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As cocaine COC is not only incorporated into hair via blood following ingestion but also by external contamination, hair samples are commonly tested for COC metabolites to prove ingestion. Analytical results of hair samples were interpreted using an established decision model for the differentiation between actual use and external contamination using metabolic ratios metabolite to COC. They were further examined concerning background of request, hair color, body site of sample collection, sex, and metabolic ratios of the main metabolites \[benzoylecgonine BE , norcocaine NC , and cocaethylene CE \]. Analytical results of hair samples Metabolic ratios of minor OH-COC metabolites were significantly higher in hair samples interpreted as originating from use than in samples interpreted as caused by contamination. However, this was not the case for OH-BE metabolic ratios. The study demonstrates that OH-COC metabolites including thresholds for their metabolic ratios must be used for a reliable interpretation of positive COC results in hair samples. Hair analysis provides the opportunity to monitor drug use retrospectively over several months \[ 1 \]. Individuals may claim or deny drug use. Therefore, forensic toxicologists are confronted with decision-making between actual use of drug or potential external contamination of hair. This particularly accounts for cocaine COC , which is often used via snorting. Moreover, COC is the most frequently used stimulant in Europe with an increasing trend in offenses for use, possession, and trafficking \[ 3 , 4 \]. However, hair testing guidelines also recommend the inclusion of the following COC metabolites to identify drug use: benzoylecgonine BE , norcocaine NC , cocaethylene CE , and ecgonine methyl ester \[ 2 \]. BE and NC are also degradation products during the preparation and purification of COC \[ 13 \] and also possibly storage of the drug. As a consequence, guidelines for hair testing have been adapted. On the one hand, this most probably arises from COC itself, which contains these by-products. On the other hand, Scholz et al. An initial approach for distinction of drug use, as opposed to the drug being present in the hair from contamination, was the introduction of cut-off values; however, later with growing experience, it was concluded that contamination can occur at any COC level. Some laboratories use extensive wash protocols to help identify contamination \[ 6 , 9 \]. In fact, it was shown that COC contamination of hair could be localized in the superficial compartments of hair, and application of multistage wash protocols resulted in a reproducible removal of the main contamination \[ 16 \]. Another approach for differentiation between contamination and incorporation from the circulatory system is the application of threshold ratios for metabolite to COC concentrations \[ 6 , 8 , 9 , 12 \]. While in the first report on OH-COC metabolites in the hair samples, the threshold concentrations for them have been proposed \[ 6 \]; later studies focused on metabolic ratios. Hill et al. Scholz et al. This decision model was established and statistically validated by investigating two sample cohorts: in vivo externally contaminated hair with COC vs hair from self-reported COC users who were submitted to detailed anamnesis, including personal interviews on psychotropic drug consumption. Higher metabolic ratios arise from higher metabolite incorporation following COC use. If COC by-products were present, their concentration ratios were determined. Furthermore, the decision model was applied to the interpretation of a large cohort of COC-positive hair samples. Additionally, we have investigated whether OH-COC metabolites are produced in vitro during the applied hair sample preparation. Details of the chemicals and reagents used for the analysis of the hair samples can be obtained from the paper by Scholz et al. A total of powdered COC samples, seized between and by the police in the Canton of Zurich, Switzerland, were obtained. They varied from Between 1. Samples were diluted and analyzed in summer The metabolic ratios were calculated as the quotient of metabolite and COC concentration. Postmortem hair samples were not included due to possible contamination by blood. In the majority of scalp hair samples, the proximal 5-cm segment was analyzed unless the hair was shorter. Body hair was analyzed in total. All hair samples were stored under dry conditions, at room temperature, and in the dark according to the SoHT guidelines \[ 18 \]. In brief, the hair was decontaminated successively with water, acetone, and hexane. After each extraction step, the samples were centrifuged and the extraction solvent was removed for evaporation. The concentration ratios were evaluated by box-and-whisker plots to determine their concentration quartiles. Statistical evaluation of more than two parameters was performed using the Kruskal—Wallis test as the data did not show normal distribution. Consequently, if this metabolic ratio alone is considered for distinction of COC use vs contamination in hair testing, there may be a risk of falsely interpreting a COC-positive analytical result in a sample as being indicative of COC use. Our results are in line with those by Hoelzle et al. Therefore, the results emphasize that the sole positivity of a hair sample for COC in addition to one of these metabolites or one metabolic ratio is not sufficient to confirm COC use. To date, OH-BE has not been detected in seized samples. The COC concentrations in the routine hair samples ranged from i. All samples were positive for BE and NC. The CE prevalence is in line with the study by Musshoff et al. The detection frequencies of OH-COC in the present evaluation are slightly higher compared to those in the study by Musshoff et al. This is again probably a matter of method sensitivity. All hair testing results were evaluated according to the decision model by Scholz et al. The evaluation resulted in The percentages of head and body hair in the two groups were Compared to the overall percentage of head to body hair samples of The present cohort comprised two samples 0. The OH-COC metabolite concentrations and their metabolic ratios in our study are within the range of those reported by Hill et al. Compared to our study, the authors found a slightly lower percentage of samples failing to match their criterion for use with Similarly, Franz et al. Likewise, Scholz et al. Overall, the variability among metabolic ratios in our study is high. It has to be noted that most probably every hair sample from an actual COC user is additionally contaminated due to handling powder and snorting being the most common administration route in Switzerland. Our hypothesis was to observe lower metabolic ratios in forensic cases because these cases mostly derive from drug traffic offenses in which high contamination of hair by COC is likely to occur due to portioning of large quantities of drug powder. As stated, The individuals on an abstinence program may still be in a COC-affine environment leading to potential contamination of hair. Schaffer et al. Overall, our data show that different metabolic COC ratios can be expected depending on the case circumstances. Hair color is defined by the melanin content. Therefore, the incorporation of p - and m -OH-COC seems to increase more with increasing melanin content of the hair compared to COC, probably due to their higher basicity. This finding is consistent with the general assumption that basic compounds can be better incorporated into dark hair \[ 1 \]. It is therefore possible that OH-BE cannot be effectively incorporated into darker hair, which contains more melanin compared to lighter hair. Contrary to study results obtained by Hill et al. We do not have an explanation for these different observations. A significant decreasing trend in all metabolic ratios was observed with decreasing content of CE and NC. The authors detected an increasing trend in all three isomer concentrations with increasing CE concentrations, irrespective of the COC concentrations. Our results corroborate these findings. On the other hand, our study reveals that the levels of COC hydroxy metabolites increase with increasing concentrations of both CE and NC. Therefore, these observations could be explained by the assumption that the higher the COC proportion incorporated into the hair from ingestion, the more metabolite is formed and incorporated. Our aim was to investigate a potential influence of sex on incorporation rates of COC and its metabolites only following use. It can be assumed that leg hair may be less exposed to COC in the environment compared to other body sites, leading to tentatively higher metabolic ratios. Otherwise, the results indicate that hair from all the investigated body sites is equally useful for hair analysis. Leg hair may be preferably collected from individuals in a COC-affiliated environment. Interestingly, Hill et al. Furthermore, their data did not reveal significant differences among different body sites concerning concentrations and percentages, which is mainly in accordance with our data \[ 9 \]. Evidently, the size of the cohorts was different. To our knowledge, this comparison is not yet documented in the literature. This solution was used to produce artificial COC-positive hair samples by soaking. They were significantly lower in the not decontaminated samples with ratios of 0. Nevertheless, all ratios were below the decision threshold of 0. In contrast, p - and m -OH-BE were not detected in any hair extract. However, all ratios were still far below the decision threshold for COC use. Similarly, Scholz et al. During soaking of hair, substances are only absorbed to the outer layer and not incorporated into the inner hair compartments. Therefore, they can be washed out more easily. The metabolic ratios in the hair samples were examined depending on the background of request, hair color, body site of collection, and sex. The decision model by Scholz et al. Overall, the data corroborate that solely the presence of COC together with one metabolite in a hair sample—as stated in the current consensus by the SoHT—does not indicate COC ingestion, as all tested metabolites have been detected in street COC samples. CE appears to be a valuable marker for COC use in light of the lower abundance in the seized COC material and high frequency of detection in the hair samples. Analytical results of a high percentage of hair samples The authors express their gratitude to Emma Louise Kessler, MD, for the generous legacy she donated to the Institute of Forensic Medicine at the University of Zurich, Switzerland, for research purposes. Supplementary data is available at Journal of Analytical Toxicology online. Pragst F , Balikova MA. State of the art in hair analysis for detection of drug and alcohol abuse. Clin Chim Acta ; : 17 — Google Scholar. Society of hair testing guidelines for drug testing in hair. Forensic Sci Int ; : 20 — European Drug Report —trends and developments. Luxembourg : Publications Office of the European Union , World Drug Report —drug market trends: cocaine, amphetamine-type stimulants, new psychoactive substances. United Nations Publication , Society of Hair Testing. Analysis of extensively washed hair from cocaine users and drug chemists to establish new reporting criteria. J Anal Toxicol ; 38 : — Determination of hydroxy metabolites of cocaine in hair samples for proof of consumption. Drug Test Anal ; 10 : — Determination of hydroxy metabolites of cocaine from hair samples and comparison with street cocaine samples. Forensic Sci Int ; : — Hydroxycocaines as metabolic indicators of cocaine ingestion. Forensic Sci Int ; J Pharm Biomed Anal ; : — Analysis of cocaine and metabolites in hair: validation and application of measurement of hydroxycocaine metabolites as evidence of cocaine ingestion. Anal Bioanal Chem ; : — Cocaine hydroxy metabolites in hair: indicators for cocaine use versus external contamination. J Anal Toxicol ; 43 : — Application of discriminant analysis to differentiate between incorporation of cocaine and its congeners into hair and contamination. Forensic Sci Int ; : 13 — European guidelines for workplace drug and alcohol testing in hair. Drug Test Anal ; 8 : — Passive exposure to cocaine on a professional level: importance of considering relevant interpretation parameters. Paris , Analyst ; : — A comprehensive multi-analyte method for hair analysis: substance-specific quantification ranges and tool for task-oriented data evaluation. Forensic Sci Int ; 84 : 3 — 6. Development and validation of an HPLC-DAD method for simultaneous determination of cocaine, benzoic acid, benzoylecgonine and the main adulterants found in products based on cocaine. Forensic Sci Int ; : 32 — Cocaine analytes in human hair: evaluation of concentration ratios in different cocaine sources, drug-user populations and surface-contaminated specimens. J Anal Toxicol ; 36 : — Performance of hair testing for cocaine use-comparison of five laboratories using blind reference specimens. J Anal Toxicol ; 47 : — Chemical Book: M-Hydroxycocaine. Chemical Book: Cocain. Oxford University Press is a department of the University of Oxford. It furthers the University's objective of excellence in research, scholarship, and education by publishing worldwide. Sign In or Create an Account. Sign in through your institution. Advanced Search. Search Menu. Article Navigation. Close mobile search navigation Article Navigation. Article Contents Abstract. Materials and methods. Results and discussion. Supplementary data. Data availability. Journal Article Corrected proof. Comprehensive evaluation of cocaine and its hydroxy metabolites in seized cocaine and a large cohort of hair samples. E-mail: milena. Oxford Academic. Teresa Denifle. Tina M Binz. Christian Bogdal. Analytics, Zurich Forensic Science Institute. Thomas Kraemer. Markus R Baumgartner. Revision received:. Editorial decision:. Corrected and typeset:. Select Format Select format. Permissions Icon Permissions. Abstract As cocaine COC is not only incorporated into hair via blood following ingestion but also by external contamination, hair samples are commonly tested for COC metabolites to prove ingestion. Table 1. Open in new tab. Overview of literature on hydroxycocaine metabolites in hair samples. Sample size and type. COC cut-off. Proposed criteria for indicating COC use. Morris-Kukoski et al. Figure 1. Open in new tab Download slide. Figure 2. Figure 3. Figure 4. Figure 5. Figure 6. Figure 7. Google Scholar Crossref. Search ADS. Ramirez Fernandez. Published by Oxford University Press. For commercial re-use, please contact reprints oup. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals. Download all slides. Views More metrics information. Total Views Month: Total Views: August 59 September Email alerts Article activity alert. Advance article alerts. New issue alert. Receive exclusive offers and updates from Oxford Academic. Citing articles via Google Scholar. Four-year evaluation of drug-impaired driving drug concentrations. Development and method validation of a sampling technique for a reproducible detection of synthetic cannabinoids in exhaled breath using an in vitro pig lung model. Identification of 5-aminometonitazene and 5-acetamidometonitazene in a post-mortem case: Are nitro-nitazenes unstable? More from Oxford Academic. Biological Sciences. Clinical Medicine. Medical Toxicology. Medicine and Health. Science and Mathematics. Toxicology Non-medical. Authoring Open access Purchasing Institutional account management Rights and permissions. Get help with access Accessibility Contact us Advertising Media enquiries. Musshoff et al. Minoli et al. Positivity according to criteria by Franz et al. Hart et al. Positivity according to criteria by Scholz et al.

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