Antioxidant Activity and Effectiveness of Fig Extract in Counteracting Carbon Tetrachloride-Induced Oxidative Damage in Rats.

Figs are the edible fruits of the fig tree, Ficus carica L., that have been used for centuries for human consumption and in traditional medicine, to treat skin problems, inflammation, and gastrointestinal disorders. Our previous study investigated the presence of phenolic compounds in aqueous extracts of two Algerian popular fig varieties, azendjar (Az) and taamriouth (Ta), as well as their in vitro antioxidant activity. In this study, we assessed hydroethanolic extracts of these fig varieties. The total phenolic content was measured, along with the phenolic profile. Rutin was determined to be the dominant phenolic compound, followed by vanillic acid, 3,4-dihydroxybenzoic acid, quercetin, 4-hydroxybenzoic acid, rosmarinic acid (in Az only), and cinnamic acid. The antioxidant activity of the extracts was evaluated both in vitro (DPPH and FRAP assays) and in vivo, in rats intoxicated with carbon tetrachloride. In all assays, the fig extract-especially the dark-peeled fig variety azendjar-showed antioxidant potency. The administration of fig extract resulted in a reduction in liver damage, expressed by both different biochemical markers and histopathological study (less degraded liver architecture, reduced fibrosis, and only mild inflammation). A dose-dependent therapeutic effect was observed. The extract from the dark-peeled fig variety, Az, was characterized by a higher phenolic content and a stronger antioxidant activity than the extract from the light-peeled variety-Ta. Our study justifies the use of figs in traditional healing and shows the potential of using fig extracts in natural medicines and functional foods.

Lavandula × intermedia-A Bastard Lavender or a Plant of Many Values? Part I. Biology and Chemical Composition of Lavandin.

This review article is the first in a series that provides an overview of the biology, chemistry, biological effects, and applications of Lavandula × intermedia (lavandin, LI). Despite its prevalence in cultivation and on the essential oil market, lavandin has received limited attention from the scientific community. Remarkably more attention is paid to Lavandula angustifolia (LA), which is commonly regarded as the superior lavender and has been extensively researched. Our goal is to provide a comprehensive review of LI, as none currently exists, and assess whether its inferior status is merited. In the first part, we outline the biological and chemical characteristics of the plant and compare it to the parent species. The chemical composition of lavandin oil is similar to that of LA but contains more terpenes, giving camphor notes that are less valued in perfumery. Nevertheless, lavandin has some advantages, including a higher essential oil yield, resulting in reduced production cost, and therefore, it is a preferred lavender crop for cultivation.

Lavandula x intermedia-A Bastard Lavender or a Plant of Many Values? Part II. Biological Activities and Applications of Lavandin.

This review article is the second in a series aimed at providing an in-depth overview of Lavandula x intermedia (lavandin). In part I, the biology and chemistry of lavandin were addressed. In part II, the focus is on the functional properties of lavandin and its applications in industry and daily life. While reviewing the biological properties, only original research articles employing lavandin were considered. Lavandin essential oil has been found to have antioxidant and biocidal activity (antimicrobial, nematicidal, antiprotozoal, insecticidal, and allelopathic), as well as other potential therapeutic effects such as anxiolytic, neuroprotective, improving sleep quality, antithrombotic, anti-inflammatory, and analgesic. Other lavandin preparations have been investigated to a much lesser extent. The research is either limited or inconsistent across all studies, and further evidence is needed to support these properties. Unlike its parent species-Lavandula angustifolia (LA)-lavandin essential oil is not officially recognized as a medicinal raw material in European Pharmacopeia. However, whenever compared to LA in shared studies, it has shown similar effects (or even more pronounced in the case of biocidal activities). This suggests that lavandin has similar potential for use in medicine.

HPLC-DAD profile of phenolic compounds and In vitro antioxidant activity of Ficus carica L. fruits from two Algerian varieties.

Ficus carica L., commonly known as the fig tree, is a plant belonging to the Moraceae family whose fruits are traditionally used for edible and therapeutic purposes. The study aimed to investigate the lyophilized aqueous extracts of two native Algerian fig varieties, azendjar (Az) and taamriouth (Ta), as a potential source of antioxidant compounds for possible use as ingredients in pharmaceuticals or nutraceuticals. The HPLC-DAD analysis revealed the presence of two phenolic acids (3,4-dihydroxybenzoic acid and vanillic acid) and two flavonoids (rutin and quercetin) at levels 3.67, 4.80, 84.16, and 6.87 µg/g respectively for Az variety extract, and 6.90, traces, 7.46 and 3.37 µg/g respectively for Ta variety extract. Total phenolic content was determined using the Folin-Ciocalteu method at levels 951.06 ± 61.08 and 730.88 ± 45.25 GAE mg/100 g of the dry extract. In contrast, the total flavonoid content was determined using Christ-Müller's method at levels 428.34 ± 15.42 and 307.63 ± 7.94 QE mg/100 g of dry extract in the Az and Ta varieties, respectively. The total polyphenolic content of the extract may be responsible for its antioxidant action. The gathered results indicate that the extracts from the dark peel fig variety - azendjar, are characterized by a higher content of phenolic and flavonoid compounds and antioxidant activity than the extract from the light peel variety - taamriouth. In conclusion, the conducted studies and in vitro assays indicate that the studied extracts are a source of natural antioxidants and can be considered functional raw materials for producing food supplements and pharmaceuticals.

Polyphenols and Pharmacological Screening of a Monarda fistulosa L. dry Extract Based on a Hydrodistilled Residue By-Product.

This study aimed to determine the composition and content of polyphenols in the dry extract obtained from the hydrodistilled residue by-product of the wild bergamot (Monarda fistulosa L., Lamiaceae Martinov family) herb (MFDE) and to evaluate its safety and pharmacological properties. The total phenolic content (TPC) in the MFDE was 120.64 mg GAE/g. The high-performance liquid chromatography (HPLC) analysis showed the presence of a plethora of phenolic compounds, including hydroxycinnamic acids and flavone derivatives in the MFDE, with rosmarinic acid and luteolin-7-O-glucoside being the main components. With an IC50 value of 0.285 mg/mL, it was found to be a strong DPPH radical scavenger. The acute toxicity study results indicate that the oral administration of MFDE to rats at the doses of 500-5,000 mg/kg did not produce any side effects or death in animals which indicates its safety. The results of the in vivo assay showed that the MFDE dose-dependently inhibited paw oedema and significantly reduced the number of writings in mice induced by the acetic acid injection suggesting its potent anti-inflammatory and analgesic activities, respectively. The conducted studies revealed that M. fistulosa hydrodistilled residue by-product could be regarded as a new natural source of polyphenols with valuable pharmacological properties.

Determination of Glyphosate and AMPA in Food Samples Using Membrane Extraction Technique for Analytes Preconcentration.

The method for determining glyphosate (NPG) and its metabolite AMPA (aminomethyl phosphonic acid) in solid food samples using UAE-SLM-HPLC-PDA technique was developed. Firstly, ultrasonic-assisted solvent extraction (UAE) and protein precipitation step were used for the analyte isolation. Then, the supernatant was evaporated to dryness and redissolved in distilled water (100 mL). The obtained solution was alkalized to pH 11 (with 1 M NaOH) and used directly as donor phase in SLM (supported liquid membrane) extraction. The SLM extraction was performed using 2 M NaCl (5 mL) as an acceptor phase. The flow rate of both phases (donor and acceptor) was set at 0.2 mL/min. The membrane extraction took 24 h but did not require any additional workload. Finally, the SLM extracts were analyzed using the HPLC technique with photo-diode array detector (PDA) and an application of pre-column derivatization with p-toluenesulfonyl chloride. Glyphosate residues were determined in food samples of walnuts, soybeans, barley and lentil samples. The LOD values obtained for the studied food were 0.002 µg g-1 and 0.021 µg g-1 for NPG and AMPA, respectively. Recoveries values ranged from 32% to 69% for NPG, 29% to 56% for AMPA and depended on the type of sample matrix. In the case of buckwheat and rice flour samples, the content of NPG and AMPA was below the detection level of a used analytical method.

Development of high-performance thin layer chromatography method for identification of phenolic compounds and quantification of rosmarinic acid content in some species of the Lamiaceae family.

AIM: Representatives of Nepetoideae Burnett subfamily are promising species of the Lamiaceae Martinov family because of accumulating such valuable groups of secondary metabolites as terpenoids and polyphenols. A high-performance thin layer chromatography (HPTLC) fingerprint method for the qualitative determination of phenolic compounds and for the quantification of rosmarinic acid (RA) content in methanol extracts of five species of this subfamily was developed for the first time. MATERIALS AND METHODS: Dried aerial parts of Dracocephalum moldavica L., Ocimum americanum L., Lophanthus anisatus (Nutt.) Benth., Monarda fistulosa L., and Satureja hortensis L. collected in flowering period were macerated with methanol. The HPTLC analysis was conducted using the CAMAG analytical system (Muttenz, Switzerland). The comparative analysis of RA contents was performed by HPTLC densitometric detection at λ = 366nm. RESULTS: Identification of polyphenols in the investigated herbs was performed by comparison of a color and R f of the chromatographic zones with six reference standards: rutin, apigenin, luteolin, caffeic acid, chlorogenic acid, and RA. HPTLC method was also validated for the quantification of RA in the extracts of investigated herbs. RA contents decreased in such a sequence: D. moldavica (24.83 ± 0.78mg/g) > M. fistulosa (20.32 ± 0.64mg/g) > O. americanum (19.59 ± 0.61mg/g) > S. hortensis (18.77 ± 0.52mg/g) > L. anisatus (12.61 ± 0.43mg/g). CONCLUSION: Obtained data can facilitate the differentiation of investigated species using the chromatographic fingerprints of their phenolic compounds. Developed and validated HPTLC method provides an approach to estimate RA content as a common marker of investigated herbs.

Direct Analysis of Psilocin and Muscimol in Urine Samples Using Single Drop Microextraction Technique In-Line with Capillary Electrophoresis.

The fully automated system of single drop microextraction coupled with capillary electrophoresis (SDME-CE) was developed for in-line preconcentration and determination of muscimol (MUS) and psilocin (PSC) from urine samples. Those two analytes are characteristic active metabolites of Amanita and Psilocybe mushrooms, evoking visual and auditory hallucinations. Study analytes were selectively extracted from the donor phase (urine samples, pH 4) into the organic phase (a drop of octanol layer), and re-extracted to the acidic acceptor (background electrolyte, BGE), consisting of 25 mM phosphate buffer (pH 3). The optimized conditions for the extraction procedure of a 200 µL urine sample allowed us to obtain more than a 170-fold enrichment effect. The calibration curves were linear in the range of 0.05-50 mg L-1, with the correlation coefficients from 0.9911 to 0.9992. The limit of detections was determined by spiking blank urine samples with appropriate standards, i.e., 0.004 mg L-1 for PSC and 0.016 mg L-1 for MUS, respectively. The limits of quantification varied from 0.014 mg L-1 for PSC and 0.045 mg L-1 for MUS. The developed method practically eliminated the sample clean-up step, which was limited only to simple dilution (1:1, v/v) and pH adjustment.

What is the form of muscimol from fly agaric mushroom (Amanita muscaria) in water? An insight from NMR experiment supported by molecular modeling.

The biologically active alkaloid muscimol is present in fly agaric mushroom (Amanita muscaria), and its structure and action is related to human neurotransmitter γ -aminobutyric acid (GABA). The current study reports on determination of muscimol form present in water solution using multinuclear 1 H and 13 C nuclear magnetic resonance (NMR) experiments supported by density functional theory molecular modeling. The structures of three forms of free muscimol molecule both in the gas phase and in the presence of water solvent, modeled by polarized continuous model, and nuclear magnetic isotropic shieldings, the corresponding chemical shifts, and indirect spin-spin coupling constants were calculated. Several J-couplings observed in proton and carbon NMR spectra, not available before, are reported. The obtained experimental spectra, supported by theoretical calculations, favor the zwitterion form of muscimol in water. This structure differs from NH isomer, previously determined in dimethyl sulfoxide (DMSO) solution. In addition, positions of signals C3 and C5 are reversed in both solvents.

Study of biocompatibility of peritoneal dialysis solutions measured as in vitro cells viability.

This paper presents the comparable viability study results of the HepG2 and Vero cells in the presence of traditional peritoneal dialysis (PD) solutions determined by three methods (3-[4,5-dimethylthiazol]-2-yl-2,5-diphenyl tetrazolium bromide (MTT), neutral red (NR) and sulforhodamine B assays) with establishing different correlations between viability and quality indexes of the tested PD solutions. The obtained results confirmed cytotoxicity of the PD solutions even compared with an isotonic solution of sodium chloride. PD solutions action resulted in a similar reduction in the HepG2 and Vero cells. Moreover, this research found that metabolic cellular activity is more vulnerable to the action of PD solutions measured in the MTT-test. One more point is that cytotoxicity is related to pH of a solution and other unknown mechanisms, while glucose degradation products, glucose or lactate did not exert an exceptional negative action on PD solutions cytotoxicity. It is concluded that MTT-test is the best suitable for comparative studies of PD solutions which differ in pH values.

Chemical Composition of Two Different Lavender Essential Oils and Their Effect on Facial Skin Microbiota.

Lavender oil is one of the most valuable aromatherapy oils, its anti-bacterial and anti-fungal activities can be explained by main components such as linalool, linalyl acetate, lavandulol, geraniol, or eucalyptol. The aim of the study was to assess the anti-microbial effects of two different lavender oils on a mixed microbiota from facial skin. The commercial lavender oil and essential lavender oil from the Crimean Peninsula, whose chemical composition and activity are yet to be published, were used. Both oils were analysed by gas chromatography coupled to mass spectrometry. The composition and properties of studied oils were significantly different. The commercial ETJA lavender oil contained 10% more linalool and linalyl acetate than the Crimean lavender oil. Both oils also had different effects on the mixed facial skin microbiota. The Gram-positive bacilli were more sensitive to ETJA lavender oil, and Gram-negative bacilli were more sensitive to Crimean lavender oil. However, neither of the tested oils inhibited the growth of Gram-positive cocci. The tested lavender oils decreased the cell number of the mixed microbiota from facial skin, but ETJA oil showed higher efficiency, probably because it contains higher concentrations of monoterpenoids and monoterpenes than Crimean lavender oil does.

The application of the supported liquid membrane and molecularly imprinted polymers as solid acceptor phase for selective extraction of biochanin A from urine.

An efficient sample clean-up and preconcentration procedure for phytoestrogens analysis in urine has been developed. It was based on a combination of solid phase extraction with hollow-fiber supported liquid membrane and molecularly imprinted beads (MIPs-HF-SLM-SPE). The molecularly imprinted polymers (MIPs) were synthesized by precipitation polymerization technique with biochanin A (BCA) as a template, giving narrowly dispersed microspheres with a regular shape. As the functional monomer, (dimethylamino)ethyl methacrylate (-DEM) turned out to be better than methacrylic acid (MAA) to get the best-imprinted effects. The MIPs used as sorbents in the MIPs-HF-SLM-SPE extraction process exhibited excellent binding selectivity for BCA, in comparison to non-imprinted polymers as well as its structural analogs (genistein and daidzein). Finally, the developed method was used to detect BCA in urine. Under optimal extraction conditions, the recovery of BCA in urine samples (using 4.5 mL sample spiked with 10 µg L-1) was over 41%, with a coefficient of variation (CV) < 6.6% (n = 5). The detection limit (LOD) and quantification limit (LOQ) for BCA analysis in urine were 0.41 and 1.36 µg L-1, respectively.

Secondary metabolites from the aerial parts of Cytisus villosus Pourr.

Phytochemical investigation of the aerial parts of Cytisus villosus Pourr. resulted in the isolation and characterization of a new isoflavan, (3S, 4S)-2',4'-dihydroxy-3'-methoxy-6,7-methylenedioxyisoflavan- 4-ol (1), and a new monoterpene, (4R,6S)-4-hydroxy-2,2,6-trimethyl-9-oxabicyclo [4.2.1] non-1(8)-en-7-one (2), together with four known flavonoids: geinstein (3), chrysin (4), chrysin -7-O-ß-D-glucopyranoside (5) and 2″-O-α-L-rhamnosylorientin (6). The structures of the new compounds were elucidated on the basis of extensive spectroscopic analysis, including 1D, 2D NMR (1H, 13C, COSY, TOCSY, HMBC and HSQC) and HRESIMS. The absolute configurations of 1 and 2 were established by the comparison of experimental and calculated electronic circular dichroism (ECD) spectra.

Chemical Composition of Herbal Macerates and Corresponding Commercial Essential Oils and Their Effect on Bacteria Escherichia coli.

This study addresses the chemical composition of some commercial essential oils (clove, juniper, oregano, and marjoram oils), as well as appropriate herbal extracts obtained in the process of cold maceration and their biological activity against selected Escherichia coli strains: E. coli ATTC 25922, E. coli ATTC 10536, and E. coli 127 isolated from poultry waste. On the basis of the gas chromatography-mass spectrometry (GCMS) analysis, it was found that the commercial essential oils revealed considerable differences in terms of the composition and diversity of terpenes, terpenoids and sesquiterpenes as compared with the extracts obtained from plant material. The commercial clove, oregano, and marjoram oils showed antibacterial properties against all the tested strains of E. coli. However, these strains were not sensitive to essential oils obtained from the plant material in the process of maceration. The tested strains of E. coli show a high sensitivity, mainly against monoterpenes (α-pinene, ß-pinene, α,ß,γ-terpinene, limonene) and some terpenoids (thymol, carvacrol). The commercial juniper oil contained mainly monoterpenes and monoterpenoids, while the extracts contained lower amounts of monoterpenes and high amounts of sesquiterpenes-the anti-microbiotic properties of the juniper herbal extract seem to be caused by the synergistic activity of mono- and sesquiterpenes.

Simultaneous determination of nine phytohormones in seaweed and algae extracts by HPLC-PDA.

An RP-HPLC-PDA method for the simultaneous analysis of 9 compounds deriving from the phytohormones class was developed and optimized, namely indoleacetic acid (IAA), indolebutyric acid (IBA), phenyleacetic acid (PAA), naphtyleacetic acid (NAA), trans-zeatin (TZ), kinetin (KA), isopentenyladenine (IA), 6-benzylaminopurine (6-BA) and abscisic acid (ABA). Validation of the method was performed on the SFE-CO2 extract made out of the mixture of Baltic algae. The regression coefficients for plant hormones were in the range from 0.997 to 0.999. The LOD and LOQ were on the levels from 0.05-0.29 and 0.15-0.88mg/L, respectively. Developed method was used for the separation and determination plant hormones in extracts obtained by SFE-CO2 (supercritical fluid extraction) made out of the mixture of Baltic algae, Cladophora glomerata and Spirulina sp. In the extract of Baltic seaweed 2 of tested compounds were present in the concentration of 154,45±20,63µg/g for TZ and 362,47±13,00µg/g for PAA, whereas in Cladophora glomerata extract contained PAA and IAA in the concentration of 229,30±7,90µg/g and 23,91±0,80µg/g, respectively (all values per g of extract). The differences in the hormones levels may occur due to the different scale of extract preparation (laboratory or industrial) and other factors like the place of algae collection, year of collection or the way of biomass preparation.

Theoretical and experimental NMR studies on muscimol from fly agaric mushroom (Amanita muscaria).

In this article we report results of combined theoretical and experimental NMR studies on muscimol, the bioactive alkaloid from fly agaric mushroom (Amanita muscaria). The assignment of (1)H and (13)C NMR spectra of muscimol in DMSO-d6 was supported by additional two-dimensional heteronuclear correlated spectra (2D NMR) and gauge independent atomic orbital (GIAO) NMR calculations using density functional theory (DFT). The effect of solvent in theoretical calculations was included via polarized continuum model (PCM) and the hybrid three-parameter B3LYP density functional in combination with 6-311++G(3df,2pd) basis set enabled calculation of reliable structures of non-ionized (neutral) molecule and its NH and zwitterionic forms in the gas phase, chloroform, DMSO and water. GIAO NMR calculations, using equilibrium and rovibrationally averaged geometry, at B3LYP/6-31G* and B3LYP/aug-cc-pVTZ-J levels of theory provided muscimol nuclear magnetic shieldings. The theoretical proton and carbon chemical shifts were critically compared with experimental NMR spectra measured in DMSO. Our results provide useful information on its structure in solution. We believe that such data could improve the understanding of basic features of muscimol at atomistic level and provide another tool in studies related to GABA analogs.

Research on acute toxicity and the behavioral effects of methanolic extract from psilocybin mushrooms and psilocin in mice.

The pharmacological activities and acute toxicity of the psilocin (PC) and dried residues of the crude extracts of psychotropic mushrooms were investigated in mice. The hallucinogenic substances were effectively isolated, by using methanol, from the species of Psilocybe semilanceata and Pholiotina cyanopus, that were collected in the north-east region of Poland. The chemical analysis of these extracts, which was performed by liquid chromatography with mass spectrometry detection (LC-MS), indicated the presence of psilocin and other hallucinogenic substances, including indolealkylamines and their phosphorylated analogues. When the pure psilocin or fungal extracts were used, slight differences in determined LD50 values were observed. However, the application of PC evoked the highest level of toxicity (293.07 mg/kg) compared to the activity of extracts from Ph. cyanopus and P. semilanceata, where the level of LD50 was 316.87 mg/kg and 324.37 mg/kg, respectively. Furthermore, the behavioral test, which considered the head-twitching response (HTR), was used to assess the effects of the studied psychotropic factors on the serotonergic system. Both, the fungal extracts and psilocin evoked characteristic serotoninergic effects depending on the dose administered to mice, acting as an agonist/partial agonist on the serotonergic system. A dose of 200 mg/kg 5-hydroxytryptophan (5-HTP) induced spontaneous head-twitching in mice (100% effect), as a result of the formation of 5-hydroxytryptamine (5-HT) in the brain. Compared to the activity of 5-HTP, the intraperitoneal administration of 1mg/kg of psilocin or hallucinogenic extracts of studied mushrooms (Ph. cyanopus and P. semilanceata) reduced the number of head-twitch responses of about 46% and 30%, respectively. In contrast, the administration of PC exhibited a reduction of about 60% in HTR numbers.

Surface molecularly imprinted silica for selective solid-phase extraction of biochanin A, daidzein and genistein from urine samples.

Selective molecularly imprinted silica polymer (SiO2MIP) for extraction of biochanin A, daidzein and genistein was synthesized using the surface molecular imprinting technique with the silica gel as a support. Biochanin A (BCA) was used as a template, 3-aminopropyltriethoxysilane (APTES) as a functional monomer, and tetraethoxysilicane (TEOS) as a cross-linker. Non-imprinted polymer with the sol-gel process (SiO2NIP) was also prepared for comparison. The synthesized polymers were characterized by Fourier transform infrared spectrometry (FTIR), scanning electron microscopy (SEM) and a standard Brunauer-Emett-Teller (BET) and Barret-Joyner-Halenda (BJH) analysis. The obtained results indicated the structural differences between imprinted and non-imprinted polymers. Finally, the SiO2MIP and SiO2NIP were adopted as the adsorbents of solid phase extraction for isolation and preconcentration of biochanin A and its structural analogues-daidzein and genistein from aqueous and urine samples. The performance analysis revealed that SiO2MIP displayed better affinity to the three investigated isoflavones compared with SiO2NIP. The recoveries of spiked samples for studied analytes ranged from 65.7% to 102.6% for molecularly imprinted silica polymer and 8.9-16.0% for non-imprinted sorbents.

Characterization of particle morphology of biochanin A molecularly imprinted polymers and their properties as a potential sorbent for solid-phase extraction.

Molecularly imprinted polymers (MIPs) with biochanin A as a template were obtained using a bulk polymerization with non-covalent imprinting approach. The polymers were prepared in acetonitrile as porogen, using ethylene glycol dimethacrylate (EDMA) as cross-linking agent. The synthesis, with an application of 1',1'-azobis(cyclohexanecarbonitrile) (ACHN) as an initiator, has been performed thermally. During the synthesis process the effect of different functional monomers such as methacrylic acid (MAA), acrylamide (AA) and 4-vinylpyridine (4-VP) was investigated. The application of nitrogen sorption porosimetry, scanning electron microscopy (SEM), and Fourier transform infrared spectroscopy (FTIR) permitted the characterization and evaluation of synthesized polymers. The adsorption capacity of obtained MIPs was checked by using the binding testing. All synthesized polymers were evaluated as solid-phase extraction (SPE) sorbents for isolation and preconcentration of biochanin A and its analogues, daidzein and genistein. The MIPs exhibited higher affinity for biochanin A over competitive compounds.

Do differences in chemical composition of stem and cap of Amanita muscaria fruiting bodies correlate with topsoil type?

Fly agaric (Amanita muscaria) was investigated using a 1H NMR-based metabolomics approach. The caps and stems were studied separately, revealing different metabolic compositions. Additionally, multivariate data analyses of the fungal basidiomata and the type of soil were performed. Compared to the stems, A. muscaria caps exhibited higher concentrations of isoleucine, leucine, valine, alanine, aspartate, asparagine, threonine, lipids (mainly free fatty acids), choline, glycerophosphocholine (GPC), acetate, adenosine, uridine, 4-aminobutyrate, 6-hydroxynicotinate, quinolinate, UDP-carbohydrate and glycerol. Conversely, they exhibited lower concentrations of formate, fumarate, trehalose, α- and ß-glucose. Six metabolites, malate, succinate, gluconate, N-acetylated compounds (NAC), tyrosine and phenylalanine, were detected in whole A. muscaria fruiting bodies but did not show significant differences in their levels between caps and stems (P value>0.05 and/or OPLS-DA loading correlation coefficient <0.4). This methodology allowed for the differentiation between the fruiting bodies of A. muscaria from mineral and mineral-organic topsoil. Moreover, the metabolomic approach and multivariate tools enabled to ascribe the basidiomata of fly agaric to the type of topsoil. Obtained results revealed that stems metabolome is more dependent on the topsoil type than caps. The correlation between metabolites and topsoil contents together with its properties exhibited mutual dependences.