Do shrooms show up in a hair follicle test

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Clean-up of the hair extracts is performed with solid phase extraction using a mixed-mode cation exchanger. Extracts are measured with liquid chromatography coupled with electrospray tandem mass spectrometry. These fortified hair samples were used for method development and can be employed as quality controls.

Hair analysis is a useful tool for retrospective drug screening, for example in the context of workplace drug testing or tests of driving ability. In Germany and Italy, it is part of the procedure for granting or re-granting the driver's because drug abuse is incompatible with the ability to drive 1 , 2. Hair analysis can also be used if an incident happened some time ago and no other sample material other than hair is available. Furthermore, the time and length of drug intake can be estimated by correlating the position of the analytes in the hair samples with the growth rate of hair 1.

Lysergic acid diethylamide LSD and psilocin are hallucinogenic substances that are consumed mostly for recreational purposes. In some countries, it is consumed as a drug 5. LSD, psilocin and bufotenine are controlled substances in many countries. Up until now no methods have been published for analyzing psilocin or bufotenine in hair and it is therefore not known if these substances are incorporated in hair at all. Furthermore, LSD was found in pubic hair two times with concentrations of 0. This article describes a method to extract psilocin, bufotenine and LSD as well as some of its metabolites from hair.

Furthermore, hair was soaked with the analytes to incorporate them into the hair and thus prepare fortified hair that was used for method development and quality control. Then, 1 g dark blond, blank hair was stored in 20 mL of this solution under a nitrogen atmosphere protected from light for 7 days. Then, the solution was decanted and the hair was rinsed nine times with 20 mL methanol. Finally, the hair was dried at room temperature. The mobile phase consisted of acetonitrile A and 2 mM ammonium acetate buffer B , both with 0.

For decontamination hair samples were washed with 20 mL methanol. It was decanted and the hair was washed in 20 mL acetone, which was decanted as well. After drying at room temperature, the hair was cut in pieces of 1—3 mm length. Extraction and clean-up were performed and protected from light. The samples were centrifuged at 4, rpm for 2 min, and the supernatant was decanted into a new test tube. Furthermore, 1 mL of 0. The test tube was centrifuged for 2 min, the supernatants were combined and 4 mL phosphate buffer 0. Clean-up of the hair extracts was performed with SPE. After sample application, the columns were washed with 2 mL water and 2 mL acetonitrile, followed by a drying step of 5 min in a stream of nitrogen.

Then, the columns were washed with 2 mL methanol and 2 mL ethyl acetate and dried again for 5 min with nitrogen. It was essential to use polypropylene needles instead of stainless steel needles for elution. For validation, drug-free, dark blond hair was used. Five calibration curves consisting of seven calibrators were prepared to determine the linearity by spiking 50 mg blank hair with analyte at concentrations of 0.

Blank hair was spiked with a high and a low concentration of all analytes see Table I. In addition, precision was determined by analyzing the fortified hair samples. Quality controls of each concentration were extracted in duplicate on five different days. In hair analysis, it is more difficult to find suitable quality controls to check the extraction efficiency than in the analysis of homogenous body fluids.

One possibility is to use pools of authentic, positive samples of drug users. Because the consumption of LSD, psilocin and bufotenine is rare, no hair samples are available. Water was substituted by methanol because psilocin is unstable in aqueous solutions, besides methanol swells hair. DMSO is able to penetrate the cell membrane, so the analytes can migrate into the hair. All analytes had been incorporated into the hair.

The concentrations measured with the developed method are shown in Table II. This is the first time that the preparation of reference material for psilocin, bufotenine and LSD and metabolites thereof in hair has been published. The fortified hair was subsequently used for method development and precision studies. Method development in hair analysis is more challenging than that in body fluids because the extraction efficiency cannot be determined by spiking blank samples.

In preliminary tests with spiked hair, the stability of the analytes under different extraction conditions was checked, because it is known that LSD and especially psilocin are relatively labile analytes. Common approaches in hair analysis were tested and evaluated Table III. However, the extractions with 0. Although the more polar analytes bufotenine, psilocin and O-H-LSD were extracted better with hydrochloric acid whereas methanol was better for LSD Figure 1 , a mixture of 0.

Furthermore, the addition of ascorbic acid prior to the extraction stabilized psilocin and bufotenine. Comparison of different extraction solutions. MeOH, methanol; HCl, 0. The duration of the extraction in the ultrasonic bath was tested from 1 to 6 h. Extraction times longer than 6 h were not tested as this is the longest period with that the extraction and the following SPE of the extracts can be done during one working day.

This is essential as psilocin and LSD are labile analytes and the risk of degradation is too high if the extracts have to be stored overnight. When comparing extraction times from 1 to 6 h, the highest amounts for all analytes were obtained after 6 h Figure 2.

Further clean-up was required due to impurities present in the hair extracts. For this purpose, the SPE procedure for the analytes in body fluids recently published was modified As matrix components extracted from hair are mostly lipophilic, the hydrophilic washing step with phosphate buffer was replaced by a lipophilic washing step with acetonitrile to reduce matrix effects. Bias ranged from —0. The precision data of the spiked blank hair and the fortified hair are comparable. This means that the fortified hair represents a homogenous sample pool and the analytes can be extracted reproducibly.

A possible epimerization of LSD to iso -LSD and vice versa during the extraction procedure was examined by extracting samples that contained only one of the analytes and checking whether the other one had been formed. No epimerization occurred. This is in accordance with the literature where it has been reported that epimerization occurs at elevated temperatures only under basic conditions 15 , whereas the extraction of hair samples was under acidic conditions.

We present a method to analyze psilocin, bufotenine, LSD and its metabolites in hair samples after extraction with methanol and hydrochloric acid and purification of the extract with SPE. This method might be a useful tool in the analysis of hallucinogens in hair. For the first time, hair fortified with these analytes has been prepared, which can be used as a quality control. Google Scholar. Google Preview. 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.

In. Advanced Search. Search Menu. Skip Nav Destination Article . Close mobile search Article . Volume Article Contents Abstract. and discussion. Conflict of interest. Article . Institute of Legal Medicine. : rafaela. Oxford Academic. Angela Gasse. Heidi Pfeiffer. Select Format Select format. Permissions Icon Permissions. Bufotenine Low 0. Open in new tab. Bufotenine 1.

Extraction method. Phosphate buffer Figure 1. Open in new tab Download slide. Figure 2. Bufotenine 10 16 Psilocin 10 22 LSD 0. State of the art in hair analysis for detection of drug and alcohol abuse. Google Scholar Crossref. Search . Google Scholar PubMed. Potentially hallucinogenic 5-hydroxytryptamine receptor ligands bufotenine and dimethyltryptamine in blood and tissues. Interlaboratory comparison studies on the analysis of hair for drugs of abuse. Two new standard reference materials for the determination of drugs of abuse in human hair. Published by Oxford University Press. All rights reserved.

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Do shrooms show up in a hair follicle test

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