Segmental Hair Analysis Research Articles

Analyses of fecal and hair glucocorticoids to evaluate short- and long-term stress and recovery of Asiatic black bears (Ursus thibetanus) removed from bile farms in China

Demand for traditional Chinese medicines has given rise to the practice of maintaining Asiatic black bears (Ursus thibetanus) in captivity to harvest bile. We evaluated hypothalamic–pituitary–adrenal (HPA) activity in Asiatic black bears on a bile farm in China by measuring cortisol in hair. We also monitored hair and fecal glucocorticoid metabolites as bears acclimated to improved husbandry at the Animals Asia Foundation China Bear Rescue Center (CBRC) after removal from other bile farms. Fecal samples were collected twice weekly for ∼1year, and hair was obtained from bears upon arrival at the CBRC and again ⩾163days later. Paired hair samples showed declines in cortisol concentrations of 12–88% in 38 of 45 (84%, p<0.001) bears after arrival and acclimation at the rehabilitation facility. Concentrations of cortisol in hair from bears on the bile farm were similar to initial concentrations upon arrival at the CBRC but were higher than those collected after bears had been at the CBRC for ⩾163days. Fecal glucocorticoid concentrations varied across months and were highest in April and declined through December, possibly reflecting seasonal patterns, responses to the arrival and socialization of new bears at the CBRC, and/or annual metabolic change. Data from segmental analysis of hair supports the first of these explanations. Our findings indicate that bears produced elevated concentrations of glucocorticoids on bile farms, and that activity of the HPA axis declined following relocation. Thus, hair cortisol analyses are particularly well suited to long-term, retrospective assessments of glucocorticoids in ursids. By contrast, fecal measures were not clearly associated with rehabilitation, but rather reflected more subtle endocrine changes, possibly related to seasonality.

Determination of bromadiolone and brodifacoum in human hair by liquid chromatography/tandem mass spectrometry and its application to poisoning cases

Bromadiolone and brodifacoum, two common anticoagulant rodenticides, are involved in the majority of anticoagulant rodenticide poisoning cases in humans in China. Hair analysis can provide long-term information on drug exposure. A method using liquid chromatography coupled with tandem mass spectrometry (LC/MS/MS) was developed and validated for the measurement of bromadiolone and brodifacoum in human hair. A 1 mL aliquot of a phosphate buffer solution (pH 6.8) was added to 20 mg of pulverized hair followed by ultrasonication and liquid-liquid extraction. Liquid chromatography was performed using a C(18) column with a mobile phase gradient of ammonium acetate (10 mM) and methanol. A tandem mass spectrometer in multiple reaction monitoring mode with a negative electrospray ionization source was employed for detection. Warfarin-d(5) was used as an internal standard for both analytes. The limits of detection (LODs) for bromadiolone and brodifacoum were 0.010 and 0.025 ng/mg, respectively. The calibration curves for both analytes were linear from 0.025 to 1 ng/mg. The accuracy ranged from 90.3 to 109.3%, and the intra-day and inter-day imprecisions were less than 15%. The established method was found effective when applied to the analyses of bromadiolone or brodifacoum in five cases, indicating that segmental hair analysis could be useful for clinical and forensic purposes by identifying the time of ingestion.

Segmental hair analysis and estimation of methamphetamine use pattern

The aim of this study was to investigate whether the results of segmental hair analysis can be used to estimate patterns of methamphetamine (MA) use. Segmental hair analysis for MA and amphetamine (AP) was performed. Hair was cut into the hair root, consecutive 1cm length segments and 1-4cm length segments. Whole hair was also analyzed. The hair samples were incubated for 20h in 1mL methanol containing 1% hydrochloric acid after washing the hair samples. Hair extracts were evaporated and derivatization was performed using trifluoroacetic anhydride in ethylacetate at 65°C for 30min. Derivatized extract was analyzed by gas chromatography/mass spectrometry. The 15 subjects consisted of 13 males and two females and their ages ranged from 25 to 42 (mean, 32). MA and AP concentrations in the whole hair ranged from 3.00 to 105.10ng/mg (mean, 34.53) and from 0.05 to 4.76ng/mg (mean, 2.42), respectively. Based on the analysis of the 1cm length segmental hair, the results were interpreted in a way to distinguish between continuous use of MA (n = 10), no recent but previous use of MA (n = 3), and recent but no previous use of MA (n = 2). Furthermore, the individuals were interpreted as light, moderate, and heavy users based on concentration ranges previously published.

GC‐MS and GC‐MS/MS in PCI Mode Determination of Mescaline in Peyote Tea and in Biological Matrices

Peyote, a cactus containing the hallucinogen mescaline, is used to induce altered states of consciousness in religious ceremonies or for recreational purpose. This study reports a case of an underage boy suspected of mescaline abuse. For this purpose, we analyzed a dark green liquid sample found in the bedroom of the boy whose urine and hair samples were collected shortly after the drink was found. A method by gas chromatography-mass spectrometry tandem mass spectrometry (GC-MS/MS) in positive chemical ionization mode was developed and validated in terms of linearity, specificity, accuracy, and sensitivity for mescaline determination at the low concentrations present in hair. GC-MS analysis of the liquid identified mescaline, while urine was negative; GC-MS/MS segmental hair analysis identified mescaline in the proximal segment (root to 2 cm), while the distal segments were negative. Although peyote was uncommonly encountered, its use was confirmed by segmental hair analysis that can provide long-term information about drugs use.

Determination of 22 synthetic cannabinoids in human hair by liquid chromatography–tandem mass spectrometry

Herbal mixtures of the “Spice“-type contain a variety of synthetic cannabinoids. To prove the contact of a person with synthetic cannabinoids in a previous period of up to several months, hair testing is ideally suited. A rapid, simple and sensitive liquid chromatography–tandem mass spectrometry (LC–MS/MS) assay was developed to determine 22 synthetic cannabinoids in human hair. The synthetic cannabinoids JWH-007, JWH-015, JWH-018, JWH-019, JWH-020, JWH-073, JWH-081, JWH-122, JWH-200, JWH-203, JWH-210, JWH-250, JWH-251, JWH-398, AM-694, AM–2201, methanandamide, RCS-4, RCS-4 ortho isomer, RCS-8, WIN 48,098 and WIN 55,212-2 were extracted from 50mg hair by 3-h ultrasonification in ethanol. The extracts were analysed on a triple-quadrupole linear ion trap mass-spectrometer in scheduled multiple reaction monitoring mode (sMRM). The method was fully validated and proved to be accurate, precise, selective and specific with satisfactory linearity within the calibrated range and a lower limit of quantification of 0.5pg/mg for 20 compounds. Authentic hair samples from chronic consumers showed the presence of two to six synthetic cannabinoids in the same segment. In the first segment, concentrations of up to 78pg/mg JWH-081 were present. In segmented hair, the concentrations of most substances increased from the first (proximal) to the third segment. The highest concentration was ca. 1100pg/mg JWH-081. The results of segmental hair analysis in chronic users suggest incorporation of the drugs in head hair via side-stream smoke condensation as a major route. In summary, the method can be used to prove the contact with herbal mixtures containing synthetic cannabinoids and thus contributes to an efficient abstinence control.

Segmental Hair Analysis for 11-Nor- 9-Tetrahydrocannabinol-9-Carboxylic Acid and the Patterns of Cannabis Use

Cannabis is the most widely abused drug in the world. The purpose of this study is to detect 11-nor-9-carboxy-Δ⁹-tetrahydrocannabinol (THCCOOH) in segmental hair and to evaluate the patterns of cannabis use. We investigated the relationship between the concentrations of THCCOOH in hair and the self-reported use data and the route of administration. For this purpose, the hair samples were washed, digested with 1 mL of 1 M NaOH at 85°C for 30 min along with the internal standard, THCCOOH-d₃ (2.5 pg/mg) and extracted in 2 mL of n-hexane-ethyl acetate (9:1) twice after adding 1 mL of 0.1N sodium acetate buffer (pH = 4.5) and 200 µL of acetic acid. The organic extract was transferred and evaporated and the mixture was derivatized with 50 µL of pentafluoropropionic anhydride and 25 µL of pentafluoropropanol for 30 min at 70°C. Reconstituted final extract was injected into the gas chromatography-tandem mass spectrometer operating in the negative chemical ionization mode. In segmental hair analysis, the concentrations of THCCOOH decreased from the proximal to distal segments. The concentrations of THCCOOH in hair and the self-reported dose and frequency of administration from cannabis users were not well correlated because of the low accuracy and reliability of the self-reported data. However, this study provides preliminary information on the dose and frequency of administration among cannabis users in our country.

Hair: An Evidence for Drug of Abuse

Hair analysis for drug abuse has been gaining increasing significance in forensic sciences. Hair is a special evidence used for the investigation of chronic drug of abuse or poisoning in criminal cases. It provides results with sensitive methods even if the drug is present in low amount. Segmental hair analysis can yield information about the time course of the substance used.

Maternal hair testing for the assessment of fetal exposure to drug of abuse during early pregnancy: Comparison with testing in placental and fetal remains

Drug use by pregnant women in the first trimester of pregnancy and subsequent fetal exposure during early gestation can be assessed only by repetitive/systematic maternal blood/urine analysis or segmental hair analysis. No evidence of any relationship between maternal/fetal exposure during this specific period of gestation has been demonstrated to date in a human model. To clarify drugs toxicokinetics and transplacental passage during early pregnancy, the presence of the most widely used recreational drugs of abuse and metabolites was investigated in the proximal 4cm hair segments of women undergoing voluntary termination of pregnancy (n=280) during the 12th week of gestation and the results were compared to those from placenta and fetal tissue samples in order to verify whether maternal hair testing can reflect fetal exposure and, if so, to what extent. Hair, placenta and fetal remains were analyzed by validated gas chromatography mass spectrometry assays. Eighty one positive hair samples were identified: 60 were positive for cannabis (74.1%), 28 for cocaine (34.6%), 7 for opiates (8.6%), 3 for MDMA (3.7%) and 18.5% were positive for more than one drug. The positive hair test results were confirmed in placenta/fetal tissues in 10 cases out of 60 for cannabis (16. 7%); in 7 out of 28 for cocaine (25%); and none for the 6 opiates positive cases and 3 MDMA cases, respectively. Drugs/metabolites in hair of pregnant women can be used as biomarkers of past drug use (repetitive or sporadic), although the use is not always reflected in fetal/placental tissues. There are several possible hypotheses to explain the results: (1) the use occurred before the start of pregnancy, (2) past sporadic consumption which could be measured in hair but not in fetal and placental remains because of the narrow window of drug detection in placental/fetal tissues; (3) the sensitivity of the analytical methods was not high enough for the detection of the minute amount of drugs of abuse and metabolites which reached these tissues (4) there is a large variability in the transplacental passage of drugs of abuse and in the placenta's metabolizing capacity.

Hair analysis of drugs involved in drug-facilitated sexual assault and detection of zolpidem in a suspected case

In drug-facilitated crimes, victims are subjected to nonconsensual acts while they are incapacitated by the effects of a drug. A specific LC-MS/MS protocol for determining benzodiazepines and hypnotics at low concentration in hair specimens was developed and validated in order to target the allegedly administered drugs on a chronological basis. In the case hereby reported, a 26-year-old woman claimed to have been sexually assaulted after being administered an allegedly drugged coffee, but toxicological analysis of urine and blood provided no evidence of any drug intake. Subsequently, a second woman accused the same man of sexual abuse. Hence, the suspect was prosecuted. Specimens were collected from four subjects (two alleged victims, the suspect and his wife) and segmental hair analysis was performed. The results revealed that zolpidem was present at low picogram per milligram concentration in three out of eleven segments of hair specimen obtained from the first of the alleged victims, offering plain evidence of single or sporadic exposure, whereas the agent was detected in the high picogram per milligram range in the hair collected from suspect's wife, coherently with therapeutic administration. The presence of interfering signals typical of the keratin-containing matrix was found and possible hair degradation by cosmetic treatments was investigated by electron microscopy, so as to obtain a judicious interpretation of the analytical findings.

Is hair analysis for dialkyl phosphate metabolites a suitable biomarker for assessing past acute exposure to organophosphate pesticides?

In the present paper, the possibility to use dialkyl phosphate metabolites (DAPs) hair segmental analysis as a biomarker of past acute exposure to organophosphates is examined. Hair samples of four acute poisoning survivors were collected and segmental hair analysis was performed. The total hair samples were divided to 1 cm segments and analysed by gas chromatography-mass spectrometry (GC-MS) for the presence of four DAP metabolites, dimethyl phosphate (DMP), diethyl phosphate (DEP), diethyl thiophosphate (DETP) and diethyl dithiophosphate (DEDTP). Results were examined under the light of pesticide type and time of hair sample collection. Although DAPs were detected all along the hair shaft, higher concentrations (peaks) were detected in the segments proximate to the suicide period. It was also observed that the elevated concentrations of the present metabolites corresponded to the ones produced by the ingested parent compound. Conclusively, measurements of DAPs in the appropriate hair segments of OP-poisoned patients can be used for assessing past acute exposure to organophosphates in certain cases.

The role of variations in growth rate and sample collection on interpreting results of segmental analyses of hair

Segmental analysis of hair for drugs, metabolites, and poisons has been widely reported in the scientific literature over the past two decades. Two fundamental assumptions in interpreting results of such analyses are (1) an average linear growth rate of head hair of 1cm/month and (2) that sample collections occur with the hair being cut directly next to the scalp. The purpose of this study was to evaluate the variability associated with growth rate of human head hair, as well as the ability to uniformly collect hair next to the scalp. The results were used to determine how these factors affect the interpretation of results generated in segmental analysis of hair. A thorough literature review was conducted to assess the range of linear growth of human head hair from the vertex posterior and occipital regions. The results were compiled to establish the average (1.06cm/month), as well as the range of possible growth rates of head hair. The range was remarkable and suggests that conclusions based on the 1-cm/month growth rate could be significantly skewed. A separate study was undertaken to evaluate collection of hair next to the scalp. Fourteen individuals were provided oral instructions, as well as a written standard collection procedure for head hair. The experience levels among the collectors varied from novice to expert. Each individual collected hair from dolls with short- and long-hair. Immediately following each collection, the sampling area was evaluated to determine how close to the scalp the cuts were made, as well as the variability in the lengths of hair remaining at the sampled area. From our collection study, we determined that 0.8±0.1cm of hair was left on the scalp after cutting. When taking into account the amount of hair left on the scalp after collecting, the use of a growth rate of 1.06cm/month, and the assumption that it takes two weeks for newly formed hair in the follicle to reach the scalp, we find that the first 1-cm segment of hair typically corresponds to hair formed 1.3±0.2 to 2.2±0.4 months (95% confidence) earlier. The impact of these findings as it relates to the corresponding time for each additional segment is demonstrated. As a result, we recommend that hair collection be delayed 8 weeks after a suspected ingestion to ensure that the sample fully represents the exposure period. The results of this study suggest that the variability in the growth rate of human head hair, as well as the inconsistent collection of hair, significantly affect the interpretation of results from segmental analysis of hair.

Evidence of Haldol (haloperidol) long-term intoxication

A case of intoxication by haloperidol is reported. Haloperidol is a butyrophenone derivative commonly used in many hospital units as an antipsychotic agent. Adverse reactions due to haloperidol intoxication include drowsiness, blurred vision, extrapyramidal effects, tardive dyskinesia, tachycardia, hypotension and muscular rigidity. In August 2008, a 49 year-old female nurse started feeling various symptoms such as muscular rigidity, drowsiness and buccal dyskinesia. After 3 months, she was hospitalized for the worsening of these symptoms. Four months later, she showed once more the same symptoms. Two open water bottles from which the nurse used to drink in the hospital were confiscated and analyzed. Moreover, the nurse was asked to give a sample of her hair for executing the inherent toxicological analyses. Haloperidol was found in both bottles 1 and 2 at a concentration of 31.5 μg/mL and 43.6 μg/mL, respectively. Based on segmental hair analysis, it was deduced that the nurse consumed haloperidol in the approximate period from August 2008 to March 2009. The higher levels of haloperidol in hair were found in accordance with the periods of most severe appearance of symptoms, requiring the hospitalization of the nurse. The analysis of preservatives and excipients led us to conclude that the pharmaceutical drug was probably added to the water bottles as "Haldol 2mg/mL oral solution".

Segmental hair analysis can demonstrate external contamination in postmortem cases

Excluding laboratory mistakes, a false positive hair result can be observed in case of contamination from environmental pollution (external contamination) or after drug incorporation into the hair from the individual body fluids, such as sweat or putrefactive fluid (post mortem artifact). From our 20 years experience of hair testing, it appears that artifact(s) cannot be excluded in some post mortem cases, despite a decontamination procedure. As a consequence, interpretation of the results is a challenge that deserves particular attention. Our strategy will be reviewed in this paper, based on six cases. In all cases, a decontamination procedure with two washes of 5 ml of dichloromethane for 5 min was performed and the last dichloromethane wash was negative for each target drug. From the histories, there was no suspicion of chronic drug use. In all six cases, the concentrations detected were similar along the hair shaft, irrespective of the tested segment. We have considered this as indicative of external contamination and suggested to the forces or the judges that it is not possible to indicate exposure before death. In contrast to smoke, it seems that contamination due to aqueous matrices (sweat, putrefactive fluid, blood) is much more difficult to remove. To explain potential incorporation of 7-aminoflunitrazepam via putrefactive material, the author incubated negative hair strands in blood spiked at 100 ng/ml and stored at +4°C, room temperature and +40 °C for 7, 14 and 28 days. After routine decontamination, 7-aminoflunitrazepam tested positive in hair, irrespective of the incubation temperature, as early as after 7 days (233-401 pg/mg). In all periods, maximum concentrations were observed after incubation at room temperature. The highest concentration (742 pg/mg) was observed after 28 days incubation at room temperature. It is concluded that a standard decontamination procedure is not able to completely remove external contamination in case of post mortem specimens. Homogenous segmental analyses can be probably indicative of external contamination and therefore a single hair result should not be used to discriminate long-term exposure to a drug. Nor should the presence of a metabolite be considered as a discrimination tool, as it can also be present in putrefactive material.

Hair analysis and self-report of methamphetamine use by methamphetamine dependent individuals

The questions of whether the dose of drug that is consumed corresponds to drug concentration levels in hair and how results of hair analyses can be interpreted are still debated. The aim of this study was to investigate (1) whether there is a correlation between doses of Methamphetamine (MA) use and MA concentration levels in hair and (2) whether results of hair analyses can be used to estimate dose, frequency, and patterns of MA use. In this study, segmental hair analysis was performed through consecutive 1 cm as well as 1–4 cm (=3 cm) segmental hair lengths. MA dependent individuals ( n = 9) provided information on doses (0.25–4 g/day) of MA use as well as the frequency of MA use. The concentrations of MA and its metabolite amphetamine (AP) in hair were determined using gas chromatography/mass spectrometry (GC/MS). One-way analysis of variance (ANOVA) test was performed to evaluate whether MA and AP concentrations in consecutive 1 cm length segmental hair were consistent with the history of MA use. The cumulative doses of MA use calculated from the daily dose and the frequency during 1–4 months were well correlated to the concentrations of MA and AP in 1–4 cm segmental hair length (correlation coefficient, r = 0.87 for MA and r = 0.77 for AP). The results from this study show the patterns and histories of MA use from MA dependent individuals and could assist in the interpretation of hair results in forensic toxicology as well as in rehabilitation and treatment programs.

Segmental hair analysis using liquid chromatography–tandem mass spectrometry after a single dose of benzodiazepines

In China, benzodiazepines are the most frequently observed compounds in cases of drug-facilitated crime. Sensitive, specific, and reproducible methods for the quantitative determination of 18 benzodiazepines in hair have been developed using LC–MS/MS. Fourteen volunteers had ingested a single 1–6 mg estazolam tablet. Hair was collected 1 month after administration. All the proximal segments were positive for estazolam. With increased dosage, estazolam can be detected in the 2–4 cm segments in some subject's hair. Even some of 4–6 cm segments were positive. Hair analysis was applied to two authentic criminal cases. Full-length hair samples collected 5 weeks after the offense were cut into segments of 2 cm from the root, analyzed and quantified. The clonazepam concentrations measured in the first two segments for V#1 and V#2 were 15.47 and 11.93 pg/mg, respectively. However, both the 4–6 cm and the 6–8 cm segment of V1# remained positive, while those of V#2 were negative. It needs more substantial guidelines to use segmental hair analysis in drug-facilitated crime.

Higher cortisol content in hair among long-term unemployed individuals compared to controls

Unemployment and financial strain are chronic stressors that have been shown to be associated with an increase in mean salivary and serum cortisol levels. Hair analysis for cortisol content is a new promising tool by which hair segmental analysis may provide a retrospective calendar of cumulative cortisol exposure over time rather than momentary assessments. Participants of this study were 31 unemployed and 28 employed individuals (46 women). Hair segmental analysis was conducted using 3-cm long segments starting with the scalp-near segment. Due to differing hair length, 52 individuals had values for the second segment and n=33 individuals had values for the third segment. Univariate analysis of variance indicated that unemployed individuals had higher cortisol content in the first (p<0.05, eta(2)=0.071) and second (p<0.05, eta(2)=0.085) hair segment (a total of 6 cm long hair representing the preceding 6 months of collection). Consistent with other data from our laboratory, there was a wash-out effect for the third segment (p<0.05 for segment 3 vs. segment 1 and 2). Unemployed individuals indicated increased levels of perceived stress and impairments in subjective well-being compared to employed individuals. These subjective measures of perceived stress and well-being were unrelated to cortisol content in hair. We conclude that hair analysis for cortisol content may be a valid method to detect differences in cumulative cortisol exposure between chronically stressed individuals and healthy controls. Due to a wash-out effect, retrospective ascertainment of cortisol exposure may be limited to the preceding 6 months of specimen collection.

A tendency for re-offending in drug-facilitated crime

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 20 mg of decontaminated and cut hair, 100 pg/mg of clonazepam-d4 was added as internal standard. Hair specimens were extracted with CH 2Cl 2/ether after incubation overnight at 56 °C in pH 7.6 buffer. Extractions were performed on blood and urine using Toxi-tube A ® with 5 ng/mL of clonazepam-d4. The residues were analyzed by LC–ESI-MS/MS. Calibration curves in blood and urine (0.5–500 ng/mL) were prepared by spiking aliquots of blank fluids ( r 2 > 0.9816 for all drugs). LOD in body fluids ranged 0.5–10 ng/mL. Calibration curves in hair (0.5–100 pg/mg) were prepared by spiking aliquots of blank hair ( r 2 > 0.9877 for all drugs). LOD in hair ranged 0.5–5 pg/mg. Case #1: Two young women were raped with an interval of approximately 1 year between the incidents. Lorazepam (present, <2 pg/mg) was detected in hair obtained from the first victim, and zolpidem (19 pg/mg) in hair of the second one. The offender was in jail between the two offences. Case #2: The offender approached a total of 8 men and women who were aged over 50 years. The offender was in jail between the two series of respectively 3 and 5 victims. Zopiclone was detected in victims’ hair ( n = 7) at concentrations 13–42 pg/mg. Case #3: The offender stole thousands of Euros using credit cards obtained from 22 different wealthy victims. He employed a cocktail of up to 6 drugs made up of: flunitrazepam, clonazepam, doxylamine, cyamemazine, zolpidem and lorazepam. Drugs were detected in all victims’ hair ( n = 18) at concentrations in the range 1–81 pg/mg for all drugs. Between the two series (of respectively 4 and 16 victims) the offender spent 6 months in jail, and then police spent 6 months looking for him while he was under judiciary control prior to his judgment. 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.

Hair analysis provides a historical record of cortisol levels in Cushing's syndrome.

The severity of Cushing's Syndrome (CS) depends on the duration and extent of the exposure to excess glucocorticoids. Current measurements of cortisol in serum, saliva and urine reflect systemic cortisol levels at the time of sample collection, but cannot assess past cortisol levels. Hair cortisol levels may be increased in patients with CS, and, as hair grows about 1 cm/month, measurement of hair cortisol may provide historical information on the development of hypercortisolism. We attempted to measure cortisol in hair in relation to clinical course in six female patients with CS and in 32 healthy volunteers in 1 cm hair sections. Hair cortisol content was measured using a commercially available salivary cortisol immune assay with a protocol modified for use with hair. Hair cortisol levels were higher in patients with CS than in controls, the medians (ranges) were 679 (279-2500) and 116 (26-204) ng/g respectively (P<0.001). Segmental hair analysis provided information for up to 18 months before time of sampling. Hair cortisol concentrations appeared to vary in accordance with the clinical course. Based on these data, we suggest that hair cortisol measurement is a novel method for assessing dynamic systemic cortisol exposure and provides unique historical information on variation in cortisol, and that more research is required to fully understand the utility and limits of this technique.

Effect of a controlled dietary change on carbon and nitrogen stable isotope ratios of human hair

The carbon ((13)C/(12)C) and nitrogen ((15)N/(14)N) stable isotope ratios of human hair can be used for the interpretation of dietary habits and nutritional status in contemporary or past populations. Although the results of bulk or segmental isotope ratio analysis of human hair have been used for the reconstruction of an individual's diet for years, only limited data of controlled dietary changes on the carbon and nitrogen isotopic composition of human hair are available. Hair of four individuals, two males and two females, who participated in a dietary change experiment for 28 days was segmentally analysed for delta(13)C and delta(15)N. The dietary change included a change from C3 to C4 plant enriched diets and a simultaneous replacement of terrestrial animal products by marine products. This resulted in an increase in delta(13)C(diet) of +8.5 to +9.9 per thousand and in delta(15)N(diet) of +1.5 to +2.2 per thousand. All subjects showed significant increases in delta(13)C(hair) and delta(15)N(hair) during the dietary change period, although no subject reached a new steady state for either carbon or nitrogen. The change in delta(15)N(hair) was faster than the change in delta(13)C(hair) for all individuals. The magnitude of change of the isotopic composition during the dietary change period could be attributed to the degree of physical activity of the individuals, with a higher physical activity resulting in a faster change.

Higher cortisol content in hair among long-term unemployed individuals compared to controls

Background: Unemployment and financial strain are chronic stressors that have been shown to be associated with an increase in mean salivary and serum cortisol levels. The impact of chronic stress on cortisol excretion is best measured using a summary index of cortisol excretion over longer periods of time rather than momentary assessments. Hair analysis for cortisol content is a new promising tool by which hair segmental analysis may provide a retrospective calendar of cumulative cortisol exposure. Methods: Participants of this study were 31 unemployed and 28 employed individuals (n=46 women). Hair segmental analysis was conducted using 3cm -long segments starting with the scalp-near segment. Due to differing hair length, n=52 individuals had values for the second segment and n=33 individuals had values for the third segment. Results: Univariate analysis of variance indicated that unemployed individuals had higher cortisol content in the first (p&lt;0.05, eta2=0.071) and second (p&lt;0.05, eta2=0.085) hair segment (a total of 6cm long hair representing the preceding 6 months of collection). Duration of unemployment was related to hair cortisol content (r=0.415 and r=0.430 for segment 1 and 2, respectively). Interestingly, BMI was related to hair cortisol content (r=0.430) but was not different between groups. Consistent with other data from our laboratory, there was a wash-out effect for the third segment (p&lt;0.05 for segment 3 vs. segment 1 and 2). Conclusions: We conclude that hair analysis for cortisol content may be a valid method to detect differences in cumulative cortisol exposure between chronically stressed individuals and healthy controls. Due to a wash-out effect, retrospective ascertainment of cortisol exposure may be limited to the preceding 6 months of specimen collection.