest and axillary hair) to evaluate the inter- and intra-individual distribution of the EtG concentration in the different keratin matrices. The data were interpreted on a statistical basis, on the assumption that large population data-sets will level off the average alcohol consumption of each group. From both inter- and intra-individual distribution data, significant differences were observed in EtG concentrations recorded in head, axillary and pubic hair samples. It is concluded that pubic hair cannot be utilized alternatively to head hair to prove chronic alcohol abuse, nor is axillary hair, since positive and negative biases respectively affect these determinations. In contrast, for chest hair, EtG distributions similar to head hair were found, although the large discrepancy between the examined population dimensions presently prevents any definitive conclusion. Thus, chest hair represents a promising alternative to head hair for EtG determinations, deserving further investigation on samples collected from the same individuals, in order to establish a clear correlation between their respective EtG concentrations.
... The substantial matching of concentration ranges among the scalp, chest and pubic hair constitutes an important finding. Indeed, these matrices present specific characteristics and furthermore are subject to different hygienic treatments affecting the detectable levels of many substances [44,45]. First of all, the growth rate for body hair is lower than scalp hair (anagen and telogen phases are considered to be of equal length) which may yield to higher concentrations; moreover, pubic hair is exposed to urinary contamination. ...
... This phenomenon may entail an overestimation of substance's levels in this keratin matrix. Overall, urine presents higher concentrations and can release high amount of substance on the pubic hair surface [45]. For this reason, the preanalytical washing step plays a key role when this matrix is analysed and it should be deeply optimized during the method validation; an effective procedure should be able to remove the contaminant avoiding the extraction of the analytes from the sample. ...
Article
Endogenous nature of GHB represents a critical issue for forensic toxicologists, especially in alleged sexual assaults. Therefore, discrimination between physiologically and additional amounts from exogenous sources of such a substance must be effective and reliable in order to avoid severe misinterpretation. This study aimed to quantify the GHB baseline concentrations in chest and pubic hairs collected from 105 healthy volunteers, non-consumers of any drugs of abuse. The final scope was to investigate if these keratin matrices could represent valid alternative to scalp hair when not available. Moreover, we also evaluated the age and gender influences on the GHB baseline levels. 25 mg of hair were incubated overnight with NaOH at 56°C. After acidification with H2SO4, the solution was liquid-liquid extracted with ethyl acetate and a trimethylsilyl derivatization was then achieved. Analysis was performed in gas chromatography-mass spectrometry in single ion monitoring mode (m/z 233, 234, 147 for GHB; m/z 239, 240 and 147 for GHB-d6). The endogenous amount in “blank” hair was estimated by the standard addition method (0.301 for chest hair and 0.235 ng/mg for pubic hair). GHB concentration ranged from 0.205 to 1.511 ng/mg for chest hair and from 0.310 to 1.913 ng/mg for pubic hair. These values were consistent with previous studies on scalp hair and on pubic hair. Unfortunately, research on chest hair is not available in literature. T-Test and Linear Regression highlighted no statistically significant differences for the two matrices and for all age/gender sub-groups. However, further studies are required to estimate a reliable cut-off value for these keratin matrices. For the first time, we demonstrated the suitability of chest and pubic hair to detect endogenous levels of GHB.
... Another limitation of beard hair is that it is only available from men adult population. Different studies have demonstrated that chest, leg and arm hair are the best secondary hair matrix in toxicological investigations, as they would be less exposed to cosmetic treatments, environmental contamination or less influenced by contamination from other matrices (urine and sweat) [6][7][8][9][10][11][12]. Due to the high proportion of telogen hair in body hair, they are considered to be unsuitable to determine the time frame within the consumption was monitored. ...
... During the last years different studies have been published concerning the analysis and comparison of drug concentrations found in different body hair regions (pubic and beard hair mostly), where samples were usually from chronic drug users [7][8][9][10][11][12], and only a small fraction of these investigations were adequately controlled studies (unknown daily dose and duration of drug intake [17][18][19][20]. The aim of this study wasto determine and compare: (A) the time course of a basic drug, dihydrocodeine (pKa 9.3, logP 1.5), after a single dose in two subjects, man and woman, in different suitable and body hair areas: frequently shaved regions in western countries as beard and legs, (B) in unshaved hair from head hair, chest hair, leg hair, and/or arm hair, as these results would give an indication of the time course of dihydrocodeine in body hair, and if body hair would be an interesting tool, in particular to single dose cases. ...
... Another limitation of beard hair is that it is only available from men adult population. Different studies have demonstrated that chest, leg and arm hair are the best secondary hair matrix in toxicological investigations, as they would be less exposed to cosmetic treatments, environmental contamination or less influenced by contamination from other matrices (urine and sweat) [6][7][8][9][10][11][12]. Due to the high proportion of telogen hair in body hair, they are considered to be unsuitable to determine the time frame within the consumption was monitored. ...
... During the last years different studies have been published concerning the analysis and comparison of drug concentrations found in different body hair regions (pubic and beard hair mostly), where samples were usually from chronic drug users [7][8][9][10][11][12], and only a small fraction of these investigations were adequately controlled studies (unknown daily dose and duration of drug intake [17][18][19][20]. The aim of this study wasto determine and compare: (A) the time course of a basic drug, dihydrocodeine (pKa 9.3, logP 1.5), after a single dose in two subjects, man and woman, in different suitable and body hair areas: frequently shaved regions in western countries as beard and legs, (B) in unshaved hair from head hair, chest hair, leg hair, and/or arm hair, as these results would give an indication of the time course of dihydrocodeine in body hair, and if body hair would be an interesting tool, in particular to single dose cases. ...
Article
Background When head hair is not suitable or not available, body hair, such as leg or beard hair might be the most suitable sample for drug hair analysis. Drugs may be detected in beard, from the first day up to more than one week. Specific information about the time course detection of drugs, after a single intake, in body hair has not been yet well documented for the different body hair samples. Aim The aim of this study was to determine and compare the detection window of dihydrocodeine in frequently shaved legs and beard after a single intake. Unshaved hair from head hair, chest hair, leg hair, and/or arm hair were also analysed. Method Before a single intake of 12 mg dihydrocodeine by test subject 1 (female), the leg hair was shaved in the morning. The second test subject 2 (male) shaved his beard in the morning and 30 min later he had a dose of 10 mg of dihydrocodeine. The samples were washed with water and shampoo, dried and collected as follows: subject 1: every 3-days shaved leg hair (n = 10) and 1-month-later head hair (n = 1). Subject 2: daily shaved beard hair (n = 15), 2 months later head hair (n = 50, different regions), and every 20 days unshaved arm, leg and chest hair (from different regions) (n = 4/area). The samples were analysed for dihydrocodeine using a validated liquid chromatography tandem mass spectrometry method with a limit of quantification (LOQ) of 16 pg/mg for dihydrocodeine. About 20 mg of hair samples were weighted, washed with dichloromethane, centrifuged, dried, and pulverized in the same disposable tubes. Then the samples were incubated with methanol (under sonication at 45 °C) during 4 h. After centrifugation, the supernatant was evaporated and a cation exchange solid phase extraction followed by separation and quantification using ultra performance liquid chromatography tandem mass spectrometry (UHLC-MS/MS) was carried out. Chromatographic separation was achieved using a BEH phenyl column eluted with 0.1% formic acid: methanol (0.1% formic acid). The UPLC-MS/MS method was validated and used in routine for drug hair analysis for already several years [1]. Results and discussion In the present study leg hair was collected every 3 days, as an average of frequent shaved hair in western woman population. Leg hair was very limited and only one hair sample from both woman legs was available per analysis. Beard was collected daily and in a higher amount. Dihydrocodeine was detected in leg hair from the first sample (3 days after the intake). Maximum concentration at 68 pg/mg for the single intake was obtained after 15 days (± 2 days), decreasing later to the LOQ from the 21st day. Beard hair was already positive from the first day sample, and the maximum concentration was observed at 66 pg/mg, 6 days after the intake, decreasing later to the LOQ from day 10. This may be explained by the ‘Richards–Meharg body and head hair growth table, where it indicates that legs have only 20% of growing hairs in anagen phase, while beard goes from 50 to 70%. In other body hair samples, dihydrocodeine was negative or detected from 1 month after the intake. Conclusion Body hair presents different time course window detection due to the different growth rates. Frequently shaved leg and beard hair may be suitable samples for recent single dihydrocodeine dose detection from the first days up to 21 or 13 days after the intake, respectively, when an LOQ of 16 pg/mg is applied.
... Moreover, an important limitation associated with hair dipping in aqueous solutions is the possible irreversible penetration of the compounds within the hair shaft. [6] In the last decade, more 'realistic' artificial contamination procedures were also used: hair incubation with spiked blood to demonstrate postmortem external contamination, [24] hair briefly dipped in authentic urine from a methamphetamine user to investigate the contamination/decontamination of pubic hair, [25] hair coating with drugs followed by sweat conditioning using synthetic sweat to study drug contamination/decontamination in the analysis of human hair, [6] and subjects hair own contamination using their hands powdered with a drug mixture. [26] Although such procedures might be relevant to clinical or forensic contexts, they are not representative of hair surface contamination by organic pollutants present in air or dust; specific approaches have to be developed for the latter purpose. ...
Article
Although increasing interest is being observed in hair analysis for the biomonitoring of human exposure to pesticides, some limitations still have to be addressed for optimum use of this matrix in that specific context. One main possible issue concerns the need to differentiate chemicals biologically incorporated into hair from those externally deposited on hair surface from contaminated air or dust. The present study focuses on the development of a washing procedure for the decontamination of hair before analysis of pesticides from different chemical classes. For this purpose, three different procedures of artificial contamination (with silica, cellulose, and aqueous solution) were used to simulate pesticides deposition on hair surface. Several washing solvents (four organic: acetone, dichloromethane, methanol, acetonitrile; and four aqueous: water, phosphate buffer, shampoo, sodium dodecylsulfate) were evaluated for their capacity to remove artificially deposited pesticides from hair surface. The most effective washing solvents were sodium dodecylsulfate and methanol for aqueous and organic solvents, respectively. Moreover, after a first washing with sodium dodecylsulfate or methanol, the majority of externally deposited pesticides was removed and a steady-state was reached since significantly lower amounts were removed by additional second and third washings. Finally, the effectiveness of a decontamination procedure comprising washing with sodium dodecylsulfate and methanol was successively demonstrated. In parallel, it was determined that the final procedure did not affect the chemicals biologically incorporated, as hair strands naturally containing pesticides were used. Such a procedure appears to remove in one-shot the fraction of chemicals located on hair surface and does not require repeated washing steps. Copyright © 2014 John Wiley & Sons, Ltd.
... It should be noted that not all kind of body hair are comparably suitable. Axillary hair seems -possibly owing to the application of astringent deodorants -to be less suitable than other hair and pubic hair is usually not used for that purpose owing to contamination issues [34][35][36]. It has been shown that the analysis of beard hair is even suitable for a time-resolved detection after a single intake [37]. ...
Article
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The use of alternative matrices such as oral fluid and hair has increased in the past decades because of advances in analytical technology. However, there are still many issues that need to be resolved. Standardized protocols of sample pretreatment are needed to link the detected concentrations to final conclusions. The development of suitable proficiency testing schemes is required. Finally, interpretation issues such as link to effect, adulteration, detection markers and thresholds will hamper the vast use of these matrices. Today, several niche areas apply these matrices with success, such as drugs and driving for oral fluid and drug-facilitated crimes for hair. Once those issues are resolved, the number of applications will markedly grow in the future.
... Pubic hair represents a reliable and widely accepted alternative matrix, often employed to ascertain or exclude drug abuse. 14,23,24 This matrix could also be contaminated, such as by an addict partner or, in case of non-addict pushers, by ''doses'' often hidden inside underwear. ...
Article
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Even if pubic hair represents a reliable and widely accepted alternative hair matrix to identify drug abusers, it might produce false positive results due to external contamination. The aim of this study was to verify whether the external contamination of pubic hair with cocaine could influence the discrimination between active users and false positive subjects. The analysis was performed on in vivo and in vitro samples; the contamination was carried out by rubbing pubic hair with cocaine hydrochloride contaminated hands for three consecutive days. Five days after the beginning of the contamination, the pubic hair was collected and analysed at different times for two months. Data from our studies show that all in vivo samples yielded false positives; the in vitro samples were negative only for 10 days and then yielded false positives.
... The scatter of elements in pubic hair is lower than that in scalp hair, but their concentrations correlate strongly with the concentrations in the latter [2, 3]. The possibility for contaminating pubic hair with metals present in urine, indicated by some analyses of drug concentrations in pubic hair, does not apply herein since their concentrations in urine are considerably lower than in hair [4]. ...
Article
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Our paper contains analytical results of zinc and lead concentrations in the pubic hair of women living in areas with different contamination degrees. Women living in the area of Knurów, Silesia (a high degree of industrialization) and female residents of the area of Łańcut (Subcarpathian region), where the level of urban- ization is much smaller, constituted the respondent group. Overall, the hair of 159 women aged 22 up to 75 was tested. Washed hair was mineralized by a microwave method with HNO 3 . Metal concentrations were deter- mined using the AAS technique and SpectrAA 880 as well as Spectra 880Z spectrophotometers by the Varian Company. It has been found that the average lead concentration in the pubic hair of the women from the Subcarpathian region was significantly higher than the concentration of this element in the group of women from Silesia, which was probably related to the influence of low emissions in the area. Zinc concentrations in the pubic hair of the women in both treatment groups did not differ. The concentration of lead in the pubic hair of the women from the Subcarpathian region increased with age.
... This difference can be observed also for other drugs and it is mainly due to a lower growth rate of pubic hair and to the contamination of urine. [26] Therefore, it is important to keep these aspects in mind when estimating a substance use history: high concentrations in pubic hair are not necessarily caused by heavy intake. ...
Article
Propofol is a short-acting intravenous hypnotic-amnesiac agent used in the general anesthesia. In recent years propofol has gained importance for recreational use (being able to give euphoria, relaxation, sexual hallucinations and disinhibition) and for suicidal purposes. For these reasons, forensic toxicology interest in this substance has increased as well, taking into account the chance of its detection in hair. In this study, a new method in liquid chromatography-tandem mass spectrometry for hair detection of propofol was applied in two real cases for clinical and forensic purposes. This method consists in propofol derivatization with a diazonium salt from aniline and a liquid-liquid extraction with dichloromethane and ethyl acetate. Lower limit of quantification in negative ionization mode is 0.1 pg/mg. In Case 1, a segmental analysis was carried out on a sample collected from a female declaring self-administration. Propofol was found in the second and third segment at concentration of 0.21 and 0.11 ng/mg. In case 2, propofol was detected at 0.50 ng/mg in pubic hair from a male underwent a gastroscopy. In this study we validated and successfully applied a new versatile, sensitive and efficient method for hair detection of propofol even after a single administration.
... In addition, in the context of alleged repeated abuse, LSD hair concentrations seem disparate: the concentrations observed by Nakahara Y. et al. in head hair of 2 LSD users (12,8, and 17 pg/mg in case #1, #2 and #3, respectively) [41] are drastically higher than those observed in other head hair samples of regular users (1 and 1.27 pg/mg in case #4 and #7, respectively) [32,42]. It is noteworthy that pubic hair is likely to be, in one hand, contaminated by urine or sweat, depending on individual hygiene habits, and, in another hand, less exposed than head hair to other external environments such as light, weather or cosmetic treatments [50]. As a consequence, pubic hair drug concentrations are generally higher than those in head hair due to a major sweat and/or urine "contamination", as well as a better stability in this matrix due to lower elimination or degradation. ...
Article
Lysergic acid diethylamide (LSD) is a powerful hallucinogen, active at very low dosages, with, as a direct consequence, potential difficulties to be detected and quantified in a clinical or forensic context, in body fluids and even more in hair. The aim of this work is to review literature data related to hair analysis of LSD with a particular focus on the main issues encountered in LSD detection in hair. Results of LSD investigation in hair remain difficult to interpret regarding the very sparse data available on LSD concentrations in hair (n=10). The possibility of pubic hair contamination by urine, as well as the lack of data about LSD incorporation and stability in pubic and head hair, further challenges the interpretation of negative or positive results. The absence of LSD in head hair should be carefully considered, as it does not formally exclude LSD consumption. In all cases of positive results, the interpretation of LSD concentrations in hair remains uncertain and it seems utopian to distinguish repeated intake from single exposure using LSD hair concentration values. Furthermore, a positive result in pubic hair cannot be used to formally prove repeated use of LSD, even in the case of a documented recent use of LSD.
... A scalp hair matrix can be used to derive a chronological history of drug use, with an extended detection window of approximately 1 month per half-inch of hair, while pubic hair grows more slowly [16,17]. On the other hand, studies suggest that pubic hair can offer an alternative way of proving previous drug use, but it should be avoided when estimating drug use history, and higher quantitative results in pubic hair do not represent heavier drug use [18]. Furthermore, pubic hair does not grow continually like scalp hair; it has a longer resting phase and a different anagen to telogen ratio [19]. ...
Article
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This study aimed to investigate the detection of morphine in fingernails from forensic autopsies using immunohistochemistry (IHC), with confirmation by ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-HRMS). A primary antibody specific to morphine and a secondary antibody conjugated to horseradish peroxidase (HRP) was used. IHC on specimens of Subjects A and B (both drug addicts) resulted in the detection of morphine on a cell layer of the nail plate matrix. UHPLC-HRMS and GC-MS analysis showed that Subject A had a morphine concentration of 0.35 ng/mg in the fingernail and 472 ng/mL in the blood, while Subject B reached 1.23 ng/mg in the fingernail and 360 ng/ml in the blood. Most of those matrices were positive for codeine, methadone, EDDP, and 6-MAM. The use of IHC in Subject C (a former addict) showed no positivity for morphine in the fingernail, while the UHPLC-HRMS analysis confirmed its absence in the fingernail and blood. Additionally, an analysis of the scalp or pubic hair of the subjects was carried out using UHPLC-HRMS. The results suggest that IHC can be used to establish the site of accumulation of morphine in the nail matrix; for postmortem diagnosis; and that basic substances can be detected by UHPLC-HRMS. There are no previous studies on the use of IHC as a technique for forensic purposes in unconventional matrices, such as nails.
... Scientific RepoRtS | (2018) 8:11420 | DOI: 10.1038/s41598-018-29772-1 long detection window (months to years), for toxicological analysis 14 . Interestingly, a recent report suggests that scalp hair follicles can be used to find genes associated with brain diseases because the brain is an ectodermal tissue and shares the developmental origins with scalp hair follicle 15 . ...
Article
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Methamphetamine (MA) is a highly addictive psychostimulant that disturbs the central nervous system; therefore, diagnosis of MA addiction is important in clinical and forensic toxicology. In this study, a MA self-administration rat model was used to illustrate the gene expression profiling of the rewarding effect caused by MA. RNA-sequencing was performed to examine changes in gene expression in rat whisker follicles collected before self-administration, after MA self-administration, and after withdrawal sessions. We identified six distinct groups of genes, with statistically significant expression patterns. By constructing the functional association network of these genes and performing the subsequent topological analysis, we identified 43 genes, which have the potential to regulate MA reward and addiction. The gene pathways were then analysed using the Reactome and Knowledgebase for Addiction-Related Gene database, and it was found that genes and pathways associated with Alzheimer's disease and the heparan sulfate biosynthesis were enriched in MA self-administration rats. The findings suggest that changes of the genes identified in rat whisker follicles may be useful indicators of the rewarding effect of MA. Further studies are needed to provide a comprehensive understanding of MA addiction.
... Sweat or sebum secretion, as a mechanism of drug incorporation, increases in axillary or pubic hair, compared with scalp hair. Moreover, there is a higher possibility of contamination depending on individual hygiene habits and lower elimination due to exposure to other external environmental conditions such as light, weather or cosmetic treatments in axillary or pubic hair [40]. ...
Article
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Metabolomics is a powerful tool used to understand comprehensive changes in the metabolic response and to study the phenotype of an organism by instrumental analysis. It most commonly involves mass spectrometry followed by data mining and metabolite assignment. For the last few decades, hair has been used as a valuable analytical sample to investigate retrospective xenobiotic exposure as it provides a wider window of detection than other biological samples such as saliva, plasma, and urine. Hair contains functional metabolomes such as amino acids and lipids. Moreover, segmental analysis of hair based on its growth rate can provide information on metabolic changes over time. Therefore, it has great potential as a metabolomics sample to monitor chronic diseases, including drug addiction or abnormal conditions. In the current review, the latest applications of hair metabolomics in animal studies and clinical settings are highlighted. For this purpose, we review and discuss the characteristics of hair as a metabolomics sample, the analytical techniques employed in hair metabolomics and the consequence of hair metabolome alterations in recent studies. Through this, the value of hair as an alternative biological sample in metabolomics is highlighted.
Article
Increasing numbers of drug-facilitated sexual assaults (DFSAs) have been reported recently and have become a social concern. In DFSAs, biological specimens (urine, blood, and hair) are analyzed to prove the victim's drug exposure, which can lead to proof of the sexual crime. Hair is the only specimen that can provide firm evidence of drug ingestion in cases of long delays (more than a week) in reporting the crime. Furthermore, detailed sectional hair analysis of a single hair strand enables to estimate the victim's drug-use history (date and amount of intake). Several recent studies have demonstrated high-sensitivity methods using mass spectrometry to detect sub-pg/mg concentrations of hypnotics in hair, and additionally illustrated the incorporation pathways of drugs and detailed distribution patterns in the hair after intake. Based on these findings, hair testing for hypnotics has been put to practical applications since 2016. In this paper, we review the practical concepts and usefulness of hair testing for hypnotics while introducing the current situation of DFSAs.
Research
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This represents one of several sections of "A Bibliography Related to Crime Scene Interpretation with Emphases in Geotaphonomic and Forensic Archaeological Field Techniques, Nineteenth Edition" (The complete bibliography is also included at ResearchGate.net.). This is the most recent edition of a bibliography containing resources for multiple areas of crime scene, and particularly outdoor crime scene, investigations. It replaces the prior edition and contains approximately 10,000 additional citations. As an ongoing project, additional references, as encountered, will be added to future editions. References in this section, like fingerprints, ballistics, and blood evidence, are most often thought of in traditional criminalistics or police sciences. Some of the references below date to the early days of forensics and criminalistics. This section is included because it represents one of the six areas comprising Geotaphonomy: Stratification, Tool Marks, Bioturbation, Sedimentation, Compression/Depression (foot and shoe prints), and Internal Compaction. In addition to the recovery and interpretation of impression evidence, other trace evidence which might be found on surface and subsurface death scenes are cited. Entomological and botanical evidence also constitute trace items left or taken from crime scenes. Because of the number of works referencing those topics, they are addressed in respective sections in this bibliography. Some citations in this section could also be cross-referenced with those in Taphonomy given their discussion of the detioration, or decomposition of hairs and fibers among victims' remains or contamination of impression evidence through prolonged exposure at crime scenes. When this compiler first became interested in forensic science, one of the first references he read, and which most influenced subsequent processing of scenes and research, was Crime Investigation by Paul Kirk (1974). In that paper the words of Kirk serve as the foundation for collecting trace evidence at any scene regardless its age or condition: "Wherever he steps, whatever he touches, whatever he leaves, even unconsciously, will serve as silent witness against him. Not only his fingerprints or his foot prints, but his hair, the fibers from his clothing, the glass he breaks, the tool mark he leaves, the paint he scratches, the blood or semen he deposits or collects. All of these and more bear mute witness against him. This evidence does not forget. It is not confused by the excitement of the moment. It is not absent because human witnesses are. It is factual evidence. Pysical evidence cannot be wrong. It cannot perure itself. It cannot be wholly absent. Only its interpretation can err. Only human failure to fint it, study and understand it, can diminish its value."
Article
Recently, confusion has arisen on the interpretation of the results of toxicological analyses of illicit drugs in human hair. The reason for this is that external contamination of hair with cocaine may lead to absorption into hair to a higher extent than had previously been expected. The extent of absorption seems to be largely dependent on hair porosity. Therefore, adequate washing of the hair sample is needed before the chemical analysis. Scientists, however, do not agree on the best washing procedure to be used. It is clear, however, that decontamination of hair with an organic solvent alone is insufficient and that additional aqueous washings should be included to allow swelling of the hair and removal of contaminating drugs that have entered the hair.
Article
The effectiveness of decontamination procedures used for the removal of external drug contamination in forensic hair analysis is an ongoing debate. This investigation evaluated wash methods complying with Society of Hair Testing (SoHT) guidelines and their capacity to remove cocaine (COC) and methamphetamine (MA) from artificially contaminated hair. The most effective decontamination method was determined using a systematic approach, involving: (1) an initial washing solvent screen; (2) optimisation of wash duration; (3) comparison of sequential wash methods; and (4) reanalysis of clinical hair samples. For analysis, hair was subjected to micropulverised methanolic extraction prior to quantitation by UHPLC‐MSMS. Methanol (MeOH) and 0.1M phosphate buffer (pH 6) were the most effective organic and aqueous solvents, respectively, removing 28‐38% of COC and 16‐31% of MA. Wash durations longer than 30‐60 min did not remove additional amounts, and a more efficient sequential wash method was subsequently developed. Despite this, the interpretation of reportable results relative to the SoHT cut‐off levels was unchanged for most clinical hair samples reanalysed after washing by agitation for 30 min with MeOH. These findings highlight the inability of decontamination solvents to completely remove external COC and MA contamination from hair, including wash methods adhering to SoHT guidelines.
Article
In forensic toxicology drugs of abuse are routinely analyzed due to its legal statues in medical or legal investigation, such as death cases, poisoning cases and drug misuse, determining potential exposure to controlled substances. Despite the wide-spread use of blood and urine as matrices for forensic toxicological analysis, the use of hair samples has grown as the limita-tions associated with this matrix are overcome and new areas of application emerge. Hair provides several advantages over urine and blood, such as the easiness of sample collection and the ability to provide a retrospective analysis of drug con-sumption. Drug analysis in hair is following a path where less amount of sample has been needed to detect drugs of abuse. Improvement in sample preparation methods and lowering sensitivity in instrumental analysis is providing forensic scientist with the tools to use drug analysis in hair as a routine approach in forensic laboratory. Actually, beside the chronological exposition to a drug, hair may provide information about the day a chemical was intake. This should become part of the standard information sent by forensic scientists to criminal investigators and courts. The aim of this review is to summon the extraction procedures used to perform forensic toxicological analysis in hair as well as the major advances in this field once sample preparation step is critical and takes up most of the total analysis time.
Article
In this study, the relationships between the concentrations of R/S-methamphetamine (R/S-MA) and its metabolite R/S-amphetamine (R/S-AP), the AP/MA ratio in hair samples, and MA dependence were investigated by performing segmental hair analysis in MA users. Authentic hair samples collected from ten chronic MA abusers were cut into 1-cm sections (total of 120 segments). The concentrations of MA and AM enantiomers were quantitatively measured by the LC-MS/MS method. The S-MA concentrations ranged from 1.17 to 256.41 ng/mg and the S-AP concentrations ranged from 0.11 to 23.31 ng/mg in the 120 segments. S-MA and S-AP were the most common analytes identified in hair; no R-MA or R-AP was found. The S-AP/S-MA ratios ranged from 0.03 to 0.32, indicating that the subjects primarily consumed S-MA rather than R-MA or AP. The S-AP/S-MA ratios in the long hair of all chronic MA abusers showed some variation, but there was an overall trend of gradual increase from the distal to the proximal end. This trend was independent of the drug concentrations. Therefore, we could conclud that the AP/MA ratios increased with the duration of MA abuse, and a higher AP/MA ratio suggested high MA dependence There was no chiral conversion of MA or AP in the hair matrix. The segmental hair analysis showed that all subjects continuously used S-MA, and some users showed an increase in drug dose or frequency of use.
Article
An important goal of forensic and clinical toxicology is to identify biological markers of ethanol consumption that allow an objective diagnosis of chronic alcohol misuse. Blood and head hair samples were collected from 175 subjects—objectively classified as non-drinkers (N = 65), social drinkers (N = 51) and active heavy drinkers (N = 59)—and analyzed to determine eight traditional indirect biomarkers of ethanol consumption [aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyltransferase (γ-GT), alkaline phosphatase (ALP), mean corpuscular volume (MCV), carbohydrate-deficient transferrin (CDT), and cholesterol and triglycerides in blood] and one direct biomarker [ethyl glucuronide (EtG) in head hair]. The experimental values obtained from these determinations were submitted to statistical evaluations. In particular, Kruskal–Wallis, Mann–Whitney and ROC curve analyses, together with principal component analysis (PCA), allowed the diagnostic performances of the various biomarkers to be evaluated and compared consistently. From these evaluations, it was possible to deduce that EtG measured in head hair is the only biomarker that can conclusively discriminate active heavy drinkers from social and non-drinkers, using a cut-off value of 30 pg/mg. In contrast, a few indirect biomarkers such as ALP, cholesterol, and triglycerides showed extremely low diagnostic abilities and may convey misleading information. AST and ALT proved to be highly correlated and exhibited quite low sensitivity and specificity. Consequently, either of these parameters can be discarded without compromising the classification efficiency. Among the indirect biomarkers, γ-GT provided the highest diagnostic accuracy, while CDT and MCV yielded high specificity but low sensitivity. It was therefore concluded that EtG in head hair is the only biomarker capable of supporting a confirmatory diagnosis of chronic alcohol abuse in both forensic and clinical practice, while it was found that γ-GT, CDT, MCV, and AST—whether used alone or in combination—do not allow the conclusive classification of subjects according to ethanol consumption. However, a diagnostic strategy combining these four parameters could be formulated in order to create a multivariate model capable of screening suspected active heavy drinkers. Figure PCA of 175 patients clinically classified as non-drinkers (green dots, N = 65), social drinkers (yellow dots, N = 51) and active heavy drinkers (red dots, N = 59). Score and loading (six parameters) bi-plot of PC1 and PC2.
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Hair differs from other materials used for toxicological analysis because of its unique ability to serve as a long-term storage of foreign substances with respect to the temporal appearance in blood. Over the last 20 years, hair testing has gained increasing attention and recognition for the retrospective investigation of chronic drug abuse as well as intentional or unintentional poisoning. In this paper, we review the physiological basics of hair growth, mechanisms of substance incorporation, analytical methods, result interpretation and practical applications of hair analysis for drugs and other organic substances. Improved chromatographic-mass spectrometric techniques with increased selectivity and sensitivity and new methods of sample preparation have improved detection limits from the ng/mg range to below pg/mg. These technical advances have substantially enhanced the ability to detect numerous drugs and other poisons in hair. For example, it was possible to detect previous administration of a single very low dose in drug-facilitated crimes. In addition to its potential application in large scale workplace drug testing and driving ability examination, hair analysis is also used for detection of gestational drug exposure, cases of criminal liability of drug addicts, diagnosis of chronic intoxication and in postmortem toxicology. Hair has only limited relevance in therapy compliance control. Fatty acid ethyl esters and ethyl glucuronide in hair have proven to be suitable markers for alcohol abuse. Hair analysis for drugs is, however, not a simple routine procedure and needs substantial guidelines throughout the testing process, i.e., from sample collection to results interpretation.
Article
In post-mortem work, blood is a potential source of external contamination of hair. The present study was carried out to investigate the amount of drug absorbed into hair which has been contaminated with blood containing either cocaine or BE. Solutions were prepared containing 0.05, 0.1, 0.2, 0.5 and 3.0 microg/mL of either cocaine or BE in human blood. Samples of approximately 3.2g of drug-free hair were contaminated by soaking in the blood solutions for 5 min. They were then removed and left at room temperature. Approximately 0.5 g of hair was collected from each of the blood soaked hair samples at 6h, 1, 2, 4 and 7 days after contamination. As each hair sample was collected it was shampoo-washed to prevent further drug absorption. Hair samples were analysed in triplicate using a fully validated method described previously. EME and cocaethylene were also measured in order to find out if cocaine or BE was breaking down to these compounds. Both cocaine and BE were absorbed into hair in significant concentrations when the concentration in the blood was 0.5 microg/mL or greater; cocaine was more readily absorbed than BE. Cocaine broke down to EME (<LOQ) at 0.5 microg/mL and to EME (>LOQ) and BE (<LOQ) at 3.0 microg/mL. When the blood concentration of cocaine was 0.5 microg/mL or less, there was no evidence of it breaking down to form BE. From the samples soaked in blood containing BE, there was no evidence of the BE breaking down. The absorption of drug into hair did not increase as the contamination period increased from 6h to 7 days.
Article
To validate information on cannabis use, we investigated human hair and pubic hair for cannabinoids (THC and THC-COOH) by gas chromatography/mass spectrometry. Samples (100 mg approximately) were decontaminated with methylene chloride, then pulverized and dissolved in 1 ml 1 N NaOH for 10 min at 95 degrees C in the presence of 200 ng of deuterated standards. After cooling, samples were extracted by n-hexane/ethyl acetate after acidification with acetic acid. After derivatization of the dry extract by PFPA/PFP-OH, the drugs were separated on a 30-m capillary column and detected using selected-ion monitoring (m/z 377 and 459 for THC and THC-COOH, respectively). Forty-three hair samples were obtained from fatal heroin overdose cases. Among them, 35% tested positive for cannabinoids. Hair concentrations ranged from 0.26 to 2.17 ng/mg (mean, 0.74 ng/mg) and 0.07 to 0.33 ng/mg (mean, 0.16 ng/mg) of THC and THC-COOH, respectively. As is generally the case for other drugs detected in hair, metabolite concentration was always lower when compared to the parent drug concentration. In public hair, THC concentrations ranged from 0.34 to 3.91 ng/mg (mean, 1.35 ng/mg) and THC-COOH concentrations from 0.07 to 0.83 ng/mg (mean, 0.28 ng/mg). In most cases, the highest cannabinoid concentration was found in pubic hair, suggesting that this sample may be the more suitable for cannabis testing.
Article
The concentrations of morphine and codeine were investigated in hair from the head, axillary and pubic regions obtained from 20 fatal heroin cases. Hair preparation involves decontamination procedure in dichloromethane at 37 degrees C for 15 min, solubilization in sodium hydroxide at 100 degrees C for 5 min, neutralization with hydrochloric acid and centrifugation. After extraction in chloroform/isopropanol/n-heptane (50:17:33; v/v) at pH 9.2, drugs were derivatized with BSTFA + 1% TMCS and separated on a 12-m BP-5 capillary column. Quantification was done by GC/MS using selected ion monitoring. The highest morphine concentrations were found in public hair (0.80-41.34 ng/mg), followed by hair of the head (0.62-27.10 ng/mg), and axillary hair (0.40-24.20 ng/mg). Codeine was also detected in all samples, and the codeine/morphine ratios ranged from 0.54 to 0.273. The differences observed in drug concentration in the three kinds of hair are discussed in the light of the existing literature.
Article
This study investigates the distribution of some drugs in hair samples taken from different parts of the body (head, pubis and axillae). Samples drawn from 15 subjects who died from drug overdose were analysed. The concentrations of the drugs detected in the biological fluids did not appear to be correlated with those present in hair. The highest drug levels were observed in pubic hair. The concentration differences observed in the various types of hair can hypothetically be ascribed to a likely incorporation of the drugs from the outside.
Article
Laboratory examination of hair from drug users has been employed for the last 5 years in Crete, in addition to the psychiatric or other forensic clinical examinations necessary for the confirmation of a person's use of drugs. The present study reports results of total and sectional head, axillary and pubic hair analysis of imprisoned abusers under interrogation and awaiting presentation at the Crime Court. Morphine levels in total head hair samples 1, 2 and 4 months after preliminary imprisonment ranged from 1.2 to 38.2, 0.5 to 22.5 and 0.1 to 4.9 ng/mg of hair respectively, while the maximum morphine levels amongst all studied groups of those arrested were found in the sections of axillary and pubic hair. Notably high morphine levels in head, axillary and pubic hair sections (13.7, 8.4 and 18.1) ng/mg respectively) were measured, even in abusers who had been imprisoned between 2 and 4 months. Hair tests may considerably assist to evaluate the systematic present and past abuse of heroin and other drugs. Consequently, it may be used as valuable expertise evidence during questioning and in court.
Article
This study was designed to compare the qualitative results and concentrations of methamphetamine (MA) and its metabolite amphetamine (AP) in head hair and hair collected from different parts of the body (axillae and pubis). Hair from subjects (N = 14) suspected MA users was simultaneously collected. Hair preparation involved washing step, fine cutting, overnight extraction, derivatization by the trifluoroacetic anhydride, and gas chromatography/mass spectrometry (GC/MS) using selective ion monitoring. In this study, we found a good correlation of the qualitative results for MA between head hair and hair on other parts of the body, but there were some differences in concentrations of MA and AP. Namely, the concentrations of MA and AP were higher in axillary and pubic hair than in head hair.
Article
In the present study, we developed a reference material (RM) using authentic hair samples for the determination of methamphetamine (MA) and its main metabolite, amphetamine (AP) in human hair. MA abusers' hair samples were collected, homogenized and finally bottled. The concentration of each bottle was determined using two extraction methods, agitation with 1% HCl in methanol at 38 degrees C and ultrasonication with methanol/5M HCl (20:1), followed by gas chromatography/mass spectrometry (GC-MS) after derivatization with trifluoroacetic anhydride (TFAA). Both analytical procedures were fully validated and their extraction efficiency was compared. The homogeneity of analytes was evaluated and their property values were determined with their uncertainties. The two methods were acceptable to analyze MA and AP in human hair through the validation and comparative studies using spiked and authentic hair samples as well as NIST SRM 2379 certified reference material. Satisfying homogeneity was reached for MA and AP in the prepared RM. Finally, a human hair RM containing MA and AP is prepared at the level of 7.64+/-1.24 and 0.54+/-0.07 ng/mg, respectively. This material can be useful in forensic laboratories for internal quality control and external quality assurance.