Article

Morphological and chemical analysis of magic mushrooms in Japan

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  • 高知県警察本部
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Abstract

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.

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... Current techniques for the analysis of magic mushrooms can be based on morphological characteristics [4,11], DNA [12][13][14], immunoassay [12] or chemical composition, mainly by chromatographic techniques [15][16][17][18][19][20][21]. ...
... However, analysis of the compounds of magic mushrooms requires sample preparation, namely extraction of the compounds, which may increase the time between harvest and analysis [22][23][24]. Also, the average content of hallucinogenic compounds is usually not higher than 2%, which requires advanced techniques [11,21,25,26]. ...
... While the caps and stalks spectra had more similarities, the gills spectra seemed to be more distinctive. In magic mushrooms, the hallucinogenic compounds are expected to be in higher content in the cap and gills than in stalks [11]. ...
Article
Magic mushrooms are naturally occurring fungi that are considered hallucinogenic drugs because they contain psilocybin and psilocin. These substances are controlled in almost every country in the world, so the use, possession, cultivation, and sale of magic mushrooms are prohibited in whole or in part. Despite this, the abuse of magic mushrooms continues and can put at risk the life of the consumer and society in general if the consumer behaves in an unsafe manner. The number of mushroom species is very high, making it difficult to correctly identify them based only on physical and morphological characteristics. Therefore, there is a need to develop non-destructive mushrooms analysis methods that have less response time and higher discrimination ability. The present work used Attenuated Total Reflectance Fourier Transform Infrared (FTIR-ATR) Spectroscopy to study 64 mushroom samples from different genera including hallucinogenic, edible, and toxic species. In addition, this study used Orthogonal Partial Least Squares - Discriminant Analysis (OPLS-DA), using SIMCA chemometric software to analyse the obtained infrared (IR) spectra. The main molecular vibrations of the components of the fungus were successfully identified by IR spectroscopy. Although the specific bands corresponding to psilocybin or psilocin could not be assigned in the spectra, the regression method was able to discriminate the various species. Hallucinogenic mushrooms were well separated from other species, allowing the method to be used as an initial screening technique to determine whether or not the seized mushrooms are of forensic interest.
... In P cubensis, the psilocin content range is 0.17-0.78% in the cap and 0.09-0.30% in the stem while the psilocybin content range is 0.44-1.35% in the cap and 0.05-1.27% in the stem [4]. Moreover, other authors also reported a higher content of psilocybin when compared to psilocin in P. cubensis [4,5]. Psilocybin, which has entheogenic properties, is found mainly in Psilocybe fungi [5]. ...
... Moreover, other authors also reported a higher content of psilocybin when compared to psilocin in P. cubensis [4,5]. Psilocybin, which has entheogenic properties, is found mainly in Psilocybe fungi [5]. One classic method to produce fungi is through fungal culture [6]. ...
... Scanning electron microscopy images of the spores showed a generally smooth surface and an oval shape, maximum length 10.1 μm and width 6.4 μm (Fig. 2). These values coincide with those reported for the fungus Psilocybe cubensis [5]. ...
Article
In nature, there are >200 species of fungi with hallucinogenic properties. These fungi are classified as Psilocybe, Gymnopilus, and Panaeolus which contain active principles with hallucinogenic properties such as ibotenic acid, psilocybin, psilocin, or baeocystin. In Chile, fungi seizures are mainly of mature specimens or spores. However, clandestine laboratories have been found that process fungus samples at the mycelium stage. In this transient stage of growth (mycelium), traditional taxonomic identification is not feasible, making it necessary to develop a new method of study. Currently, DNA analysis is the only reliable method that can be used as an identification tool for the purposes of supporting evidence, due to the high variability of DNA between species. One way to identify the species of a distinctive DNA fragment is to study PCR products analyzed by real time PCR and sequencing. One of the most popular sequencing methods of forensic interest at the generic and intra-generic levels in plants is internal transcribed spacer (ITS). With real time PCR it is possible to distinguish PCR products by differential analysis of their melting temperature (Tm) curves. This paper describes morphological, chemical, and genetic analysis of mycelia of psychedelic fungi collected from a clandestine laboratory. The fungus species were identified using scanning electron microscopy (SEM), mass spectrometry, HRM analysis, and ITS sequencing. The sporological studies showed a generally smooth surface and oval shape, with maximum length 10.1 μm and width 6.4 μm. The alkaloid Psilocyn was identified by mass spectrometry, while HRM analysis and ITS sequencing identified the species as Psilocybe cubensis. A genetic match was confirmed between the HRM curves obtained from the mycelia (evidence) and biological tissue extracted from the fruiting bodies. Mycelia recovered from the evidence and fruiting bodies (control) were genetically indistinguishable.
... The mushrooms are sold via the internet or in so-called "smart shops". They are supplied in a fresh or air-dried state or as powder incapsules for later use [1,2]. Additionally, "grow-kits" can be purchased which consist of inoculated substrate in plastic boxes. ...
... Most of the hallucinogenic fungi on the market belong to the Psilocybe genus. They mainly contain two hallucinogenic alkaloids, psilocin and its phosphorylated derivative psilocybin [1][2][3][4][5][6][7][8][9] which are controlled substances in many countries. These alkaloids are present at total concentrations of approximately 1-2% in dried mushrooms and the amount of the phosphorylated compound psilocybin usually exceeds the amount of psilocin [1][2][3][4][5][6][7][8][9]. ...
... They mainly contain two hallucinogenic alkaloids, psilocin and its phosphorylated derivative psilocybin [1][2][3][4][5][6][7][8][9] which are controlled substances in many countries. These alkaloids are present at total concentrations of approximately 1-2% in dried mushrooms and the amount of the phosphorylated compound psilocybin usually exceeds the amount of psilocin [1][2][3][4][5][6][7][8][9]. ...
... The content of psilocybin found within commonly used cultivar P. cubensis varies but is on average 1% dry weight. 2,9,10 There has yet to be a validated clinical study focused specifically on the clinical effects of microdosing with psilocybin; published work to date is limited to surveys. [6][7][8]11,12 However, the importance of recreational psilocybin user surveys is not to be discounted. ...
... mushrooms contain psilocybin and its related compounds at approximately 0.2-1.78% of its dry weight. 2,3,9 In a commercial sense, this is a relatively low yield necessitating a significant amount of mushroom biomass in order to extract an economically viable amount of active compound. ...
Article
Psilocybin is a psychoactive alkaloid that is produced naturally by approximately 200 species of mush- rooms. The potential medical use of this molecule for the treatment of mental illness is gaining renewed momentum. As demand grows and clinical trials progress, appropriate methods for producing a quality pharmaceutical product are needed. This review highlights the methods currently available, such as the prominent synthetic method and its biosyn- thetic alternatives, as well as others on the near horizon. This article further seeks to discuss the rapid and evolving nature of the psilocybin industry in the 21st century.
... and Br.) Singer + – USA (Hawaii) [28] Copelandia tropicalis (Ola'h) Singer + – USA (Hawaii) [28] Copelandia sp. + + Japan [46] Galerina Galerina steglichii Besl spec. nov + + Germany [47] Gerronema Gerronema fibula (Bull) Singer + – Germany [48] Gerronema swartzii (Fr.) ...
... + + USA [17, 24, 68] Psilocybe bohemica Sebek + + Czech Republic [18, 19, 57,697071 Psilocybe bohemica Sebek + + Switzerland [21] Psilocybe bonetii Guzman + – Mexico [60] Psilocybe caeruleoannulata Sing + + Brazil [20] Psilocybe caerulescens Murrill + + Brazil [20] Psilocybe caerulipes Peck + + USA [72] Psilocybe candidipes Singer and A.H.Sm + – Mexico [60] Psilocybe copninifacies (Roll). Pouz + – Czech Republic, Slovenia [73] Psilocybe cubensis (Earle) Singer + + Japan, Germany, Mexico, Brazil [20, 46, 61, 74] Psilocybe cyanescens Wakef + + Czech Republic, Switzerland, USA [17, 21, 57, 75] Psilocybe cyanofibrillosa Guzman and Stamets + + USA [22] Psilocybe fimetaria (P.D. Orton) Watling + – United Kingdom [43] Psilocybe hoogshagenii Heim + + Brazil [20] Psilocybe liniformans Guzman and Bas + – USA [21, 22] (continued) converted by the mechanism of dephosphorylation into psilocin (another pharmacologically active compound). Psilocin has the molecular formula C 12 H 16 N 2 O (mol. ...
Chapter
Full-text available
Studies on bioactive alkaloids have gained considerable importance in recent years and have been given prominent position in the field of medicine, both with respect to their biological activity and role played in the introduction of new pharmaceuticals. The biotechnological approach has further enhanced their industrial applications. Some specific groups of bioactive alkaloids have been ignored and not been given much attention because of various reasons, including their legal status. The tryptophan-based indole alkaloids, psilocybin and psilocin, are the best examples in this category which have been identified as most attractive bioactive alkaloids in large number of mushrooms, especially of the genus Psilocybe, but waiting since long for more control studies to ascertain their therapeutic role in some other conditions, apart from psychiatry. These alkaloids have long history of association with mankind for their use as sacraments in religious ceremonies along with medical and recreational purposes and thus need more attention to explore their therapeutic role.
... The two most commonly reported substances used for microdosing among our sample were LSD (median dose 13 mcg LSD; 11.3% of one tab) and psilocybin (median dose 0.3 g of psilocybin mushrooms, presumed to be dried). The psilocybin content of dried Psilocybe cubensis, a common strain of psilocybin-containing mushroom, has been shown to be in the range of 0.37-1.30% of the whole mushroom (Tsujikawa et al., 2003). Taking a value of 1% psilocybin per gram of dried mushroom, we estimate a mean psilocybin dose of approximately 3 mg reported by our sample. ...
... We expect this issue to be present with psilocybin as well, as microdosers generally consume fractions of dried psilocybin mushrooms. Psilocybin content is likely to vary by mushroom species, and the psilocybin content of dried mushroom caps may be greater than in stems (Tsujikawa et al., 2003). This speaks to the importance of dose standardization and the necessity of controlled clinical trials to test the safety and potential benefits of microdosing psychedelics. ...
Article
Rationale Microdosing psychedelics – the practice of consuming small, sub-hallucinogenic doses of substances such as LSD or psilocybin – is gaining attention in popular media but remains poorly characterized. Contemporary studies of psychedelic microdosing have yet to report the basic psychiatric descriptors of psychedelic microdosers. Objectives To examine the practices and demographics of a population of psychedelic microdosers – including their psychiatric diagnoses, prescription medications, and recreational substance use patterns – to develop a foundation on which to conduct future clinical research. Methods Participants ( n = 909; M age = 26.9, SD = 8.6; male = 83.2%; White/European = 79.1%) recruited primarily from the online forum Reddit completed an anonymous online survey. Respondents who reported using LSD, psilocybin, or both for microdosing were grouped and compared with non-microdosing respondents using exploratory odds ratio testing on demographic variables, rates of psychiatric diagnoses, and past-year recreational substance use. Results Of microdosers, most reported using LSD (59.3%; M dose = 13 mcg, or 11.3% of one tab) or psilocybin (25.9%; M dose = 0.3 g of dried psilocybin mushrooms) on a one-day-on, two-days-off schedule. Compared with non-microdosers, microdosers were significantly less likely to report a history of substance use disorders (SUDs; OR = 0.17 (95% CI: 0.05–0.56)) or anxiety disorders (OR = 0.61 (95% CI: 0.41–0.91)). Microdosers were also more likely to report recent recreational substance use compared with non-microdosers (OR = 5.2 (95% CI: 2.7–10.8)). Conclusions Well-designed randomized controlled trials are needed to evaluate the safety and tolerability of this practice in clinical populations and to test claims about potential benefits.
... and Br.) Singer + – USA (Hawaii) [28] Copelandia tropicalis (Ola'h) Singer + – USA (Hawaii) [28] Copelandia sp. + + Japan [46] Galerina Galerina steglichii Besl spec. nov + + Germany [47] Gerronema Gerronema fibula (Bull) Singer + – Germany [48] Gerronema swartzii (Fr.) ...
... + + USA [17, 24, 68] Psilocybe bohemica Sebek + + Czech Republic [18, 19, 57,697071 Psilocybe bohemica Sebek + + Switzerland [21] Psilocybe bonetii Guzman + – Mexico [60] Psilocybe caeruleoannulata Sing + + Brazil [20] Psilocybe caerulescens Murrill + + Brazil [20] Psilocybe caerulipes Peck + + USA [72] Psilocybe candidipes Singer and A.H.Sm + – Mexico [60] Psilocybe copninifacies (Roll). Pouz + – Czech Republic, Slovenia [73] Psilocybe cubensis (Earle) Singer + + Japan, Germany, Mexico, Brazil [20, 46, 61, 74] Psilocybe cyanescens Wakef + + Czech Republic, Switzerland, USA [17, 21, 57, 75] Psilocybe cyanofibrillosa Guzman and Stamets + + USA [22] Psilocybe fimetaria (P.D. Orton) Watling + – United Kingdom [43] Psilocybe hoogshagenii Heim + + Brazil [20] Psilocybe liniformans Guzman and Bas + – USA [21, 22] (continued) converted by the mechanism of dephosphorylation into psilocin (another pharmacologically active compound). Psilocin has the molecular formula C 12 H 16 N 2 O (mol. ...
Chapter
Full-text available
Studies on bioactive alkaloids have gained considerable importance in recent years and have been given prominent position in the field of medicine, both with respect to their biological activity and role played in the introduction of new pharmaceuticals. The biotechnological approach has further enhanced their industrial applications. Some specific groups of bioactive alkaloids have been ignored and not been given much attention because of various reasons, including their legal status. The tryptophan-based indole alkaloids, psilocybin and psilocin, are the best examples in this category which have been identified as most attractive bioactive alkaloids in large number of mushrooms, especially of the genus Psilocybe, but waiting since long for more control studies to ascertain their therapeutic role in some other conditions, apart from psychiatry. These alkaloids have long history of association with mankind for their use as sacraments in religious ceremonies along with medical and recreational purposes and thus need more attention to explore their therapeutic role.
... In this regard, MAE achieved in our study a maximum yield of 1.609 ± 0.081 mg/g relative alkaloids (1.255 ± 0.098 mg/g psilocin and 0.354 ± 0.085 mg/g psilocybin). This extraction yield is greater than the mean yields reported by previous studies where UAE was the extraction method employed; Gotvaldová et al. obtained 0.54 mg/g psilocybin and 0.25 mg/g psilocin by vortexing dried mushroom samples [28], Laussmann et al. reported 1.151 ± 0.228 mg/g psilocybin and 0.126 ± 0.066 mg/g psilocin by UAE [35] and Tsujikawa et al. reported even lower yields of 0.835 ± 0.465 mg/g psilocybin and 0.280 ± 0.140 mg/g psilocin [36] when two UAE cycles were employed. ...
Article
Full-text available
Hallucinogenic fungi, mainly those from the Psilocybe genus, are being increasingly consumed even though there is no control on their culture conditions. Due to the therapeutic potential as antidepressants and anxiolytics of the alkaloids that they produce (psilocin and psilocybin), some form of control on their production would be highly recommended. Prior to identifying their optimal culture condition, a methodology that allows their study is required. Microwave-assisted extraction method (MAE) is a technique that has proven its efficiency to extract different compounds from solid matrices. For this reason, this study intends to optimize a MAE method to extract the alkaloids found in Psylocibe cubensis. A surface-response Box–Behnken design has been employed to optimize such extraction method and significantly reduce time and other resources in the extraction process. Based on the Box–Behnken design, 50 °C temperature, 60% methanol as extraction solvent, 0.6 g:10 mL sample mass:solvent ratio and 5 min extraction time, were established as optimal conditions. These mild conditions, combined with a rapid and efficient UHPLC analysis result in a practical and economical methodology for the extraction of psilocin and psilocybin from Psylocibe cubensis.
... Transverse sections of the mushroom cap along with the gills were cut into several sections and were mounted in a glass slide with both Lactophenol cotton blue and Melzer's solution (chloral hydrate, potassium iodide and iodine) (Tsujikawa et al., 2003). Sections were viewed in Leica Microsystems (CH-9435, Heerbrugg) after covering the section with a cover slip. ...
Article
Full-text available
Earthball mushrooms or ‘Puffball’ mushroom from subtropical forest of lower belt of the Himalayan region of India, regularly consumed in eastern India, was identified based on fungal ITS molecular barcode. The phylogenetic analysis revealed that it clades under the genus Scleroderma of order Boletales which was verified by Neighbor Joining (NJ) tool. The detail analysis of the gill section and the spore morphotype revealed lack of gills with abundant veins like connections. Spore morphotype in the scanning electron microscope (SEM) revealed a globular structure with an articulate covering. The morphology, microscopy and molecular data revealed the puffball structure to be of Sclerodermataceae family under the Agaricomycetes group. This study reports the characterization of irregular potato shaped earthball mushroom, Scleroderma from northeast India.
... Homer and Sperry [89] have recently reviewed on the isolation of mushroomderived indole alkaloids, along with their associated biological activities. The alkaloid compounds can be found in different quantities based on mushroom species, their developmental stages, climatic conditions, and the availability of soluble nitrogen and phosphorous in the soil [90]. ...
... Homer and Sperry [89] have recently reviewed on the isolation of mushroomderived indole alkaloids, along with their associated biological activities. The alkaloid compounds can be found in different quantities based on mushroom species, their developmental stages, climatic conditions, and the availability of soluble nitrogen and phosphorous in the soil [90]. ...
... Methods applied in toxicological analysis include highperformance liquid chromatography with diode array detection, gas chromatography with mass spectrometry, and liquid chromatography with mass detection (13,14). These methods enable not only identification of a specific active substance, but also quantitative analysis, provided appropriate standards are used. ...
Article
Full-text available
In forensic practice, mushroom poisonings - both accidental and those resulting from crimes and suicide attempts - are a frequently encountered type of case. In recent years, there has also been an increased interest in hallucinogenic mushrooms. Under the Act on Counteracting Drug Addiction dated July 29, 2005, possession of and trade in wild mushrooms which contain narcotic substances is a crime. Therefore, precise identification of species of fungi that contain illegal and toxic substances is very important for the purposes of judicial proceedings. Morphological analysis, a standard method for fungi species identification, does not always yield satisfactory results. Due to the reliability, speed and decreasing cost of DNA analysis, genetic methods are an interesting alternative for determining the species of biological material. Markers frequently employed in species identification of fungi are internal transcribed spacers regions (ITS1 and ITS2). The objective of a project that is being implemented at the Institute of Forensic Research is the preparation of an ITS1 and ITS2 regions sequence database of the analyzed fungal species and the validation of methods for DNA sequence analysis of these regions in the case of processed samples.
... Silica and silica based sorbents can be damaged due to partial dissolution of the silica at alkaline condition. The majority of HPLC procedures available in the scientific literature is based on reversed phase [2][3][4][5][6] separation which disadvantageous particularly for the psilocybin because of its insufficient retention caused by the high polarity of the compound. The flow-through may even hinder the mass spectrometric detection of psilocybin because of the quenching of formed ions by other unretained co-eluting matrix components. ...
... In this case, the suspect Table 2: DNA analysis technique used for analysis of different types of fungi . [10], [42][43][44] [45][46][47][48], [24] S.No Type of fungi Technique denied the claim of vic m and said that they had consensual sexual rela ons in a local park, away from the alleged crime scene. Inves gators collected samples from each place, clothing, footwear from each party and all the places that were considered to be relevant to the case were visited, and flora was recorded. ...
... Otras veces se analiza la presencia de toxinas o se emplean marcadores genéticos. En España el laboratorio del Instituto Nacional de Toxicología y Ciencias Forenses (INTCF) es el centro técnico y de investigación en temas toxicológicos por ingestión de setas (ITURRAL-DE & al., 2002, 2003. El conocimiento de la especie causante de la intoxicación es fundamental porque condiciona el protocolo terapéutico a seguir. ...
Article
Full-text available
... Methods applied in toxicological analysis include highperformance liquid chromatography with diode array detection, gas chromatography with mass spectrometry, and liquid chromatography with mass detection (13,14). These methods enable not only identification of a specific active substance, but also quantitative analysis, provided appropriate standards are used. ...
Article
Full-text available
Aim: To assess the usefulness of a DNA-based method for identifying mushroom species for application in forensic laboratory practice. Methods: Two hundred twenty-one samples of clinical forensic material (dried mushrooms, food remains, stomach contents, feces, etc) were analyzed. ITS2 region of nuclear ribosomal DNA (nrDNA) was sequenced and the sequen-ces were compared with reference sequences collected from the National Center for Biotechnology Information gene bank (GenBank). Sporological identification of mushrooms was also performed for 57 samples of clinical material. Results: Of 221 samples, positive sequencing results were obtained for 152 (69%). The highest percentage of positive results was obtained for samples of dried mushrooms (96%) and food remains (91%). Comparison with GenBank sequences enabled identification of all samples at least at the genus level. Most samples (90%) were identified at the level of species or a group of closely related species. Sporological and molecular identification were consistent at the level of species or genus for 30% of analyzed samples. Conclusion: Molecular analysis identified a larger number of species than sporological method. It proved to be suitable for analysis of evidential material (dried hallucinogenic mushrooms) in forensic genetic laboratories as well as to complement classical methods in the analysis of clinical material.
... Observation of the spore characteristics is inevitably important for generating a correct taxonomic profile of each specimen from the relatively unexplored mushrooms of the region. Although there have been a few reports on scanning electron microscopic studies of mushroom spores [14,[30][31][32], to the best of our knowledge this is the first detailed report on spore morphology of wild edible fungi involving SEM from this part of the world. The information generated in the present study is expected to serve as a taxonomic reference point for further studies on wild edible mushrooms. ...
Article
Full-text available
Wild edible mushrooms consumed by the ethnic tribes of India were collected from the local markets and forests and identified based on their morphology. The edible fungi collected were affiliated to 10 different species under genera and 8 families. The fruiting bodies of the wild edible mushroom species generated unique colour change profiles upon reaction with 3 % aqueous solution of phenol, 10 % solution of potassium hydroxide and 10 % solution of ferrous sulphate. A detailed study on the spore morphology 16 of the specimens was carried out using staining techniques and Scanning Electron Microscopy. The complementary use of Scanning electron microscope (SEM) with an optical microscope offered characteristic features on spore morphology with SEM providing a three-dimensional microarchitecture of the mushroom spores. The present study will, therefore, serve as a taxonomic reference point for further studies on mushrooms of the region.
... The identity of these derivatives was confirmed by the presence of the characteristic fragments m/z 58, 189 and 261 for bis-trimethyl-silyl-psilocybin and 58, 290 and 348 for bistrimethyl-silyl-psilocin . Differences in the psilocin and psilocybin contents in hallucinogenic mushrooms depends on factors such as developmental stage, weather conditions, and the availability of soluble nitrogen and phosphorous in the soil (Tsujikawa et al. 2003). These results confirm the presence of psilocin and psilocybin for the species and, consequently, the hallucinogenic potential of Psilocybe wrightii. ...
Article
Full-text available
Psilocybe wrightii (Strophariaceae, Agaricales), a subtropical and hallucinogenic species known only from southern Brazil and northern Argentina, is confirmed to contain psilocybin and psilocin. Detailed description of southern Brazilian specimens, chemical analysis using gas chromatography of the basidiomes, and discussion of its taxonomy are presented.
... Transverse sections of the mushroom cap along with the gills were cut into several sections and were mounted in a glass slide with both Lactophenol cotton blue and Melzer's solution (chloral hydrate, potassium iodide and iodine) (Tsujikawa et al., 2003). Sections were viewed in Leica Microsystems (CH-9435, Heerbrugg) after covering the section with a cover slip. ...
... In this case, the suspect Table 2: DNA analysis technique used for analysis of different types of fungi . [10], [42][43][44] [45][46][47][48], [24] S.No Type of fungi Technique denied the claim of vic m and said that they had consensual sexual rela ons in a local park, away from the alleged crime scene. Inves gators collected samples from each place, clothing, footwear from each party and all the places that were considered to be relevant to the case were visited, and flora was recorded. ...
... (psilocybin) for Copelandia, respectively. Thus, the former is more psilocybin-rich than the latter, and the latter contains more psilocin compared to the former (Tsujikawa et al., 2003). The Psilocybe semilanceata is the most common British species. ...
Article
Full-text available
Background In the past few years, the issue of ‘microdosing’ psychedelics has been openly discussed in the public arena where claims have been made about their positive effect on mood state and cognitive processes such as concentration. However, there are very few scientific studies that have specifically addressed this issue, and there is no agreed scientific consensus on what microdosing is. Aim This critique paper is designed to address questions that need to be answered by future scientific studies and to offer guidelines for these studies. Approach Owing to its proximity for a possible approval in clinical use and short-lasting pharmacokinetics, our focus is predominantly on psilocybin. Psilocybin is allegedly, next to lysergic acid diethylamide (LSD), one of the two most frequently used psychedelics to microdose. Where relevant and available, data for other psychedelic drugs are also mentioned. Conclusion It is concluded that while most anecdotal reports focus on the positive experiences with microdosing, future research should also focus on potential risks of (multiple) administrations of a psychedelic in low doses. To that end, (pre)clinical studies including biological (e.g. heart rate, receptor turnover and occupancy) as well as cognitive (e.g. memory, attention) parameters have to be conducted and will shed light on the potential negative consequences microdosing could have.
... [28][29][30] In our study, ultra-high performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry (UHPLC-MS/MS) was employed to investigate the stability of five tryptamine alkaloids in the biomass of cultivated fruiting bodies of the psychoactive species Psilocybe cubensis. [31][32][33] 2 | MATERIALS AND METHODS ...
Article
Psilocybin, psilocin, baeocystin, norbaeocystin, and aeruginascin are tryptamines structurally similar to the neurotransmitter serotonin. Psilocybin and its pharmacologically active metabolite psilocin in particular are known for their psychoactive effects. These substances typically occur in most species of the genus Psilocybe (Fungi, Strophariaceae). Even the sclerotia of some of these fungi known as “magic truffles” are of growing interest in microdosing due to them improving cognitive function studies. In addition to microdosing studies, psilocybin has also been applied in clinical studies, but only its pure form has been administrated so far. Moreover, the determination of tryptamine alkaloids is used in forensic analysis. In this study, freshly cultivated fruit bodies of Psilocybe cubensis were used for monitoring stability (including storage and processing conditions of fruiting bodies). Furthermore, mycelium and the individual parts of the fruiting bodies (caps, stipes, and basidiospores) were also examined. The concentration of tryptamines in final extracts was analyzed using ultra-high-performance liquid chromatography coupled with mass spectrometry. No tryptamines were detected in the basidiospores, and only psilocin was present at 0.47 wt.% in the mycelium. The stipes contained approximately half the amount of tryptamine alkaloids (0.52 wt.%) than the caps (1.03 wt.%); however, these results were not statistically significant, as the concentration of tryptamines in individual fruiting bodies is highly variable. The storage conditions showed that the highest degradation of tryptamines was seen in fresh mushrooms stored at −80°C, and the lowest decay was seen in dried biomass stored in the dark at room temperature.
... The arrow indicates the germ pores. Reproduced with permission from Elsevier, 2003(Tsujikawa et al. 2003). d SEM of a yeast cell. ...
Chapter
Green approach is an advance technique of synthetic chemistry. It opens up a new vista of modified version of known synthetic reactions in a newer way with associated enlarged potentiality accompanied by sustainability. Revolutionary measures against solvent-based reaction have been invited in green routes. Organic solvents have been substituted by non-organic media in organic synthetic reactions to overcome the hazard of volatile organic solvents and to preserve the greenness of the reactions, following green principles, supported by green matrices. This chapter illustrates the successive green revolutionary measures in solvent-based organic chemistry followed by solvent-free synthetic routes and their future trends in a nut shell.
... The data in the previously published literature showed that the contents of psilocybin in the canopies and stems of hallucinogenic mushrooms were higher than those of psilocin, which was exactly the opposite of our experimental results [12,15,16]. However, the literature published by Tsujikawa et al. [27] [12] showed that the contents of psilocin in the canopies and stems of Copelandia were higher than those of psilocybin (0.64-0.74%/0.02-0.22% (psilocin/psilocybin) in the canopies and 0.31-0.78%/0.01-0.39% ...
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Purpose In this study, we developed a very sensitive method for quantitative analysis of psilocin and psilocybin in hair samples of magic mushroom consumers. Methods The analyses were performed with pretreatments of samples, followed by ultra-high pressure liquid chromatography (LC) connected to a Q-Trap type tandem mass spectrometry (MS/MS). For LC, mobile phase (A) consisted of 0.1% formic acid in water, and mobile phase (B) was acetonitrile for gradient elution using a Acquity™ UPLC HSS T3 column. For MS/MS, electrospray ionization measurements in positive selected reaction monitoring mode were used. Results The calibration curves were linear from 5 to 500 pg/mg ( r > 0.99) and no selectivity problems occurred. The limit of detection was 1 pg/mg, and the lower limit of quantitation was 5 pg/mg. The ranges of the matrix effects and recovery rates were 90.4–107% and 76.0–102%, respectively. Conclusions The concentrations of psilocin in two authentic hair were 161 and 150 pg/mg, respectively, and psilocybin was not detected from both samples. This method was also used to analyze the distribution of psilocin and psilocybin in seven hallucinogenic mushrooms. To our knowledge, this is the first demonstration of psilocin concentrations in hair samples of hallucinogenic mushroom consumers, and also our method is most sensitive for quantitative analysis of psilocin and psilocybin in hair samples.
... Mushroom is a general layman term commonly used to describe the sporocarps of Ascomycetous and Basidiomycetous fungi (Tsujikawa et al., 2003;Adejumo and Awosanya, 2005;Atri et al., 2010). These are mostly saprophytic; however, many of them form mycorrhizal association with plants and a few of them including Armillaria mellea (Vahl.) ...
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The chapter embodies nutritional profi le of 20 wild edible mushrooms of North (Scop.: Fr.) Sing., M. rhacodes (Vitt.) Sing., Pleurotus cystidiosus O.K. Miller, P. fl oridanus Singer, P. pulmonarius (Fr.) Quél, P. sajor-caju (Fr.) Sing. and P. sapidus Quél., with respect to their carbohydrates, crude fat, proteins, fi bers, ash, minerals, heavy metals and vitamins contents. Carbohydrate constitutes the largest fraction of mushroom dry matter ranging from 33.3% in Termitomyces medius to 89.10% in Lentinus cladopus. Maximum percentage of crude fat (3.4%) has been documented in Macrolepiota procera, while minimum amount (0.48%) was observed in Lentinus connatus. Protein percentage is maximum in Termitomyces medius (46.2) and minimum in Lentinus connatus (0.525). The percentage of fi ber ranged from 1.83 in L. squarrosulus to 8.0 in Termitomyces mammiformis. The ash content was
... Mushroom is a general layman term commonly used to describe the sporocarps of Ascomycetous and Basidiomycetous fungi (Tsujikawa et al., 2003;Adejumo and Awosanya, 2005;Atri et al., 2010). These are mostly saprophytic; however, many of them form mycorrhizal association with plants and a few of them including Armillaria mellea (Vahl.) ...
... Many genera containing hallucinogenic compounds include the Amanita, Conocybe, Galerina, Gymnopilus, Inocybe, Panaeolus, Pluteus, and among all these, Psilocybe and Panaeolus are the most commonly used hallucinogenic mushrooms. All these members are known to synthesize the controlled substances such as psilocin and psilocybin, and in the illegal market, magic mushrooms are often seized in intact or fresh state (Linacre et al. 2002;Stra et al. 2014;Tsujikawa et al. 2003Tsujikawa et al. , 2007Nagy and Veress 2016). Cultivation and possession of mushrooms that contain psilocin and/or psilocybin is illegal in many countries (Gonmori et al. 2011;Margarida et al. 2015;Zuber et al. 2011). ...
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In forensic investigations, mycological evidence can be an important marker for ascertaining the cause of death, time since death, and location of the corpse. Moreover, many hallucinogenic mushrooms are also encountered in illegal black markets. In both the scenarios, it is of utmost significance to identify the fungal species in question. Species identification can be performed using conventional (taxonomic) methods as well as by using DNA barcoding. Conventional methods such as morphological identification suffer from serious limitations in both poisoning and illegal trade cases. In mushroom poisoning cases, degradation and contamination occurs due to cooking, gastric juices, vomit, and fecal matter, whereas in cases of illegal trade, confiscated samples are found in powdered form. This leads to the absence of identification features which are required for the taxonomic identification. One prevalent alternative to counter this problem is “DNA barcoding,” which unlike taxonomic identification does not depend on sample morphology. In this chapter, an attempt has been made to discuss various facets of DNA barcoding of fungal evidence along with the recent advancements made in this field in the past few decades. In spite of wide applications of DNA barcoding in other fields, this technique has largely remained underexplored in forensic mycology due to the lack of skilled mycologists and limited awareness of current methods among the criminal investigators. Recent advancements in fungal identification on the cadavers have preferred DNA-based methods over conventional techniques to determine time since death by analyzing the fungal growth rate. High-throughput sequencing technology (HTS) has also improved the identification capabilities of DNA barcoding in fungal species identification.
... Transverse sections of the mushroom cap along with the gills were viewed in Leica Microsystems (CH-9435; Leica, Heerbrugg, Germany) using Melzer's solution [12]. ...
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... Mushroom is a general layman term commonly used to describe the sporocarps of Ascomycetous and Basidiomycetous fungi (Tsujikawa et al., 2003;Adejumo and Awosanya, 2005;Atri et al., 2010). These are mostly saprophytic; however, many of them form mycorrhizal association with plants and a few of them including Armillaria mellea (Vahl.) ...
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The chapter embodies nutritional profi le of 20 wild edible mushrooms of North (Scop.: Fr.) Sing., M. rhacodes (Vitt.) Sing., Pleurotus cystidiosus O.K. Miller, P. fl oridanus Singer, P. pulmonarius (Fr.) Quél, P. sajor-caju (Fr.) Sing. and P. sapidus Quél., with respect to their carbohydrates, crude fat, proteins, fi bers, ash, minerals, heavy metals and vitamins contents. Carbohydrate constitutes the largest fraction of mushroom dry matter ranging from 33.3% in Termitomyces medius to 89.10% in Lentinus cladopus. Maximum percentage of crude fat (3.4%) has been documented in Macrolepiota procera, while minimum amount (0.48%) was observed in Lentinus connatus. Protein percentage is maximum in Termitomyces medius (46.2) and minimum in Lentinus connatus (0.525). The percentage of fi ber ranged from 1.83 in L. squarrosulus to 8.0 in Termitomyces mammiformis. The ash content was
... Morphology is a common method based on surface features from fruiting body such as color, shape, size, or reticulate pattern (Tsujikawa et al., 2003). However, this method did neglect the phenotypic variability of mushroom. ...
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The plateau specialty agricultural products, wild porcini mushrooms, have great value both as a superb cuisine and as a potential medication. Due to quality different between species added with the fraud behavior in sales process, make poor quality or poisonous sample inflow into the market, which pose a health risk for consumers, but also disrupted the mushroom market. Traditional analysis way is time‐consuming and laborious. Therefore, the aim of this study is to develop a way using fourier transform mid‐infrared (FT‐MIR) spectrometry and data fusion strategies for the fast and accurate species discrimination and predict amount of total polyphenol in four porcini mushrooms. The t‐distributed stochastic neighbor embedding based on mid‐level data fusion showed two species of Boletus edulis and B. umbriniporus have been identified. The order of correct rate of PLS‐DA models was mid‐level data fusionq (100%) > mid‐level data fusione (97.06%) = mid‐level data fusionv (97.06%) = stipes (97.06%) > low‐level data fusion (94.12%) > caps (91.18%). The order of correct rate of grid‐search support vector machine models was low‐level data fusion (100%) > caps (94.12%) > stipes (91.18%), and the order of particle swarm optimization support vector machine was low‐level data fusion (100%) > caps (97.06%) > stipes (88.24%). The mid‐level data fusionq and low‐level data fusion had best discrimination accuracy (100%) allowing each mushroom classed into its real species, which could be used for accurate discrimination of samples. B. edulis mushrooms had highest total polyphenol, with 14.76 mg/g dw and 17.33 in caps and stipes mg/g dw, respectively. The phenols were easier to accumulate in the caps in Leccinum rugosiceps (1.03) and B. tomentipes (1.19), and the opposite phenomenon is observed in B. edulis (0.85) and B. umbriniporus (0.95). The correlation coefficient and residual predictive deviation of best prediction model were 86.76% and 2.40%, respectively, indicating that that there is good relevance between FT‐MIR and total polyphenol content, which could be used to predict roughly polyphenols content in mushrooms. The current study developed a way with fast, cheap and reliable for discriminating mushroom species and a way to roughly predict polyphenol content in mushrooms. Besides, the study applied three ways of feature extraction and provided data of content and enrich ability of total polyphenol in different morphological part of mushroom for the first time.
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Psilocybe cubensis, or “magic mushroom,” is the most common species of fungus with psychedelic characteristics. Two primer sets were designed to target Psilocybe DNA using web-based software and NBCI gene sequences. DNA was extracted from eighteen samples, including twelve mushroom species, using the Qiagen DNeasy® Plant Mini Kit. The DNA was amplified by the polymerase chain reaction (PCR) using the primers and a master mix containing either a SYBR® Green I, Radiant™ Green, or LCGreen Plus® intercalating dye; amplicon size was determined using agarose gel electrophoresis. The PCR assays were tested for amplifiability, specificity, reproducibility, robustness, sensitivity, and multiplexing with primers that target marijuana. The observed high resolution melt (HRM) temperatures for primer sets 1 and 7 were 78.85 ± 0.31°C and 73.22 ± 0.61°C, respectively, using SYBR® Green I dye and 81.67 ± 0.06°C and 76.04 ± 0.11°C, respectively, using Radiant™ Green dye.
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History Identifying Characteristics Exposure Dose Effect Toxicokinetics Histopathology and Pathophysiology Clinical Response Diagnostic Testing Treatment References
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The alkaloid psilocybin (4-phosphoryloxy-N,N-dimethyltryptamine) and the neurologically active psilocin (4-hydroxy-N,N-dimethyltryptamine) are the foremost compounds of pharmaceutical interest in Psilocybe mushrooms. As these compounds are infrequently analyzed in analytical labs, validated methods for rapid purity analysis are lacking. Newfound therapeutic use has invigorated academic and commercial interests in the molecules and new methods of production and available products are expanding. As a result, high-throughput methods of analysis for psilocybin must be improved to promptly determine chemical differences between mushroom genera or other sources of psilocybin and psilocin, as well as refined product purity. To address this, we developed an inexpensive HPLC technique for the efficient quantification of psilocybin and psilocin by using readily available equipment and dilute reagents. Aqueous ammonium formate (0.143 mM) was found to be preferable over techniques with much higher buffer concentrations or stronger acids for controlling psilocybin Zwitterion resolution. The chromatographic run time satisfied high-throughput analytical requirements with an efficient total runtime under 2 minutes. A standard octadecyl silica (C18) column provided excellent resolution between psilocybin and psilocin signals. The quality of the method was validated using certified analytical reference standards and was found to be accurate (3.5% bias, Psilocybin), reliable (0.32% RSD), and efficient (Psilocybin k’ = 1.78).
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A new method for in-cellular staining of yeast like fungi with Oregon Green and SYTOX Green is presented enabling their detection as well as the observation of cellular details via confocal laser scanning microscopy. Fluorochromes play an important role in many scientific disciplines including medicine, cell biology and botany. For the visualisation of fungal cell walls Calcofluor White is the flourochrome of choice. The necessity of an UV laser for its excitation makes it unpracticable for daily use. Safranin O, DAPI, 2NBDG, Ethidium Bromide and Acridin-orange are commonly used stains for nuclei in fugal microscopy. The attention was given to the possibility of using the differences in staining patterns to distinguish certain pathogenic yeast species e.g. Candida albicans and Candida krusei. Our results show that high quality microscopy of yeast like organisms can readily be achieved by the use of two suitable fluorochromes.
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Psilocybin, a drug most commonly recognized as a recreational psychedelic, is quickly gaining attention as a promising therapy for an expanding range of neurological conditions, including depression, anxiety, and addiction. This growing interest has led to many recent advancements in psilocybin synthesis strategies, including multiple in vivo fermentation-based approaches catalyzed by recombinant microorganisms. In this work, we show that psilocybin can be produced in biologically relevant quantities using a recombinant E. coli strain in a homebrew style environment. In less than 2 days, we successfully produced approximately 300 mg/L of psilocybin under simple conditions with easily sourced equipment and supplies. This finding raises the question of how this new technology should be regulated as to not facilitate clandestine biosynthesis efforts, while still enabling advancements in psilocybin synthesis technology for pharmaceutical applications. Here, we present our homebrew results, and suggestions on how to address the regulatory concerns accompanying this new technology.
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In this chapter, a few interesting findings on renewable carbons derived from various biomass precursors and their electrochemical applications, especially supercapacitors and lithium-ion batteries were discussed. Electrochemical energy storage devices are progressively crucial since it helps in a significant reduction in the use of different fossils-based resources. The use of energy storage devices in a particular system rests on the nature of the electrode materials. Among an extensive range of electrodes, biomass-based carbons have gained significant consideration as an electrode because of their variable physicochemical features, environmental concern, and commercial value. We have also discussed a few recent developments of biomass-derived carbons and some important techniques such as carbonization and activation conditions that control their property and performance of carbon electrodes. Besides, some of the parameters such as pore structure, surface property, and degree of graphitization, which regulate the electrochemical functioning of the device were discussed in detail. In the final section of this chapter, we included some energy storage applications of biomass-based carbons, and their effectiveness as an electrode were summarized. In brief, this chapter provides a fundamental understanding of several biomass-derived carbon materials and suggests the essential strategies for the fabrication of various energy storage systems.
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This represents one of several sections of "A Bibliography Related to Crime Scene Interpretation with Emphases in Geotaphonomic and Forensic Archaeological Field Techniques, Nineteenth Edition" (The complete bibliography is also included at ResearchGate.net.). This is the most recent edition of a bibliography containing resources for multiple areas of crime scene, and particularly outdoor crime scene, investigations. It replaces the prior edition and contains approximately 10,000 additional citations. As an ongoing project, additional references, as encountered, will be added to future editions. Botanical or plant evidence is often overlooked at crime scenes. Unless plant remains such as seeds, or “burrs” are found adhering to the victim’s hair or remains, incidental to the collection of the body, independent control and questioned samples of plant parts “from leaf to root” are typically not recognized as evidence. Perhaps the standards of scientists such as Hall (1988, 1997), Willey and Heilman (1987), Warren (1975, 1980, 1984), or Bock and Norris (1997), will continue to alert investigators and prosecutors to the value of plant and plant component evidence. Compilations such as Miller Coyle's Forensic Botany (2004) serve to reinforce and update developments in this critical component of crime scene analysis and reconstruction. Forensic botany is a relatively young application of an old science. The majority of the references below address botanical and palynological collections and analyses as they relate to archaeological site deposits. For this reason, the researcher should also be aware of archaeological site reports under Excavation and Recovery Strategies which might contain information on site specific collections and analyses of plant remains. It has long been a standard in archaeology protocol to collect soil samples or “constant volume” samples for separation and identification of small plant remains. The use of this information by archaeologists to help interpret the climate and environment of sites during particular time periods, or to determine the diet of a site’s population have direct application to forensic settings. The same “answers” are often sought by crime scene investigators seeking to validate subject or witness accounts, as well as time and location of death. A reflection of forensic botany's progression along with other forensic sciences is the increased number of resources addressing research and observations in plant DNA, (Bever, et al., [2002]; Linacre, et al., [2002]; Weising, et al., [2005]; and Craft, et al., [2007] to name a few.) This bibliography’s references in Taphonomy may also contain information/observations on the effect(s) of plant activity on disposed remains. Like entomological evidence, plant remains offer the opportunity to place remains or associated evidence in temporal and spatial contexts. Over time, plant growth may first serve to demarcate areas in which remains were disposed, and then impact the remains by utilizing them as a component of the lithosphere or biosphere. Botanical evidence also represents trace evidence which could link subject and victim and crime scene, as well as establish the movement of the subject and/or victim through a crime scene. From pollen to a rash from poison ivy, botanical clues follow Locard's principles of exchange. Trackers utilize damage to plants to find and follow paths taken by subjects and victims. The use and interpretation of botanical evidence requires holistic knowledge of crime scene environments, the accurate recording of same, and site specific conditions which might affect plant growth (see Photography, Reconnaissance, Survey, and Mapping Techniques, Excavation and Recovery Strategies, as well as Geoarchaeology and Soil Science).
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The switch of polluted soils from heavy metals is the utmost challenging mission, predominantly on a huge scale. The soil is composed of organic and inorganic solid constituents, water and mixture of different gases present in various proportions. During prospective study about the accumulation of metals in plants; the enormous growth of weeds and mushrooms attract our attention to check these in comparison with growth of plant under investigation in the contaminated soil. It was established after checking the growth of plant under study that these weeds were supportive in growth regulation via accretion of metal leaving soil for growth of regular plant. It was observed that Myco-remediation competently collapse contaminants and heavy metals by directing them to the fruit bodies of mushrooms. It was related with capability of mushrooms to absorb the metal and act as a scavenger. The function was very clear after observing the growth of the plants under study and can easily be understood that fungal mycelia grip soil organized, aid in hold water and makes the nutrient accessible for vegetation. The mechanism of mushroom action to absorb the toxins via enzymes secretion into soil was developed. Their growth in contaminated environment opens new biological remediation techniques to secure the food chain even in presence of toxins. It was suggested on the basis of naturally grown spore of wild mushroom and weeds in metal contaminated environment; that it may be powerful tool in new emerging remediation technology at domestic and global hazardous waste management sites especially in developing countries, where toxins are in action and it's a dream of safe atmosphere for just health of concerns.
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This chapter discusses the application of liquid chromatography with the conventional detection for food toxicants. Conventional ultraviolet (UV) detection has been applied for several food toxicants on various kinds of food matrices. Biogenic amines represent a group of harmful compounds that can be analyzed using UV detection. UV allows the detection of tyramine, tryptamine, phenylethylamine, and histamine. Diode array-detectors (DAD) supplement additional features to spectrophotometric detection compared with the conventional ultraviolet/ visible light (UV/VIS) detectors. In addition to the identification by the retention time, qualitative information of the analyte can be obtained with DAD, which allows the selection of the wavelength with the highest absorbance for separate analytes. Fluorescence intensity is quantitatively dependent on the same parameters as absorbance, fluorescence quantum yield of the analyte, excitation source intensity, and fluorescence collection efficiency of the instrument. Electrochemical detectors have frequently been used to quantify heterocyclic amines. It is found that electrochemical detection is especially applicable to analyze mixtures of polar and less polar amines in different matrices.
Chapter
IntroductionToxins Responsible for Plant Poisoning and Poisoning with Herbal SupplementsRole of the Clinical Laboratory in Treating Patients with Plant Poisoning and Toxicity from Using Herbal RemediesDigoxin Immunoassay in Detecting Plant Poisoning and Herbal ToxicityAbnormal Liver Function Tests as an Indication for Plant Poisoning and Herbal ToxicityOther Abnormal Laboratory Tests as Indications for Plant Poisoning and Herbal ToxicityChromatographic Methods for the Diagnosis of Plant Poisoning and Herbal ToxicityMiscellaneous Plant and Herbal Medicine PoisoningsConclusions References
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The synthesis and application of biomass-derived carbon in energy storage have drawn increasing research attention due to the ease of fabrication, cost-effectiveness, and sustainability of the meso/microporous carbon produced from various biological precursors, including plants, fruits, microorganisms, and animals. Compared to the artificial nanostructured carbons, such as fullerene, carbon nanotube and graphene, the biomass-derived carbons may obtain superior capacitance, rate performance and stability in supercapacitor applications ascribing to their intrinsic nanoporous and hierarchical structures. However, challenges remain in processing techniques to obtain biomass-derived carbons with high carbon yield, high energy density, and controllable graphitic microstructures, which may require a clear understanding over the chemical and elemental compositions, and the intrinsic microstructural characteristics of the biological precursors. Herein we present comprehensive analyses over the impacts of the chemical and elemental compositions of the precursors on the carbon yield of the biomass, as well as the mechanism of chemical activation on the nanoporous structure development of the biomass-derived carbons. The structure–property relationship and functional performance of various biomass-derived carbons for supercapacitor applications are also discussed in detail and compared. Finally, useful insights are also provided for the improvements of biomass-derived carbons in supercapacitor applications.
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• Amatoxins have been determined in untreated and heat-treated sample of Amanita phalloides by HPLC. • Sample was given various heat treatments like boiling, grilling, baking and frying to simulate real time sample condition. • In untreated sample, high amount of toxins was found in cap followed by stalk and base. • In heat-treated sample lowest amount of amatoxins were found in the boiled sample. • First study to determine amatoxins in the degraded form of poisonous mushroom.
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The main representatives of narcotic substances of natural origin are considered. Methodological solutions used in their screening, identification, and quantitative determination in raw materials, dosage forms, and biological materials are discussed. Prospects of application of state-of-the art hybrid methods combining chromatographic separation with mass spectrometric determination are noted.
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Development of rapid and reliable immunochemical methods for monitoring psilocybin (4-phosphoryloxy-N,N-dimethyltryptamine; Pyb) and psilocin (dephosporylated metabolite; Psi), the psychoactive compounds contained within hallucinogenic mushrooms (magic mushrooms), is desirable in order to identify these mushrooms and regulate their illicit use. Because no antibody was publicly available for this purpose, we generated two independent monoclonal antibodies (mAbs) against Pyb or Psi, and then developed enzyme-linked immunosorbent assays (ELISAs) by using them. To generate the specific antibodies, novel immunogenic conjugates were prepared by linking Pyb or Psi molecules to carrier proteins by modifying their 2-(N,N-dimethylamino)ethyl side chains. Spleen cells from mice immunized with these conjugates were fused with P3/NS1/1-Ag4-1 myeloma cells, and hybridoma clones secreting anti-Pyb and anti-Psi mAbs were established. These mAbs were characterized for their biochemical features and then applied to competitive ELISAs, which used microplates coated with Pyb or Psi linked with albumin. These ELISAs enabled the determination of Pyb or Psi with measurable ranges of ca. 0.20−20 or 0.040−2.0 μg/assay (limit of detection was 0.14 or 0.029 μg/assay), respectively. The related tryptamines were satisfactorily discriminated as exemplified by the cross-reactivity of the ELISA to determine Pyb (or Psi) with Psi (or Pyb) that were found to be 2.8% (or <0.5%), respectively. The Pyb and Psi contents in a dried powder of the hallucinogenic mushroom, Psilocybe cubensis, were determined to be 0.39 and 0.32 (w/w)%, respectively. The ELISAs developed using the current mAbs are promising tools for identifying illegal hallucinogenic mushrooms.
Chapter
Drug abuse is a worldwide problem. Although commonly abused drugs can be identified during routine urine drug testing, less commonly abused drugs may escape detection. These less commonly abused drugs not only include some designer drugs such as synthetic cannabinoid but also include abuse of psychedelic magic mushroom (active ingredients: psilocybin and psilocin), peyote cactus (active ingredient: mescaline), and khat plants (active ingredient: cathinone). Moreover, solvent and glue abuse is gaining popularity among teenagers and young adults which may even cause fatality. Amphetamine/methamphetamine immunoassay has a low cross-reactivity with psilocin. Cathinone, if present in the urine, can be detected by amphetamine/methamphetamine immunoassay due to cross-reactivity of cathinone with assay antibody. Currently there is one commercially available immunoassay which is capable of detecting synthetic cathinone known as bath salts as well as mescaline. However, gas chromatography combined with mass spectrometry as well as liquid chromatography combined with tandem mass spectrometry (LC/MS/MS)-based method is available for confirmation of the active ingredients present in magic mushroom, peyote cactus, and khat plant. Such chromatography-based methods also offer more sensitivity and specificity compared to an immunoassay.
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The number of hallucinogenic compounds that have been separated simultaneously by liquid chromatography is limited. This research aimed to identify a column(s) that can allow for separation of several hallucinogens. The extent of separation of seven polar hallucinogenic tryptamine and phenethylamine derived alkaloids containing a basic N atom that becomes protonated at low pH values were investigated on five reverse-based columns and one hydrophilic interaction liquid chromatography (HILIC) column. An RP-phenyl and a negatively charged fused core HILIC were identified and recommended as effective columns in this regard. This research is the first to introduce a HILIC column in separation of hallucinogenic alkaloids and to simultaneously study the separation of the two principal psychoactive agents, muscimol and psilocin. The results of this study showed that better separation in HILIC columns can be achieved if an aprotic diluent of relatively lower polarity is used to introduce analytes into a mobile phase of higher polarity. Tetrahydrofuran was recommended where acetonitrile is used as the organic component of the mobile phase.
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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 × 10−8 and 3.5 × 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.
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This paper presents the results of recent ethnomycological exploration in southern Thailand. Field observations, interviews and collection of fungi specimens were carried out primarily on two islands, Koh Samui and Koh Pha-ngan, situated in the western region of the Gulf of Siam. Some fieldwork was also conducted in the northern Thai province of Chiang Dao and in the southern Thai province of Surat Thani. During five separate excursions (1989-90), observations were made of occurrence, harvesting, use, and marketing of psychoactive fungi by local Thai natives (males and females, adults and children), foreign tourists, and German immigrants. The first records of psychoactive Psilocybe subcubensis and Copelandia dung fungi in Thailand are presented in this paper. These fungi exhibited intense bluing reactions when handled, indicating the presence of psilocybin and/or psilocin. Seven collections of Psilocybe cubensis (Earle) Singer and/or Psilocybe subcubensis Guzman and four collections of Copelandia sp. were harvested and sun-dried for herbarium deposit. These fungi are cultivated or occur spontaneously, often appearing in the decomposed manure of domesticated water buffalo (Bubalus bubalis) and at least three different species of cattle (Bos indicus, B. guarus, and B. sundaicus). The psychoactive fungi are cultivated in clandestine plots, both indoors and outdoors, in the uplands and villages on Koh Samui by both Thai natives and some foreigners. The sale of psychoactive fungi directly to tourists and to resort restaurants for use in edible food items such as omelettes and soups is discussed in detail. The preparation and sale of mushroom omelettes adulterated with artificial hallucinogens in some restaurants is also discussed. In addition, the marketing of items such as hand painted T-shirts, post cards, and posters bearing mushroom related motifs in Thailand is described.
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Analysis of Psilocybe cubensis (Earle) Singer grown in controlled culture showed that the level of psilocin was generally zero in the first (or sometimes even the second) fruiting of the mushroom from a given culture and that the level reached a maximum by the fourth flush. The level of psilocybin, which was nearly always at least twice the level of psilocin, showed no upward or downward trend as fruiting progressed, but was variable over a factor of four. Samples obtained from outside sources had psilocybin levels varying by over a factor of ten from one collection to the next.
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Several fungi species collected in the Hawaiian Islands have been reported to be psychoactive. Previous chemical analyses together with the present study indicate that 5 coprophilous and one non-coprophilous species occurring in the islands are now known to contain psychoactive alkaloids. At least some of these species are consumed in the Hawaiian Islands, as well as elsewhere, for non-traditional, recreational purposes. These include Copelandia cyanescens (Berk. et Br.) Singer, Copelandia tropicalis (Ola'h) Singer and Weeks (syn. Panaeolus tropicalis Ola'h), Copelandia anomala Murrill, and Panaeolus subbalteatus (Berk. and Br.) Sacc., which have already been described from the Hawaiian Islands. Three more mind-altering fungi and one non-psychoactive species are reported from this archipelago for the first time. These psychoactive fungi include Copelandia bispora (Malençon et Bertault) Singer and Weeks from O'ahu, Copelandia cambodginiensis (Ola'h et Heim) Singer and Weeks from O'ahu, and Amanita muscaria (L.) Hooker from Kaua'i. Panaeolus goossensiae Beeli identified from O'ahu contains tryptamine compounds; however, the psychoactive alkaloids psilocybin and psilocin were not found in this dung species.
Article
High-performance liquid chromatography (HPLC) was used for the analysis of the minor consistuents psilocybin and psilocin in fungi of the genus Psilocybe. The separation and determination of these compounds was carried out on a stationary phase of LiChrosorb RP-18. The analytical column (A) and semipreparative column (B) were eluted isocratically with water—ethanol—acetic acid (79.2:20:0.8) at flow-rates of 20 ml/h (A) and 180 ml/h (B). The compounds were detected with a UV detector at 267 nm and a fluorometric detector (excitation, 280 nm; emission, 360 nm). The UV detection limit of psilocybin was 20–40 ng (267 nm) and several ng could be detected fluorometrically. The identity of the compounds was verified by HPLC and thin-layer chromatography and by mass spectrometry and UV spectroscopy. The compounds were determined by means of a direct calibration method and by means of the method of internal normalization. The standard deviation of the determination was ±3.4% (relative). The above methods were used to determine these compounds in extracts of fruit bodies of two species of the genus Psilocybe growing at various places in Czechoslovakia, and found to contain 0.25–1.15% of psilocybin and 0.02–0.16% psilocin per dry mass.
Article
With the combined technique of GLC-mass spectrometry, psilocin and psilocybin, two hallucinogenic indoles, were analyzed as their trimethylsilyl derivatives. The method was applied to these two components in an extract of Psilocybe cubensis (Earle) Sing.
Article
Methods of examining dried fragments of hallucinogenic fungi are given; a key to the preliminary identification of suspected hallucinogenic mushrooms is offered to assist those working in this field. The key depends principally on simple microscopy.
Article
A method has been developed for the rapid isolation and identification of psilocin from psilocin/psilocybin-containing mushrooms. Based on the difference in the solubility properties in butyl chloride of psilocin and other constituents present in psilocin/psilocybin-containing mushrooms, psilocin was easily separated in pure form.
Article
A method using high performance liquid chromatography (HPLC) with an acetonitrile, water, and phosphoric acid mobile phase and a bonded cyano-amino-type polar phase column has been developed for the rapid, selective, and accurate quantitation of psilocybin in dry mushroom material. A simple one-step procedure is used for the quantitative extraction of psilocybin in under 60 min. The 267:254 nm absorbance ratio is used as a check on peak purity for the psilocybin response.
Article
The occurrence and extraction of indole derivatives in six species from four genera of higher fungi were investigated. By using pure methanol for extraction of the mushrooms analysis revealed the highest concentrations of psilocybin and baeocystin. The psilocin content of the species was higher by using aqueous solutions of alcohols than with methanol alone but was an artificial phenomenon caused by enzymatic destruction of psilocybin. The extraction with dilute acetic acid yielded better results than with the water containing alcohols. The simple one-step procedure with methanol for the quantitative extraction is still the safest method to obtain the genuine alkaloids from fungal biomass.
Article
A new method has been developed for the rapid analysis of psilocybin and/or psilocin in fungus material using ion mobility spectrometry. Quantitative analysis was performed by gas chromatography-mass spectrometry after a simple one-step extraction involving homogenization of the dried fruit bodies of fungi in chloroform and derivatization with MSTFA. The proposed methods resulted in rapid procedures useful in analyzing psychotropic fungi for psilocybin and psilocin.
Article
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.
Article
'Magic mushrooms' is the name most commonly given to psychoactive fungi containing the hallucinogenic components psilocybin and psilocin. Material confiscated by local authorities was examined using morphologic, microscopic, microchemical, and toxicological methods. Psilocybe cubensis was the most popular mushroom in the sample collective, followed by Psilocybe semilanceata, Panaeolus cyanescens and Psilocybe tampanensis. The alkaloid content was determined with <0.003-1.15% of psilocybin and 0.01-0.90% psilocin. Panaeolus cyanescens was the mushroom with highest levels of psilocybin and psilocin.
Analyisis of psilocybin and psilocin in psychotomimetic mushrooms by ion-pair liquid chromatography
  • S Kimoto
  • K Kato
  • K Yokoyama
  • M Nasu
S. Kimoto, K. Kato, K. Yokoyama, M. Nasu, Analyisis of psilocybin and psilocin in psychotomimetic mushrooms by ion-pair liquid chromatography, Jpn. J. Forensic Toxicol. 20 (2002) 192-193 (in Japanese).
Mushroom grow laboratory in a dorm room at the university of Maine/Orono
Mushroom grow laboratory in a dorm room at the university of Maine/Orono, Microgram 35 (2002) 191.
Analyisis of psilocybin and psilocin in psychotomimetic mushrooms by ion-pair liquid chromatography
  • Kimoto
Determination of tryptamine derivatives in hallucinogenic mushrooms using high-performance liquid chromatography with photodiode array detection
  • Bomer