Detection and validated quantification of 21 benzodiazepines and 3 "z-drugs" in human hair by LC-MS/MS

A method for detection and quantification of 21 benzodiazepines and the pharmacologically related "z-drugs" in human hair samples was developed and fully validated using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). After methanolic and methanolic/aqueous extraction, the analytes were separated using two different LC-MS systems (AB Sciex 3200 QTRAP and AB Sciex 5500 QTRAP). Separation columns, mobile phases and MS modes for both systems were: Phenomenex Kinetex, 2.6 μm, 50/2.1; 5mM ammonium formate buffer pH 3.5/methanol, total flow 0.75 mL/min; electrospray ionization (ESI), multiple reaction monitoring (MRM), information dependent acquisition (IDA), enhanced product ion scan (EPI). The assays were found to be selective for the tested compounds (alprazolam, 7-aminoclonazepam, 7-aminoflunitrazepam, bromazepam, chlordiazepoxide, clonazepam, N-desalkylflurazepam, diazepam, flunitrazepam, flurazepam, alpha-hydroxymidazolam, lorazepam, lormetazepam, midazolam, nitrazepam, nordazepam, oxazepam, phenazepam, prazepam, temazepam, triazolam, zaleplon, zolpidem and zopiclone), all validation criteria were in the required ranges according to international guidelines, except for bromazepam. Matrix effects, and process efficiencies were in the acceptable ranges evaluated using the post-extraction addition approach. Lower limits of quantification were between 0.6 and 16 pg/mg of hair. The LC-MS/MS assay has proven to be applicable for determination of the studied analytes in human hair in numerous authentic cases (n=175).     

Accumulation of explosives in hair

The sorption of explosives (TNT, RDX, PETN, TATP, EGDN) to hair during exposure to their vapors is examined. Three colors of hair were simultaneously exposed to explosive vapor. Following exposure of hair, the sorbed explosive was removed by extraction with acetonitrile and quantified. Results show that sorption of explosives, via vapor diffusion, to black hair is significantly greater than to blond, brown or bleached hair. Furthermore, the rate of sorption is directly related to the vapor density of the explosive: EGDN > TATP >TNT > PETN > RDX. In some cases, the explosive-containing hair was subject to repeated washings with sodium dodecylsulfate or simply left out in an open area to determine the persistence of the explosive contamination. While explosive is removed from hair with time or washing, some persists. These results indicate that hair can be a useful indicator of explosive exposure/handling.     

毛发中毒品分析

毛发分析在法庭毒品分析领域有其独特的优势,很多国家的法化学实验室,毛发分析已成为毒品检测的常规操作,并已得到了法庭的承认、采纳。本文对毒品进入毛发的机制、毛发的现场勘查、毛发中毒品分析程序、毛发中毒品分析结果的评判进行了综述。简单地介绍了毛发在现场勘查中采取、包装、送检的基本方法和技术人员在操作过程中的注意事项,以及针对毛发检验中的特殊技术处理;另外,介绍了毛发中毒品分析的特点,通过分析毛发毒品的药理机制,总结出了一些高效、便利、快速的毒品分析方法,并对各种方法进行了介绍,得出了毛发中毒品分析结果的一些特点。     

Opiate levels in hair

By means of radioimmunoassay-technique, hair samples of users, drug related fatalities, carcinoma patients receiving morphine and of experimental guinea pigs receiving codeine were investigated for opiates. The RIA-investigations require a minimum of material; our routine procedures need only 50 mg of hair. No correlation existed between administered doses of opiates and their concentrations in hair of both human and experimental animals. By sectioning the hair, the approximate period of drug use in man could be detected. However, these findings could not be confirmed by the animal experiments. The growth rate of the hair, diffusion and adhesion processes may influence the transport of drugs along the hair. External contaminations and washing procedures were shown to increase or diminish the drug concentration of the samples.     

Determination of hair group by preserved hair cell bulbs

Difficulties appearing in determining hair group were the reason for conducting this research for the purpose of finding additional possibilities to solve the mentioned problem. It was established that the preliminary cell coloring, related with determination of hair sex, does not influence a consequent detection of antigens. Methods or the fixation of material were selected. The most suitable reagents and their titers as well as different time periods of absorption for detecting antigens A and B are offered. All stages of examination are described in detail.     

Statistical examination of hair color as a potential biasing factor in hair analysis

We review eight different data sets in this paper for the purposes of assessing the possibility that reported color of hair can produce a systematic bias in the interpretation of hair assays. We review studies or data sets that include heroin and its metabolites, cocaine and its metabolites, MDMA and its analogs, and amphetamine and methamphetamine. The studies have utilized a variety of different degrees of color categorization, ranging from the simple dichotomy of brown and black, to a high of 12 categories. The mean number of categories reported approaches 6 (mean = 5.875). There are a total of 2791 data points in this analysis. We utilize two major statistical techniques for assessing significance; one-way analysis of variance, and Tukey's Honestly Significant Difference procedure. In circumstances were only dichotomous contrasts are possible, one-way analysis of variance is used. In contrasts involving three or more categorical groups, Tukey's procedure is used. In circumstances where the homogeneity of group variances is not sustained by the Levene statistic, we use the Tamahane procedure, allowing an assessment that assumes unequal variances. The analysis of this data fails to discern a significant color effect. We speculate that it may be that variance is large in many domains affecting analyte recovery from hair. In large groups these variations tend to regress towards a typical or mean value. Thus the data here show that while there are group or aggregate differences in these 'typical' values, they are not great when considered in relation to the within-group variations which exist for those values. It is our view that color may play a role in the accumulation of drugs in hair, however it is likely to account for only a very small part of the complex process of drug accumulation.     

Methadone concentrations in human hair of the head, axillary and pubic hair

The concentrations of methadone and its metabolites in the hair of the head, axillary and pubic hair obtained from patients receiving a daily maintenance doses, were determined. Comparison of the concentrations provides the highest values in the axillary hair, followed by pubic hair and the hair of the head.     

Tetrahydrocannabinols in hair of hashish smokers

The study investigates the presence of tetrahydrocannabinols in the head hair and the pubic and axillary hair. The hair samples were obtained from hashish smokers. The concentrations were determined by radioimmunoassay and reflect total tetrahydrocannabinols and metabolites. The values found ranged from 0.4 ng/mg hair up to 3.8 ng/mg hair. The presence of the drug in the hair samples was also demonstrated by GC/MS.     

Theoretical limits of the evaluation of drug concentrations in hair due to irregular hair growth

When examining concentration relationships of doses it must be taken into account that hair growth is irregular. Hair growing from the shaved skin after a single dose of a certain drug cannot possibly contain the same concentration as hair after the same dose that has not been cut over a long period. Concentrations can even change during the hair growth in cases where the hair had been cut a couple of months before the hair sample was taken. The variations in the expected concentrations can exceed 20%. On the other hand, the evaluation of a hair tuft which has grown after the last drug consumption may be important in forensic cases where the hair which has grown earlier is not available. This may lead to misinterpretations at low concentrations. Expected concentrations are calculated assuming a telogen part of 10%.     

Effect of hair care and hair cosmetics on the concentrations of fatty acid ethyl esters in hair as markers of chronically elevated alcohol consumption

Fatty acid ethyl esters (FAEE) can be used as alcohol markers in hair. It was investigated in this study whether this diagnostic method is disturbed by hair care and hair cosmetics. Traces of ethyl myristate, ethyl palmitate, ethyl oleate and ethyl stearate were detected in all of 49 frequently applied hair care products by headspace solid phase microextraction (HS-SPME) and gas chromatography-mass spectrometry (GC-MS). The highest concentration was 0.003% in a hair wax. From experiments with separated hair samples of alcoholics as well as from the evaluation of the FAEE concentrations and the data about hair care of 75 volunteers (alcoholics, social drinkers and teetotalers) follows that usual shampooing, permanent wave, dyeing, bleaching or shading are of minor importance as compared to the drinking amount and other individual features. However, false positive results were found after daily treatment with a hair lotion containing 62.5% ethanol, with a deodorant and with a hair spray. As an explanation, it is assumed that FAEE are formed in the sebum glands also after regular topical application of products with a higher ethanol content.     

Value of hair analysis in postmortem toxicology

It is generally accepted that chemical testing of biological fluids is the most objective means of diagnosis of drug use. The presence of a drug analyte in a biological specimen can be used to document exposure. The standard in postmortem drug testing is a general unknown screening, followed by the gas chromatographic/mass spectrometric confirmation conducted on a whole blood sample. In recent years, remarkable advances in sensitive analytical techniques have enabled the analysis of drugs in unconventional biological specimens such as hair. The advantages of this sample over traditional media, like urine and blood, are obvious: collection is almost non-invasive, relatively easy to perform, and in forensic situations it may be achieved under close supervision of law enforcement officers to prevent adulteration or substitution. Moreover, the window of drug detection is dramatically extended to weeks, months or even years. The aim of this review is to document the current status of hair analysis in postmortem toxicology.     

度冷丁在豚鼠毛发中的分布研究

目的探讨度冷丁在毛发中的分布状况。方法通过动物实验-给豚鼠持续腹腔注射度冷丁,探讨度冷丁在豚鼠毛发中出现与消失的时间过程及注射剂量与毛发中的含量关系。结果给豚鼠隔日连续腹腔注射度冷丁7次,第4天可在豚鼠毛发中检出度冷丁,第8天达浓度高峰,停止注射后第5天检不出度冷丁;豚鼠毛发中度冷丁含量与度冷丁的注射剂量有关,但无明显的线性关系。结论研究结果为实际案例分析结果的评价提供依据。     

Determination of chloroquine and monodesethylchloroquine in hair

Using thin-layer and gas chromatography and mass spectrometry, chloroquine and its major metabolite (monodesethylchloroquine) were identified in hair samples of numerous patients who received this antimalarial drug for several months. In two patients the amounts of chloroquine were, respectively, 310 and 145 mg/kg hair and those of the monodesethylchloroquine 23 and 11 mg/kg. The respective proportions (93 and 7%) are the same in the two subjects. The chloroquine percentage was near those in the spleen or stomach wall after poisoning. Other metabolites in hair are being identified. Hair analysis may provide a good toxicologic and forensic science complement to the blood, urine, and tissues. It may be useful for the control of chloroquine therapy.     

Weather-induced changes in cannabinoid content of hair

Authentic hair samples from Cannabis users and a drug free hair sample which was separately spiked with tetrahydrocannabinol (THC), cannabidiol (CBD) or cannabinol (CBN) were exposed outside as well as to natural sunlight at prevailing and elevated humidity in quartz glass tubes during 8 weeks. In addition, authentic and spiked hair samples were exposed to xenon arc radiation in a light exposure cabinet for 24 hours. Stability of THC, CBD and CBN in authentic samples differed from that of the spiked hair. The radiation experiment revealed that CBN could not be measured in hair which had been spiked with THC. Under all conditions chosen the concentrations of THC, CBD and CBN decreased. At high humidity the concentrations declined more rapidly. In both authentic and spiked samples THC was most unstable compared to CBD and CBN. Therefore, in hair analysis determination of CBD and CBN seems promising to detect Cannabis exposure even under unfavorable conditions.     

Structure of the hair in thallium poisoning

The alterations in the hair roots discovered by Widy in 1956 were interpreted as accumulations of pigment which form as a consequence of a catalytic action of the poison. The goal of the present investigation was to clarify the structure of these inclusions. Hair from the heads of six victims who were involved in the poisoning of Würzburg medical students in January 1983 was available as investigation material. In the investigation in transmitted light and in polarized light, the black zones typical for thallium intoxication were found filling to varying extents the root and hair shaft near the root. Their intensity corresponded to the degree of severity of the intoxication. When examined under reflected light, the inclusions were shown up with a white color. They thus showed the same optical behavior as the air-filled medullary strand of normal hairs. This indicates that gaseous constituents are involved. This hypothesis could be confirmed by further investigations. After mechanical damage to the hair (pressing under high pressure), the gaseous inclusions disappeared and with them the "thallium strip." The same effect was attained by the chemical action of various acids, embedding agents, and dye solutions. This process was especially rapid after exposure to thioglycolic acid; the escape of the gas bubbles can be directly observed here. Scanning electron microscopic investigations on transverse sections of hair revealed a loosening of the spindle-shaped elements of the fiber layer as signs of structural disturbance. The gaseous constituents in thallium hair arise as the result of a trophic disorder in keratin formation. The structural alteration due to this leads to alopecia.