Psychotropic drugs in developmental mushrooms: a case study review

Psilocyn and psilocybin can be identified in different stages of developing psilocybe mushrooms. Knowing the various stages of the mushroom development can be useful when receiving evidence from illicit mushroom growing operations. Exhibits from three separate cases were submitted to the drug analysis section of the Minnesota Bureau of Criminal Apprehension Forensic Science Laboratory. Each case contained different stages of developing mushrooms. This report describes the evidence in each case, the sample preparation, the sample analysis and the final report that was written.     

Detecting psychoactive drugs in the developmental stages of mushrooms

The following questions regarding the detection of psychoactive drugs in mushrooms are addressed: At what stage of the mushroom development can the psychoactive drugs psilocyn and psilocybin be identified, and what effect does light have on the growth of these mushrooms. To answer these questions, Psilocybe cyanescens Wakefield mushrooms were grown from their spores in a controlled setting. At various times of their development, samples were taken and analyzed for psilocyn and psilocybin. Knowing what stage of development the psychoactive drugs can be identified may be useful to law enforcement personnel and forensic chemists. Methanolic extracts of various samples were analyzed by TLC and by GC/MS. It was determined that the mycelium knot stage of the mushroom was the earliest stage at which the psychoactive drugs could be detected. It was observed that light affected the time of development and the appearance of these mushrooms.     

Unpredictable Behavior Under the Influence of “Magic Mushrooms”: A Case Report and Review of the Literature

Fatalities implicating psychedelic mushrooms are not a common clinical situation in everyday forensic medicine. Despite classification as an illegal drug in many countries, psilocybin mushrooms have the reputation of being safe. We report the case of a young man who jumped from a second story balcony under the influence of psilocybin mushrooms. The psilocin assay was performed by gas chromatography coupled to an electron‐impact ionization time‐of‐flight detector (GC‐EI‐TOF) after solid‐phase extraction. Total psilocin was quantified in peripheral and cardiac blood as 60 and 67 ng/mL, respectively, and in urine (2230 ng/mL), bile (3102 ng/mL), and vitreous humor (57 ng/mL). This case report and review of literature highlights the danger of psilocybin mushrooms. Isolated use of psilocybin mushrooms by a regular consumer without psychiatric history, even under “safe” circumstances, can lead to a fatal outcome.     

Amanita phalloides mushroom poisoning: a cluster of four fatalities

A group of four illegal aliens had been without food for several days when they found a group of wild mushrooms growing in a field in Southern California. Each man consumed a meal of one to six fried mushrooms. Two days after eating the mushrooms, all four men developed abdominal pain, nausea, diarrhea, and intractable vomiting. Three days after consuming the mushrooms, all four were hospitalized and their clinical courses rapidly deteriorated to refractory hepatorenal failure and coma. Three of the victims died three days after admission to the hospital and the fourth died eight days after admission. The autopsy findings are presented and the mechanism of Amanita phalloides mushroom poisoning is discussed.     

赛洛新和赛洛西宾分析方法研究进展

致幻剂是一类低剂量就使人意识、知觉和视觉异常,引起幻觉和情绪障碍的精神活性物质.赛洛新和赛洛西宾是一类具有致幻作用的色胺类化合物,存在于裸盖伞属和斑褶菇属的致幻蘑菇中.当前滥用及娱乐性使用含此类物质的致幻蘑菇的现象日趋严重,由此引起的危害公共安全案(事)件不断增多.因此,对于致幻蘑菇滥用的鉴定是当前司法鉴定领域的重要需...     

A concept study on identification and attribution profiling of chemical threat agents using liquid chromatography-mass spectrometry applied to Amanita toxins in food

Accidental or deliberate poisoning of food is of great national and international concern. Detecting and identifying potentially toxic agents in food is challenging due to their large chemical diversity and the complexity range of food matrices. A methodology is presented whereby toxic agents are identified and further characterized using a two-step approach. First, generic screening is performed by LC/MS/MS to detect toxins based on a list of selected potential chemical threat agents (CTAs). After identifying the CTAs, a second LC/MS analysis is performed applying accurate mass determination and the generation of an attribution profile. To demonstrate the potential of the methodology, toxins from the mushrooms Amanita phalloides and Amanita virosa were analyzed. These mushrooms are known to produce cyclic peptide toxins, which can be grouped into amatoxins, phallotoxins and virotoxins, where α-amanitin and β-amanitin are regarded as the most potent. To represent a typical complex food sample, mushroom stews containing either A. phalloides or A. virosa were prepared. By combining the screening method with accurate mass analysis, the attribution profile for the identified toxins and related components in each stew was established and used to identify the mushroom species in question. In addition, the analytical data was consistent with the fact that the A. virosa specimens used in this study were of European origin. This adds an important piece of information that enables geographic attribution and strengthens the attribution profile.     

Analysis of hallucinogenic constituents in Amanita mushrooms circulated in Japan

The constituents of seven mushrooms sold as Amanita muscaria or Amanita pantherina (five A. muscaria and two A. pantherina) and four "extracts purported to contain A. muscaria" products that are currently circulated in Japan were determined. All mushroom samples were identified as A. muscaria or A. pantherina by macroscopic and microscopic observation. The dissociative constituents, ibotenic acid (IBO) and muscimol (MUS), were extracted with 70% methanol twice and determined by gas chromatography/mass spectrometry. The IBO (as the hydrate)/MUS contents were in the range of <10-2845ppm/46-1052ppm in the cap of A. muscaria and 188-269ppm/1554-1880ppm in the cap of A. pantherina. In the caps, these compounds had a tendency to be more concentrated in the flesh than in the cuticle. On the other hand, the IBO/MUS contents in the stem were far lower than in the caps. In the "extracts purported to contain A. muscaria" products, IBO/MUS were detected below the lower limit of calibration curve (<10ppm/<25ppm) or not detected. However, these samples contained other psychoactive compounds, such as psychoactive tryptamines (5-methoxy-N,N-diisopropyltryptamine and 5-methoxy-N,N-dimethyltryptamine), reversible monoamine oxidase inhibitors (harmine and harmaline) and tropane alkaloids (atropine and scopolamine), which were not quantified. This is the first report of the chemical analysis of Amanita mushrooms that are circulated in the drug market.     

Morphological and chemical analysis of magic mushrooms in Japan

Morphological and toxicological analyses were performed on hallucinogenic mushrooms that are currently circulated in Japan. Scanning electron microscope (SEM) indicated a three-dimensional microstructures in the mushrooms. The complementary use of SEM with an optical microscope was effective for observing characteristic tissues, such as basidiomycetes, spores, cystidia and basidia. Hallucinogenic alkaloids were extracted with methanol and determined by high performance liquid chromatography (HPLC) with a UV detector set at 220 nm. The psilocin/psilocybin contents in Psilocybe cubensis were in the range of 0.14-0.42%/0.37-1.30% in the whole mushroom (0.17-0.78%/0.44-1.35% in the cap and 0.09-0.30%/0.05-1.27% in the stem), respectively. The hallucinogenic alkaloids in Copelandia were 0.43-0.76%/0.08-0.22% in the whole mushroom (0.64-0.74%/0.02-0.22% in the cap and 0.31-0.78%/0.01-0.39% in the stem). It thus appears that P. cubensis is psilocybin-rich, whereas Copelandia is psilocin-rich.     

Desorption atmospheric pressure photoionization-mass spectrometry in routine analysis of confiscated drugs

A comprehensive study was made, where desorption atmospheric pressure photoionization (DAPPI) was applied to the direct analysis of confiscated drugs and pharmaceuticals of various forms and matrices. The analyzed samples included herbal products [Catha edulis (khat), Psilocybe mushrooms, opium and Spice], designer drugs in tablet and powder form [e.g. meta-chlorophenylpiperazine (mCPP), 3-fluoromethamphetamine (3-FMA), methylenedioxypyrovalerone (MDPV) and methylone], and anabolic steroids in oil and tablets. The analyses were performed with ion trap mass spectrometer in MS and MS(2) modes and the obtained spectra were compared with GC-MS results. Contamination of the mass spectrometer was avoided by careful adjustment of the distance of the sample from the mass spectrometer inlet. DAPPI proved to be a fast and specific analysis technique, which does not require any sample preparation, and which therefore suits well to this type of forensic analysis.     

An aqueous-organic extraction method for the isolation and identification of psilocin from hallucinogenic mushrooms

A simple aqueous extraction method for the isolation and identification of psilocin from Psilocybe cubensis mushrooms is reported. This method employs a dephosphorylation of the phosphate ester to psilocin, which facilitates a greater product yield and simplifies identification. Psilocin extracted by this method is sufficiently concentrated and free of cocontaminants to allow identification by infrared spectroscopy and gas chromatography/mass spectrometry.     

The determination of psilocin and psilocybin in hallucinogenic mushrooms by HPLC utilizing a dual reagent acidic potassium permanganate and tris(2,2'-bipyridyl)ruthenium(II) chemiluminescence detection system

This paper describes a procedure for the determination of psilocin and psilocybin in mushroom extracts using high-performance liquid chromatography with postcolumn chemiluminescence detection. A number of extraction methods for psilocin and psilocybin in hallucinogenic mushrooms were investigated, with a simple methanolic extraction being found to be most effective. Psilocin and psilocybin were extracted from a variety of hallucinogenic mushrooms using methanol. The analytes were separated on a C12 column using a (95:5% v/v) methanol:10 mM ammonium formate, pH 3.5 mobile phase with a run time of 5 min. Detection was realized through a dual reagent chemiluminescence detection system of acidic potassium permanganate and tris(2,2'-bipyridyl)ruthenium(II). The chemiluminescence detection system gave improved detectability when compared with UV absorption at 269 nm, with detection limits of 1.2 x 10(-8) and 3.5 x 10(-9) mol/L being obtained for psilocin and psilocybin, respectively. The procedure was applied to the determination of psilocin and psilocybin in three Australian species of hallucinogenic mushroom.     

Detection of psilocin in body fluids

Active compounds of some mushrooms e.g. Psilocybe cubensis, Paneolus subalteatus or Stropharia coronilla, the psychotropic agents psilocybin and psilocin, have hallucinogenic effects. In one case of 'magic mushroom' intake, we had to analyse blood and urine. Psilocin was detected in the urine with REMEDi HS. Most of the psilocin was excreted as the glucuronide. Therefore an enzymatic hydrolysis should be the first step in analysis. Free psilocin was determined at a concentration of 0.23 mg/l while the total amount was 1.76 mg/l urine. The concentration of psilocin in serum was too low for detection with REMEDi HS. We proved a GC-MS-method with d(3)-morphine as internal standard and silylation with MSTFA. Similarly to urine, most of the psilocin in serum was found in the conjugated form. The concentration of free psilocin was 0.018 mg/l, that of total psilocin, 0.052 mg/l serum.     

Chemical and toxicological studies in mushroom poisoning

Two accessory diagnostic tables reflecting the relationship between toxic effect of mushrooms and the nature and chemical structure of toxin have been developed for improving the diagnosis of mushroom poisoning. New color tests have been developed and conditions for isolation and chromatography in a thin adsorbent layer determined. The Schiff's test is specific for Amanita phalloides. This mushroom's toxins can be detected in the viscera after the maximal survival of 50 h, other mushrooms' toxins after 26-30 h.     

The Special Forces Unembellished

"Alpha," "Vitiaz'," "Grad" … Light blue, spotted, and green berets … New special units for carrying out intelligence and sabotage activities and combating terrorism are springing up like mushrooms after a rain.     

Changes in adrenergic and cholinergic innervation of the external genitalia in alcohol and drug intoxication

Eighteen cadavers of persons, who died of acute and chronic alcohol and drug intoxication (ephedron-addiction), were used to study the changes in the spinal nerve plexuses and external genitals. The activity of acetylcholinesterase and of NADPH-diaphorase was determined histochemically; the adrenergic nerve fibers were detected by a glyoxyl-acid technique. Rectified changes in the relative density of adrenal-cholinergic- and nitroxydergic structures and in the activity of mediators in different intoxications were found. The chronic intoxication by alcohol and ephedron was accompanied by a reduced activity of enzymes involved in the metabolism of acetylcholine, nitric oxide and catecholamines. A lower density was observed in the nerve fibers that contained the examined substances. The results can be used as an additional criterion in evaluating the male sexual-function status.     

A shot through the window

At issue in this case was whether an unusual window defect seen in two of the crime scene photographs was due to a bullet and if so, if that same bullet fatally wounded the victim. The window appeared to have been cracked prior to the apparent shot through it. A .22 bullet recovered from autopsy, when examined only by light microscopy, failed to show associated glass fragments. A previously cracked test window was shot a number of times with .22 caliber bullets near the cracks in an effort to simulate the window defect seen in the crime scene photographs. Several of the defects produced by the test window shots appeared similar to the crime scene window defect. The .22 bullet taken from the victim and several of the test bullets (collected by a cotton box) were examined by scanning electron microscopy/energy dispersive X-ray spectroscopy. The test bullets showed glass particles on and embedded in their surfaces. Particles of similar size and composition were found embedded in the surface of the bullet from the victim. The bullet likely struck the window prior to hitting the victim. It was apparent by the morphology of some of the mushroomed test .22 bullets that they hit the window crack. These bullets showed that the glass on one side of a crack often fails before the other side during the strike. Aggregations of powdered glass on many of the mushroomed surfaces of the .22 bullets suggest that as the bullet mushrooms during impact on the window surface, the glass in contact with the bullet powderizes and coats the mushroomed surface of the bullet with a layer of fine glass particles.     

Establishing paternity using minisatellite DNA probes when the putative father is unavailable for testing

A paternity case involving a putative father who had died a few years earlier in an automobile accident was referred to the laboratory for testing. The child and his mother, the deceased's parents, and nine of the deceased's siblings were available for analysis. As previously reported, paternity testing using red blood cell groups, human leukocyte antigens (HLA), red blood cell enzymes, serum proteins, and immunoglobulin allotypes gave a cumulative paternity index of 43,300 and a combined probability of paternity equal to 99.998%. RFLP analysis using Hinf I and Sau 3A single digests and the minisatellite deoxyribonucleic acid (DNA) probes 15.1.11.4 and 6.3 showed no exclusion of paternity and gave nearly conclusive evidence that the putative father was the biological father of the child.     

Variability of cannabinoid findings in blood

Cannabis products have been administered for many centuries; today, cannabis is the most widely used illegal drug all over the world. Nevertheless, the interpretation of cannabis findings in blood with regard to consumption behaviour and/or estimating the elapsed time since the last cannabis use is still a very challenging task. A wide variation of pharmacokinetic parameters has been observed even in experimental studies. Different chemical structures of precursors, smoking dynamics, pyrolysis of phytocannabinoids and frequency of drug use affect the amount of THC absorbed. Polymorphic enzymes are involved in phase-I-metabolism of THC. Pharmacological effects of other cannabinoids or medication on the pharmacokinetics of THC have not yet been studied in detail. Hydrolysis of cannabis conjugates may occur during storage of blood samples and processing of specimens for analysis; knowledge on the stability of cannabinoids in forensic specimens is still poor. Whether determination of cannabinoid conjugates may be useful is a matter of further consideration. At present, the broad variation of pharmacokinetic parameters and the limiting factors discussed in the present paper should be taken into account when using data from experimental studies for interpretation of analytical results in forensic case work.     

美国马里兰州涉及电击枪死亡案例分析

目的 研究美国马里兰州涉及电子武器(电击枪)死亡案件的特点,探讨其法医学鉴定要点.方法 收集2004-2009年美国马里兰州法医局鉴定的涉及电击枪死亡案件13例,案件均经详细的现场调查,尸体进行系统解剖,并进行全面的毒(药)物检测及组织病理学检验.从一般情况、电击枪类型及接触方式、毒物检测结果、死亡方式、死亡原因等方面...     

高校司法鉴定机构应对市场化的初探

随着司法鉴定制度改革的逐步推进,新的司法鉴定机构如雨后春笋般纷纷设立,司法鉴定机构也必将被我国特有的市场经济所覆盖,司法鉴定机构市场化在所难免。高校司法鉴定机构是否能够不被这股强劲的司法鉴定制度改革浪潮所淹没,甚至脱颖而出,取决于高校司法鉴定机构能否在其市场化的过程中整合特有资源,顺应市场,克服难点,发挥优势。