Studies on the persistence of estradiol benzoate and nortestosterone decanoate in hair of cattle following treatment with growth promoters,determined by ultra-high-performance liquid chromatography–tandem mass spectrometry

Measurement of steroid esters in bovine hair samples, using sensitive liquid chromatography-tandem mass spectrometry (LC–MS/MS), provides a powerful tool for identifying animals treated illicitly with growth promoters. The successful application of such testing requires appropriate sampling of hair from treated animals. This paper describes the results of hair analysis by LC–MS/MS for two animal studies in which animals were treated with estradiol-3-benzoate and nortestosterone decanoate. The results from the first animal study indicate that animals treated with these anabolic steroids may not always be identified from analysis of hair samples; positive test results occur sporadically and only for some of the treated animals. The results from the second animal study identify conditions attaching to positive hair samples, such as, that concentrations of steroid esters in hair are related to distance of sampling from point of injection and to time post-treatment, that concentrations of steroid esters in hair are related to dose given to the animal but that this relationship may vary over time post-treatment, and that steroid esters may be measured in regrowth hair taken some weeks after treatment. Steroid esters are determined along the length of the hair, confirming that accumulation of steroid esters into hair occurs from various sources, including blood, sweat and sebum. The reported research provides some useful insights into the mechanisms governing the persistence of steroid esters in bovine hair following illicit treatment with growth promoters.     

Neutron activation analysis and X-ray Rayleigh and Raman scattering of hair and nail clippings as noninvasive bioindicators for Cu liver status in Labrador Retrievers

The heritability of chronic hepatitis in the Labrador Retriever is studied with the aim of identifying the related gene mutation. Identification of cases and controls is largely based on instrumental neutron activation analysis (INAA) Cu determination in liver biopsies. The burden for these companion animals may be reduced if nail clippings and hair (fur) could serve as a noninvasive indicator for the hepatic Cu concentrations. No correlation was found between hepatic Cu concentrations and Cu concentrations in hair and nail samples. However, hair and nail samples were also analyzed by X-ray tube excitation, taking advantage of the X-ray Compton, Rayleigh, and Raman scattering which reflects the organic components such as the type of melanin. Principal component analysis provided first indications that some differentiation between healthy and sick dogs could indeed be obtained from hair and nail analysis. Figure Principal component analysis of scattered region of x-ray fluorescence spectra of Labrador dog nails, demonstrating the differentiation towards dogs with high and low Cu liver levels (respectively positive and negative PC2 values) reflecting hepatitis, as well as gender (PC1: negative values for female and positive values for males)     

Determination of Mercury and Selenium in Hair Samples of Brazilian Indian Populations Living in the Amazonic Region by NAA

Biomonitoring of mercury contamination of Brazilian Indian population groups living in the Xingu Park, a reservation situated in the Amazonic region, has revealed very high levels of mercury in hair samples as compared to controls. Total mercury was determined by INAA in most of the tribes living in the Park and methylmercury was determined by CVAAS in samples with total mercury above 10 mg/kg. Due to the fact that selenium seems to protect animals against the toxic effects of methylmercury, it was considered also of interest to determine its concentrations in the hair samples with very high mercury levels. Selenium was determined by INAA via the short-lived radionuclide ⁷⁷mSe (T _1/2 = 17.45 s). The correlations between selenium and mercury concentrations in Brazilian controls and in the Indian population groups are discussed.     

A study of trace elements in scalp hair and fingernails of industrial workers of Ontario,Canada

Instrumental neutron activation analysis was performed on scalp hair and fingernail samples collected randomly from a group of heavily exposed healthy male industrial workers living in the cities of Hamilton and Toronto (Ont.), Canada. The concentrations of trace elements (Ag, Au, Br, Cd, Co, Cr, Cs, Fe, Hg, Lu, Mn, Mo, Rb, Ru and T₃) were evaluated and compared for scalp hair and fingernails. Various trace elements show similar variations in the two kinds of samples. The results have also been compared with those for normal subjects from USA (scalp hair) and Canada (fingernails).     

Analysis of ketamine and norketamine in hair samples using molecularly imprinted solid-phase extraction (MISPE) and liquid chromatography–tandem mass spectrometry (LC-MS/MS)

An anti-ketamine molecularly imprinted polymer (MIP) was synthesized and used as the sorbent in a solid-phase extraction protocol to isolate ketamine and norketamine from human hair extracts prior to LC-MS/MS analysis. Under optimised conditions, the MIP was capable of selectively rebinding ketamine, a licensed anaesthetic that is widely misused as a recreational drug, with an apparent binding capacity of 0.13 μg ketamine per mg polymer. The limit of detection (LOD) and lower limit of quantification (LLOQ) for both ketamine and norketamine were 0.1 ng/mg hair and 0.2 ng/mg hair, respectively, when 10 mg hair were analysed. The method was linear from 0.1 to 10 ng/mg hair, with correlation coefficients (R ²) of better than 0.99 for both ketamine and norketamine. Recoveries from hair samples spiked with ketamine and norketamine at a concentration of 50 ng/mg were 86% and 88%, respectively. The method showed good intra- and interday precisions (<5%) for both analytes. Minimal matrix effects were observed during the LC-MS/MS analysis of ketamine (ion suppression −6.8%) and norketamine (ion enhancement +0.2%). Results for forensic case samples demonstrated that the method successfully detected ketamine and norketamine concentrations in hair samples with analyte concentrations ranging from 0.2 to 5.7 ng/mg and from 0.1 to 1.2 ng/mg, respectively.     

Neutron activation analysis of organohalogens in Chinese human hair

To effectively extract organohalogens from human hair, two factors, the extracting time and hair length on the extraction efficiency of organohalogens were studied by neutron activation analysis (NAA) and gas chromatograph-electron capture detector (GC-ECD), respectively. Furthermore, the concentrations of extractable organohalogens (EOX) and extractable persistent organohalogens (EPOX) in hair samples from angioma and control babies were also measured by the established method. The results indicated that the optimal Soxhlet-extraction time for EOX and EPOX in hair was from 8 to 11 hours, and the extraction efficiencies for organochlorine pesticides in hair were in the order of powder >2 mm>5 mm. Also, the mean levels of EOC1 and EPOC1 in hair of the angioma babies were significantly higher than those in the control babies (P _EOC1<0.01; P _EPOC1<0.05), which implied the possible relationship between the environmental pollution and angioma.     

Indication of environmental pollution by means of INAA of the hair of some free living mammals

Hair samples collected from free living mammals common hare (Lepus europaeus Pall.), common vole (Microtus arvallis Pall.) and wood mouse (Apodemus sylvaticus L.) have been subjected to instrumental neutron activation analysis (INAA). up to 18 elements As, Au, Br, Ce, Co, Cr, Cu, Fe, Hg, K, La, Na, Sb, Sc, Se, Sm, Th and Zn have been determined in each hair sample. Animal hair samples from areas polluted by thermal power plants burning coal were taken and compared with hair samples from the animals living in relatively non-polluted control areas. The results show that animal hair samples from areas with higher level of pollution contain usually higher concentrations of toxic and essential elements as As, Co, Cr, Fe and Se. Muride rodents can be used for more detailed monitoring of environmental exposure than hare. Moreover, hair of common vole shows usually higher levels of contamination as compared with wood mouse, which could be explained by different compositions of feed. Animal hair has been shown to be a rather sensitive indicator of environmental exposure and INAA has proved to be a suitable analytical tool for this purpose.     

火焰原子吸收光谱法测定高血压患者头发中锌铜钙镁

用火焰原子吸收光谱法测定高血压患者45例及健康对照者34例头发中锌、铜、钙、镁的含量。结果表明,高血压患者头发中锌、铜、钙含量显著低于健康对照组（p〈0．05）,镁含量极显著低于健康对照组,揭示体内锌、铜、钙、镁元素含量与高血压发病存在一定关系。     

Were Chinchorros exposed to arsenic? Arsenic determination in Chinchorro mummies' hair by laser ablation inductively coupled plasma-mass spectrometry (LA-ICP-MS)

Chinchorros, a fishermen culture, who lived about 7000 years ago in the coastal region of the Atacama Desert in the northern outpost of present-day Chile, practiced an intricate system of mummification of their dead. The drinking water in this region is rich with arsenic, and the mummies were found in these arsenic endemic areas. Well preserved mummy hair samples provided a unique opportunity to explore the ancient arsenic exposure of the Chinchorros by laser ablation inductively coupled plasma-mass spectrometry (LA-ICP-MS) using a single hair strand without any elaborate sample preparation. Forty-six hair samples from mummies found in five burial sites around the Atacama Desert, Chile, were used for this study. After cleaning, hair strands were placed on mounting tape and ablated using a Nd-YAG UV laser coupled to ICP-MS. A suite of contemporary human hair from the same region with known arsenic concentrations was used for calibration of LA-ICP-MS. Satisfactory linear calibration functions were obtained for arsenic in hair. The method detection limit was 0.8 µg/g and the sample throughput for this method is ∼ 10 samples per hour. It appears that mummies from the Morro (Arica), Iquique and Camarones had the elevated concentration of arsenic in hair (AsH > 10 µg/g) in this sub-set of samples, where Morro had the broad distribution of As concentrations.     

Simultaneous LC-HRMS determination of 28 benzodiazepines and metabolites in hair

A liquid chromatography–high resolution mass spectrometry (LC-HRMS) method for the simultaneous identification and quantification of 28 benzodiazepines, including 6 metabolites, in 50 mg of hair has been validated. Positive ion electrospray ionization and HRMS determination in full-scan mode were realized on an Orbitrap mass spectrometer at a nominal resolving power of 60,000. In-source collisional experiments were conducted to obtain additional information for a more reliable identification of the investigated drugs. HRMS in full-scan mode allowed the exact determination of molecular masses of all analytes eluting in the HPLC run, so that both the immediate and retrospective screening of results for drugs and their metabolites were available. Sample preparation consisted of an overnight incubation in phosphate buffer pH 8.4 and a subsequent liquid/liquid extraction with methylene chloride/diethyl ether (90:10). Gradient elution was performed on a Luna C18 analytical column and four deuterated analogues were used as internal standards (IS). Validation was performed using both spiked hair samples and hair samples from subjects treated with benzodiazepines. Selectivity was evaluated by analysis of 20 certified blank hair samples. Extraction efficiency and matrix effects were evaluated by analysis of true positive samples. The lowest limits of quantification (LLOQs) ranged from 1 to 10 pg/mg. Linearity was investigated in the range from LLOQ to 1,000 pg/mg, for each compound (R ² 0.998–0.999). Mean relative errors, calculated at three concentration levels, ranged from 1 to 20% (absolute value). Precision, at concentrations higher than the LLOQs, was always less than 15% expressed as percentage relative standard deviation. After validation, the procedure was applied to real samples collected for clinical and forensic toxicology purposes from subjects who were assumed to have taken benzodiazepines.     

Arsenic, antimony and bismuth in human hair from potentially exposed individuals in the vicinity of antimony mines in Southwest China

To study the effect of the environmental pollution in exposed population, human hair samples of residents were collected from two typical antimony mines (Xikuangshan antimony mine and Qinglong antimony mine, Southwest China) and one non-mining city (Guiyang, Southwest China), and the concentrations of arsenic, antimony and bismuth in these samples were analyzed by hydride generation-atomic fluorescence spectrometry. Arsenic concentrations for Xikuangshan, Qinglong, and Guiyang ranged 0.236-48.4 (mean 4.21), 0.130-16.1 (mean 2.96), and 0.104-0.796 (mean 0.280) μg/g, respectively. Antimony concentrations for Xikuangshan, Qinglong, and Guiyang ranged 0.250-82.4 (mean 15.9), 0.060-45.9 (mean 5.15), and 0.065-2.87 (mean 0.532) μg/g, respectively. Bismuth contents were found to be greater than the limit of detection (LOD > 0.016 μg/g) in all the human hair samples collected from residents from Qinglong antimony mine, 95.5% samples from Xikuangshan mine and only 22.7% samples from Guiyang. There were no significant differences in both arsenic and antimony concentrations between hair samples from male and female individuals in the same area (P > 0.05). Arsenic and bismuth were mainly present in samples from children (5-9 years) and adults aged 41-51 years. Relatively high antimony contents (≥ 3 μg/g) were detected mainly in samples from children and adults aged ≥ 41 years. Significant correlation was found between the concentrations of arsenic and antimony in the human hair samples (r = 0.523, P < 0.05). The results indicate that arsenic and antimony in antimony mining area may significantly affect human health.     

Microanalysis of the antiretroviral nevirapine in human hair from HIV-infected patients by liquid chromatography-tandem mass spectrometry

Sufficient drug exposure is crucial for maintaining durable responses to HIV treatments. However, monitoring drug exposure using single blood samples only provides short-term information and is highly subject to intra-individual pharmacokinetic variation. Drugs can accumulate in hair over a long period of time, so hair drug levels can provide drug exposure information over prolonged periods. We now report on a specific, sensitive, and reproducible liquid chromatography-tandem mass spectrometry method for measuring nevirapine (NVP), a widely used antiretroviral drug, levels in human hair using even a single short strand of hair. Hair samples are cut into small segments, and the drug is extracted in methanol/trifluoroacetic acid (v/v, 9:1) shaken at 37 °C in a water bath overnight, followed by liquid–liquid extraction under alkaline conditions. The extracted samples are then separated on a BDS-C₁₈ column with a mobile phase composed of 50% acetonitrile containing 0.15% acetic acid and 4 mM ammonium acetate with an isocratic elution for a total run time of 3 min and detected by triple quadrupole electrospray multiple reaction mode at precursor/product ion at 267.0 > 225.9 m/z. Deuterated nevirapine-d5 was used as an internal standard. This method was validated from 0.25 to 100 ng/mg using 2 mg hair samples. The accuracies for spiked NVP hair control samples were 98–106% with coefficients of variation (CV) less than 10%. The CV for incurred hair control samples was less than 7%. The extraction efficiency for incurred control hair samples was estimated at more than 95% by repeated extractions. This method has been successfully applied to analyze more than 1,000 hair samples from participants in a large ongoing cohort study of HIV-infected participants. We also showed that NVP in human hair can easily be detected in a single short strand of hair. This method will allow us to identify drug non-adherence using even a single strand of hair.     

Limitations of the scalp-hair biologic monitor in assessing selenium status in epidemiological investigations

Scalp hair is routinely used to assess exposure to toxic trace elements and nutritional status of some required trace elements. The advantages and disadvantages of hair as a biologic monitor have been comprehensively discussed in the literature for many years. Among the concerns is distinguishing between exogenous and endogenous contributions. Nested in this issue is the longitudinal distribution of a trace element along the hair strand. The typical observation for many elements of interest is that the element concentration increases from the root end to the distal end; and this is attributed to continuing contamination from exogenous sources. In this study we used neutron activation analysis to measure 14 trace elements in 6 mm segments of full-length scalp hair from three healthy members of the same household having light-urban environmental exposure. To extend the data set for selenium, we included three adult female subjects with longer than average scalp hair. From these trace-element concentrations we calculated the root-to-distal end ratios as a profile diagnostic of trace-element distributions. Ratios fall into three diagnostic categories, >1, ≈1, and <1 corresponding to profiles having decreasing root-to-distal concentrations, unchanging concentrations, and increasing concentrations, respectively. Of the 14 elements measured, only Se has R > 1, Zn and S have R ≈ 1, and the remaining 11 elements all have R < 1 in the order: As > I > Hg ≈ Au ≈ Mg ≈ Mn ≈ Sb ≈ Ca > Cu > Al ≈ Ag. R _Se is greater than 1 and increases with hair length (P = 0.02) corresponding to a continuous longitudinal loss of Se in stark and puzzling contrast to the other elements measured. An analogous loss of Se in the nail monitor was not observed leading us to conclude that the nail is less prone to misclassification of selenium status in epidemiological studies.     

Exploring lead exposure in ancient Chilean mummies using a single strand of hair by laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS)

Lead exposure has received increased attention over the past few decades since it has been shown to have adverse effects on physical and intellectual development in humans. The use of biological tissues such as blood, teeth, hair, and bone for assessment of lead exposure has been previously demonstrated. While analysis of blood for trace metals provides information concerning recent exposure, hair offers insight into a period of several months, and is preferable since it is non-invasively collected and easily stored. The present study analyzed total of 49 ancient hair samples for lead (Pb_H) using LA-ICP-MS. Samples belonged to an ancient fisher hunter–gatherer culture called the “Chinchorro,” and who occupied regions of the Atacama Desert on the northern coast of Chile from approximately 5000–1500 B.C. and practiced the first-known form of artificial mummification. Several samples from a post-Chinchorro agricultural community (n = 12) ca. 1000–1400 A.D. were also analyzed. A suite of hair standards was developed using contemporary hair from the same region and was subsequently used to make linear calibration functions for lead determination in single strands of hair using LA-ICP-MS. Three linear scans ranged from 500 to 1000 μm were performed for each sample and signal intensities were normalized over ¹³C. The distribution of lead in the central medulla in a 100 μm cross-section scan of hair strand demonstrated minimal exogenous contamination. Hair lead (Pb_H) concentrations ranged between 2.2 μg/g and 12.8 μg/g could be accurately quantified with these standards. Twenty one out of 49 samples (43%) showed Pb_H concentrations higher than the average value of 5 μg/g for unexposed individuals (range 1.1–228.0 μg/g). Median hair lead concentrations by burial sites and are shown in order of decreasing concentration: Morro (13.8 μg/g) > Iquique (6.6 μg/g) > Azapa (4.5 μg/g) > Yungay (4.1 μg/g) > Camarones (2.7 μg/g). Most of the burial sites showed Pb_H concentrations greater than the normal value for unexposed individuals and outliers heavily influenced average concentrations. The results suggest that the Chinchorro and later agro-pastoral populations were not widely exposed to naturally elevated lead.     

Determination of trace elements in scalp hair of an elderly population by neutron activation analysis

Neutron activation analysis was applied to assess trace elements concentrations in head hair from healthy elderly people living in the São Paulo metropolitan area. Concentrations of As, Br, Ca, Cl, Co, Cr, Cu, Fe, K, La, Mn, Na, Sb, Se and, Zn were determined. Comparisons were made between the results obtained for dyed and non-dyed hair as well as for hair from females and males of two different age groups. The results were also compared with range values established by clinical laboratories and published data.     

Determination of trace element levels in human scalp hair in occupationally exposed subjects by XRF

Trace element levels in hair of individuals living in urban areas were determined by energy dispersive XRF. Two groups of subjects were investigated, the first group was assumed to be from a healthy environment, the other one was exposed to a high level of contamination due to working conditions. The results were compared to data reported in the literature. The concentrations of Ca, Fe, Cu, Zn and Pb in the scalp hair were determined and the correlation between hair trace element levels and environmental effects was discussed. The results given by the second group show that environmental exposure effects hair trace element levels which are related to body trace element concentrations.     

Fast ultrasound-assisted extraction of copper,iron, manganese and zinc from human hair samples prior to flow injection flame atomic absorption spectrometric detection

A dynamic ultrasound-assisted extraction procedure utilizing diluted nitric acid was developed for the determination of copper, iron, manganese and zinc in human hair taken from workers in permanent contact with a polluted environment. The extraction unit of the dynamic ultrasound-assisted extraction system contains a minicolumn into which a specified amount of hair (5–50 mg) is placed. Once inserted into the continuous manifold, trace metals were extracted at 3 mL min⁻¹ with 3 mol L⁻¹ nitric acid under the action of ultrasound for 2 min for zinc and 3 min for copper, iron and manganese determination, and using an ultrasonic water-bath temperature of 70 °C for zinc and 80 °C for copper, iron and manganese determination. The system permits the direct analysis of hair and yields concentrations with relative standard deviations of <3% (n = 11). The applicability of the procedure was verified by analysing human hair samples from workers exposed to welding fumes, and its accuracy was assessed through comparison with a conventional sample dissolution procedure and the use of a certified reference material (BCR 397, human hair).     

Quantification of testosterone undecanoate in human hair by liquid chromatography–tandem mass spectrometry

Testosterone undecanoate (T‐C11) can be used by athletes in order to improve performance. After oral intake, T‐C11 is rapidly metabolized, hampering discrimination between exogenous and endogenous testosterone. A possible alternative is to detect the intact ester in hair. A method based on liquid chromatography–tandem mass spectrometry was developed for the determination of T‐C11 in hair. The sample procedure consisted of digestion of 200 mg of pulverized hair with tris(2‐carboxyethyl)phosphine hydrochloride and liquid–liquid extraction with n‐pentane. Several parameters such as the mobile phase, the ionization source and the washing step were optimized. The method was validated at different spiked levels obtaining satisfactory values for accuracy (between 92 and 102%) with relative standard deviations lower than 7% and a limit of detection of 0.2 ng/g. The applicability of the method was checked by the analysis of three samples from patients using T‐C11. A peak for the analyte was detected in all samples with concentrations between 0.4 and 8.4 ng/g. Copyright © 2009 John Wiley & Sons, Ltd.     

Chiral analyses of dextromethorphan/levomethorphan and their metabolites in rat and human samples using LC-MS/MS

In order to develop an analytical method for the discrimination of dextromethorphan (an antitussive medicine) from its enantiomer, levomethorphan (a narcotic) in biological samples, chiral analyses of these drugs and their O-demethyl and/or N-demethyl metabolites in rat plasma, urine, and hair were carried out using LC-MS/MS. After the i.p. administration of dextromethorphan or levomethorphan to pigmented hairy male DA rats (5 mg/kg/day, 10 days), the parent compounds and their three metabolites in plasma, urine and hair were determined using LC-MS/MS. Complete chiral separation was achieved in 12 min on a Chiral CD-Ph column in 0.1% formic acid–acetonitrile by a linear gradient program. Most of the metabolites were detected as being the corresponding O-demethyl and N, O-didemethyl metabolites in the rat plasma and urine after the hydrolysis of O-glucuronides, although obvious differences in the amounts of these metabolites were found between the dextro and levo forms. No racemation was observed through O- and/or N-demethylation. In the rat hair samples collected 4 weeks after the first administration, those differences were more clearly detected and the concentrations of the parent compounds, their O-demethyl, N-demethyl, and N, O-didemethyl metabolites were 63.4, 2.7, 25.1, and 0.7 ng/mg for the dextro forms and 24.5, 24.6, 2.6, and 0.5 ng/mg for the levo forms, respectively. In order to fully investigate the differences of their metabolic properties between dextromethorphan and levomethorphan, DA rat and human liver microsomes were studied. The results suggested that there might be an enantioselective metabolism of levomethorphan, especially with regard to the O-demethylation, not only in DA rat but human liver microsomes as well. The proposed chiral analyses might be applied to human samples and could be useful for discriminating dextromethorphan use from levomethorphan use in the field of forensic toxicology, although further studies should be carried out using authentic human samples.     

Trace elements in the scalp hair of patients with alcohol induced psychosis

A number of essential trace elements play a major role in various metabolic pathways and in many diseases like autoimmune, neurological and psychiatric. This study is undertaken with an aim to evaluate the levels of different trace elements in the scalp hair samples of patients suffering from alcohol induced psychosis by particle induced X-ray emission technique (PIXE). It is observed that Fe (p?<?0.0005), Cu (p?<?0.001) are significantly higher in the hair samples of alcohol induced psychosis patients compared to those in normals while concentrations of Mn (p?<?0.005) and Zn (p?<?0.0001) are lower. The concentrations of Co and Ni in the hair samples are found to be in consonance with the concentrations in the normals.