Exploration of the chemical constituents of Tricholomopsis aurea (Beeli) Desjardin using UHPLC-ESI-MS/MS and NMR spectroscopy

Exploration of the chemical constituents of Tricholomopsis aurea (Beeli) Desjardin using UHPLC-ESI-MS/MS and NMR spectroscopy | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Exploration of the chemical constituents of Tricholomopsis aurea (Beeli) Desjardin using UHPLC-ESI-MS/MS and NMR spectroscopy Ache Roland Ndifor, Ngnintedo Dominique, Mossebo Dominique Claude, and 8 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4585453/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Tricholomopsis aurea (Beeli) Desjardin & B.A. Perry is a wild edible fungus that is scarce in Cameroon, but common in other parts of the world. The metabolites of this food matrix have not been extensively analyzed thus far. This work aimed to establish the metabolic profile of Tricholomopsis aurea by UHPLC-ESI-MS/MS and to isolate some of the major compounds using chromatographic separation techniques. Overall, 65 compounds were identified as belonging to 18 classes, including amino acids, carbohydrates, vitamins, aminoglycolipids, phosphoglycolipids, fatty acids, phenolics, nucleosides, steroids, fatty acid amides, acyl glycerides, sphingolipids, alkaloids, terpenes, carotenoids, saponins, aromatic amines, and peptides. Six of the isolated compounds were characterized as oleic acid, elaidic acid petroselinic acid, ergosterol, ergosterol 5α,8α-endoperoxide and adenosine using 1D and 2D NMR spectroscopy. Fatty acids, steroids, amino acids and peptides were the main components of the mushroom. T. aurea has been proven to be a valuable source of chemically diverse compounds and to contain indispensable nutrients, such as amino acids, fats, carbohydrates and vitamins, which are essential for promoting the health span. Analytical Biochemistry Analytical Chemistry Organic Chemistry Food Chemistry Tricholomopsis aurea phytochemical profiling UHPLC-ESI-MS/MS NMR spectroscopy Figures Figure 1 Figure 2 INTRODUCTION The species Tricholomopsis aurea (Beeli) Desjardin & B.A. Perry is a wild edible fungus 1 that is scarce in Cameroon but common in the Democratic Republic of Congo, Burundi, Tanzania, Zimbabwe, São Tomé, Togo, Uganda, Martinique, Guadeloupe, and Trinidad2. The specimens of T. aurea collected in Cameroon are characterized by a bright to sulphur yellow basidiomata measuring 25–53 mm in diameter, a conspicuously rather flattened, fistulose to hollow and centrally depressed (from top to bottom) stipe measuring 35–55 mm long and 6.0–10 mm in diameter, thin flesh, and white spore-prints. Its basidia include four ( 4 ) and two ( 2 ) but also one ( 1 ) sterigmata, the presence of a cheilo-type and absence of pleurocystidia, and cutis-type pileipellis with erected hyphae that are subcylindrical to subclavate at the apex, some of which are incrusted 2 . In recent years, ultrahigh-performance liquid chromatography (UHPLC) and high-pressure liquid chromatography (HPLC) combined with tandem mass spectrometry (MS/MS) methods, have been used effectively in the chemical profiling of food matrices and natural extracts. This is because of their high sensitivity in obtaining accurate molecular masses to generate empirical formulas, retention times (tR) and fragment ions of the complexed samples analyzed 3,4 . Additionally, electrospray ionization mass spectrometry (ESI-MS) has been used for decades to characterize bioactive compounds, including fatty acids and other classes of compounds from mushrooms 5 . The metabolic profile of T. aurea has not been extensively analyzed thus far. Hence, the objective of the present study was to identify the phytochemical profile of T. aureus by UHPLC-ESI-MS/MS and to isolate and characterize some of the major compounds using nuclear magnetic resonance (NMR) spectroscopy. MATERIALS AND METHODS Sample collection and identification The sample was collected on April 10th, 2022, in Bambui (northwestern region of Cameroon) on a dead cypress stump. These species were subsequently identified as Tricholomopsis aurea (Beeli) Desjardin & B.A. Perry by Professor MOSSEBO Dominique Claude, a Mycologist of the Mycological Laboratory at the Department of Plant Biology and Physiology, University of Yaoundé 1 in Cameroon, by comparing the macro- and micromorphological features of the sample with those described in the most relevant literature 2 . The identified samples were thereafter preserved in the Mycological Herbarium of the University of Yaoundé 1 under voucher specimen number HUY1-DM 1937. UHPLC-ESI-MS/MS conditions For chromatographic separation, a Waters® ACQUITY UPLC® I-Class System equipped with a Binary Solvent Manager and FTN Sample Manager (Waters GmbH, Eschborn, Germany) was used. Eluents A and B were 0.3 mmol/L ammonium formate (adjusted to pH 3.5 with formic acid) and acetonitrile, respectively. The gradient profile for eluent B was as follows: 0.0–2 min, 5%; 2–19 min, 95%; 19–21 min, 95%; and 21–21.5 min, 5%. For re-equilibration of the UHPLC column, the gradient was set to 3% eluent B for 6 min. The column oven was set at 40°C, and the autosampler was cooled to 5°C. The flow rate was kept constant at 0.4 mL/min. A volume of 5 µL of the sample was injected onto an EC 150/2 Nucleoshell RP18 column (endcaped C18 phase, ID 2 mm, length 150 mm, particle size 2.7 µm; Macherey Nagel, Düren, Germany). Mass spectrometric detection in positive and negative ion modes was performed using a QqTOF MS instrument (TripleTOF 6600, Sciex, Concord, Ontario, Canada) equipped with an ESI-DuoSpray-Ion-Source and resolving power (full width at half-maximum (FWHM), at m/z 400) set of 34,000 in MS and 30,000 in data-dependent acquisition mode (DDA). All the above steps were performed for ions greater than m/z 100 and with an intensity greater than 500, with dynamic background subtraction (DBS) and exclusion criteria of 3 s after 1 occurrence. The automated calibration device system performed an external calibration. The Turbo V ion drive source equipped with a stainless-steel electrode (100 µm internal diameter) was operated under the following MS conditions: nebulizing gas (gas 1), nitrogen (60 psi), drying gas (gas 2), nitrogen (70 psi), ion spray voltage, 5500 V (positive ion mode), 4500 V (negative mode), ion-source temperature, 450°C, and curtain gas, nitrogen (55 psi). The TOF MS accumulation time was set to 50 ms, and the mass range was 65 to 1250 m/z. A 100 ms accumulation time, a collision energy (CE) of 45 V with a collision energy spread (CES) of 35 V and a declustering potential (DP) of 35 V were used for the TOF MS/MS experiments in positive ion mode. In negative ion mode, the CE, CES and DP were − 45 V, 35 V and − 35 V, respectively. All MS parameters were controlled by Analyst TF 1.7.1 software (Sciex). Data evaluation was performed using the software PeakView Version 2.2 (Sciex). With respect to the retrieval of chemical structure information, databases such as Chem-Spider ( http://www.chemspider.com ), SciFinder ( https://scifinder.cas.org ), NIST/EPA/NIH Mass Spectral Library, PubChem ( http://pubchem.ncbi.nlm.nih.gov ), MassBank ( https://massbank.jp ), LipidBank ( https://lipidbank.jp ), and the LIPID MAPS Structure Database (LMSD) ( https://www.lipidmaps.org ) were explored, as were published studies. Additional analytical methods The mass spectra were registered on a Waters ZQ-200 in electrospray ionization mode. NMR spectra were recorded on an AVANCE-NEO Bruker spectrometer equipped with a magnetic field of 14T operating at a proton frequency of 600 MHz. Low-resolution electrospray ionization mass spectrometry (ESI-MS) experiments were carried out on a Micromass Quattro Micro mass spectrometer. The LCMS used were a UHPLC (Waters) and a QqToF-MS/MS instrument (TriTOF 6600; Sciex). Column chromatography was performed on silica gel (0.04–0.063 nm; 230–400 mesh, ASTM; Merck, Germany). Thin layer chromatography (TLC) was performed on a silica gel 60 F254 (0.1 mm thick; Merck) with a size of 20 × 20 cm, and spots were detected by fluorescence at 254 nm or 366 nm on a UV-85/L basis. The sample was sprayed with 10% H 2 SO 4 , and then heated at 70°C. The solvents used were of analytical grade. Melting points were recorded using a Stuart SMP3 melting point apparatus. Extraction and isolation of compounds A total of 250 g of the air-dried basidiocarp was blended and extracted with ethanol and water (EtOH-H2O, 7:3 v/v) for 72 hours, after which the filtrate was evaporated at reduced pressure to yield a brown paste (25 g). A small amount of the paste (0.5 mg) was dissolved in ammonium formate (adjusted to pH 3.5 with formic acid) and acetonitrile and subjected to UHPLC analysis, while 16 g of the paste was chromatographed using increasing proportions of hexane (Hex), ethyl acetate (EtOAc) and methanol (MeOH), affording 157 fractions. Based on thin layer chromatography (TLC) profiles, matching similar fractions of F50-60 (Hex-EtOAc, 76:24) and F69-72 (Hex-EtOAc, 68:32) and washing the precipitates with a mixture of Hex-EtOAc 3% afforded ergosterol (18.5 mg) and ergosterol 5α,8α-endoperoxide (8 mg), respectively. Fraction F38-40 (Hex-EtOAc, 88:12) was rechromatographed over a silica gel column using an increasing gradient system of Hex-EtOAc to yield petroselinic acid (10 mg, Rf: 0.60) and (Z)-oleic acid (12 mg, Rf: 0.73). The purification of fraction F41-43 (Hex-EtOAc, 82:18) using an increasing gradient system of Hex-EtOAc gave subfraction SF14-16, affording elaidic acid (15 mg). A portion of fraction F136-138 (22 mg) was purified on a silica gel column using an isocratic system of EtOAc-MeOH-H2O-pyridine (10:4:1:0.5) to yield adenosine (12 mg). Results Metabolite profiling of the 30% hydroethanolic (HEE) extract of T. aurea was performed via UHPLC-ESI-MS/MS. Figure 1 shows the total ion chromatograms (TICs) analyzed in both positive and negative ionization modes, while Table 1 summarizes the experimental and observed mass‒charge ratios (m/z) of the precursor ions, mass errors (ppm), molecular formulas, plausible fragments, retention times (rt), and tentatively characterized compounds. The accurate masses of the positive ions and negative ions detected by QqTOF MS were converted into a molecular formula, such as CxHyNzOwS k P L , with errors presented below 3 ppm 6 . A total of 65 compounds were tentatively identified as belonging to 18 different classes, such as amino acids ( 7 ), carbohydrates ( 2 ), vitamins ( 2 ), aminoglycolipids ( 1 ), phosphoglycolipids ( 2 ), fatty acids ( 14 ), phenolics ( 3 ), nucleosides ( 2 ), steroids ( 12 ), fatty acid amides ( 2 ), acyl glycerides ( 1 ), sphingolipids ( 2 ), alkaloids ( 1 ), terpenes ( 3 ), carotenoids ( 2 ), saponins ( 1 ), aromatic amines ( 1 ), and peptides ( 7 ). Of these, 6 compounds were isolated and described as oleic acid, elaidic acid petroselinic acid, ergosterol, ergosterol 5α,8α-endoperoxide and adenosine using 1D and 2D NMR spectroscopy. Characterization of the hydroethanolic extracts of T. aurea by UHPLC-ESI-MS/MS Discussion The ingestion of peptides, amino acids and proteins from different food sources is essential for maintaining health 7,8 . In fact, in this study, seven amino acids and seven peptides were tentatively detected in both the positive and negative ion modes. The positive ion mode showed the precursor ion [M + H] + of valine (peak 59, C 5 H 12 NO 2 ) at m/z 118.0868. The peak at 72.08132 [M + H-CO 2 ] could be attributed to the loss of carbon dioxide. Valine has been quantified in many mushroom species 9,10 and is needed by animals for the development of mammary glands and ovaries 11 . Additionally, in positive ion mode, arginine (peak 49, C 6 H 14 N 4 O 2 ) was observed at m/z 174.11119 in wild edible mushrooms 9 . In negative ionization mode, isomers of the (R)-pyroglutamic acid (peak 2, C 5 H 7 NO 3 ) precursor [M-H] − were identified at 128.03502 in shiitake mushrooms 10 , and (S)-pyroglutamic acid (peak 7, C 5 H 7 NO 3 ) was identified at m/z 128.035 in Morchella sp 12 . Both of these compounds presented fragment ions at 84.04494 (M-H-CO 2 ) − , indicating decarboxylation, as did 56.02621 [M-H-CO 2 -CH 2 NH] − , which was attributed to decarboxylation and loss of methanimine. Other amino acids identified were leucine (peak 4, C 6 H 13 NO 2 ) at m/z 130.08715, which was detected in shiitake mushrooms 10 ; phenylalanine (peak 10, C 9 H 11 NO 2 ) at m/z 164.07131, which was reported in Arabidopsis thaliana 13 ; and α-hydroxymethylserine (peak 18, C 4 H 9 NO 4 ) at m/z 134.04572, which was isolated from the seeds of Vicia pseudo-Orobus 14 . The tripeptide Gln-Trp-Arg 15 , which eluted at a retention time of 18.427 min, was detected as an [M-H] − ion at m/z 487.24178 (peak 63). This was attributed to the molecular formula C 22 H 32 N 8 O 5 . The fragment ion at m/z 442.24409 [M -H − 44] − could be due to the loss of carbon dioxide molecules. Peak 62 (rt: 18.284 min) was detected as an [M + H] + ion at m/z 471.35491, and its molecular formula was assigned as C 24 H 46 N 4 O 5 due to mass; thus, this peak was attributed to the tetrapeptide Leu-Leu-Leu-Leu 16 . Its fragment at 426.35699 [M + H-44] + suggested the loss of a carbon dioxide molecule from the terminal carboxyl group. Peak 65 (rt: 12.894 min) was observed as an [M-H] − ion at m/z 611.31979 and has a molecular formula of C31H44N6O7. This was attributed to cyclo (-d-glu-ala-d-allo-ile-leu-d-trp), commonly known as BE-18257B 17 . L-365,209, peak 64 (rt: 12.836 min) was detected as an [M-H]- ion at m/z 665.38151 with the molecular formula C 39 H 50 N 6 O 6 . It was previously reported as a class of cyclic hexapeptide oxytocin antagonists from Streptomyces silvensis 18 . Muracein A (peak 15, C 26 H 44 N 6 O 14 ) at m/z 663.28431 was detected in Nocardia orientalis 19 ; pandanamide A (peak 19, C 31 H 47 N 7 O 9 ) at m/z 660.33697 was isolated from Streptomyces sp . 20 ; and the tripeptide Leu-His-Ala peak 20 (C 15 H 25 N 5 O 4 ) at m/z 338.18271 was reported in dry-cured Jinhua ham 21 . Sphingolipids and glycerophospholipids participate in a variety of important neurological, metabolic and intracellular signaling processes 22 . Sphingolipids serve as building blocks of the plasma membrane of eukaryotic cells. They anchor lipid-bound carbohydrates to cell surfaces and construct the epidermal water permeability barrier 23 . As shown in Table 1, the protonated ion [M + H] + at m/z 302.30516 (peak 31, C 18 H 40 NO 2 ) was attributed to sphinganine detected previously in Arabidopsis thaliana roots 24 , while that at m/z 298.27453 (peak 44, C 18 H 35 NO 2 ) was attributed to sphinga-4E,8E-dienine, which was revealed in oyster mushrooms 25 . Deinococcucin D displayed a precursor ion [M-H] − at m/z 543.36482 (peak 26, C 28 H 52 N 2 O 8 ). It was isolated from the gut of queen carpenter ants 26 , while mannosyl (4S)-4-methylheptacosyl phosphate, a synthetic analog of β-D-mannosyl C32-phosphomycoketide27, was detected at m/z 651.46027 (peak 60, C 34 H 69 O 9 P). 1-(9Z-Octadecenoyl)-2-(5-hydroxy-7-carboxy-6E-heptenoyl)-sn-glycero-3-phosphate (peak 17, C 31 H 54 O 13 P) at m/z 665.33064 was described previously as an oxidized phospholipid 28 . Table 1: Based tentative identification of compounds from the hydroethanolic extract of T. aurea by UHPLC-ESI-MS/MS Terpenes were identified in the total ion chromatogram (retention time range 12.449–18.427 min). Peak 45, with a protonated molecular ion [M-H] + at m/z 329.16008 (C 16 H 24 O 7 ), was assigned to zataroside B, described in Zataria multiflora 29 . The fragment ion at 165.09155 [M-162-H] − could be due to the loss of a hexose unit. Trichodermanin D (peak 50, C 20 H 34 O 3 ), precursor ion [M + H] + at m/z 323.25831, was detected in the fungus Trichoderma harzianum 30 . The analog of nodulisporic acid A (peak 22, C45H57NO7) and the pseudomolecular ion [M-H] − at m/z 722.40634 were isolated from an endophytic fungus 31 . As shown in Table 1, the deprotonated molecular ion of Aspacoside D (peak 56, C 38 H 60 O 16 ) was observed at 771.38034 [M-H] − in negative ion mode and could afford fragment ions at 609.32749 [M-H-162] − , 476.27740 [M-H-162-132] − and 315.23241 [M-H-162-162] − , corresponding to the loss of one pentose and two hexose units. This molecule was isolated from Asparagus cochinchinensis32. The protonated molecular ion of the phenolic de-O-methyllasiodiplodin was observed at m/z 279.15965 (peak 32, C 16 H 22 O 4 ) in positive ion mode (Table 1). This compound has been described in Gouania longipetala and Glyphaea brevis 33 . Two additional phenolic compounds were characterized in negative ionization mode, with pseudomolecular ions at m/z 331.11842 (peak 14, C 18 H 20 O 6 ) and 151.03924 (peak 12, C 8 H 8 O 3 ), attributed to 3',5'-dihydroxy-2',4',6'-trimethoxydihydrochalcone, which was isolated from the twigs of Lindera lucida 34 , and anisic acid or 4-methoxybenzoic acid, which was isolated from Rhododendron 35 . The aromatic amine 2,4-diaminophenol observed at m/z 125.07146 (peak 61, C 6 H 9 N 2 O) is a known cosmetic ingredient 36 . The exact mass of uridine at m/z 243.06126 (peak 3, C 9 H 11 N 2 O 6 ) was obtained in negative ionization mode. The presence of uridine in the mushroom Pleurotus giganteu reportedly increases the phosphorylation of extracellular signal-regulated kinases and protein kinase B 37 . The precursor ion of glucose was observed at m/z 179.05523 (peak 1, C 6 H 12 O 6 ) in negative mode (Table 1). Glucose has been detected in mushrooms and is a substrate for energy-producing processes 38 . In the positive ion mode, peak 8 (C 6 H 13 NO 5 ) at m/z 180.08637 and peak 9 (C 5 H 10 O 5 ) at m/z 163.06027 were attributed to nojirimycin 39 and 1,5-anhydrofructose 40 , respectively. The precursor ion [M-H] − for xanthine was observed at m/z 151.02575 (peak 5, C 5 H 4 N 4 O 2 ). Tea, cocoa seeds, and coffee are the natural sources of xanthine 41 . The acyl glyceride 1-palmitoyl glycerol (peak 39, C 19 H 38 O 4 ) protonated molecular ion [M + H] + at 331.28455 could yield a fragment ion at 239.23749 [M-C 3 H 7 O 3 ] + as a result of the loss of the glycerol moiety. It serves as an emulsifying agent and has been used to increase the viscoelasticity of processed cheese 42 . The precursor ion [M + H] + at m/z 396.34738 was attributed to semiplenamide E (peak 55, C 24 H 45 NO 3 ), which was detected in the marine cyanobacterium Lyngbya semiplena 43 , whereas the natural product N-palmitoyl-D-leucine (Peak 61, C 22 H 43 NO 3 ) precursor ion [M + H] + at m/z 370.33209 was identified in the extracts of rat brains or bovine spinal cords 44 . Fourteen fatty acids were detected in the positive and negative ion modes, among which (11E,13E)-9,10-dihydroxy-11,13-octadecadienoic acid (peak 24, C 18 H 32 O 4 ) at m/z 313.23759 was detected in M. giganteus 45 , and palmitoleic acid (peak 25, C 16 H 30 O 2 ) was detected at m/z 255.23172 in T. rutilans 46 . Cis-linoleic acid (peak 27, C18H32O2) at m/z 281.24815 was described in Lactarius salmonicolor , Polyporus squamosus , Pleurotus ostreatus and Flammulina velutipes 47 ; linolenic acid (peak 28, C 18 H 32 O 2 ) at m/z 279.23241 was detected in Helichrysum pedunculatum , Polyporus squamosus , and Pleurotus ostreatus 48 . Trans-linoleic acid (peak 30, C 18 H 32 O 2 ) at m/z 281.24783 has been described in F. hepatica and T. equestre 47 . Caproliec acid, also known as 9-decenoic acid (peak 47, C 10 H 18 O 2 ), was detected at m/z 171.13875 in beer and yeast 49 ; octadecanedioic acid (peak 46, C 18 H 34 O 4 ), at m/z 315.25343, was detected in Arabidopsis thaliana 50 . 9-Octadecynoic acid or stearolic acid (peak 43, C 28 H 32 O 2 ) at m/z 281.2482 was detected in plant oil seeds 51 , while 𝛾-linolenic acid (peak 38, C 28 H 30 O 2 ) at m/z 279.23195 was determined from Zygomycetous Fungi 52 . Suberic acid (peak 13, C 8 H 14 O 4 ) at m/z 173.0813 was detected in urine 53 , and 13-hydroxy-9Z,11E,15E-octadecatrienoic acid (peak 51, C 18 H 30 O 3 ) [M + H] + at m/z 295.22709 was isolated from the leaves of Cucurbita moschata 54 . As enumerated in Table 1, the protonated molecular ion of the isomeric compound 6β-hydroxy-ergosta-4,7,22-trien-3-one (peak 40, C 28 H 42 O 2 ) was at m/z 411.32631 [M + H] + , and its dehydration of C6-OH with B-ring hydrogen could yield the fragment ion at m/z 392.30792 [M–18] + . In addition, the peak at m/z 268.1872 [M + H-18-C 9 H 17 ] + was attributed to dehydration and loss of the side chain on the D ring 55 . Its isomer 6α-hydroxy-ergosta-4,7,22-trien-3-one (peak 42, C 28 H 42 O 2 ) at m/z 411.32675 presented a similar fragmentation pattern. These compounds were all described in G. lucidum 55 . Gagarlol B (peak 35, C 28 H 42 O 4 ) was detected at m/z 443.31606 and was isolated from the edible mushroom Grifola gargal 56 ; ganodermaside C (peak 48, C 28 H 38 O 3 ) at m/z 423.28935 was isolated from Ganoderma lucidum 57 ; ergosta-4,6,8( 14 ),22‐tetraen‐3‐one (peak 54, C 28 H 40 O) at m/z 393.31563 was noted in Porodaedalea pini 58 ; 5α-8α-epidioxy-24-methylcholesta-6,9( 11 ),24( 28 )-trien-3β-ol (peak 34, C 28 H 42 O 3 ) at m/z 427.32115 was identified in the marine sponge Coscinoderma sp . 59 ; and stigmasta-3,5-diene (peak 57, C 29 H 48 ) at m/z 397.38317 was detected in Cuscuta reflexa Roxb 60 . The negative mode ESI enabled the identification of 12β-acetoxy-3,7,11,15,23-pentaoxo-5α-lanosta-8-en-26-oic acid ethyl ester (peak 16, C 34 H 46 O 9 ) at m/z 597.30632, reported in the fruiting body of Ganoderma lucidum 61 ; peak 23 (C 34 H 46 O 8 ) at m/z 581.31188, ascribed to a helvolic acid methyl ester derivative from the endophytic Ficus carica 62 ; and 3β-amino-5-spirosolene (peak 58, C 27 H 44 N 2 O) at m/z 411.33725, which was isolated from Solanum triste 63 . The precursor ion [M-H] − at m/z 122.02425 was attributed to niacin (peak 6, C 6 H 5 NO 2 ), a water soluble vitamin that has been found in different kinds of plant-based foods64. In positive ionization mode, the precursor ion at m/z 429.33619 (peak 52, C 28 H 44 O 3 ) was attributed to the fat soluble vitamin 1α,25-dihydroxy-24-epivitamin D2 65 . The carotenoid 7,8-dihydroparasiloxanthinin (peak 37, C 40 H 60 O 2 ) at m/z 573.466628 was reported in Pleurotus mushrooms 66 , while α-citraurin (peak 29, C 30 H 40 O 2 ) and the pseudomolecular ion [M-H] − at m/z 431.29492 were isolated previously from the sea hare 67 . The nucleoside adenosine was observed at m/z 268.10442 (peak 11, C 10 H 13 N 5 O 4 ). The fragment ion visible at m/z 249.150 [M-H 2 O] + was attributed to the loss of water molecules 68 . ESIMS (positive mode) showed adducts at m/z 305.9 [M + K] + and 290.6 [M + Na] + (supplementary material and Fig. 2 ). In addition, its 1 H NMR spectrum presented signals corresponding to two singlets at δ 8.62 (s, H-2) and 8.37 (s, H-8), with corresponding carbons at δ 153.7 (C-2) and 141.0 (C-8), respectively. The HMBC spectrum showed correlations between the proton singlets and the carbons at δ 158.2 (C-6), 150.6 (C-4) and 121.9 (C-5), thus suggesting a purine ring. The 1 H signals at δ 6.70 (d, J = 6.0 Hz, H-1'); 5.52 (t, J = 4.8 Hz, H-2'); 5.08 (brs, H-3'); 4.78 (q, J = 5.4 Hz, H-4'); 4.38 (dd(distorted), J = 11.4, 3.0 Hz, H-5'a); 4.17 (dd(distorted), J = 11.4, 3.0 Hz, H-5'b)) in conjunction with their carbons at δ153.7 (C-2), 150.6 (C-4), 121.9 (C-5), 158.2 (C-6), 141.0 (C-8), 91.3 (C-1'), 76.2 (C-2), 72.9 (C-3'), 88.3 (C-4'), 63.5 (C5'), and 30.6 (C-14) suggested the presence of a ribose unit, thus corroborating its identification as adenosine. This compound was reported previously in Euryale ferox Salisb 69 . Ergosterol (peak 53, C 28 H 44 O) at m/z 437.36533 was isolated as a white powder. The 1 H NMR spectrum showed signals at δ 3.96 (1H, m, H-3), 5.52 (1H, q, J = 3.6, 6.6 Hz, H-7), 5.32 (1H, m, H-22) and 5.29 (1H, m, H-23), with carbons appearing at δ 70.6 (C-3), 117.8 (C-7), 136.8 (C-22) and 132.8 (C-23), respectively (Supplementary material). In addition, signals corresponding to six methyl groups were observed at δ 1.06 (3H, s, H-18), 0.91 (3H, d, J = 4.2 Hz, H-26), 0.89 (3H, d, J = 4.2 Hz, H-27), 0.70 (3H, s, H-19), 0.99 (3H, d, J = 8.4 Hz, H-28) and 1.12 (3H, d, J = 8.4 Hz, H-21). Its positive ESI mode displayed an adduct at m/z 790.9 [2M + H] + and molecular ions at m/z 396.8 [M] + and 395.8 [M – H] + , respectively. The dehydration of C3-OH led to the fragment ion at m/z 377.7 [M + H – 18] + , while the fragment ion at m/z 271.4 [M + H – 125] + was attributed to the loss of the lateral chain. Ergosterol was previously extracted from the mushroom Sparassia crispa 70 . Ergosterol 5α,8α-endoperoxide (peak 54, C 28 H 42 O 3 ) at m/z 429.33649 was obtained as a white needle. ESIMS (positive mode) showed adducts at m/z 882.1 [2M + Na] + and 451.8 [M + Na] + . 1 H signals were observed at δ 3.78 (1H, m, H-3), 6.28 (1H, d, J = 8.1 Hz, H-6), 6.57 (1H, d, J = 8.1 Hz, H-7), 5.22 (1H, dd, J = 4.5, 9.3 Hz, H-22) and 5.28 (1H, dd, J = 4.5, 9.3 Hz, H-23), with corresponding carbons at δ 66.9 (C-3), 136.8 (C-6), 131.8 (C-7), 136.8 (C-22) and 133.5 (C-23), respectively. Furthermore, signals corresponding to six methyl groups were observed at δ 0.85 (3H, s, H-18), 0.94 (3H, d, J = 3.9 Hz, H-26), 1.03 (3H, d, J = 3.9 Hz, H-27), 0.92 (3H, s, H-19), 0.86 (3H, d, J = 1.8 Hz, H-28) and 2.01 (3H, d, J = 3.0 Hz, H-21). The carbon signals at δ 83.4 (C-5) and 80.7 (C-8) were established using the HMBC spectrum (supplementary material). Ergosterol 5α,8α-endoperoxide was previously described in Desmodium uncinatum 71 . (Z)-oleic acid (peak 33, C18H34O2) at m/z 324.29008 was isolated as a white oil. Its carbon spectrum showed an acid functional group at δ181.3 (C-1). In its 1 H NMR spectrum, the signals at δ 5.36 (m, H-9, H-10), 0.92 (t, J = 7.2 Hz, H-18) and 1.29–1.33 (m, H-12 to H-16, H-3 to H-7) were assigned to ethylinic, terminal methyl and long chain units, respectively (supplementary material). The negative mode ESI spectrum of oleic acid showed a molecular ion peak at m/z 282.0 [M] − . ; hydrated fragment ions at m/z 256 [M + H 2 O-C 3 H 7 ] − ; 171.7 [M + H 2 O-129] − . A prominent fragment ion at m/z 127.4 [C 7 H 11 O 2 ] − resulted from the McLafferty rearrangement. The adducts in the positive ESI mode at 305.8 [M + Na] + , 321.8 [M + K] + and 301.7 [M + H 2 O] + as well as the fragment ion at 129.5 [M-C 11 H 21 ] + supported its characterization as oleic acid, previously recognized in T. rutilans 46 . Elaidic acid or trans-9-octadecenoic acid (peak 36, C 18 H 34 O 2 ) at m/z 300.28993 was isolated as a white oil. The spectrum displayed 1 H signals at δ 5.34 (m, H-9, H-10), 0.89 (t, J = 6.0 Hz, H-18) and 2.26 (t, J = 6.0 Hz, H-2), assigned to vinylic groups, methyl groups and methylene groups, respectively. The corresponding carbon signals were observed at δ 130.8 (C-8), 130.9 (C-10), 14.4 (C-18) and 35.0 (C-2), while the acid functional group appeared at δ 177.7 (C-1). The positive ESI mode presented an adduct at m/z 321.7 [M + K] + (Supplementary material). Elaidic acid is an isomer of oleic acid, is one of the major trans fatty acids (TFAs) in foods and accounts for 40–60% of the TFAs found in humans72. Petroselinic acid (peak 21, C 18 H 34 O 2 ) at m/z 300.28992 is a positional isomer of oleic acid. 1 H signals were observed at 5.34 (m, H-6, H-7); 0.90 (t, J = 6.0 Hz, H-18); 1.59 (m, H-17); 2.18 (t, J = 6.0 Hz, H-2); 2.02 (m, H-5, to H-8); and carbon at 130.9 (C-6), 130.8 (C-7), 14.4 (C-18), 23.7 (C-17), 37.8 (C-2), 28.1 (C-5) and 28.2 (C-8). The carbon spectrum showed a signal at δ 179.4 (C-1), attributed to an acid functional group (Supplemental material). In negative ESI mode, a molecular ion peak at 282.0 [M] − was observed, and an intense fragment ion at m/z 113.5 [C 6 H 9 O 2 ] − formed through McLafferty rearrangement 73 . This compound has been detected in the vegetable oil of Coriandrum sativum fruits 74 . Conclusion This work tentatively identified 65 known compounds belonging to 18 chemical classes. Among these, 6 of the compounds were isolated and characterized as petroselinic acid, oleic acid, elaidic acid, ergosterol, ergosterol-5α,8α-endoperoxide and adenosine. This is the first report of provisional and isolated compounds from this species. However, some of the compounds were previously identified in the literature as vital dietary supplements necessary for promoting healthspan. Based on our findings, the mycelia of T. aurea should be collected and cultivated by mushroom farmers to ensure their availability as a steady food supplement in our communities. Declarations ACKNOWLEDGMENTS The authors are thankful to the bioprofiling platform, supported by the European Regional Development Fund and the Walloon Region Belgium, as well as to YaBiNaPA and Cameroon for the facilities that enabled the realization of this work. The Leibniz Institute of Plant Biochemistry in Halle (Saale), Germany, is acknowledged for supporting the UHPLC-ESI-MS/MS analyses. Author contributions Ache R.N: Study design, methodological experiments and writing of the main manuscript. Ngnintedo D: Interpretation of the results and editing of the prepared manuscript. Ambassa P: Interpretation of the results and editing of the prepared manuscript. Mossebo D. C: Identification of the mushroom specimen and revision of the manuscript. Yanick K. M: Interpretation of the results and final editing of the manuscript. Njinga N. S: designed and revised the manuscript. Céline H: Analytical experiments and editing of the prepared manuscript. Sophie L: Analytical experiments and editing of the prepared manuscript. Sonchieu J: Revising and final editing. Ngameni B: Revising and final editing. Fotso W.G: Analytical experiments and revision of the final manuscript. Competing interests The authors declare no competing interests. Data Availability Statement The data that support the findings of this study are available from the corresponding author upon reasonable request at https://getshared.com/UpuXwUU2 Funding This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors. References Eyi-Ndong H, Degreef J, De Kesel A (2011) Champignons comestibles des forêts denses d’Afrique centrale. Taxonomie et identification. Abc Taxa. 10 Desjardin DE, Perry BA (2017) The gymnopoid fungi (Basidiomycota, Agaricales) from the Republic of São Tomé and Príncipe, West Africa. Mycosphere 8(9):1317–1391 Zeng Y, Lu Y, Chen Z, Tan J, Bai J, Li P, Wang Z, Du S (2018) Rapid characterization of components in Bolbostemma paniculatum by UPLC/LTQ-Orbitrap MSn analysis and multivariate statistical analysis for herb discrimination. Molecules 23:1155 Eric B, Sandor S, Ce´line B, Sebastian B, Juho R (2018) Liquid-chromatography retention order prediction for metabolite identification. Bioinformatics 34:i875–i883 Oni JO, Ferdinand A, Akomaye A-A, Markson A, Augustine CE (2020) GC–MS Analysis of Bioactive Compounds in Some Wild-Edible Mushrooms from Calabar, Southern Nigeria. EJBIO 1(6):1–8 Pedro ES, do N et al (2023) Characterization of Flavonoids Profiles in Polar Extracts from Croton grewioides Baill. Using Ultra-High Resolution Mass Spectrometry. J Braz Chem Soc 34(7):937–948 Marshall WE (1994) Amino Acids, Peptides, and Proteins. In: Goldberg I (ed) Functional Foods. Springer, Boston, MA, pp 242–260 Castro GA (1991) Digestion and absorption. In: Johnson LR (ed) Gastrointestinal Physiology, 4th edn. Mosby Year Book, St. Louis, pp 108–130 Thalisa Y-A, Luchai B, Pa-Nag Y (2020) Amino acids and antioxidant activities of extracts from wild edible mushrooms from a community forest in the Nasrinual District, Maha Sarakham, Thailand. Food Sci Technol Campinas 40(3):712–720 Meng D et al (2018) Simultaneous quantification of free amino acids and 5′-nucleotides in shiitake mushrooms by stable isotope labeling-LC–MS/MS analysis. Food Chem 268:57–65 Nisreen H, Touseef HT, Lata M (2019) Amino acids as Medical food and their Therapeutic uses. IJSRR 8(2):579–585 Dong H et al (2022) Metabolomics Analysis of Morchella sp. From Different Geographical Origins of China Using UPLC-Q-TOF-MS. Front Nutr 9:1–11 Cho M-H et al (2007) Phenylalanine Biosynthesis in Arabidopsis thaliana : identification and characterization of arogenate dehydratases. JBC 282(42):30827–30835 Koshi S, Jun F, Shigenobou O, Shin-Ichi H (1985) α-(Hydroxymethyl) serine from Vicia pseudo-orobus. Phytochemistry 24(4):853–854 Ayabe T et al (2015) A novel Alaska pollack-derived peptide, which increases glucose uptake in skeletal muscle cells, lowers the blood glucose level in diabetic mice. Food Funct 6(8):2749–2757 Bonnamour J, Métro T-X, Martinez J, Lamaty F (2013) Environmentally benign peptide synthesis using liquid-assisted ball-milling: application to the synthesis of Leu-enkephalin. Green Chem 15(5):1116 Katsuhisa K et al (1991) Endothelin-binding inhibitors, BE-18257A and Be-18257B. I. Taxonomy, fermentation, isolation and characterization. J Antibiot 44(12):1342–1347 Freidinger RM et al (1990) Cyclic hexapeptide oxytocin antagonists. Potency-, selectivity-, and solubility-enhancing modifications. J Med Chem 33(7):1843–1845 Singh PD, Johnson JH (1984) Muraceins–muramyl peptides produced by Nocardia orientalis as angiotensin-converting enzyme inhibitors. II. Isolation and structure determination. J Antibiot 37(4):336–343 John WB et al (2017) Marine natural products. Nat Prod Rep 34:235–294 Zhu C-Z, Tian W, Li M-Y, Liu Y-X, Zhao G-M (2018) Separation and identification of peptides from dry-cured Jinhua ham. Int J Food Prop. 1–10 Castro-Gómez P, Garcia-Serrano A, Visioli F, Fontecha J (2015) Relevance of dietary glycerophospholipids and sphingolipids to human health. Prostaglandins, Leukotrienes and Essential Fatty Acids (PLEFA). 101:45–51. http://dx.doi.org/10.1016/j.plefa.2015.07.004 Kolter T, Sandhoff K (1999) Sphingolipids–Their Metabolic Pathways and the Pathobiochemistry of Neurodegenerative Diseases. Angew Chem Int Ed 38:153–1568 Chao D-Y et al (2011) Sphingolipids in the Root Play an Important Role in Regulating the Leaf Ionome in Arabidopsis thaliana . Plant Cell 23(3):1061–1081 Hayashi A, Matsubara T (1970) On the occurrence of sphinga-4,8-dienine in oyster glycolipid. Biochim Biophys Acta 202(1):228–230 Bora S et al (2017) Deinococcucins A-D, Aminoglycolipids from Deinococcus sp., a Gut Bacterium of the Carpenter Ant Camponotus japonicas. J Nat Prod 80(11):2910–2916 Ly D et al (2013) CD1c tetramers detect ex vivo T-cell responses to processed phosphomycoketide antigens. JEM 210(4):729–741 Eugene A et al (2002) A Novel Family of Atherogenic Oxidized Phospholipids Promotes Macrophage Foam Cell Formation via the Scavenger Receptor CD36 and Is Enriched in Atherosclerotic Lesions. J Biol Chem 277(41):38517–38523 Ali MS et al (1999) Three p-cymene derivatives from Zataria multiflora. Phytochemistry 52:685–688 Takeshi Y, Mayo S, Takanobu A, Takashi K, Reiko T (2017) Trichodermanins C–E, New Diterpenes with a Fused 6-5-6-6 Ring System Produced by a Marine Sponge-Derived Fungus. Mar Drugs 15(6):169, 2–7 Meinke PT, Smith M-HM, Shoop WL (2002) Nodulisporic Acid: Its Chemistry and Biology. Curr Top Med Chem 2:655–674 Tenji K, Junzo S (1979) Studies on the constituents of Asparagi kadix. I.1) on the structures of Furostanol Oligosides of Asparagus cochinchinensis (Loureio) Merrill. Chem Pharm Bull 27(12):3086–3094 Ekuadzi E et al (2014) Chemical constituents from Gouania longipetala and Glyphaea brevis Nat. Prod Res 28(15):1210–1213 Leong Y-W, Harrison LJ, Bennett GJ, Kadir AA, Connolly (1998) J.D. A dihydrochalcone from Lindera lucida. Phytochemistry 47(5):891–894 Olennikov DN, Tankhaeva LM (2010) Phenolic compounds from Rhododendron dauricum from the Baikal region. Chem Nat Compd 46(3):471–473 Andersen FA (1994) Cosmetic Ingredient Review, 1101 17th Street NW, Suite 310, Washington, DC 20036, U.S.A. Journal of the American College of Toxicology. 13(5), 330–343, Raven Press, Ltd. New York Phan C-W, David P, Wong K-H, Naidu M, Sabaratnam V (2015) Uridine from Pleurotus giganteus and Its Neurite Outgrowth Stimulatory Effects with Underlying Mechanism. PLoS ONE 10(11):e0143004 Vetter. J (2023) The Mushroom Glucans: Molecules of High Biological and Medicinal Importance. Foods 12:1–19 Inouye S, Tsuruoka T, Ito T, Niida T (1968) Structure and synthesis of nojirimycin. Tetrahedron 24(5):2125–2144 Mami F, Kazuhiro Y, Kenko M, Jun-ichi A, Susumu H (1999) Preparation and Antioxidative Activity of 1,5-Anhydrofructose. J Appl Glycosci 46(4):439–444 Singh N, Shreshtha AK, Thakur MS, Patra S (2018) Xanthine scaffold: scope and potential in drug development. Heliyon e 00829 Bunka F et al (2007) Effect of 1-monoglycerides on viscoelastic properties of processed cheese. Int J Food Prop 10(4):819–828 Bingnan H, Kerry LMP, Alessia L, Vincenzo DM, William HG, Semiplenamides A-G (2003) Fatty Acid Amides from a Papua New Guinea Collection of the Marine Cyanobacterium Lyngbya semiplena. J Nat Prod 66:1364–1368 Tan B et al (2010) Identification of endogenous acyl amino acids based on a targeted lipidomics approach. JLR 51(1):112–119 András S et al (2020) Cerebrosides and Steroids from the Edible Mushroom Meripilus giganteus with Antioxidant Potential. Molecules 25:1395, 1–8 Hakan I, İbrahim T, Aydın ŞB, Handan ÇY (2019) Determination of fatty acid profile and mineral contents of Tricholomopsis rutilans collected from Yozgat. Ant J Bot 3(2):64–68 Bárbara R, Paula GdP, Paula BA, Paula B (2009) Patrícia, V. Fatty acid composition of wild edible mushrooms species: A comparative study. Microchem J 93:29–35 Dilika F, Bremner PD, Meyer JJM (2000) Antibacterial activity of linoleic and oleic acids isolated from Helichrysum pedunculatum: a plant used during circumcision rites. Fitoterapia 71(4):450–452 Chen C-HE, Ho C-T (1981) Identification of 9-Decenoic Acid in Beer and Yeast. JASBC 39(2):70–71 Guelette UFBS (2012) Benning and Susanne Hoffmann-Benning. Identification of lipids and lipid-binding proteins in phloem exudates from Arabidopsis thaliana . JXB 63(10):3603–3616 Avato P, Tava A (2022) Rare fatty acids and lipids in plant oilseeds: occurrence and bioactivity. Phytochem Rev 21:401–428 Weete JD, Shewmaker F, Gandhi SR (1998) γ-Linolenic Acid in Zygomycetous Fungi: Syzygites megalocarpus. JAOCS 75(10):1367–1372 Katsunobu Y, Nobuo S, Toyofumi N, Keiichi N, Keiichi N (1994) Measurements of urinary adipic acid and suberic acid using high-performance liquid chromatography. J Chromatogr B Biomed App 655(2):189–193 Bang MH et al (2002) 13-hydroxy-9Z,11E,15E-octadecatrienoic acid from the leaves of Cucurbita moschata. Arch Pharm Res 25:438–440 Nishitoba T, Sato H, Oda K, Sakamura S (1988) Novel Triterpenoids and a Steroid from the Fungus Ganoderma lucidum. Agric Biol Chem 52(1):211–216 Wu J et al (2011) Osteoclast-forming suppressing compounds, gargalols A, B, and C, from the edible mushroom Grifola gargal. Tetrahedron 67:6576–6581 Weng Y et al (2011) Ganodermasides C and D, two new anti-aging ergosterols from spores of the medicinal mushroom Ganoderma lucidum. Biosci Biotechnol Biochem 75:800–803 Deveci E, Tel-Çayan G, Duru ME, Öztürk M (2019) Chemical constituents of Porodaedalea pini mushroom with cytotoxic, antioxidant and anticholinesterase activities. J Food Meas Charact, 1–10 Jeong H et al (2019) Isolation and characterization of antiplasmodial constituents from the marine sponge Coscinoderma sp. Z für Naturforschung C 74(11–12):313–318 Riaz M et al (2017) Natural products from Cuscuta reflexa Roxb. with antiproliferation activities in HCT116 colorectal cell lines. Nat Prod Res 31(5):583–587 Cheng CR et al (2010) Cytotoxic triterpenoids from Ganoderma lucidum. Guo Phytochemistry 71(13):1579–1585 Xi-Ai L, Yang-Min M, Hong-Chi Z, Rui L (2016) A new helvolic acid derivative from an endophytic Fusarium sp. of Ficus carica. Nat Prod Res 30(21):2407–2412 Maxwell A, Seepersaud M, Pingal R 3 P-aminospirosolane steroidal alkaloids from Solanum triste. J Nat Prod 58 (4), 625–628 (I995). Pirjo M et al (2001) Contents of Vitamins, Mineral Elements, and Some Phenolic Compounds in Cultivated Mushrooms. Agric Food Chem 49:2343–2348 Rafal RS, Yoko T, Heinrich KS, Hector FD (1985) Synthesis of 1α,25-dihydroxyvitamin D2, its 24 epimer and related isomers, and their binding affinity for the 1,25-dihydroxyvitamin D3 receptor. Bioorg Chem 13:158–169 Strapáč, Kuruc M1, Baranová M (2017) Determination of antioxidant parameters of Pleurotus mushrooms growing on different wood substrates. Folia veterinaria. 61, 4: 53–58, 2017 Yamashita E, Matsuno T (1990) A new apocarotenoid from the sea hare Aplysia kurodai. Comp Biochem Physiol Part B: Biochem Mol Biology 96B(3):465–470 Hong W, Qinan W, Chengying W, Zheng J (2015) Simultaneous Determination of 16 Nucleosides and Nucleobases in Euryale ferox Salisb. By Liquid Chromatography Coupled with Electro Spray Ionization Tandem Triple Quadrupole Mass Spectrometry (HPLC-ESI-TQ-MS/MS) in Multiple Reaction Monitoring (MRM) Mode. JCS 53:1386–1394 Zhixin W et al (2019) Rapid Characterization of Chemical Components in Edible Mushroom Sparassis crispa by UPLC-Orbitrap MS Analysis and Potential Inhibitory Effects on Allergic Rhinitis. Molecules 24(16):3014, 1–17 Borice TT et al (2017) A New Tetracyclic Triterpenoid from Desmodium uncinatum (Jacq.) DC. (Fabaceae). NPS 23(2):113–118 Murphy RC (2014) Tandem Mass Spectrometry of Lipids. Royal Soc Chem. 1–280 Gebauer SK, Psota TL, Kris-Etherton PM (2007) The diversity of health effects of individual trans fatty acid isomers. Lipids 42(9):787–799 Shigeo T, Kayoko T, Takuhei Y, Nobuyuki S (2020) Positional determination of the carbon–carbon double bonds in unsaturated fatty acids mediated by solvent plasmatization using LC–MS. Sci Rep 10:12988 Delbeke EIP et al (2016) Petroselinic acid purification and its use for the fermentation of new sophorolipids. AMB Expr 6:28 Table Table 1 is available in the Supplementary Files section Additional Declarations The authors declare no competing interests. Supplementary Files Supplementarymaterial.docx Table1.docx Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4585453","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":314743466,"identity":"acc7bd95-c76b-4c1b-b2fb-0952dab19527","order_by":0,"name":"Ache Roland 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I","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Fotso","middleName":"Wabo","lastName":"Ghislain","suffix":""}],"badges":[],"createdAt":"2024-06-15 07:53:26","currentVersionCode":1,"declarations":{"humanSubjects":false,"vertebrateSubjects":false,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":false,"coiExplicitlySet":false},"doi":"10.21203/rs.3.rs-4585453/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4585453/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":58587263,"identity":"64f80ce8-a770-4adb-ac7c-4ab5b8fcaadc","added_by":"auto","created_at":"2024-06-18 14:34:22","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":105963,"visible":true,"origin":"","legend":"\u003cp\u003eTotal ion chromatogram (TIC) of \u003cem\u003eT. aurea\u003c/em\u003ehydroethanolic extract by UHPLC-ESI-MS/MS in positive and negative ionization modes\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-4585453/v1/0b289618533aebe13aee041d.png"},{"id":58587267,"identity":"1463a4f0-52eb-4c91-b181-086fa44a6c9f","added_by":"auto","created_at":"2024-06-18 14:34:22","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":32594,"visible":true,"origin":"","legend":"\u003cp\u003eStructures of the isolated compounds from \u003cem\u003eT. aurea\u003c/em\u003e\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-4585453/v1/4a3258558ab35189adf2dc5f.png"},{"id":58587853,"identity":"17866aad-4e28-423b-820c-b4ddd7fd03e9","added_by":"auto","created_at":"2024-06-18 14:42:26","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":775441,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4585453/v1/d70d6b49-06f5-4782-9d4e-2883b4da3e62.pdf"},{"id":58587269,"identity":"deb2d78a-9e8c-4eaa-9bba-da165f9e4609","added_by":"auto","created_at":"2024-06-18 14:34:22","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":2701223,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cbr\u003e\u003c/p\u003e","description":"","filename":"Supplementarymaterial.docx","url":"https://assets-eu.researchsquare.com/files/rs-4585453/v1/5a6a1074e9af370960808bd9.docx"},{"id":58587264,"identity":"dd02b93d-b06f-4d59-bc5c-9214a9588967","added_by":"auto","created_at":"2024-06-18 14:34:22","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":29817,"visible":true,"origin":"","legend":"","description":"","filename":"Table1.docx","url":"https://assets-eu.researchsquare.com/files/rs-4585453/v1/493081f71ee69395fbc5cd34.docx"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"\u003cp\u003e\u003cstrong\u003eExploration of the chemical constituents of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eTricholomopsis aurea\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e (Beeli) Desjardin using UHPLC-ESI-MS/MS and NMR spectroscopy\u003c/strong\u003e\u003c/p\u003e","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eThe species \u003cem\u003eTricholomopsis aurea\u003c/em\u003e (Beeli) Desjardin \u0026amp; B.A. Perry is a wild edible fungus\u003csup\u003e1\u003c/sup\u003e that is scarce in Cameroon but common in the Democratic Republic of Congo, Burundi, Tanzania, Zimbabwe, S\u0026atilde;o Tom\u0026eacute;, Togo, Uganda, Martinique, Guadeloupe, and Trinidad2. The specimens of \u003cem\u003eT. aurea\u003c/em\u003e collected in Cameroon are characterized by a bright to sulphur yellow basidiomata measuring 25\u0026ndash;53 mm in diameter, a conspicuously rather flattened, fistulose to hollow and centrally depressed (from top to bottom) stipe measuring 35\u0026ndash;55 mm long and 6.0\u0026ndash;10 mm in diameter, thin flesh, and white spore-prints. Its basidia include four (\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e) and two (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e) but also one (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e) sterigmata, the presence of a cheilo-type and absence of pleurocystidia, and cutis-type pileipellis with erected hyphae that are subcylindrical to subclavate at the apex, some of which are incrusted\u003csup\u003e2\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eIn recent years, ultrahigh-performance liquid chromatography (UHPLC) and high-pressure liquid chromatography (HPLC) combined with tandem mass spectrometry (MS/MS) methods, have been used effectively in the chemical profiling of food matrices and natural extracts. This is because of their high sensitivity in obtaining accurate molecular masses to generate empirical formulas, retention times (tR) and fragment ions of the complexed samples analyzed\u003csup\u003e3,4\u003c/sup\u003e. Additionally, electrospray ionization mass spectrometry (ESI-MS) has been used for decades to characterize bioactive compounds, including fatty acids and other classes of compounds from mushrooms\u003csup\u003e5\u003c/sup\u003e. The metabolic profile of \u003cem\u003eT. aurea\u003c/em\u003e has not been extensively analyzed thus far. Hence, the objective of the present study was to identify the phytochemical profile of \u003cem\u003eT. aureus\u003c/em\u003e by UHPLC-ESI-MS/MS and to isolate and characterize some of the major compounds using nuclear magnetic resonance (NMR) spectroscopy.\u003c/p\u003e"},{"header":"MATERIALS AND METHODS","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eSample collection and identification\u003c/h2\u003e \u003cp\u003eThe sample was collected on April 10th, 2022, in Bambui (northwestern region of Cameroon) on a dead cypress stump. These species were subsequently identified as \u003cem\u003eTricholomopsis aurea\u003c/em\u003e (Beeli) Desjardin \u0026amp; B.A. Perry by Professor MOSSEBO Dominique Claude, a Mycologist of the Mycological Laboratory at the Department of Plant Biology and Physiology, University of Yaound\u0026eacute; 1 in Cameroon, by comparing the macro- and micromorphological features of the sample with those described in the most relevant literature\u003csup\u003e2\u003c/sup\u003e. The identified samples were thereafter preserved in the Mycological Herbarium of the University of Yaound\u0026eacute; 1 under voucher specimen number HUY1-DM 1937.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eUHPLC-ESI-MS/MS conditions\u003c/h2\u003e \u003cp\u003eFor chromatographic separation, a Waters\u0026reg; ACQUITY UPLC\u0026reg; I-Class System equipped with a Binary Solvent Manager and FTN Sample Manager (Waters GmbH, Eschborn, Germany) was used. Eluents A and B were 0.3 mmol/L ammonium formate (adjusted to pH 3.5 with formic acid) and acetonitrile, respectively. The gradient profile for eluent B was as follows: 0.0\u0026ndash;2 min, 5%; 2\u0026ndash;19 min, 95%; 19\u0026ndash;21 min, 95%; and 21\u0026ndash;21.5 min, 5%. For re-equilibration of the UHPLC column, the gradient was set to 3% eluent B for 6 min. The column oven was set at 40\u0026deg;C, and the autosampler was cooled to 5\u0026deg;C. The flow rate was kept constant at 0.4 mL/min. A volume of 5 \u0026micro;L of the sample was injected onto an EC 150/2 Nucleoshell RP18 column (endcaped C18 phase, ID 2 mm, length 150 mm, particle size 2.7 \u0026micro;m; Macherey Nagel, D\u0026uuml;ren, Germany).\u003c/p\u003e \u003cp\u003eMass spectrometric detection in positive and negative ion modes was performed using a QqTOF MS instrument (TripleTOF 6600, Sciex, Concord, Ontario, Canada) equipped with an ESI-DuoSpray-Ion-Source and resolving power (full width at half-maximum (FWHM), at m/z 400) set of 34,000 in MS and 30,000 in data-dependent acquisition mode (DDA). All the above steps were performed for ions greater than m/z 100 and with an intensity greater than 500, with dynamic background subtraction (DBS) and exclusion criteria of 3 s after 1 occurrence. The automated calibration device system performed an external calibration. The Turbo V ion drive source equipped with a stainless-steel electrode (100 \u0026micro;m internal diameter) was operated under the following MS conditions: nebulizing gas (gas 1), nitrogen (60 psi), drying gas (gas 2), nitrogen (70 psi), ion spray voltage, 5500 V (positive ion mode), 4500 V (negative mode), ion-source temperature, 450\u0026deg;C, and curtain gas, nitrogen (55 psi). The TOF MS accumulation time was set to 50 ms, and the mass range was 65 to 1250 m/z. A 100 ms accumulation time, a collision energy (CE) of 45 V with a collision energy spread (CES) of 35 V and a declustering potential (DP) of 35 V were used for the TOF MS/MS experiments in positive ion mode. In negative ion mode, the CE, CES and DP were \u0026minus;\u0026thinsp;45 V, 35 V and \u0026minus;\u0026thinsp;35 V, respectively. All MS parameters were controlled by Analyst TF 1.7.1 software (Sciex). Data evaluation was performed using the software PeakView Version 2.2 (Sciex). With respect to the retrieval of chemical structure information, databases such as Chem-Spider (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://www.chemspider.com\u003c/span\u003e\u003cspan address=\"http://www.chemspider.com\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e), SciFinder (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://scifinder.cas.org\u003c/span\u003e\u003cspan address=\"https://scifinder.cas.org\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e), NIST/EPA/NIH Mass Spectral Library, PubChem (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://pubchem.ncbi.nlm.nih.gov\u003c/span\u003e\u003cspan address=\"http://pubchem.ncbi.nlm.nih.gov\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e), MassBank (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://massbank.jp\u003c/span\u003e\u003cspan address=\"https://massbank.jp\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e), LipidBank (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://lipidbank.jp\u003c/span\u003e\u003cspan address=\"https://lipidbank.jp\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e), and the LIPID MAPS Structure Database (LMSD) (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.lipidmaps.org\u003c/span\u003e\u003cspan address=\"https://www.lipidmaps.org\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) were explored, as were published studies.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eAdditional analytical methods\u003c/h2\u003e \u003cp\u003eThe mass spectra were registered on a Waters ZQ-200 in electrospray ionization mode. NMR spectra were recorded on an AVANCE-NEO Bruker spectrometer equipped with a magnetic field of 14T operating at a proton frequency of 600 MHz. Low-resolution electrospray ionization mass spectrometry (ESI-MS) experiments were carried out on a Micromass Quattro Micro mass spectrometer. The LCMS used were a UHPLC (Waters) and a QqToF-MS/MS instrument (TriTOF 6600; Sciex). Column chromatography was performed on silica gel (0.04\u0026ndash;0.063 nm; 230\u0026ndash;400 mesh, ASTM; Merck, Germany). Thin layer chromatography (TLC) was performed on a silica gel 60 F254 (0.1 mm thick; Merck) with a size of 20 \u0026times; 20 cm, and spots were detected by fluorescence at 254 nm or 366 nm on a UV-85/L basis. The sample was sprayed with 10% H\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u003c/sub\u003e, and then heated at 70\u0026deg;C. The solvents used were of analytical grade. Melting points were recorded using a Stuart SMP3 melting point apparatus.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eExtraction and isolation of compounds\u003c/h2\u003e \u003cp\u003eA total of 250 g of the air-dried basidiocarp was blended and extracted with ethanol and water (EtOH-H2O, 7:3 v/v) for 72 hours, after which the filtrate was evaporated at reduced pressure to yield a brown paste (25 g). A small amount of the paste (0.5 mg) was dissolved in ammonium formate (adjusted to pH 3.5 with formic acid) and acetonitrile and subjected to UHPLC analysis, while 16 g of the paste was chromatographed using increasing proportions of hexane (Hex), ethyl acetate (EtOAc) and methanol (MeOH), affording 157 fractions. Based on thin layer chromatography (TLC) profiles, matching similar fractions of F50-60 (Hex-EtOAc, 76:24) and F69-72 (Hex-EtOAc, 68:32) and washing the precipitates with a mixture of Hex-EtOAc 3% afforded ergosterol (18.5 mg) and ergosterol 5α,8α-endoperoxide (8 mg), respectively. Fraction F38-40 (Hex-EtOAc, 88:12) was rechromatographed over a silica gel column using an increasing gradient system of Hex-EtOAc to yield petroselinic acid (10 mg, Rf: 0.60) and (Z)-oleic acid (12 mg, Rf: 0.73). The purification of fraction F41-43 (Hex-EtOAc, 82:18) using an increasing gradient system of Hex-EtOAc gave subfraction SF14-16, affording elaidic acid (15 mg). A portion of fraction F136-138 (22 mg) was purified on a silica gel column using an isocratic system of EtOAc-MeOH-H2O-pyridine (10:4:1:0.5) to yield adenosine (12 mg).\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003eMetabolite profiling of the 30% hydroethanolic (HEE) extract of \u003cem\u003eT. aurea\u003c/em\u003e was performed via UHPLC-ESI-MS/MS. Figure\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e shows the total ion chromatograms (TICs) analyzed in both positive and negative ionization modes, while Table\u0026nbsp;1 summarizes the experimental and observed mass‒charge ratios (m/z) of the precursor ions, mass errors (ppm), molecular formulas, plausible fragments, retention times (rt), and tentatively characterized compounds. The accurate masses of the positive ions and negative ions detected by QqTOF MS were converted into a molecular formula, such as CxHyNzOwS\u003csub\u003ek\u003c/sub\u003eP\u003csub\u003eL\u003c/sub\u003e, with errors presented below 3 ppm\u003csup\u003e6\u003c/sup\u003e. A total of 65 compounds were tentatively identified as belonging to 18 different classes, such as amino acids (\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e), carbohydrates (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e), vitamins (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e), aminoglycolipids (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e), phosphoglycolipids (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e), fatty acids (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e), phenolics (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e), nucleosides (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e), steroids (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e), fatty acid amides (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e), acyl glycerides (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e), sphingolipids (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e), alkaloids (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e), terpenes (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e), carotenoids (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e), saponins (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e), aromatic amines (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e), and peptides (\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e). Of these, 6 compounds were isolated and described as oleic acid, elaidic acid petroselinic acid, ergosterol, ergosterol 5α,8α-endoperoxide and adenosine using 1D and 2D NMR spectroscopy.\u003c/p\u003e \u003cp\u003e \u003cb\u003eCharacterization of the hydroethanolic extracts of\u003c/b\u003e \u003cb\u003eT. aurea\u003c/b\u003e \u003cb\u003eby UHPLC-ESI-MS/MS\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe ingestion of peptides, amino acids and proteins from different food sources is essential for maintaining health\u003csup\u003e7,8\u003c/sup\u003e. In fact, in this study, seven amino acids and seven peptides were tentatively detected in both the positive and negative ion modes. The positive ion mode showed the precursor ion [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003e of valine (peak 59, C\u003csub\u003e5\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eNO\u003csub\u003e2\u003c/sub\u003e) at m/z 118.0868. The peak at 72.08132 [M\u0026thinsp;+\u0026thinsp;H-CO\u003csub\u003e2\u003c/sub\u003e] could be attributed to the loss of carbon dioxide. Valine has been quantified in many mushroom species\u003csup\u003e9,10\u003c/sup\u003e and is needed by animals for the development of mammary glands and ovaries\u003csup\u003e11\u003c/sup\u003e. Additionally, in positive ion mode, arginine (peak 49, C\u003csub\u003e6\u003c/sub\u003eH\u003csub\u003e14\u003c/sub\u003eN\u003csub\u003e4\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e) was observed at m/z 174.11119 in wild edible mushrooms\u003csup\u003e9\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eIn negative ionization mode, isomers of the (R)-pyroglutamic acid (peak 2, C\u003csub\u003e5\u003c/sub\u003eH\u003csub\u003e7\u003c/sub\u003eNO\u003csub\u003e3\u003c/sub\u003e) precursor [M-H]\u003csup\u003e\u0026minus;\u003c/sup\u003e were identified at 128.03502 in shiitake mushrooms\u003csup\u003e10\u003c/sup\u003e, and (S)-pyroglutamic acid (peak 7, C\u003csub\u003e5\u003c/sub\u003eH\u003csub\u003e7\u003c/sub\u003eNO\u003csub\u003e3\u003c/sub\u003e) was identified at m/z 128.035 in \u003cem\u003eMorchella sp\u003c/em\u003e\u003csup\u003e\u003cem\u003e12\u003c/em\u003e\u003c/sup\u003e. Both of these compounds presented fragment ions at 84.04494 (M-H-CO\u003csub\u003e2\u003c/sub\u003e)\u003csup\u003e\u0026minus;\u003c/sup\u003e, indicating decarboxylation, as did 56.02621 [M-H-CO\u003csub\u003e2\u003c/sub\u003e-CH\u003csub\u003e2\u003c/sub\u003eNH]\u003csup\u003e\u0026minus;\u003c/sup\u003e, which was attributed to decarboxylation and loss of methanimine. Other amino acids identified were leucine (peak 4, C\u003csub\u003e6\u003c/sub\u003eH\u003csub\u003e13\u003c/sub\u003eNO\u003csub\u003e2\u003c/sub\u003e) at m/z 130.08715, which was detected in shiitake mushrooms\u003csup\u003e10\u003c/sup\u003e; phenylalanine (peak 10, C\u003csub\u003e9\u003c/sub\u003eH\u003csub\u003e11\u003c/sub\u003eNO\u003csub\u003e2\u003c/sub\u003e) at m/z 164.07131, which was reported in \u003cem\u003eArabidopsis thaliana\u003c/em\u003e\u003csup\u003e\u003cem\u003e13\u003c/em\u003e\u003c/sup\u003e; and α-hydroxymethylserine (peak 18, C\u003csub\u003e4\u003c/sub\u003eH\u003csub\u003e9\u003c/sub\u003eNO\u003csub\u003e4\u003c/sub\u003e) at m/z 134.04572, which was isolated from the seeds of \u003cem\u003eVicia pseudo-Orobus\u003c/em\u003e\u003csup\u003e14\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eThe tripeptide Gln-Trp-Arg\u003csup\u003e15\u003c/sup\u003e, which eluted at a retention time of 18.427 min, was detected as an [M-H]\u003csup\u003e\u0026minus;\u003c/sup\u003e ion at m/z 487.24178 (peak 63). This was attributed to the molecular formula C\u003csub\u003e22\u003c/sub\u003eH\u003csub\u003e32\u003c/sub\u003eN\u003csub\u003e8\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e. The fragment ion at m/z 442.24409 [M -H \u0026minus;\u0026thinsp;44]\u003csup\u003e\u0026minus;\u003c/sup\u003e could be due to the loss of carbon dioxide molecules. Peak 62 (rt: 18.284 min) was detected as an [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003e ion at m/z 471.35491, and its molecular formula was assigned as C\u003csub\u003e24\u003c/sub\u003eH\u003csub\u003e46\u003c/sub\u003eN\u003csub\u003e4\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e due to mass; thus, this peak was attributed to the tetrapeptide Leu-Leu-Leu-Leu\u003csup\u003e16\u003c/sup\u003e. Its fragment at 426.35699 [M\u0026thinsp;+\u0026thinsp;H-44]\u003csup\u003e+\u003c/sup\u003e suggested the loss of a carbon dioxide molecule from the terminal carboxyl group. Peak 65 (rt: 12.894 min) was observed as an [M-H]\u003csup\u003e\u0026minus;\u003c/sup\u003e ion at m/z 611.31979 and has a molecular formula of C31H44N6O7. This was attributed to cyclo (-d-glu-ala-d-allo-ile-leu-d-trp), commonly known as BE-18257B\u003csup\u003e17\u003c/sup\u003e. L-365,209, peak 64 (rt: 12.836 min) was detected as an [M-H]- ion at m/z 665.38151 with the molecular formula C\u003csub\u003e39\u003c/sub\u003eH\u003csub\u003e50\u003c/sub\u003eN\u003csub\u003e6\u003c/sub\u003eO\u003csub\u003e6\u003c/sub\u003e. It was previously reported as a class of cyclic hexapeptide oxytocin antagonists from \u003cem\u003eStreptomyces silvensis\u003c/em\u003e\u003csup\u003e18\u003c/sup\u003e. Muracein A (peak 15, C\u003csub\u003e26\u003c/sub\u003eH\u003csub\u003e44\u003c/sub\u003eN\u003csub\u003e6\u003c/sub\u003eO\u003csub\u003e14\u003c/sub\u003e) at m/z 663.28431 was detected in \u003cem\u003eNocardia orientalis\u003c/em\u003e\u003csup\u003e19\u003c/sup\u003e; pandanamide A (peak 19, C\u003csub\u003e31\u003c/sub\u003eH\u003csub\u003e47\u003c/sub\u003eN\u003csub\u003e7\u003c/sub\u003eO\u003csub\u003e9\u003c/sub\u003e) at m/z 660.33697 was isolated from \u003cem\u003eStreptomyces sp\u003c/em\u003e.\u003csup\u003e20\u003c/sup\u003e; and the tripeptide Leu-His-Ala peak 20 (C\u003csub\u003e15\u003c/sub\u003eH\u003csub\u003e25\u003c/sub\u003eN\u003csub\u003e5\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e) at m/z 338.18271 was reported in dry-cured Jinhua ham\u003csup\u003e21\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eSphingolipids and glycerophospholipids participate in a variety of important neurological, metabolic and intracellular signaling processes\u003csup\u003e22\u003c/sup\u003e. Sphingolipids serve as building blocks of the plasma membrane of eukaryotic cells. They anchor lipid-bound carbohydrates to cell surfaces and construct the epidermal water permeability barrier\u003csup\u003e23\u003c/sup\u003e. As shown in Table\u0026nbsp;1, the protonated ion [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003e at m/z 302.30516 (peak 31, C\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e40\u003c/sub\u003eNO\u003csub\u003e2\u003c/sub\u003e) was attributed to sphinganine detected previously in \u003cem\u003eArabidopsis thaliana\u003c/em\u003e roots\u003csup\u003e24\u003c/sup\u003e, while that at m/z 298.27453 (peak 44, C\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e35\u003c/sub\u003eNO\u003csub\u003e2\u003c/sub\u003e) was attributed to sphinga-4E,8E-dienine, which was revealed in oyster mushrooms\u003csup\u003e25\u003c/sup\u003e. Deinococcucin D displayed a precursor ion [M-H]\u003csup\u003e\u0026minus;\u003c/sup\u003e at m/z 543.36482 (peak 26, C\u003csub\u003e28\u003c/sub\u003eH\u003csub\u003e52\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e8\u003c/sub\u003e). It was isolated from the gut of queen carpenter ants \u003csup\u003e26\u003c/sup\u003e, while mannosyl (4S)-4-methylheptacosyl phosphate, a synthetic analog of β-D-mannosyl C32-phosphomycoketide27, was detected at m/z 651.46027 (peak 60, C\u003csub\u003e34\u003c/sub\u003eH\u003csub\u003e69\u003c/sub\u003eO\u003csub\u003e9\u003c/sub\u003eP). 1-(9Z-Octadecenoyl)-2-(5-hydroxy-7-carboxy-6E-heptenoyl)-sn-glycero-3-phosphate (peak 17, C\u003csub\u003e31\u003c/sub\u003eH\u003csub\u003e54\u003c/sub\u003eO\u003csub\u003e13\u003c/sub\u003eP) at m/z 665.33064 was described previously as an oxidized phospholipid\u003csup\u003e28\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eTable\u0026nbsp;1: Based tentative identification of compounds from the hydroethanolic extract of \u003cem\u003eT. aurea\u003c/em\u003e by UHPLC-ESI-MS/MS\u003c/p\u003e \u003cp\u003eTerpenes were identified in the total ion chromatogram (retention time range 12.449\u0026ndash;18.427 min). Peak 45, with a protonated molecular ion [M-H]\u003csup\u003e+\u003c/sup\u003e at m/z 329.16008 (C\u003csub\u003e16\u003c/sub\u003eH\u003csub\u003e24\u003c/sub\u003eO\u003csub\u003e7\u003c/sub\u003e), was assigned to zataroside B, described in \u003cem\u003eZataria multiflora\u003c/em\u003e\u003csup\u003e29\u003c/sup\u003e. The fragment ion at 165.09155 [M-162-H]\u003csup\u003e\u0026minus;\u003c/sup\u003e could be due to the loss of a hexose unit. Trichodermanin D (peak 50, C\u003csub\u003e20\u003c/sub\u003eH\u003csub\u003e34\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e), precursor ion [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003e at m/z 323.25831, was detected in the fungus \u003cem\u003eTrichoderma harzianum\u003c/em\u003e\u003csup\u003e30\u003c/sup\u003e. The analog of nodulisporic acid A (peak 22, C45H57NO7) and the pseudomolecular ion [M-H]\u003csup\u003e\u0026minus;\u003c/sup\u003e at m/z 722.40634 were isolated from an endophytic fungus\u003csup\u003e31\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eAs shown in Table\u0026nbsp;1, the deprotonated molecular ion of Aspacoside D (peak 56, C\u003csub\u003e38\u003c/sub\u003eH\u003csub\u003e60\u003c/sub\u003eO\u003csub\u003e16\u003c/sub\u003e) was observed at 771.38034 [M-H]\u003csup\u003e\u0026minus;\u003c/sup\u003e in negative ion mode and could afford fragment ions at 609.32749 [M-H-162]\u003csup\u003e\u0026minus;\u003c/sup\u003e, 476.27740 [M-H-162-132]\u003csup\u003e\u0026minus;\u003c/sup\u003e and 315.23241 [M-H-162-162]\u003csup\u003e\u0026minus;\u003c/sup\u003e, corresponding to the loss of one pentose and two hexose units. This molecule was isolated from Asparagus cochinchinensis32. The protonated molecular ion of the phenolic de-O-methyllasiodiplodin was observed at m/z 279.15965 (peak 32, C\u003csub\u003e16\u003c/sub\u003eH\u003csub\u003e22\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e) in positive ion mode (Table\u0026nbsp;1). This compound has been described in \u003cem\u003eGouania longipetala\u003c/em\u003e and \u003cem\u003eGlyphaea brevis\u003c/em\u003e\u003csup\u003e33\u003c/sup\u003e. Two additional phenolic compounds were characterized in negative ionization mode, with pseudomolecular ions at m/z 331.11842 (peak 14, C\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e20\u003c/sub\u003eO\u003csub\u003e6\u003c/sub\u003e) and 151.03924 (peak 12, C\u003csub\u003e8\u003c/sub\u003eH\u003csub\u003e8\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e), attributed to 3',5'-dihydroxy-2',4',6'-trimethoxydihydrochalcone, which was isolated from the twigs of \u003cem\u003eLindera lucida\u003c/em\u003e\u003csup\u003e34\u003c/sup\u003e, and anisic acid or 4-methoxybenzoic acid, which was isolated from Rhododendron\u003csup\u003e35\u003c/sup\u003e. The aromatic amine 2,4-diaminophenol observed at m/z 125.07146 (peak 61, C\u003csub\u003e6\u003c/sub\u003eH\u003csub\u003e9\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eO) is a known cosmetic ingredient\u003csup\u003e36\u003c/sup\u003e. The exact mass of uridine at m/z 243.06126 (peak 3, C\u003csub\u003e9\u003c/sub\u003eH\u003csub\u003e11\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e6\u003c/sub\u003e) was obtained in negative ionization mode. The presence of uridine in the mushroom \u003cem\u003ePleurotus giganteu\u003c/em\u003e reportedly increases the phosphorylation of extracellular signal-regulated kinases and protein kinase B\u003csup\u003e37\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eThe precursor ion of glucose was observed at m/z 179.05523 (peak 1, C\u003csub\u003e6\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eO\u003csub\u003e6\u003c/sub\u003e) in negative mode (Table\u0026nbsp;1). Glucose has been detected in mushrooms and is a substrate for energy-producing processes\u003csup\u003e38\u003c/sup\u003e. In the positive ion mode, peak 8 (C\u003csub\u003e6\u003c/sub\u003eH\u003csub\u003e13\u003c/sub\u003eNO\u003csub\u003e5\u003c/sub\u003e) at m/z 180.08637 and peak 9 (C\u003csub\u003e5\u003c/sub\u003eH\u003csub\u003e10\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e) at m/z 163.06027 were attributed to nojirimycin\u003csup\u003e39\u003c/sup\u003e and 1,5-anhydrofructose\u003csup\u003e40\u003c/sup\u003e, respectively. The precursor ion [M-H]\u003csup\u003e\u0026minus;\u003c/sup\u003e for xanthine was observed at m/z 151.02575 (peak 5, C\u003csub\u003e5\u003c/sub\u003eH\u003csub\u003e4\u003c/sub\u003eN\u003csub\u003e4\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e). Tea, cocoa seeds, and coffee are the natural sources of xanthine\u003csup\u003e41\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eThe acyl glyceride 1-palmitoyl glycerol (peak 39, C\u003csub\u003e19\u003c/sub\u003eH\u003csub\u003e38\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e) protonated molecular ion [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003e at 331.28455 could yield a fragment ion at 239.23749 [M-C\u003csub\u003e3\u003c/sub\u003eH\u003csub\u003e7\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e]\u003csup\u003e+\u003c/sup\u003e as a result of the loss of the glycerol moiety. It serves as an emulsifying agent and has been used to increase the viscoelasticity of processed cheese\u003csup\u003e42\u003c/sup\u003e. The precursor ion [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003e at m/z 396.34738 was attributed to semiplenamide E (peak 55, C\u003csub\u003e24\u003c/sub\u003eH\u003csub\u003e45\u003c/sub\u003eNO\u003csub\u003e3\u003c/sub\u003e), which was detected in the marine cyanobacterium \u003cem\u003eLyngbya semiplena\u003c/em\u003e\u003csup\u003e43\u003c/sup\u003e, whereas the natural product N-palmitoyl-D-leucine (Peak 61, C\u003csub\u003e22\u003c/sub\u003eH\u003csub\u003e43\u003c/sub\u003eNO\u003csub\u003e3\u003c/sub\u003e) precursor ion [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003e at m/z 370.33209 was identified in the extracts of rat brains or bovine spinal cords\u003csup\u003e44\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eFourteen fatty acids were detected in the positive and negative ion modes, among which (11E,13E)-9,10-dihydroxy-11,13-octadecadienoic acid (peak 24, C\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e32\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e) at m/z 313.23759 was detected in \u003cem\u003eM. giganteus\u003c/em\u003e\u003csup\u003e45\u003c/sup\u003e, and palmitoleic acid (peak 25, C\u003csub\u003e16\u003c/sub\u003eH\u003csub\u003e30\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e) was detected at m/z 255.23172 in \u003cem\u003eT. rutilans\u003c/em\u003e\u003csup\u003e46\u003c/sup\u003e. Cis-linoleic acid (peak 27, C18H32O2) at m/z 281.24815 was described in \u003cem\u003eLactarius salmonicolor\u003c/em\u003e, \u003cem\u003ePolyporus squamosus\u003c/em\u003e, \u003cem\u003ePleurotus ostreatus\u003c/em\u003e and \u003cem\u003eFlammulina velutipes\u003c/em\u003e\u003csup\u003e47\u003c/sup\u003e; linolenic acid (peak 28, C\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e32\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e) at m/z 279.23241 was detected in \u003cem\u003eHelichrysum pedunculatum\u003c/em\u003e, \u003cem\u003ePolyporus squamosus\u003c/em\u003e, and \u003cem\u003ePleurotus ostreatus\u003c/em\u003e\u003csup\u003e48\u003c/sup\u003e. Trans-linoleic acid (peak 30, C\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e32\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e) at m/z 281.24783 has been described in \u003cem\u003eF. hepatica\u003c/em\u003e and \u003cem\u003eT. equestre\u003c/em\u003e\u003csup\u003e47\u003c/sup\u003e. Caproliec acid, also known as 9-decenoic acid (peak 47, C\u003csub\u003e10\u003c/sub\u003eH\u003csub\u003e18\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e), was detected at m/z 171.13875 in beer and yeast\u003csup\u003e49\u003c/sup\u003e; octadecanedioic acid (peak 46, C\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e34\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e), at m/z 315.25343, was detected in \u003cem\u003eArabidopsis thaliana\u003c/em\u003e\u003csup\u003e50\u003c/sup\u003e. 9-Octadecynoic acid or stearolic acid (peak 43, C\u003csub\u003e28\u003c/sub\u003eH\u003csub\u003e32\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e) at m/z 281.2482 was detected in plant oil seeds\u003csup\u003e51\u003c/sup\u003e, while \u0026#120574;-linolenic acid (peak 38, C\u003csub\u003e28\u003c/sub\u003eH\u003csub\u003e30\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e) at m/z 279.23195 was determined from Zygomycetous Fungi\u003csup\u003e52\u003c/sup\u003e. Suberic acid (peak 13, C\u003csub\u003e8\u003c/sub\u003eH\u003csub\u003e14\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e) at m/z 173.0813 was detected in urine\u003csup\u003e53\u003c/sup\u003e, and 13-hydroxy-9Z,11E,15E-octadecatrienoic acid (peak 51, C\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e30\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e) [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003e at m/z 295.22709 was isolated from the leaves of \u003cem\u003eCucurbita moschata\u003c/em\u003e\u003csup\u003e54\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eAs enumerated in Table\u0026nbsp;1, the protonated molecular ion of the isomeric compound 6β-hydroxy-ergosta-4,7,22-trien-3-one (peak 40, C\u003csub\u003e28\u003c/sub\u003eH\u003csub\u003e42\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e) was at m/z 411.32631 [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003e, and its dehydration of C6-OH with B-ring hydrogen could yield the fragment ion at m/z 392.30792 [M\u0026ndash;18]\u003csup\u003e+\u003c/sup\u003e. In addition, the peak at m/z 268.1872 [M\u0026thinsp;+\u0026thinsp;H-18-C\u003csub\u003e9\u003c/sub\u003eH\u003csub\u003e17\u003c/sub\u003e]\u003csup\u003e+\u003c/sup\u003e was attributed to dehydration and loss of the side chain on the D ring\u003csup\u003e55\u003c/sup\u003e. Its isomer 6α-hydroxy-ergosta-4,7,22-trien-3-one (peak 42, C\u003csub\u003e28\u003c/sub\u003eH\u003csub\u003e42\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e) at m/z 411.32675 presented a similar fragmentation pattern. These compounds were all described in \u003cem\u003eG. lucidum\u003c/em\u003e\u003csup\u003e55\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eGagarlol B (peak 35, C\u003csub\u003e28\u003c/sub\u003eH\u003csub\u003e42\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e) was detected at m/z 443.31606 and was isolated from the edible mushroom \u003cem\u003eGrifola gargal\u003c/em\u003e\u003csup\u003e56\u003c/sup\u003e; ganodermaside C (peak 48, C\u003csub\u003e28\u003c/sub\u003eH\u003csub\u003e38\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e) at m/z 423.28935 was isolated from \u003cem\u003eGanoderma lucidum\u003c/em\u003e\u003csup\u003e57\u003c/sup\u003e; ergosta-4,6,8(\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e),22‐tetraen‐3‐one (peak 54, C\u003csub\u003e28\u003c/sub\u003eH\u003csub\u003e40\u003c/sub\u003eO) at m/z 393.31563 was noted in \u003cem\u003ePorodaedalea pini\u003c/em\u003e\u003csup\u003e58\u003c/sup\u003e; 5α-8α-epidioxy-24-methylcholesta-6,9(\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e),24(\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e)-trien-3β-ol (peak 34, C\u003csub\u003e28\u003c/sub\u003eH\u003csub\u003e42\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e) at m/z 427.32115 was identified in the marine sponge \u003cem\u003eCoscinoderma sp\u003c/em\u003e.\u003csup\u003e59\u003c/sup\u003e; and stigmasta-3,5-diene (peak 57, C\u003csub\u003e29\u003c/sub\u003eH\u003csub\u003e48\u003c/sub\u003e) at m/z 397.38317 was detected in \u003cem\u003eCuscuta reflexa\u003c/em\u003e Roxb\u003csup\u003e60\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eThe negative mode ESI enabled the identification of 12β-acetoxy-3,7,11,15,23-pentaoxo-5α-lanosta-8-en-26-oic acid ethyl ester (peak 16, C\u003csub\u003e34\u003c/sub\u003eH\u003csub\u003e46\u003c/sub\u003eO\u003csub\u003e9\u003c/sub\u003e) at m/z 597.30632, reported in the fruiting body of \u003cem\u003eGanoderma lucidum\u003c/em\u003e\u003csup\u003e61\u003c/sup\u003e; peak 23 (C\u003csub\u003e34\u003c/sub\u003eH\u003csub\u003e46\u003c/sub\u003eO\u003csub\u003e8\u003c/sub\u003e) at m/z 581.31188, ascribed to a helvolic acid methyl ester derivative from the endophytic \u003cem\u003eFicus carica\u003c/em\u003e\u003csup\u003e62\u003c/sup\u003e; and 3β-amino-5-spirosolene (peak 58, C\u003csub\u003e27\u003c/sub\u003eH\u003csub\u003e44\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eO) at m/z 411.33725, which was isolated from \u003cem\u003eSolanum triste\u003c/em\u003e\u003csup\u003e63\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eThe precursor ion [M-H]\u003csup\u003e\u0026minus;\u003c/sup\u003e at m/z 122.02425 was attributed to niacin (peak 6, C\u003csub\u003e6\u003c/sub\u003eH\u003csub\u003e5\u003c/sub\u003eNO\u003csub\u003e2\u003c/sub\u003e), a water soluble vitamin that has been found in different kinds of plant-based foods64. In positive ionization mode, the precursor ion at m/z 429.33619 (peak 52, C\u003csub\u003e28\u003c/sub\u003eH\u003csub\u003e44\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e) was attributed to the fat soluble vitamin 1α,25-dihydroxy-24-epivitamin D2\u003csup\u003e65\u003c/sup\u003e. The carotenoid 7,8-dihydroparasiloxanthinin (peak 37, C\u003csub\u003e40\u003c/sub\u003eH\u003csub\u003e60\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e) at m/z 573.466628 was reported in \u003cem\u003ePleurotus mushrooms\u003c/em\u003e\u003csup\u003e66\u003c/sup\u003e, while α-citraurin (peak 29, C\u003csub\u003e30\u003c/sub\u003eH\u003csub\u003e40\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e) and the pseudomolecular ion [M-H]\u003csup\u003e\u0026minus;\u003c/sup\u003e at m/z 431.29492 were isolated previously from the sea hare\u003csup\u003e67\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eThe nucleoside adenosine was observed at m/z 268.10442 (peak 11, C\u003csub\u003e10\u003c/sub\u003eH\u003csub\u003e13\u003c/sub\u003eN\u003csub\u003e5\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e). The fragment ion visible at m/z 249.150 [M-H\u003csub\u003e2\u003c/sub\u003eO]\u003csup\u003e+\u003c/sup\u003e was attributed to the loss of water molecules\u003csup\u003e68\u003c/sup\u003e. ESIMS (positive mode) showed adducts at m/z 305.9 [M\u0026thinsp;+\u0026thinsp;K]\u003csup\u003e+\u003c/sup\u003e and 290.6 [M\u0026thinsp;+\u0026thinsp;Na]\u003csup\u003e+\u003c/sup\u003e (supplementary material and Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). In addition, its \u003csup\u003e1\u003c/sup\u003eH NMR spectrum presented signals corresponding to two singlets at δ 8.62 (s, H-2) and 8.37 (s, H-8), with corresponding carbons at δ 153.7 (C-2) and 141.0 (C-8), respectively. The HMBC spectrum showed correlations between the proton singlets and the carbons at δ 158.2 (C-6), 150.6 (C-4) and 121.9 (C-5), thus suggesting a purine ring. The \u003csup\u003e1\u003c/sup\u003eH signals at δ 6.70 (d, J\u0026thinsp;=\u0026thinsp;6.0 Hz, H-1'); 5.52 (t, J\u0026thinsp;=\u0026thinsp;4.8 Hz, H-2'); 5.08 (brs, H-3'); 4.78 (q, J\u0026thinsp;=\u0026thinsp;5.4 Hz, H-4'); 4.38 (dd(distorted), J\u0026thinsp;=\u0026thinsp;11.4, 3.0 Hz, H-5'a); 4.17 (dd(distorted), J\u0026thinsp;=\u0026thinsp;11.4, 3.0 Hz, H-5'b)) in conjunction with their carbons at δ153.7 (C-2), 150.6 (C-4), 121.9 (C-5), 158.2 (C-6), 141.0 (C-8), 91.3 (C-1'), 76.2 (C-2), 72.9 (C-3'), 88.3 (C-4'), 63.5 (C5'), and 30.6 (C-14) suggested the presence of a ribose unit, thus corroborating its identification as adenosine. This compound was reported previously in \u003cem\u003eEuryale ferox\u003c/em\u003e Salisb\u003csup\u003e69\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eErgosterol (peak 53, C\u003csub\u003e28\u003c/sub\u003eH\u003csub\u003e44\u003c/sub\u003eO) at m/z 437.36533 was isolated as a white powder. The \u003csup\u003e1\u003c/sup\u003eH NMR spectrum showed signals at δ 3.96 (1H, m, H-3), 5.52 (1H, q, J\u0026thinsp;=\u0026thinsp;3.6, 6.6 Hz, H-7), 5.32 (1H, m, H-22) and 5.29 (1H, m, H-23), with carbons appearing at δ 70.6 (C-3), 117.8 (C-7), 136.8 (C-22) and 132.8 (C-23), respectively (Supplementary material). In addition, signals corresponding to six methyl groups were observed at δ 1.06 (3H, s, H-18), 0.91 (3H, d, J\u0026thinsp;=\u0026thinsp;4.2 Hz, H-26), 0.89 (3H, d, J\u0026thinsp;=\u0026thinsp;4.2 Hz, H-27), 0.70 (3H, s, H-19), 0.99 (3H, d, J\u0026thinsp;=\u0026thinsp;8.4 Hz, H-28) and 1.12 (3H, d, J\u0026thinsp;=\u0026thinsp;8.4 Hz, H-21). Its positive ESI mode displayed an adduct at m/z 790.9 [2M\u0026thinsp;+\u0026thinsp;H]\u0026thinsp;+\u0026thinsp;and molecular ions at m/z 396.8 [M]\u003csup\u003e+\u003c/sup\u003e and 395.8 [M \u0026ndash; H]\u003csup\u003e+\u003c/sup\u003e, respectively. The dehydration of C3-OH led to the fragment ion at m/z 377.7 [M\u0026thinsp;+\u0026thinsp;H \u0026ndash; 18]\u003csup\u003e+\u003c/sup\u003e, while the fragment ion at m/z 271.4 [M\u0026thinsp;+\u0026thinsp;H \u0026ndash; 125]\u003csup\u003e+\u003c/sup\u003e was attributed to the loss of the lateral chain. Ergosterol was previously extracted from the mushroom \u003cem\u003eSparassia crispa\u003c/em\u003e\u003csup\u003e70\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eErgosterol 5α,8α-endoperoxide (peak 54, C\u003csub\u003e28\u003c/sub\u003eH\u003csub\u003e42\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e) at m/z 429.33649 was obtained as a white needle. ESIMS (positive mode) showed adducts at m/z 882.1 [2M\u0026thinsp;+\u0026thinsp;Na]\u003csup\u003e+\u003c/sup\u003e and 451.8 [M\u0026thinsp;+\u0026thinsp;Na]\u003csup\u003e+\u003c/sup\u003e. \u003csup\u003e1\u003c/sup\u003eH signals were observed at δ 3.78 (1H, m, H-3), 6.28 (1H, d, J\u0026thinsp;=\u0026thinsp;8.1 Hz, H-6), 6.57 (1H, d, J\u0026thinsp;=\u0026thinsp;8.1 Hz, H-7), 5.22 (1H, dd, J\u0026thinsp;=\u0026thinsp;4.5, 9.3 Hz, H-22) and 5.28 (1H, dd, J\u0026thinsp;=\u0026thinsp;4.5, 9.3 Hz, H-23), with corresponding carbons at δ 66.9 (C-3), 136.8 (C-6), 131.8 (C-7), 136.8 (C-22) and 133.5 (C-23), respectively. Furthermore, signals corresponding to six methyl groups were observed at δ 0.85 (3H, s, H-18), 0.94 (3H, d, J\u0026thinsp;=\u0026thinsp;3.9 Hz, H-26), 1.03 (3H, d, J\u0026thinsp;=\u0026thinsp;3.9 Hz, H-27), 0.92 (3H, s, H-19), 0.86 (3H, d, J\u0026thinsp;=\u0026thinsp;1.8 Hz, H-28) and 2.01 (3H, d, J\u0026thinsp;=\u0026thinsp;3.0 Hz, H-21). The carbon signals at δ 83.4 (C-5) and 80.7 (C-8) were established using the HMBC spectrum (supplementary material). Ergosterol 5α,8α-endoperoxide was previously described in \u003cem\u003eDesmodium uncinatum\u003c/em\u003e\u003csup\u003e71\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003e(Z)-oleic acid (peak 33, C18H34O2) at m/z 324.29008 was isolated as a white oil. Its carbon spectrum showed an acid functional group at δ181.3 (C-1). In its \u003csup\u003e1\u003c/sup\u003eH NMR spectrum, the signals at δ 5.36 (m, H-9, H-10), 0.92 (t, J\u0026thinsp;=\u0026thinsp;7.2 Hz, H-18) and 1.29\u0026ndash;1.33 (m, H-12 to H-16, H-3 to H-7) were assigned to ethylinic, terminal methyl and long chain units, respectively (supplementary material). The negative mode ESI spectrum of oleic acid showed a molecular ion peak at m/z 282.0 [M]\u003csup\u003e\u0026minus;\u003c/sup\u003e. ; hydrated fragment ions at m/z 256 [M\u0026thinsp;+\u0026thinsp;H\u003csub\u003e2\u003c/sub\u003eO-C\u003csub\u003e3\u003c/sub\u003eH\u003csub\u003e7\u003c/sub\u003e]\u003csup\u003e\u0026minus;\u003c/sup\u003e; 171.7 [M\u0026thinsp;+\u0026thinsp;H\u003csub\u003e2\u003c/sub\u003eO-129]\u003csup\u003e\u0026minus;\u003c/sup\u003e. A prominent fragment ion at m/z 127.4 [C\u003csub\u003e7\u003c/sub\u003eH\u003csub\u003e11\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e]\u003csup\u003e\u0026minus;\u003c/sup\u003e resulted from the McLafferty rearrangement. The adducts in the positive ESI mode at 305.8 [M\u0026thinsp;+\u0026thinsp;Na]\u003csup\u003e+\u003c/sup\u003e, 321.8 [M\u0026thinsp;+\u0026thinsp;K]\u003csup\u003e+\u003c/sup\u003e and 301.7 [M\u0026thinsp;+\u0026thinsp;H\u003csub\u003e2\u003c/sub\u003eO]\u003csup\u003e+\u003c/sup\u003e as well as the fragment ion at 129.5 [M-C\u003csub\u003e11\u003c/sub\u003eH\u003csub\u003e21\u003c/sub\u003e]\u003csup\u003e+\u003c/sup\u003e supported its characterization as oleic acid, previously recognized in \u003cem\u003eT. rutilans\u003c/em\u003e\u003csup\u003e46\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eElaidic acid or trans-9-octadecenoic acid (peak 36, C\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e34\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e) at m/z 300.28993 was isolated as a white oil. The spectrum displayed \u003csup\u003e1\u003c/sup\u003eH signals at δ 5.34 (m, H-9, H-10), 0.89 (t, J\u0026thinsp;=\u0026thinsp;6.0 Hz, H-18) and 2.26 (t, J\u0026thinsp;=\u0026thinsp;6.0 Hz, H-2), assigned to vinylic groups, methyl groups and methylene groups, respectively. The corresponding carbon signals were observed at δ 130.8 (C-8), 130.9 (C-10), 14.4 (C-18) and 35.0 (C-2), while the acid functional group appeared at δ 177.7 (C-1). The positive ESI mode presented an adduct at m/z 321.7 [M\u0026thinsp;+\u0026thinsp;K]\u003csup\u003e+\u003c/sup\u003e (Supplementary material). Elaidic acid is an isomer of oleic acid, is one of the major trans fatty acids (TFAs) in foods and accounts for 40\u0026ndash;60% of the TFAs found in humans72.\u003c/p\u003e \u003cp\u003ePetroselinic acid (peak 21, C\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e34\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e) at m/z 300.28992 is a positional isomer of oleic acid. \u003csup\u003e1\u003c/sup\u003eH signals were observed at 5.34 (m, H-6, H-7); 0.90 (t, J\u0026thinsp;=\u0026thinsp;6.0 Hz, H-18); 1.59 (m, H-17); 2.18 (t, J\u0026thinsp;=\u0026thinsp;6.0 Hz, H-2); 2.02 (m, H-5, to H-8); and carbon at 130.9 (C-6), 130.8 (C-7), 14.4 (C-18), 23.7 (C-17), 37.8 (C-2), 28.1 (C-5) and 28.2 (C-8). The carbon spectrum showed a signal at δ 179.4 (C-1), attributed to an acid functional group (Supplemental material). In negative ESI mode, a molecular ion peak at 282.0 [M]\u003csup\u003e\u0026minus;\u003c/sup\u003e was observed, and an intense fragment ion at m/z 113.5 [C\u003csub\u003e6\u003c/sub\u003eH\u003csub\u003e9\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e]\u003csup\u003e\u0026minus;\u003c/sup\u003e formed through McLafferty rearrangement\u003csup\u003e73\u003c/sup\u003e. This compound has been detected in the vegetable oil of \u003cem\u003eCoriandrum sativum\u003c/em\u003e fruits\u003csup\u003e74\u003c/sup\u003e.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThis work tentatively identified 65 known compounds belonging to 18 chemical classes. Among these, 6 of the compounds were isolated and characterized as petroselinic acid, oleic acid, elaidic acid, ergosterol, ergosterol-5α,8α-endoperoxide and adenosine. This is the first report of provisional and isolated compounds from this species. However, some of the compounds were previously identified in the literature as vital dietary supplements necessary for promoting healthspan. Based on our findings, the mycelia of \u003cem\u003eT. aurea\u003c/em\u003e should be collected and cultivated by mushroom farmers to ensure their availability as a steady food supplement in our communities.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eACKNOWLEDGMENTS\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors are thankful to the bioprofiling platform, supported by the European Regional Development Fund and the Walloon Region Belgium, as well as to YaBiNaPA and Cameroon for the facilities that enabled the realization of this work. The Leibniz Institute of Plant Biochemistry in Halle (Saale), Germany, is acknowledged for supporting the UHPLC-ESI-MS/MS analyses.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAche R.N: Study design, methodological experiments and writing of the main manuscript. Ngnintedo D: Interpretation of the results and editing of the prepared manuscript. Ambassa P: Interpretation of the results and editing of the prepared manuscript. Mossebo D. C: Identification of the mushroom specimen and revision of the manuscript. Yanick K. M: Interpretation of the results and final editing of the manuscript. Njinga N. S: designed and revised the manuscript. C\u0026eacute;line H: Analytical experiments and editing of the prepared manuscript. Sophie L: Analytical experiments and editing of the prepared manuscript. Sonchieu J: Revising and final editing. Ngameni B: Revising and final editing. Fotso W.G: Analytical experiments and revision of the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability Statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe data that support the findings of this study are available from the corresponding author upon reasonable request at https://getshared.com/UpuXwUU2\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eEyi-Ndong H, Degreef J, De Kesel A (2011) Champignons comestibles des for\u0026ecirc;ts denses d\u0026rsquo;Afrique centrale. Taxonomie et identification. Abc Taxa. 10\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDesjardin DE, Perry BA (2017) The gymnopoid fungi (Basidiomycota, Agaricales) from the Republic of S\u0026atilde;o Tom\u0026eacute; and Pr\u0026iacute;ncipe, West Africa. Mycosphere 8(9):1317\u0026ndash;1391\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZeng Y, Lu Y, Chen Z, Tan J, Bai J, Li P, Wang Z, Du S (2018) Rapid characterization of components in Bolbostemma paniculatum by UPLC/LTQ-Orbitrap MSn analysis and multivariate statistical analysis for herb discrimination. Molecules 23:1155\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEric B, Sandor S, Ce\u0026acute;line B, Sebastian B, Juho R (2018) Liquid-chromatography retention order prediction for metabolite identification. Bioinformatics 34:i875\u0026ndash;i883\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOni JO, Ferdinand A, Akomaye A-A, Markson A, Augustine CE (2020) GC\u0026ndash;MS Analysis of Bioactive Compounds in Some Wild-Edible Mushrooms from Calabar, Southern Nigeria. EJBIO 1(6):1\u0026ndash;8\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePedro ES, do N et al (2023) Characterization of Flavonoids Profiles in Polar Extracts from Croton grewioides Baill. Using Ultra-High Resolution Mass Spectrometry. J Braz Chem Soc 34(7):937\u0026ndash;948\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMarshall WE (1994) Amino Acids, Peptides, and Proteins. In: Goldberg I (ed) Functional Foods. Springer, Boston, MA, pp 242\u0026ndash;260\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCastro GA (1991) Digestion and absorption. In: Johnson LR (ed) Gastrointestinal Physiology, 4th edn. Mosby Year Book, St. Louis, pp 108\u0026ndash;130\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eThalisa Y-A, Luchai B, Pa-Nag Y (2020) Amino acids and antioxidant activities of extracts from wild edible mushrooms from a community forest in the Nasrinual District, Maha Sarakham, Thailand. Food Sci Technol Campinas 40(3):712\u0026ndash;720\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMeng D et al (2018) Simultaneous quantification of free amino acids and 5\u0026prime;-nucleotides in shiitake mushrooms by stable isotope labeling-LC\u0026ndash;MS/MS analysis. Food Chem 268:57\u0026ndash;65\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNisreen H, Touseef HT, Lata M (2019) Amino acids as Medical food and their Therapeutic uses. IJSRR 8(2):579\u0026ndash;585\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDong H et al (2022) Metabolomics Analysis of Morchella sp. From Different Geographical Origins of China Using UPLC-Q-TOF-MS. Front Nutr 9:1\u0026ndash;11\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCho M-H et al (2007) Phenylalanine Biosynthesis in \u003cem\u003eArabidopsis thaliana\u003c/em\u003e: identification and characterization of arogenate dehydratases. JBC 282(42):30827\u0026ndash;30835\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKoshi S, Jun F, Shigenobou O, Shin-Ichi H (1985) α-(Hydroxymethyl) serine from Vicia pseudo-orobus. Phytochemistry 24(4):853\u0026ndash;854\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAyabe T et al (2015) A novel Alaska pollack-derived peptide, which increases glucose uptake in skeletal muscle cells, lowers the blood glucose level in diabetic mice. Food Funct 6(8):2749\u0026ndash;2757\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBonnamour J, M\u0026eacute;tro T-X, Martinez J, Lamaty F (2013) Environmentally benign peptide synthesis using liquid-assisted ball-milling: application to the synthesis of Leu-enkephalin. Green Chem 15(5):1116\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKatsuhisa K et al (1991) Endothelin-binding inhibitors, BE-18257A and Be-18257B. I. Taxonomy, fermentation, isolation and characterization. J Antibiot 44(12):1342\u0026ndash;1347\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFreidinger RM et al (1990) Cyclic hexapeptide oxytocin antagonists. Potency-, selectivity-, and solubility-enhancing modifications. J Med Chem 33(7):1843\u0026ndash;1845\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSingh PD, Johnson JH (1984) Muraceins\u0026ndash;muramyl peptides produced by Nocardia orientalis as angiotensin-converting enzyme inhibitors. II. Isolation and structure determination. J Antibiot 37(4):336\u0026ndash;343\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJohn WB et al (2017) Marine natural products. Nat Prod Rep 34:235\u0026ndash;294\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhu C-Z, Tian W, Li M-Y, Liu Y-X, Zhao G-M (2018) Separation and identification of peptides from dry-cured Jinhua ham. Int J Food Prop. 1\u0026ndash;10\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCastro-G\u0026oacute;mez P, Garcia-Serrano A, Visioli F, Fontecha J (2015) Relevance of dietary glycerophospholipids and sphingolipids to human health. Prostaglandins, Leukotrienes and Essential Fatty Acids (PLEFA). 101:45\u0026ndash;51. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://dx.doi.org/10.1016/j.plefa.2015.07.004\u003c/span\u003e\u003cspan address=\"10.1016/j.plefa.2015.07.004\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKolter T, Sandhoff K (1999) Sphingolipids\u0026ndash;Their Metabolic Pathways and the Pathobiochemistry of Neurodegenerative Diseases. Angew Chem Int Ed 38:153\u0026ndash;1568\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChao D-Y et al (2011) Sphingolipids in the Root Play an Important Role in Regulating the Leaf Ionome in \u003cem\u003eArabidopsis thaliana\u003c/em\u003e. Plant Cell 23(3):1061\u0026ndash;1081\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHayashi A, Matsubara T (1970) On the occurrence of sphinga-4,8-dienine in oyster glycolipid. Biochim Biophys Acta 202(1):228\u0026ndash;230\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBora S et al (2017) Deinococcucins A-D, Aminoglycolipids from Deinococcus sp., a Gut Bacterium of the Carpenter Ant Camponotus japonicas. J Nat Prod 80(11):2910\u0026ndash;2916\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLy D et al (2013) CD1c tetramers detect ex vivo T-cell responses to processed phosphomycoketide antigens. JEM 210(4):729\u0026ndash;741\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEugene A et al (2002) A Novel Family of Atherogenic Oxidized Phospholipids Promotes Macrophage Foam Cell Formation via the Scavenger Receptor CD36 and Is Enriched in Atherosclerotic Lesions. J Biol Chem 277(41):38517\u0026ndash;38523\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAli MS et al (1999) Three p-cymene derivatives from Zataria multiflora. Phytochemistry 52:685\u0026ndash;688\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTakeshi Y, Mayo S, Takanobu A, Takashi K, Reiko T (2017) Trichodermanins C\u0026ndash;E, New Diterpenes with a Fused 6-5-6-6 Ring System Produced by a Marine Sponge-Derived Fungus. Mar Drugs 15(6):169, 2\u0026ndash;7\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMeinke PT, Smith M-HM, Shoop WL (2002) Nodulisporic Acid: Its Chemistry and Biology. Curr Top Med Chem 2:655\u0026ndash;674\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTenji K, Junzo S (1979) Studies on the constituents of Asparagi kadix. I.1) on the structures of Furostanol Oligosides of Asparagus cochinchinensis (Loureio) Merrill. Chem Pharm Bull 27(12):3086\u0026ndash;3094\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEkuadzi E et al (2014) Chemical constituents from Gouania longipetala and Glyphaea brevis Nat. Prod Res 28(15):1210\u0026ndash;1213\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLeong Y-W, Harrison LJ, Bennett GJ, Kadir AA, Connolly (1998) J.D. A dihydrochalcone from Lindera lucida. Phytochemistry 47(5):891\u0026ndash;894\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOlennikov DN, Tankhaeva LM (2010) Phenolic compounds from Rhododendron dauricum from the Baikal region. Chem Nat Compd 46(3):471\u0026ndash;473\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAndersen FA (1994) Cosmetic Ingredient Review, 1101 17th Street NW, Suite 310, Washington, DC 20036, U.S.A. Journal of the American College of Toxicology. 13(5), 330\u0026ndash;343, Raven Press, Ltd. New York\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePhan C-W, David P, Wong K-H, Naidu M, Sabaratnam V (2015) Uridine from Pleurotus giganteus and Its Neurite Outgrowth Stimulatory Effects with Underlying Mechanism. PLoS ONE 10(11):e0143004\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVetter. J (2023) The Mushroom Glucans: Molecules of High Biological and Medicinal Importance. Foods 12:1\u0026ndash;19\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eInouye S, Tsuruoka T, Ito T, Niida T (1968) Structure and synthesis of nojirimycin. Tetrahedron 24(5):2125\u0026ndash;2144\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMami F, Kazuhiro Y, Kenko M, Jun-ichi A, Susumu H (1999) Preparation and Antioxidative Activity of 1,5-Anhydrofructose. J Appl Glycosci 46(4):439\u0026ndash;444\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSingh N, Shreshtha AK, Thakur MS, Patra S (2018) Xanthine scaffold: scope and potential in drug development. Heliyon e 00829\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBunka F et al (2007) Effect of 1-monoglycerides on viscoelastic properties of processed cheese. Int J Food Prop 10(4):819\u0026ndash;828\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBingnan H, Kerry LMP, Alessia L, Vincenzo DM, William HG, Semiplenamides A-G (2003) Fatty Acid Amides from a Papua New Guinea Collection of the Marine Cyanobacterium Lyngbya semiplena. J Nat Prod 66:1364\u0026ndash;1368\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTan B et al (2010) Identification of endogenous acyl amino acids based on a targeted lipidomics approach. JLR 51(1):112\u0026ndash;119\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAndr\u0026aacute;s S et al (2020) Cerebrosides and Steroids from the Edible Mushroom Meripilus giganteus with Antioxidant Potential. Molecules 25:1395, 1\u0026ndash;8\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHakan I, İbrahim T, Aydın ŞB, Handan \u0026Ccedil;Y (2019) Determination of fatty acid profile and mineral contents of Tricholomopsis rutilans collected from Yozgat. Ant J Bot 3(2):64\u0026ndash;68\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eB\u0026aacute;rbara R, Paula GdP, Paula BA, Paula B (2009) Patr\u0026iacute;cia, V. Fatty acid composition of wild edible mushrooms species: A comparative study. Microchem J 93:29\u0026ndash;35\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDilika F, Bremner PD, Meyer JJM (2000) Antibacterial activity of linoleic and oleic acids isolated from Helichrysum pedunculatum: a plant used during circumcision rites. Fitoterapia 71(4):450\u0026ndash;452\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChen C-HE, Ho C-T (1981) Identification of 9-Decenoic Acid in Beer and Yeast. JASBC 39(2):70\u0026ndash;71\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGuelette UFBS (2012) Benning and Susanne Hoffmann-Benning. Identification of lipids and lipid-binding proteins in phloem exudates from \u003cem\u003eArabidopsis thaliana\u003c/em\u003e. JXB 63(10):3603\u0026ndash;3616\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAvato P, Tava A (2022) Rare fatty acids and lipids in plant oilseeds: occurrence and bioactivity. Phytochem Rev 21:401\u0026ndash;428\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWeete JD, Shewmaker F, Gandhi SR (1998) γ-Linolenic Acid in Zygomycetous Fungi: Syzygites megalocarpus. JAOCS 75(10):1367\u0026ndash;1372\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKatsunobu Y, Nobuo S, Toyofumi N, Keiichi N, Keiichi N (1994) Measurements of urinary adipic acid and suberic acid using high-performance liquid chromatography. J Chromatogr B Biomed App 655(2):189\u0026ndash;193\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBang MH et al (2002) 13-hydroxy-9Z,11E,15E-octadecatrienoic acid from the leaves of Cucurbita moschata. Arch Pharm Res 25:438\u0026ndash;440\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNishitoba T, Sato H, Oda K, Sakamura S (1988) Novel Triterpenoids and a Steroid from the Fungus Ganoderma lucidum. Agric Biol Chem 52(1):211\u0026ndash;216\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWu J et al (2011) Osteoclast-forming suppressing compounds, gargalols A, B, and C, from the edible mushroom Grifola gargal. Tetrahedron 67:6576\u0026ndash;6581\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWeng Y et al (2011) Ganodermasides C and D, two new anti-aging ergosterols from spores of the medicinal mushroom Ganoderma lucidum. Biosci Biotechnol Biochem 75:800\u0026ndash;803\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDeveci E, Tel-\u0026Ccedil;ayan G, Duru ME, \u0026Ouml;zt\u0026uuml;rk M (2019) Chemical constituents of Porodaedalea pini mushroom with cytotoxic, antioxidant and anticholinesterase activities. J Food Meas Charact, 1\u0026ndash;10\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJeong H et al (2019) Isolation and characterization of antiplasmodial constituents from the marine sponge Coscinoderma sp. Z f\u0026uuml;r Naturforschung C 74(11\u0026ndash;12):313\u0026ndash;318\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRiaz M et al (2017) Natural products from Cuscuta reflexa Roxb. with antiproliferation activities in HCT116 colorectal cell lines. Nat Prod Res 31(5):583\u0026ndash;587\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCheng CR et al (2010) Cytotoxic triterpenoids from Ganoderma lucidum. Guo Phytochemistry 71(13):1579\u0026ndash;1585\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eXi-Ai L, Yang-Min M, Hong-Chi Z, Rui L (2016) A new helvolic acid derivative from an endophytic Fusarium sp. of Ficus carica. Nat Prod Res 30(21):2407\u0026ndash;2412\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMaxwell A, Seepersaud M, Pingal R 3 P-aminospirosolane steroidal alkaloids from Solanum triste. J Nat Prod 58 (4), 625\u0026ndash;628 (I995).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePirjo M et al (2001) Contents of Vitamins, Mineral Elements, and Some Phenolic Compounds in Cultivated Mushrooms. Agric Food Chem 49:2343\u0026ndash;2348\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRafal RS, Yoko T, Heinrich KS, Hector FD (1985) Synthesis of 1α,25-dihydroxyvitamin D2, its 24 epimer and related isomers, and their binding affinity for the 1,25-dihydroxyvitamin D3 receptor. Bioorg Chem 13:158\u0026ndash;169\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eStrap\u0026aacute;č, Kuruc M1, Baranov\u0026aacute; M (2017) Determination of antioxidant parameters of Pleurotus mushrooms growing on different wood substrates. Folia veterinaria. 61, 4: 53\u0026ndash;58, 2017\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYamashita E, Matsuno T (1990) A new apocarotenoid from the sea hare Aplysia kurodai. Comp Biochem Physiol Part B: Biochem Mol Biology 96B(3):465\u0026ndash;470\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHong W, Qinan W, Chengying W, Zheng J (2015) Simultaneous Determination of 16 Nucleosides and Nucleobases in Euryale ferox Salisb. By Liquid Chromatography Coupled with Electro Spray Ionization Tandem Triple Quadrupole Mass Spectrometry (HPLC-ESI-TQ-MS/MS) in Multiple Reaction Monitoring (MRM) Mode. JCS 53:1386\u0026ndash;1394\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhixin W et al (2019) Rapid Characterization of Chemical Components in Edible Mushroom Sparassis crispa by UPLC-Orbitrap MS Analysis and Potential Inhibitory Effects on Allergic Rhinitis. Molecules 24(16):3014, 1\u0026ndash;17\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBorice TT et al (2017) A New Tetracyclic Triterpenoid from Desmodium uncinatum (Jacq.) DC. (Fabaceae). NPS 23(2):113\u0026ndash;118\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMurphy RC (2014) Tandem Mass Spectrometry of Lipids. Royal Soc Chem. 1\u0026ndash;280\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGebauer SK, Psota TL, Kris-Etherton PM (2007) The diversity of health effects of individual trans fatty acid isomers. Lipids 42(9):787\u0026ndash;799\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShigeo T, Kayoko T, Takuhei Y, Nobuyuki S (2020) Positional determination of the carbon\u0026ndash;carbon double bonds in unsaturated fatty acids mediated by solvent plasmatization using LC\u0026ndash;MS. Sci Rep 10:12988\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDelbeke EIP et al (2016) Petroselinic acid purification and its use for the fermentation of new sophorolipids. AMB Expr 6:28\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"},{"header":"Table","content":"\u003cp\u003eTable 1 is available in the Supplementary Files section\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"None","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Tricholomopsis aurea, phytochemical profiling, UHPLC-ESI-MS/MS, NMR spectroscopy","lastPublishedDoi":"10.21203/rs.3.rs-4585453/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4585453/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cem\u003eTricholomopsis aurea\u003c/em\u003e(Beeli) Desjardin \u0026amp; B.A. Perry is a wild edible fungus that is scarce in Cameroon, but common in other parts of the world. The metabolites of this food matrix have not been extensively analyzed thus far. This work aimed to establish the metabolic profile of \u003cem\u003eTricholomopsis aurea\u003c/em\u003e by UHPLC-ESI-MS/MS and to isolate some of the major compounds using chromatographic separation techniques. Overall, 65 compounds were identified as belonging to 18 classes, including amino acids, carbohydrates, vitamins, aminoglycolipids, phosphoglycolipids, fatty acids, phenolics, nucleosides, steroids, fatty acid amides, acyl glycerides, sphingolipids, alkaloids, terpenes, carotenoids, saponins, aromatic amines, and peptides. Six of the isolated compounds were characterized as oleic acid, elaidic acid petroselinic acid, ergosterol, ergosterol 5α,8α-endoperoxide and adenosine using 1D and 2D NMR spectroscopy. Fatty acids, steroids, amino acids and peptides were the main components of the mushroom. \u003cem\u003eT. aurea\u003c/em\u003ehas been proven to be a valuable source of chemically diverse compounds and to contain indispensable nutrients, such as amino acids, fats, carbohydrates and vitamins, which are essential for promoting the health span.\u003c/p\u003e","manuscriptTitle":"Exploration of the chemical constituents of Tricholomopsis aurea (Beeli) Desjardin using UHPLC-ESI-MS/MS and NMR spectroscopy","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-06-18 14:34:17","doi":"10.21203/rs.3.rs-4585453/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"732ef0bd-6aa5-40f5-b3af-137dfe75ef22","owner":[],"postedDate":"June 18th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":33290125,"name":"Analytical Biochemistry"},{"id":33290126,"name":"Analytical Chemistry"},{"id":33290127,"name":"Organic Chemistry"},{"id":33290128,"name":"Food Chemistry"}],"tags":[],"updatedAt":"2024-06-18T14:34:17+00:00","versionOfRecord":[],"versionCreatedAt":"2024-06-18 14:34:17","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4585453","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4585453","identity":"rs-4585453","version":["v1"]},"buildId":"rHA-KDH7Qsr4HCuvH75dn","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}