毒蝇伞毒理学相关特性和中毒时的化学毒理学分析： 科学综述

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INTRODUCTION

Every year, hallucinogenic mushroom poisoning attracts more and more attention from researchers and practitioners. Amanita poisoning is a particularly pressing issue. The Department of Acute Poisoning and Somatopsychiatric Disorders at the Sklifosovsky Institute for Emergency Medicine reports an increase in cases of Amanita poisoning over the past four years, reaching 260 cases, with 94 in 2023 and 55 in the first half of 2024 [1]. A. muscaria and A. pantherina are the two Amanita species responsible for the increase in the cases of poisoning.

Since ancient times, Amanita mushrooms have been used for their psychodysleptic properties to induce an altered state of consciousness for religious and shamanic purposes. Amanita mushrooms are used for various purposes, including their psychodysleptic effects, religious rituals, research, and therapy.

In addition, cases of accidental Amanita ingestion should be mentioned [2, 3]. Poisoning cases in pets are also reported [4]. This may be largely due to a lack of awareness about mushrooms and their effects, as well as the increasing popularity of microdosing [5].

Microdosing involves taking subthreshold doses of psychedelic substances that do not cause intoxication or altered consciousness, in order to enhance creativity and physical performance, promote emotional stability, and alleviate anxiety [6, 7].

The growing popularity of microdosing is evident from the increase in advertisements and online stores offering microdosing products. The number of search queries can indirectly suggest consumer demand for Amanita-based products. From 2022 to 2023, the number of Google searches for A. muscaria increased by 114%, and this trend persisted in 2024 [5].

Various Amanita-based products are available for microdosing, including packaged dried materials, powders, capsules, tinctures, extracts, and creams. The Russian market offers a variety of certified food additives. Manufacturers state that these products improve:

• Cognitive functions
• Sleep
• Anxiety
• Improved alertness
• Physical activity.

Some products are promoted as supplements for treating alcohol addiction and attention deficit hyperactivity disorder when antidepressants are not an option [5].

Therefore, it is crucial to develop and evaluate physical and chemical tests to confirm and assess the severity of Amanita poisoning in order to deliver prompt and appropriate medical treatment.

This article provides information on the morphology and toxicology of Amanita, as well as methods for identifying their toxins. This information will be used to develop a laboratory method for diagnosing mushroom poisoning.

Amanita Species

Fig. 1a illustrates the appearance of the fruiting bodies of A. muscaria.

• The cap ranges in size from 5 to 20 cm, sometimes reaching up to 50 cm.
• The cap begins as an almost spherical shape and then turns from convex to concave.
• The caps range in color from bright red to orange or nearly yellow.
• Warts may be white or yellowish, or they may also be absent.
• The cap has the gill-bearing underside.
• The stem has warts near its basal bulb and a white ring that hangs down as the fungus grows.
• It smells and tastes pleasant.

A. muscaria is widespread in forests throughout Russia [8–10]. This ectomycorrhizal fungus forms a symbiotic relationship with both coniferous and deciduous trees [4].

Fig. 1b illustrates the appearance of the fruiting bodies of A. pantherina.

• The cap is round with a flat depression in the center.
• The cap is brown, often with a greenish tint, but is never reddish or yellowish.
• Warts are white or grayish in color and arranged in a concentric pattern.
• The stem is white and has a hanging ring that usually disappears quickly.
• A special, sack-shaped structure called a volva is located at the base of the stem.
• The fungus is tasteless and smells like raw potatoes.

A. pantherina is widespread in the northern and temperate zones of Russia, as well as in mountain forests [8–10].

 

Fig. 1. Appearance of fruiting bodies of mushrooms: a — red fly agaric (Amanita muscaria); b — panther fly agaric (Amanita pantherina). © Wikipedia, 2006. Distributed under license CC BY-SA 3.0.

 

The circulation of Amanita-based products is legal in Russia, but some other countries regulate it, particularly its primary active ingredients: muscimol and ibotenic acid. The collection and ingestion of A. muscaria are illegal in Romania, the Netherlands, Taiwan and the US state of Louisiana. However, the collection and ingestion of A. pantherina are not regulated anywhere. Australia regulates the circulation of muscimol, but not mushrooms [5, 9].

Chemical Composition

The quantity of active substances present in a fruiting body depends on the following factors:

• Geographic race
• Season
• Phases and conditions of growth
• Processing and storage options [9].

The two toxicologically significant components of Amanita mushrooms are ibotenic acid (Fig. 2a) and muscimol (Fig. 2b). Both are isoxazole derivatives. The highest concentrations of ibotenic acid and muscimol are found in the cap, followed by the volva, then the stem. These substances have relatively high boiling points of 152°C and 175°C, respectively. The acid dissociation constant (pKa) is 4.8 and 8.4 for muscimol [11] and 3.0, 5.0, and 8.2 for ibotenic acid [12], which indicates that ibotenic acid is more acidic. Pure substances are crystalline and freely soluble in water. In addition, they can pass through the blood-brain barrier by active transporters present in the brain [8]. In an aqueous medium, ibotenic acid forms a zwitterion¹, hindering sample preparation and chromatographic analysis [11]. Ibotenic acid is unstable when exposed to chemicals (including in the gastrointestinal tract), light, or temperature changes because it converts into muscimol, a primary amine, or the decarboxylated form, when dehydrated (Fig. 2b) [8–10, 13].

 

Fig. 2. Structural formulas of isoxazole derivatives: a — ibotenic acid; b — muscimol.

 

Amanita mushrooms contain another derivative of isoxazole called muscazone. It forms when ultraviolet rays act on muscimol, thereby intensifying its hallucinogenic properties. This explains why Amanita mushrooms collected in sunny meadows have a stronger psychodysleptic effect than those grown in shaded areas [5].

In addition, Amanita mushrooms contain tricholomacridic acid, stizolobic scid, stizolobinic acid, and methyl-tetrahydro-carboline-carboxylic (MTC) acid, as well as various alkaloids [8, 9, 14–16]. Stizolobic and stizolobinic acids are amino acids of non-protein origin, oxidation products of L-dihydroxyphenylalanine (3,4-dihydroxyphenylalanine). They are most often found in the fruiting bodies of Amanita pantherina. These amino acids are found in trace amounts in A. muscaria mushrooms, especially those with yellowish caps. In addition, extracts from the fruiting bodies of A. muscaria contain MTC with a structure similar to that of harmaline compounds [16]. The main alkaloids found in them are muscarine and nightshade alkaloids such as scopolamine, atropine, and hyoscyamine. The poisonous substance bufotenine, found in the skin of toads, is less common [5]. Amanita mushrooms can bioaccumulate certain metals in the form of organometallic compounds. This process can sometimes increase their toxicity. For example, in Amanita muscarica, vanadium accumulates in the form of amavadin [3].

Clinical Picture of Poisoning

The effects of different species, or even populations of the same species, may vary depending on the substances they contain. Therefore, in Russia, the effects of Amanita species can be classified as either mycoatropin syndrome or pantherin syndrome [9, 10]. Table 1 shows the main clinical features of poisoning from various species of Amanita.

 

Table 1. Main syndromes appearing in case of poisoning by different species of genus Amanita mushrooms and their characteristics

Mycoatropin	Pantherin
No clear presentation of poisoning; it can easily be confused with other conditions, such as a stroke, alcohol intoxication, or, less often, gastroenteritis.
Amanita species
A. muscaria A. regalis A. gemmata A. strobiliformis	A. pantherina
Main substances
Ibotenic acid Muscimol Muscazone	Ibotenic acid Muscimol Muscazone Nightshade alkaloids: scopolamine and atropine
Characteristics
Higher content of ibotenic acid Predominance of CNS excitation over depression Severe confusion, agitation	A 3–4 times stronger effect Higher content of muscimol Higher content of nightshade alkaloids Predominance of depression over excitation Coma is common

 

Mycoatropin syndrome is a complex of symptoms that are similar to the effects of atropine and caused by ibotenic acid derivatives found in mushrooms. This syndrome is caused by consuming the following species:

• A. muscaria
• A. regalis
• A. gemmata
• A. strobiliformis [8, 9].

Pantherin syndrome is a similar condition caused by ingestion of A. pantherina. Both syndromes develop within 15–30 minutes of Amanita ingestion due to the action of isoxazole derivatives and small amounts of alkaloids. This type of poisoning has no specific clinical presentation or typical symptoms [17]. In contrast to acute intoxication with other poisonous mushrooms, signs of toxic gastroenteropathy syndrome and gastroenteritis are not consistently observed in cases of Amanita poisoning, even during autopsy [16, 18]. Therefore, clinics often misdiagnose conditions such as alcohol intoxication and cerebrovascular disease [17]. However, pantherin syndrome is characterized by the predominance of central nervous system (CNS) depression, whereas mycoatropin syndrome is characterized by the predominance of CNS excitation. This is due to the high levels of ibotenic acid found in A. muscaria, A. regalis, A. gemmata and A. strobiliformis, and the high levels of muscimol found in A. pantherina [14, 17]. There is no specific antidote. In case of poisoning, only supportive and symptomatic therapy can be provided [3].

Amanita species that cause mycoatropin syndrome, contain the main psychodysleptic component called ibotenic acid. Ibotenic acid is a glutamate analog that can stimulate various types of glutamate receptor. Its primary target is ionotropic glutamate receptors that bind selectively to N-methyl-D-aspartate (NMDA). Their stimulation activates the nervous system [8, 15]. This is why ibotenic acid is used in rat models of Alzheimer disease [3, 12].

The first stage of the excitation phase is characterized by relatively harmless symptoms:

• Strong desire to be active
• Excitement
• Assertiveness
• Vitality
• Improved alertness [2, 17]

All of these manifestations are associated with the ability of ibotenic acid to increase serotonin levels. This is similar to the effects of lysergic acid diethylamide (LSD) [3].

The second phase manifests as follows:

• Severe psychomotor agitation with convulsions, which often develop in children [19], and ataxia
• Emotional arousal up to the point of hysteria
• Illusions such as micropsia and macropsia, as well as blurred vision and blurred outlines of objects (as in a migraine aura)
• Hallucinations involving visual, auditory, and gustatory [2, 17]

The excitement ends in the third phase, when hallucinations worsen and loss of orientation and coordination develop. The hallucinogenic effect is also caused by the effects of MTC on the CNS [16]. A patient becomes disoriented in space, time, and person. CNS depression eventually develops [2, 17].

CNS depression is mainly caused by the effects of muscimol. Muscimol is structurally similar to gamma-aminobutyric acid (GABA), the primary inhibitory neurotransmitter in the CNS. Therefore, muscimol interacts with GABA_A receptors, and partially with GABA_C receptors [8, 15]. At the same time, it provides relatively long-lasting suppression because, unlike GABA, it is not affected by GABA transaminases or their reuptake systems. In addition, CNS depression is largely the result of muscimol's ability to reduce catecholamine levels in the CNS. In contrast, ibotenic acid increases catecholamine levels [3]. This phase is characterized by:

• Depression of consciousness
• Lethargy
• Exhaustion
• Apathy
• Somnolence

The depression phase usually culminates in sopor, which is characterized by higher awakening thresholds and vivid, memorable dreams. Awakening is typically accompanied by retrograde amnesia [16]. In the case of pantherin syndrome, CNS depression prevails over excitation, and the depression phase is longer, often culminating in coma [2, 14, 17]. Ingesting a large amount of A. pantherina at once results in a rapid transition (within 30–60 min) from the initial stages to a coma. These phases may repeat and alternate many times [16].

These syndromes are also associated with:

• Mydriasis (less commonly miosis [16])
• Dyspnea
• Tachycardia (in the excitation phase) and bradycardia (in the depression phase)
• Salivation and lacrimation
• Excessive sweating
• Attacks of abdominal pain

Abdominal pain is associated with the effects of muscarine, which is present in very low amounts in both species of Amanita. Due to its poor oral absorption, however, this alkaloid plays a minimal role in toxicity [17]. The anticholinergic effects are primarily caused by stizolobic and stizolobinic acids [16]. Gastrointestinal spasms are primarily associated with the increased serotonin levels caused by muscimol [3]. In cases of severe poisoning, vomiting and diarrhea are possible [16].

Therefore, the clinical presentation varies greatly due to significant fluctuations in levels of ibotenic acid, muscimol, and other intoxicating substances. It is important to identify the most common symptoms of this syndrome when diagnosing [9, 16].

A single 50–70 gram fruiting body is sufficient to initiate nervous disorders. It may contain 0.015%–0.1% isoxazole derivatives (such as muscimol and ibotenic acid, at concentrations of approximately 6 mg and 30–60 mg, respectively) [3, 12, 16]. Notably, hallucinations occur with exposure to 10–15 mg of muscimol and 50–100 mg of ibotenic acid, respectively [11]. The most severe cases of poisoning are reported in children, older adults, and people with pre-existing health conditions [19, 20]. In most cases, however, poisoning from these Amanita species resolves favorably within 24 to 49 hours after the onset of symptoms [16]. The risk of death increases significantly when ingesting other Amanita species:

• A. phalloides
• A. verna
• A. virosa [21]

Fatal cases of poisoning from A. muscaria and A. pantherina are extremely rare. Most cases occur from acute poisoning after ingesting large quantities of the mushrooms [12, 16, 20], or after ingesting the mushrooms with tranquilizers, such as benzodiazepines [20]. Death occurs as a result of cardiac and respiratory arrest [22]. An autopsy does not reveal any typical pathological signs of Amanita poisoning [16]. In rats, the lethal dose (LD₅₀) of muscimol was found to be 4.5 mg/kg intravenously and 45 mg/kg orally.

Isoxazole derivatives are highly soluble in water, which results in the relatively rapid excretion of some unchanged compounds. Both ibotenic acid and muscimol can be detected in urine within one hour of ingestion. However, approximately 10%–20% of ibotenic acid is converted into muscimol when the mushrooms are digested in the acidic environment of the stomach [3, 12]. Ibotenic acid is also metabolized in the liver and brain [3].

Toxicological Analysis

Currently, there are many ways to detect Amanita poisoning. Previously, one way to confirm ingestion of these mushrooms was to perform macro- and microscopic examination of stomach or intestinal contents for mushroom pieces and spores. However, microscopy is a labor-intensive method that is more appropriate for postmortem identification [4, 12]. These methods are prone to error because crushed mushroom tissues lose their specific morphological characteristics when exposed to gastric and intestinal secretions. This makes visual identification impossible. Activated carbon and other sorbents are often used for self-treatment when dyspeptic symptoms of poisoning appear. This complicates identification because the chyme is stained black.

In some cases, the etiology of poisoning was established using polymerase chain reaction. However, this only identified the species of mushroom and not the amount of toxicants that entered the body. Even when gene databases (e.g., GenBank²²) and amino acid sequence search algorithms (e.g., BLAST³³) are used, inaccurate species identification is possible because many fungi grow together (for example, molds can be found in chyme) [4].

It is necessary to identify the toxicants involved in order to determine the severity of the poisoning and monitor the effectiveness of the therapy [11]. Advanced physical and chemical techniques, such as chromatography and electrophoresis, are the most reasonable methods for the qualitative and quantitative determination of ibotenic acid and muscimol. A sample of the biological object should be prepared by purifying the analytes⁴⁴ from related substances in the biological matrix.

Sample Preparation

Samples are prepared using extraction. Fresh fruiting bodies are crushed and dried samples are ground into a powder. Single-stage, single-phase extraction is primarily used to prepare samples for qualitative identification. Because of the high hydrophilicity of ibotenic acid and muscimol, various polar solvent systems are used for extraction, such as a mixture of methanol and water at 7:3 or a solution of 50% ethanol and 0.1% formic acid [15]. The use of 75% methanol as a universal extractant is considered optimal because it inhibits decarboxylase activity, which stabilizes ibotenic acid and prevents its conversion to muscimol. For the same reason, samples of fresh mushrooms are stored in methanol at a ratio of 4 mL per gram of raw material. The resulting sample is then stored at 4°C [23].

Highly purified extracts are obtained for the quantitative analysis of biological objects, such as urine and plasma. This is necessary for the optimal use of chromatographic columns. In addition, dansyl chloride, a common derivatization agent⁵⁵ in high-performance liquid chromatography (HPLC), reacts not only with ibotenic acid and muscimol but also with the amino acid residues of proteins when analyzing fruiting bodies or plasma, and this decreases the sensitivity of the method [24]. Multi-stage liquid-liquid or solid-phase extraction is typically used (Table 2).

 

Table 2. Isolation schemes of ibotenic acid and muscimol out of bioassays

Sample Preparation	Source
Plasma
Extract with a mixture of formic acid and acetonitrile. Centrifugate. Dansylation. Extract with acidic dichloromethane. Dry with nitrogen. Redissolve in a mixture of acetonitrile and water.	[24]
Urine
Perform an ion exchange. Wash with acidified ethanol (HCl), then remove the aqueous layer. Perform ethylation. Purify with pyridine. Dry with nitrogen. Redissolve in ethyl acetate.	[27]
Dilute at 1:10 or 1:100 with a mobile phase containing high concentrations of isoxazoles, then concentrate using solid-phase extraction at low concentrations. Dry with nitrogen. Re-dissolve in a mixture of mobile phase A and mobile phase B at 1:1.	[28]
Dried mushroom fruiting bodies
Extract with 50% ethanol. Filter. Perform multi-stage cleaning with organic solvents. Re-extract with methanol.	[29]
Extract with 50% ethanol. Centrifugate. Perform solid-phase extraction using formic acid in methanol as an eluent. Evaporate. Redissolve in a mixture of formic acid and methanol.	[30]
Perform double extraction using a mixture of methanol and water at 7:3. Centrifugate.	[26]
Extract in 70% methanol using an ultrasonic bath. Centrifugate. Perform trimethylsilylation.	[25]

 

Qualitative Analysis

The most common techniques for qualitative identification include thin-layer chromatography (TLC), gas-liquid chromatography (GLC), and gas-liquid chromatography-mass spectrometry (GLC-MS). TLC uses a methanol extract obtained from fresh, frozen, and dried fruiting bodies. The mobile phase can be represented by various solvent systems, for example:

• N-propanol : 10% ammonia (95:5)
• N-butanol : acetic acid : water (60:20:20) etc.

Silica gel is the most common stationary phase [23]. Ninhydrin and fluorescamine can be used as detectors, followed by ultraviolet treatment [25]. Pentacyanoferrate reagents, which produce purple spots, are less common. Note that ninhydrin interacts with not only MTC but also with stizolobic and stizolobinic acids, both of which are non-protein amino acids. However, they produce orange and yellow-brown coloration, respectively [16]. Preparative TLC can be used for purification and preparation for further GLC and HPLC. Elution is performed using n-butanol, ethanol, and aqueous acetic acid [26].

Various instrumental chromatographic techniques are available (Table 3 for general information).

 

Table 3. Schemes of chemical and toxicological analysis of blood plasma and urine for the presence of ibotenic acid and muscimol and chemical analysis of mushroom fruiting bodies via chromatographic methods

Techniques	Mobile phase	Stationary phase	Detector	Conditions	Calculation method	Source
Plasma
RP-HPLC	A: 10 mM formic acid and 6 mM ammonium formate B: acetonitrile	A non-polar C18 column with heptafluorobutyric acid	Time-of-flight mass spectrometric	Dansylation Gradient mode 30°С	Internal standard: Ibotenic acid: L-tyrosine-13C9,15N; Muscimol: tyramine-d4	[24]
Urine
GLC-MS	Helium	Semi-polar column HP–5MS UI® (Agilent Technologies, USA)	Mass spectrometric	Ethylation	Internal standard: cycloserine	[27]
HPLC-MS/MS	A: 10 mM ammonium acetate in a mixture of water : acetonitrile at 1:9 B: 0.1% aqueous formic acid	Semi-polar column: ZORBAX StableBond SB-CN® (Agilent Technologies, USA)	Quadrupole Orbitrap mass spectrometer	Gradient mode 40°С Positive ionization	External standard: ibotenic acid; muscimol	[28]
Dried mushroom fruiting bodies
HPLC, UHPLC	Acetonitrile : ammonium acetate 10 mM: 80:20 pH 6.8	Hydrophilic interaction liquid chromatography: silica gel with cross-linked triazole	Ultraviolet, 255 nm	Isocratic mode	Spike test	[29]
HPLC-MS/MS	A: 0.5% aqueous formic acid B: a mixture of 0.5% formic acid and acetonitrile	Silica gel with cross-linked carbamoyl groups: TSK-GEL Amide-80® (Tosoh Bioscience, Japan)	Mass spectrometric	Gradient mode	Internal standard: Acivicin	[30]
RP-HPLC-MS	A: 10 mM ammonium acetate B: acetonitrile	Non-polar С18 column	Mass spectrometric Diode array (256 nm)	Gradient mode Dansylation with ethylation 40°С	External standard: muscimol; ibotenic acid	[26]
GLC-MS	Helium	Semi-polar column: DB–5MS® (Agilent Technologies, USA)	Mass spectrometric	Trimethylsilylation	External standars: muscimol; ibotenic acid Internal standard: n-pentadecane	[25]

Note. RP-HPLC, reversed-phase high-performance liquid chromatography; UHLPC, ultra-high performance liquid chromatography; GLC, gas liquid chromatography; MS, mass-spectrometry.

 

GLC-MS is only performed after the derivatization stage⁵ because the substances have a fairly high boiling point and ibotenic acid is thermally unstable [25]. In addition, when analyzed directly without derivatization,⁵ ibotenic acid and muscimol typically exhibit only one characteristic transition in the mass spectrum:

• ibotenic acid: 159 → 113
• muscimol: 115 → 98

However, this is insufficient to accurately identify toxicants [24]. Trimethylsilyl and ethyl derivatives are obtained [25, 27]. Ethylation is used to identify compounds in urine, but methods for trimethylsilylated compounds are also being developed [1]. The stationary phase is usually represented by semi-polar silicon dioxide sorbents with cross-linked phenolic groups, and helium serves as the carrier gas [25, 27].

HPLC is considered the most effective technique. If the levels of ibotenic acid and/or muscimol in the biofluids are considered high, the samples are diluted 10- or 100-fold. If the levels are low, the samples are concentrated using solid-phase extraction. Acetate or formate buffers mixed with acetonitrile are most often used as mobile phases. Gradient mode is preferred due to the differences in acid-base properties between the substances being analyzed and the ability of ibotenic acid to form a zwitterion¹ in an aqueous medium [13]. More polar sorbents are the most preferable because the high polarity of substances being analyzed reduces the efficiency of non-polar C18 columns [24, 26–30]. Levels are calculated using both internal and external standards, as well as the spike test, though the latter is significantly less common (see Table 3).

Electrophoresis

Chromatographic techniques are complex, time-consuming, and relatively expensive, especially in cases of multi-stage sample preparation with derivatization.⁵ These factors limit their use when a quick result is necessary. Therefore, special attention is given to capillary electrophoresis using a diode-array or mass spectrometric detector (Table 4). The main advantages of electrophoresis include:

• Rapid test
• Easy sample preparation
• High sensitivity to ibotenic acid and muscimol

 

Table 4. Schemes of chemical and toxicological analysis of urine for the presence of ibotenic acid derivates and chemical analysis of mushroom fruiting bodies via capillary electrophoresis

Sample preparation	Injection parameters	Conditions			Detection	Testing time, min	Detection limit		Source
		pH	T, °С	Special conditions			Ibotenic acid	Muscimol
Urine
Dilute five times with deionized water. Filter through a 0.22 µm membrane microfilter.	0.1 atm 5 s	2.7	25	Create the environment using 0.2 M acetic acid. Perform electrospray ionization in the isocratic mode.	Mass spectrometric	24	0.15 ng/mL	0.05 ng/mL	[12]
Perform single-drop microextraction. Extract from the donor phase into octanol at pH 4. Re-extract into the aqueous phase (pH 3).	0.0068 atm 1 s	3	18	The creatinine appears on the electropherogram.	Diode array (214 nm)	7	—	16 µg/L	[11]
Dry fruiting bodies
Extract into a mixture of methanol and buffer solution with ultrasound. Filter and evaporate the extract. Dissolve in a mixture of methanol and buffer at 2:1.	0.034 atm 5 s	3	22	The mobile phase contains 5% acetonitrile.	Diode array (214 nm)	20	1.5 µg/mL	1.8 µg/mL	[24]

 

This technique, when used with tandem mass spectrometry, allows for the identification of isoxazole derivatives in urine at a relatively low detection threshold. For example, the threshold is 0.15 ng/mL and 0.05 ng/mL for ibotenic acid and muscimol, respectively [5]. The following characteristic transitions are detected:

• Ibotenic acid: 159 → 113 and 159 → 99
• Muscimol: 115 → 98 and 115 → 86 [12, 24]

A faster, more efficient technique has been developed to determine muscimol. This technique uses microextraction for sample preparation. A urine sample diluted with a phosphate buffer solution at pH of 4 serves as the donor phase. It contains a large number of dihydrogen phosphate anions, which form neutral ion pairs with the positively charged muscimol molecule. Ion pairs are extracted into octanol. Due to a concentration gradient, they then move into a more acidic acceptor phase (pH 3) which contains fewer dihydrogen phosphate ions and more muscimol cations. Electrophoresis is performed on the acceptor phase. This technique requires a minimal amount of chemical reagents, can be performed quickly (in 7 minutes), and allows for the analysis of only 200 µL of sample [11].

CONCLUSION

A review of publications reveals a global trend of increased use of Amanita species, particularly A. muscaria and A. pantherina via microdosing to enhance creativity, physical performance, and emotional well-being. This trend is associated with an increase in mushroom poisoning rates. This is due to the low awareness of the pharmacological effects of Amanita and the wide availability of various Amanita-based products on the market. This is why it is important to develop and implement toxicological techniques that can rapidly and accurately detect the active substances, muscimol and ibotenic acid, in the A. muscaria and A. pantherina.

GLC and HPLC are the most common chromatographic techniques. Among rapid tests, electrophoresis with mass spectrometric or diode-matrix detection stands out due to its high sensitivity. This makes it a promising approach for developing techniques in chemical toxicology to detect Amanita poisoning.

ADDITIONAL INFORMATION

Authors’ contributions: V.A. Zelenshchikova: conceptualization, collection and analysis of published data, writing—original draft; M.V. Belova, E.V. Melnik: conceptualization, collection and analysis of published data, writing—review & editing. Thereby, all authors provided approval of the version to be published and agree to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.

Acknowledgment: The authors express their gratitude to V.V. Severtsev for providing information on the statistics of poisoning by mushrooms of the genus fly agaric according to the department of acute poisoning of somatopsychiatric disorders of the Sklifosovsky Research Institute for Emergency Medicine; to V.S. Kulbatsky for providing methodological recommendations on the analysis of isoxazole derivatives.

Ethics approval: Not applicable.

Funding sources: No funding.

Disclosure of interests: The authors declare that they have no relationships, activities or interests over the past three years related to third parties (commercial and non-commercial) whose interests may be affected by the content of the article.

Statement of originality: When creating this work, the authors did not use previously published information (text, illustrations, data).

Data availability statement: The editorial policy regarding data sharing does not apply to this work.

Generative AI: Generative AI technologies were not used for this article creation.

Provenance and peer-review: This article was submitted to the Journal on an unsolicited basis and reviewed according to the usual procedure. One external reviewer, a member of the editorial board, and the scientific editor of the Journal were involved in the review process.

 

¹ A zwitterion is a molecule that is usually electrically neutral, yet contains functional groups that carry both positive and negative charges.

² GenBank® [Internet]. Bethesda (MD): National Library of Medicine (US); 2013–2024. Available at: https://www.ncbi.nlm.nih.gov/genbank/

³ Basic local alignment search tool® [Internet]. (MD): National Library of Medicine (US); 1990–2024. Available at: https://blast.ncbi.nlm.nih.gov/Blast.cgi Accessed on January 9, 2025.

⁴ An analyte is a target chemical substance for analysis.

⁵ In chemistry, derivatization is an analytical technique that converts a chemical compound into a derivative, which is a product with a similar chemical structure.

作者简介

Varvara A. Zelenshchikova

Sechenov First Moscow State Medical University (Sechenov University)

Email: barbarazelen02@mail.ru
ORCID iD: 0000-0002-9152-6379
SPIN 代码: 7929-8875
俄罗斯联邦, 27 Baryshikha st, Moscow,125368

Maria V. Belova

Sechenov First Moscow State Medical University (Sechenov University)
Sklifosovsky Research Institute for Emergency Medicine

Email: belova_m_v@staff.sechenov.ru
ORCID iD: 0000-0002-0861-5945
SPIN 代码: 6197-3906

Dr. Sci. (Biology), Professor

俄罗斯联邦, 27 Baryshikha st, Moscow,125368; Moscow

Elizaveta V. Melnik

Sechenov First Moscow State Medical University (Sechenov University)

编辑信件的主要联系方式.
Email: melnik_e_v_2@staff.sechenov.ru
ORCID iD: 0000-0002-9727-9452
SPIN 代码: 5523-1359

Cand. Sci. (Pharmacy)

俄罗斯联邦, 27 Baryshikha st, Moscow,125368

参考

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