# Rapid Identification of Chemical Constituents inHericium erinaceus Based on LC-MS/MS Metabolomics

Authors: Fei Yang; Honglin Wang; Guoquan Feng; Sulan Zhang; Jinmei Wang; Lili Cui
Journal: Journal of Food Quality
 (2021)

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## Abstract

Hericium erinaceus is a precious edible and medicinal fungus with high nutritional value. It has many functions, such as enhancing immunity, antitumor antioxidation, antihyperglycemic, antihyperlipidemic, and protecting gastric mucosa. However, there are few researches about the H. erinaceus compounds. In this paper, ultraperformance liquid chromatography tandem high-resolution mass spectrometry (UPLC-Q-exactive-MS/MS) was used to isolate and identify the compounds in H. erinaceus. 102 compounds were identified in H. erinaceus by comparing with standard databases such as MZVault, MZCloud, and BGI Library (self-built standard Library by BGI Co., Ltd), including flavonoids, terpenoids, phenolic acids, phenylpropanoids, steroids, organic acids, and amino acids.


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## Body

## 1. Introduction

Hericium erinaceus is a precious edible and medicinal fungus, and it is listed as one of the “Four Famous Cuisines” of China, together with bear’s paws, trepang, and shark’s fin [1]; it has been used for a long time in traditional Chinese medicine [2]. Researches showed that the chemical constituents of H. erinaceus include terpenoids, phenolics, steroids, pyranones, fatty acids, and alkaloids; about 80 small molecular compounds were isolated and identified from H. erinaceus [3]. Terpenoids in H. erinaceus were mainly diterpenoids with cyathane skeleton. Terpenoids in H. erinaceus were first isolated and identified by Kawagishi et al. from mycelia of H. erinaceus, named Erinacines A-C [4]. Subsequently, Kawagishi et al. isolated and identified Erinacines D-G, Erinacines J, and Erinacines K from mycelia of H. erinaceus [5–7]; Lee et al. isolated and identified Erinacines H and Erinacines I from mycelia of H. erinaceus [8]; Kenmoku et al. isolated and identified Erinacine P and Erinacine Q from mycelia of H. erinaceus [9, 10]. Most of these diterpenoids compounds were stimulators of nerve growth factor-synthesis. Phenolics were also main constituents in H. erinaceus. 8 phenolics were isolated and identified from fruiting bodies of H. erinaceus by the Kawagishi team between 1990 and 1993, named Hericenone A-H [11–13]. After that, three phenolics were isolated and identified by Arnone et al. [14] with 5′- carbonyl group replaced by 5′- methylene, named Hericenes A-C. Subsequently, two new phenolics were isolated from fruiting bodies of H. erinaceus by Ma et al. [15], named Hericenone I and Hericenone D; they have the same fatty acid chain. A new skeleton phenolic compound was isolated from fruiting bodies of H. erinaceus by Li et al. [16], named Erinacene D. In addition to terpenoids and phenolics compounds, H. erinaceus was rich in steroids, six new (erinarols A-F), and five known ergostane-type sterol fatty acid esters were isolated from the fruiting bodies of H. erinaceus; erinarols A and B significantly activated the transcriptional activity of PPARs, PPARα, and PPARγ [17]. Previous pharmacological studies showed that H. erinaceus had many pharmacological activities, such as regulating immunity [18, 19], neuroprotection [20, 21], antidepressant [22, 23], antioxidant [24], antitumor [25], antihyperglycemic [26], and antihyperlipidemic properties [27], and protecting gastric mucosa [28, 29].High-performance liquid chromatography tandem mass spectrometry (LC-MS/MS) technology has the advantage of rapid identification of compounds. With the maturity of LC-MS/MS technology, it is more and more widely used in the field of food and medicine [30–35]. In this study, ultraperformance liquid chromatography tandem high-resolution mass spectrometry (UPLC-Q-Exactive-MS/MS) combined with standard substance database was used to rapidly identify the chemical components of H. erinaceus.


## 2. Materials and Methods

### 2.1. Instrument

The LC-MS experiment was carried out by ultraperformance liquid chromatography (Waters 2D UPLC, USA) and high-resolution mass spectrometry (Q Exactive, Thermo, USA). The Hypersil GOLD aQ column (100 mm∗2.1 mm, 1.9 μm) was purchased from Thermo Fisher Scientific, USA. Low-temperature high-speed centrifuge (Centrifuge 5430) was purchased from Eppendorf. The vortex finder (QL-901) was purchased from Qilinbeier Instrument Manufacturing Co., Ltd. The pure water meter (Milli-Q) was purchased from Integral Millipore Corporation, USA.


### 2.2. Reagent

d3-leucine, 13C9-phenylalanine, d5-tryptophan, and 13C3-progesterone were used as internal standards. Methanol (A454-4) and acetonitrile (A996-4) were both in mass spectral grade, which were purchased from Thermo Fisher Scientific, USA. Ammonium formate (17843–250 G) was obtained from Honeywell Fluka, USA. Formic acid (50144–50 mL) was obtained from DIMKA, USA.


### 2.3. Materials

Fruiting bodies ofH. erinaceus were obtained from Henan Longfeng Industrial Co., Ltd. The specimens (No. 2020-09-09) were saved in National Research and Development Center of Edible Fungi Processing Technology, Henan University.


### 2.4. Preparation of Sample

According to the methods commonly used by our team, dry fruiting bodies ofH. erinaceus were crushed by the grinding machine. 100 g of H. erinaceus powder was immersed with 50% ethanol (1000 mL) for 2 times at room temperature, each time for 3 days. The filtrate was concentrated to obtain 48.6 g extract. 50 mg extract of H. erinaceus was weighed, and then 800 μL of solution with 70% methanol: internal standard (V/V = 85 : 1) was added to the sample. Two small steel beads were added to the sample and ground in a tissue grinder (50 Hz, 5 min), ultrasound for 10 min at 4°C water, and the sample was placed for 1 h at −20°C. After that, the sample was centrifuged for 15 min at 4°C, 25000 rpm, 500 μL of supernatant was filtered through 96-well filter plate, and the filtrate was used to detect.


### 2.5. Chromatographic Conditions

The LC-MS experiment was carried out on Hypersil GOLD a Q column (100 mm∗2.1 mm, 1.9 μm). The mobile phase was 0.1% formic acid-water (liquid A) and 0.1% formic acid-acetonitrile (liquid B). The elution gradient was as follows: 0–2 min 5% B; 2–22 min 5%–95% B; 22–27 min 95% B; 27.1–30 min 5% B. The flow rate was 0.3 mL/min, the column temperature was 40°C, and the injection volume was 5 μL.


### 2.6. Mass Spectrometry Conditions

The MS and MS2 data were collected by the Q Exactive mass spectrometer. The range of m/z was 150–1500 with the MS resolution 70000, and the AGC was 1 e6, and the maximum injection time was 100 ms. According to the strength of the MS ions, top3 was selected for determining MS2 fragmentation. The MS2 resolution was 35000, AGC was 2 e5, the maximum injection time was 50 ms, and the fragmentation energy was set as 20, 40, and 60 eV. Ion source (ESI) parameters are as follows: sheath gas flow rate was 40, aux gas flow rate was 10, spray voltage (|KV|) of positive ion mode was 3.80, spray voltage (|KV|) of negative ion mode was 3.20, capillary temp was 320°C, and aux gas heater temp was 350°C.


### 2.7. Data Analysis

High-resolution mass spectrometer (Q Exactive, Thermo Fisher Scientific, USA) was used to collect data in positive and negative ion modes, respectively, to improve the chemical constituent coverage. Raw mass spectrum data collected by LC-MS/MS were imported into Compound Discoverer 3.1 (Thermo Fisher Scientific, USA) for data processing. It mainly includes peak extraction, retention time correction within and between groups, additive ion merging, missing value filling, background peak labeling, and metabolite identification. Finally, the molecular weight, retention time, peak area, and identification results of the compound were derived.


## 2.1. Instrument

The LC-MS experiment was carried out by ultraperformance liquid chromatography (Waters 2D UPLC, USA) and high-resolution mass spectrometry (Q Exactive, Thermo, USA). The Hypersil GOLD aQ column (100 mm∗2.1 mm, 1.9 μm) was purchased from Thermo Fisher Scientific, USA. Low-temperature high-speed centrifuge (Centrifuge 5430) was purchased from Eppendorf. The vortex finder (QL-901) was purchased from Qilinbeier Instrument Manufacturing Co., Ltd. The pure water meter (Milli-Q) was purchased from Integral Millipore Corporation, USA.


## 2.2. Reagent

d3-leucine, 13C9-phenylalanine, d5-tryptophan, and 13C3-progesterone were used as internal standards. Methanol (A454-4) and acetonitrile (A996-4) were both in mass spectral grade, which were purchased from Thermo Fisher Scientific, USA. Ammonium formate (17843–250 G) was obtained from Honeywell Fluka, USA. Formic acid (50144–50 mL) was obtained from DIMKA, USA.


## 2.3. Materials

Fruiting bodies ofH. erinaceus were obtained from Henan Longfeng Industrial Co., Ltd. The specimens (No. 2020-09-09) were saved in National Research and Development Center of Edible Fungi Processing Technology, Henan University.


## 2.4. Preparation of Sample

According to the methods commonly used by our team, dry fruiting bodies ofH. erinaceus were crushed by the grinding machine. 100 g of H. erinaceus powder was immersed with 50% ethanol (1000 mL) for 2 times at room temperature, each time for 3 days. The filtrate was concentrated to obtain 48.6 g extract. 50 mg extract of H. erinaceus was weighed, and then 800 μL of solution with 70% methanol: internal standard (V/V = 85 : 1) was added to the sample. Two small steel beads were added to the sample and ground in a tissue grinder (50 Hz, 5 min), ultrasound for 10 min at 4°C water, and the sample was placed for 1 h at −20°C. After that, the sample was centrifuged for 15 min at 4°C, 25000 rpm, 500 μL of supernatant was filtered through 96-well filter plate, and the filtrate was used to detect.


## 2.5. Chromatographic Conditions

The LC-MS experiment was carried out on Hypersil GOLD a Q column (100 mm∗2.1 mm, 1.9 μm). The mobile phase was 0.1% formic acid-water (liquid A) and 0.1% formic acid-acetonitrile (liquid B). The elution gradient was as follows: 0–2 min 5% B; 2–22 min 5%–95% B; 22–27 min 95% B; 27.1–30 min 5% B. The flow rate was 0.3 mL/min, the column temperature was 40°C, and the injection volume was 5 μL.


## 2.6. Mass Spectrometry Conditions

The MS and MS2 data were collected by the Q Exactive mass spectrometer. The range of m/z was 150–1500 with the MS resolution 70000, and the AGC was 1 e6, and the maximum injection time was 100 ms. According to the strength of the MS ions, top3 was selected for determining MS2 fragmentation. The MS2 resolution was 35000, AGC was 2 e5, the maximum injection time was 50 ms, and the fragmentation energy was set as 20, 40, and 60 eV. Ion source (ESI) parameters are as follows: sheath gas flow rate was 40, aux gas flow rate was 10, spray voltage (|KV|) of positive ion mode was 3.80, spray voltage (|KV|) of negative ion mode was 3.20, capillary temp was 320°C, and aux gas heater temp was 350°C.


## 2.7. Data Analysis

High-resolution mass spectrometer (Q Exactive, Thermo Fisher Scientific, USA) was used to collect data in positive and negative ion modes, respectively, to improve the chemical constituent coverage. Raw mass spectrum data collected by LC-MS/MS were imported into Compound Discoverer 3.1 (Thermo Fisher Scientific, USA) for data processing. It mainly includes peak extraction, retention time correction within and between groups, additive ion merging, missing value filling, background peak labeling, and metabolite identification. Finally, the molecular weight, retention time, peak area, and identification results of the compound were derived.


## 3. Results

### 3.1. Total Ion Chromatogram

The total ion chromatogram ofH. erinaceus is shown in Figure 1.Figure 1
Total ion chromatogram ofH. erinaceus in the positive ion mode (pos) and negative ion mode (neg).


### 3.2. Results of Compound Identification

The compounds ofH. erinaceus were analyzed by LC-MS/MS; the identification of structure was based on the retention time, MS data, and MS2 data compared with the standard database. The identified compounds were classified into three grades (Level 1, Level 2, and Level 3) according to the comparison results, there into, Level 2 was confirmed on the basis of MS data, MS2 data, and properties of compounds, and Level 3 was identified on the basis of MS data and MS2 data. The credibility sequence is as follows: Level 1> Level 2> Level 3. Detailed results are shown in Table 1. 102 compounds were preliminarily identified in H. erinaceus with grade of identification as Level 1 and Level 2, including 31 organic acids, 10 nucleotides and analogues, 8 amino acids, 6 carbohydrates and derivatives, 5 flavonoids, 3 unsaturated fatty acids, 3 terpenoids, 3 phenolic acids, 1 phenylpropanoid, 1 steroid, and 32 other compounds. Most flavonoids and organic acids were easily deprotonated and responded in the negative ion mode. Most nucleoside compounds were easily protonated and responded in the positive ion mode. The identification process of some compounds was as follows.Table 1
Identification results of compounds inH. erinaceus.
NumberRetention time (min)Molecular formulaRelative molecular weight (Da)The measured values (m/z)Adduct ionsError (ppm)PeakCompound nameCompound typemzVault best matchmzCloud best matchLevel10.743C17H27N3O17P2607.08113606.07379[M − H]−−0.72143833617.1Udp-n-acetylglucosamineNucleotides and analoguesNA95.8Level 220.826C7H15NO3161.10508162.11214[M + H]+−0.70455483170.3L(-)-CarnitineOthers77.985.7Level 230.869C4H6O5134.02147133.01422[M − H]−−0.4123529567982Dl-Malic acidOrganic acidsNA98.8Level 240.87C4H4O4116.011115.00369[M − H]−0.331996525749Fumaric acidOrganic acids90.798.1Level 150.883C5H7NO3129.04263128.03534[M − H]−0.3111989167254-OxoprolineAmino acidsNA88Level 260.928C5H12O5152.06831151.0611[M − H]−−1.075339098404RibitolCarbohydrates and derivatives90.698.1Level 170.939C6H14O6182.07893181.07164[M − H]−−0.613288369180D-(-)-MannitolCarbohydrates and derivatives9398.4Level 280.94C12H22O12358.1107357.10416[M − H]−−1.1958626917.89Lactobionic acidCarbohydrates and derivatives87.8NALevel 191.026C6H14O6182.07914183.0865[M + H]+0.55272725138.5GalactitolCarbohydrates and derivatives85.693.8Level 1101.061C9H13N3O5243.08565487.17841[2M + H]+0.551565196711CytarabineNucleotides and analogues7985.6Level 2111.066C9H11NO3181.07393182.08116[M + H]+0.21226778893.2L-TyrosineAmino acids86.591.3Level 2121.079C8H14O7222.07377221.06636[M − H]−−0.81939035522.5Ethyl-β-d-glucuronideOthersNA96.2Level 2131.08C9H12N2O6244.0691243.0618[M − H]−−1.77270847192.6UridineNucleotides and analogues87.986.2Level 2141.084C6H8O7192.02693191.01961[M − H]−−0.382611016825Citric acidOrganic acids92.196.7Level 1151.088C5H8O5148.03713147.02985[M − H]−−0.31355626048.3D-α-hydroxyglutaric acidOrganic acidsNA91.2Level 2161.094C6H6O6174.0163173.00929[M − H]−−0.82116011083.2Trans-aconitic acidOrganic acids43.781.9Level 2171.095C7H11NO5189.06359188.05672[M − H]−−0.6791397041.99N-acetyl-dl-glutamic acidAmino acidsNA87.1Level 2181.099C6H10O6178.04763177.04068[M − H]−−0.59177666674.9Δ-Gluconic acidδ-lactoneCarbohydrates and derivativesNA90.6Level 2191.106C9H8O3164.04747182.0813[M + NH4]+0.74246283118.72-Hydroxycinnamic acidPhenylpropanoids77.286.1Level 2201.134C4H6O4118.02667117.01939[M − H]−0.54319647862.4Succinic acidOrganic acids91.991Level 1211.14C10H13N5O4267.09628312.09454[M + FA−H]−−1.76302054711.9AdenosineNucleotides and analogues86.185.2Level 2221.142C7H10O7206.0424205.03548[M − H]−−1.251187152213-Hydroxy-3-(methoxycarbonyl) pentanedioic acidOthers4.887.8Level 2231.143C5H5N5135.05446134.04736[M − H]−−0.25111025841.7AdenineNucleotides and analogues76.281.1Level 1241.15C10H13N5O5283.0917284.09888[M + H]+0.12843230783.7GuanosineNucleotides and analogues84.8NALevel 2251.153C5H5N5O151.04949152.05672[M + H]+0.53291864775.2GuanineNucleotides and analogues8091.6Level 1261.181C10H13N5O3251.10188252.10921[M + H]+0.17597456561.52′-DeoxyadenosineNucleotides and analogues84.593.6Level 2271.187C6H12O6180.06332161.0457[M − H − H2O]−−0.40136214268.5L-SorboseCarbohydrates and derivatives66.289.4Level 1281.47C11H15N5O4281.11214282.11905[M + H]+−0.94592442404.42′-o-MethyladenosineNucleotides and analoguesNA85.2Level 2293.116C11H9NO2187.06332188.07059[M + H]+−0.03184845897.5Indole-3-acrylic acidOthers83.990.6Level 2303.123C11H12N2O2204.08972203.08231[M − H]−−0.7546680672.16L-TryptophanAmino acids9292.9Level 1313.29C11H15N5O3S297.08932298.09692[M + H]+−0.82822526337.55′-s-Methyl-5′-thioadenosineNucleotides and analoguesNA93Level 2324.218C6H6O3126.03166125.02444[M − H]−−0.2723877907.51PyrogallolPhenolic acids85.769.8Level 2334.221C7H6O5170.02147169.01414[M − H]−−0.3371859123.52Gallic acidPhenolic acids85.584.6Level 2344.236C7H12O4160.07347159.06624[M − H]−−0.56770557559.23-Methyladipic acidOrganic acids87.196.9Level 2354.352C12H12N2O2216.08996217.09717[M + H]+0.37179365585.22,3,4,9-Tetrahydro-1h-β-carboline-3-carboxylic acidOrganic acidsNA92.9Level 2364.734C6H12O3132.07867131.07133[M − H]−0.21111408164.42-Hydroxycaproic acidOrganic acidsNA89.2Level 2375.007C8H15NO3173.10533174.11256[M + H]+0.7749085835.4N-Acetyl-l-leucineAmino acids84.288.9Level 2385.273C8H15NO3173.10522172.09795[M − H]−0.17228758811.12-(Acetylamino)hexanoic acidOrganic acidsNA94.9Level 2395.495C9H10O3166.06292165.05562[M − H]−−0.4771202767.79L-(-)-3-Phenyllactic acidOthers79.892.4Level 2405.783C11H18N2O2210.13664211.14392[M + H]+−0.87148811351.4Cyclo(leucylprolyl)Amino acidsNA86.6Level 2415.832C8H14O4174.08912173.08185[M − H]−−0.50361132054.6Suberic acidOrganic acids89.292.8Level 2425.85C11H13NO3207.08931206.08209[M − H]−−1.14138547722N-Acetyl-L-phenylalanineAmino acids89.488.5Level 2436.629C9H14O4186.08906185.0818[M − H]−−0.7868946334.821-(Carboxymethyl)cyclohexanecarboxylic acidOrganic acidsNA89.1Level 2447.097C12H10N4O2242.08046243.08763[M + H]+0.35161472292.4LumichromeOthers89.639.1Level 2457.105C9H16O4188.10468187.09747[M − H]-−0.973465873760Azelaic acidOrganic acids93.296.7Level 1467.755C12H14O2190.09933191.1066[M + H]+−0.2720998200.24LigustilideOthers87.5NALevel 2477.998C12H16O3208.10987209.11719[M + H]+−0.388947399.972Β-AsaroneOthers8285.7Level 2488.296C10H18O4202.12036201.1131[M − H]−−0.74435644695.33-Tert-butyladipic acidOrganic acidsNA86.6Level 2498.504C15H10O6286.04722285.04053[M − H]−−1.8017137849.28LuteolinFlavonoids94.386.7Level 2509.458C15H10O5270.05241269.04538[M − H]−−1.529772253.507ApigeninFlavonoids96.286.7Level 1519.756C16H12O6300.06343301.07077[M + H]+0.1517886631.77DiosmetinFlavonoids7888.5Level 2529.757C16H12O6300.06297299.05612[M − H]−−1.3910020073.46HispidulinFlavonoids9379.8Level 1539.785C15H20O3248.14096249.14859[M + H]+−1.14765330608.2(3ar,8r,8ar,9ar)-8-Hydroxy-8a-methyl-3,5-bis(methylene)decahydronaphtho[2,3-b]furan-2(3 h)-one （Others81.789.9Level 25410.161C12H18O2194.13062195.13806[M + H]+−0.32139186228.7SedanolideOthersNA88.1Level 25510.163C18H34O5330.24027329.23309[M − H]−−1.084975697767(15z)-9,12,13-Trihydroxy-15-octadecenoic acidOrganic acidsNA90.2Level 25610.459C12H22O4230.15155229.1442[M − H]−−1.1318453143.37Dodecanedioic acidOrganic acidsNA90.1Level 25711.508C20H30O4334.21425335.22144[M + H]+−0.48288459427.22-Hydroxy-4,5′,8a′-trimethyl-1′-oxo-4-vinyloctahydro-1′h-spiro[cyclopentane-1,2′-naphthalene]-5′-carboxylic acidOrganic acids68.185.2Level 25811.573C10H20O3188.14116187.13385[M − H]−−0.4661126120.0910-Hydroxydecanoic acidOrganic acidsNA85.5Level 25911.73C22H26O6386.17282387.17999[M + H]+−0.30162320645.1Bis(methylbenzylidene) sorbitolOthers15.491.5Level 26011.791C16H12O5284.06798283.06070[M − H]−−1.7519826019.94AcacetinFlavonoids94.683.2Level 16111.863C15H18O3246.12566247.13295[M + H]+0.2743672909.94ArglabinTerpenoids6586.7Level 26212.006C15H20O3248.14096249.14842[M + H]+−1.1416706391.7Atractylenolide IIITerpenoids92.992.4Level 16312.437C14H26O4258.18284257.17584[M − H]−−1.033550240.872Tetradecanedioic acidOrganic acidsNA88.7Level 26412.818C20H32O3320.23487321.24219[M + H]+−0.87240249419.63-Methyl-5-[(1 s,2 r,4ar)-1,2,4a,5-tetramethyl-7-oxo-1,2,3,4,4a,7,8,8a-octahydro-1-naphthalenyl] pentanoic acidOrganic acids78.190.5Level 26512.858C20H32O2304.23998305.24768[M + H]+−0.836899745.63Arachidonic acidOrganic acids61.989.3Level 26612.934C24H30O6414.20434415.21118[M + H]+0.241507553854Bis(4-ethylbenzylidene) sorbitolOthersNA94.9Level 26712.976C9H10O3166.06299167.07022[M + H]+−0.0292932147.44Ethyl parabenOthers8487.1Level 26812.977C11H14O3194.09427195.1017[M + H]+−0.12215420117.34-Ethoxy ethylbenzoateOthers6990.6Level 26913.247C18H34O4314.24521313.23795[M − H]−−1.601064033527(±)9,10-Dihydroxy-12z-octadecenoic acidOrganic acidsNA94.9Level 27013.379C12H26O4S266.15457265.14728[M − H]−−2.31919909720.2Dodecyl sulfateOthersNA94.3Level 27113.427C20H28O2300.20881301.21606[M + H]+−0.39260138250.1IsotretinoinOrganic acidsNA85.1Level 27213.908C18H35NO2297.26671298.27426[M + H]+−0.2257608549.562-Aminooctadec-4-yne-1,3-diolOthersNA92.4Level 27314.152C18H28O3292.20372293.21088[M + H]+−0.424772792399 s,13r-12-Oxophytodienoic acidOrganic acidsNA92.4Level 27414.574C12H16O2192.11508193.12238[M + H]+0.28134018825.8Senkyunolide AOthers85.9NALevel 27514.763C21H44NO7P453.28511452.27756[M − H]-−0.95115696629.3Glycerophospho-n-palmitoyl ethanolamineOthersNA93.5Level 27614.975C18H30O2278.22423279.23184[M + H]+−1.243057598727Α-Eleostearic acidUnsaturated fatty acids9196.7Level 27715.058C18H30O3S326.19129325.18396[M − H]−−0.84934129316.54-Dodecylbenzenesulfonic acidOthersNA92.4Level 27815.149C20H32O3320.23482321.24207[M + H]+−1.02654199927.6(3 s)-3-Methyl-5-[(1 s,8ar)-2,5,5,8a-tetramethyl-4-oxo-1,4,4a,5,6,7,8,8a-octahydro-1-naphthalenyl] pentanoic acidOrganic acids64.389.1Level 27915.292C18H30O2278.22423279.23172[M + H]+−1.261402325459Pinolenic acidOrganic acids90.295.8Level 28015.332C14H28O3244.20347243.19621[M − H]−−1.5222659032.03(r)-3-Hydroxy myristic acidOrganic acidsNA88.9Level 28115.832C18H30O3294.2195295.22685[M + H]+0.00358460636019-Oxo-10(e),12(e)-octadecadienoic acidOrganic acidsNA97.6Level 28216.357C30H46O4470.33934471.34756[M + H]+−0.5730986480.7418-β-glycyrrhetinic acidTerpenoids87.390.6Level 28317.175C20H32O3320.23482321.24222[M + H]+−1.02129134534.2Ginkgolic acidPhenolic acids6686.1Level 28417.422C19H32O2292.23998293.24725[M + H]+−0.8784449956.049(z),11(e),13(e)-Octadecatrienoic acid methyl esterOthersNA94Level 28517.727C20H34O2306.25582307.26297[M + H]+−0.2159379266.82Linolenic acid ethyl esterOthers91.196.2Level 28617.935C21H38O4354.27685355.28397[M + H]+−0.44469393052.81-Linoleoyl glycerolOthersNA94.4Level 28718.154C16H32O3272.23457271.22726[M − H]-−2.12232231687416-Hydroxyhexadecanoic acidOrganic acidsNA93.3Level 28818.184C22H36O3348.26628349.2735[M + H]+−0.4779521502.693-Methyl-5-(5,5,8a-trimethyl-2-methylene-7-oxodecahydro-1-naphthalenyl) pentyl acetateOthersNA85Level 28918.301C20H30O3318.21935319.22678[M + H]+−0.4687070678.6411-α-hydroxy-17-methyltestosteroneSteroids74.985.5Level 29018.528C16H30O2254.22444255.23161[M + H]+−0.5493625271.65Palmitoleic acidOrganic acids79.897.2Level 29118.532C16H30O2254.22421253.21733[M − H]−−1.4575050753.73Δ2-Trans-hexadecenoic acidUnsaturated fatty acidsNA86.4Level 29218.608C18H37NO2299.28227300.28955[M + H]+−0.5260691604.46Palmitoyl ethanolamideOthersNA95.2Level 29318.618C20H39NO2325.29787326.30499[M + H]+−0.64205900632.2Oleoyl ethanolamideOthersNA94.2Level 29418.94C20H32O3320.23475319.228[M − H]−−1.2211930213.55-[(3z)-5-Hydroxy-3-methyl-3-penten-1-yl]-1,4a-dimethyl-6-methylenedecahydro-1-naphthalenecarboxylic acidOrganic acids80.586.2Level 29518.948C18H32O2280.23944279.23248[M − H]−−2.8119540670539(z),11(e)-Conjugated linoleic acidUnsaturated fatty acids85.887.3Level 29619.741C18H35NO281.27169282.27902[M + H]+−0.61800775254.3OleamideOthersNA97.3Level 29720.138C16H33NO255.25604256.26334[M + H]+−0.68144475650.8HexadecanamideOthersNA90.2Level 29820.821C20H34O2306.25582307.26343[M + H]+−0.216347388.901Linolenic acid ethyl esterOthers90.296.2Level 19920.977C21H42O4358.30821359.31543[M + H]+−0.27268288479.91-StearoylglycerolOthersNA89.1Level 210021.524C18H34O2282.25578283.26306[M + H]+−0.3421237622.49Ethyl palmitoleateOthers64.893.9Level 210122.225C18H37NO283.28734284.29741[M + H]+−0.61133000171.1StearamideOthersNA90.6Level 210222.746C20H38O2310.28707311.29428[M + H]+−0.3722068099.07Ethyl oleateOthers57.896.3Level 2Compound 49, C15H10O6, was easily deprotonated in the negative ion mode to produce ion m/z 285 [M − H]− and then RDA cracking to get 2 fragment ions m/z 133 [C8H6O2 – H]− and m/z 151 [C7H4O4 – H]−. Compound 49 was identified as luteolin compared to the database and references [36]. The MS2 spectrum is shown in Figure 2.Figure 2
The MS2 spectrum of luteolin.Compound 60 produced ionm/z 283 [M − H]−, then lost the methyl group, and got ion 268 [M − H − CH3]−, combined with retention time, MS data, and MS2 data, compound 60 was identified as acacetin by comparin with the database and references [37]. The MS2 spectrum is shown in Figure 3.Figure 3
The MS2 spectrum of acacetin.Compound 62 was easily protonated in the positive ion mode to produce ionm/z 249 [M + H]+, then lost the hydroxyl group, and got ion m/z 231 [M + H – H2O]+, Compound 62 was identified as atractylenolide III by comparing with the database and references [38]. The MS2 spectrum is shown in Figure 4.Figure 4
The MS2 spectrum of acacetin atractylenolide III.


## 3.1. Total Ion Chromatogram

The total ion chromatogram ofH. erinaceus is shown in Figure 1.Figure 1
Total ion chromatogram ofH. erinaceus in the positive ion mode (pos) and negative ion mode (neg).


## 3.2. Results of Compound Identification

The compounds ofH. erinaceus were analyzed by LC-MS/MS; the identification of structure was based on the retention time, MS data, and MS2 data compared with the standard database. The identified compounds were classified into three grades (Level 1, Level 2, and Level 3) according to the comparison results, there into, Level 2 was confirmed on the basis of MS data, MS2 data, and properties of compounds, and Level 3 was identified on the basis of MS data and MS2 data. The credibility sequence is as follows: Level 1> Level 2> Level 3. Detailed results are shown in Table 1. 102 compounds were preliminarily identified in H. erinaceus with grade of identification as Level 1 and Level 2, including 31 organic acids, 10 nucleotides and analogues, 8 amino acids, 6 carbohydrates and derivatives, 5 flavonoids, 3 unsaturated fatty acids, 3 terpenoids, 3 phenolic acids, 1 phenylpropanoid, 1 steroid, and 32 other compounds. Most flavonoids and organic acids were easily deprotonated and responded in the negative ion mode. Most nucleoside compounds were easily protonated and responded in the positive ion mode. The identification process of some compounds was as follows.Table 1
Identification results of compounds inH. erinaceus.
NumberRetention time (min)Molecular formulaRelative molecular weight (Da)The measured values (m/z)Adduct ionsError (ppm)PeakCompound nameCompound typemzVault best matchmzCloud best matchLevel10.743C17H27N3O17P2607.08113606.07379[M − H]−−0.72143833617.1Udp-n-acetylglucosamineNucleotides and analoguesNA95.8Level 220.826C7H15NO3161.10508162.11214[M + H]+−0.70455483170.3L(-)-CarnitineOthers77.985.7Level 230.869C4H6O5134.02147133.01422[M − H]−−0.4123529567982Dl-Malic acidOrganic acidsNA98.8Level 240.87C4H4O4116.011115.00369[M − H]−0.331996525749Fumaric acidOrganic acids90.798.1Level 150.883C5H7NO3129.04263128.03534[M − H]−0.3111989167254-OxoprolineAmino acidsNA88Level 260.928C5H12O5152.06831151.0611[M − H]−−1.075339098404RibitolCarbohydrates and derivatives90.698.1Level 170.939C6H14O6182.07893181.07164[M − H]−−0.613288369180D-(-)-MannitolCarbohydrates and derivatives9398.4Level 280.94C12H22O12358.1107357.10416[M − H]−−1.1958626917.89Lactobionic acidCarbohydrates and derivatives87.8NALevel 191.026C6H14O6182.07914183.0865[M + H]+0.55272725138.5GalactitolCarbohydrates and derivatives85.693.8Level 1101.061C9H13N3O5243.08565487.17841[2M + H]+0.551565196711CytarabineNucleotides and analogues7985.6Level 2111.066C9H11NO3181.07393182.08116[M + H]+0.21226778893.2L-TyrosineAmino acids86.591.3Level 2121.079C8H14O7222.07377221.06636[M − H]−−0.81939035522.5Ethyl-β-d-glucuronideOthersNA96.2Level 2131.08C9H12N2O6244.0691243.0618[M − H]−−1.77270847192.6UridineNucleotides and analogues87.986.2Level 2141.084C6H8O7192.02693191.01961[M − H]−−0.382611016825Citric acidOrganic acids92.196.7Level 1151.088C5H8O5148.03713147.02985[M − H]−−0.31355626048.3D-α-hydroxyglutaric acidOrganic acidsNA91.2Level 2161.094C6H6O6174.0163173.00929[M − H]−−0.82116011083.2Trans-aconitic acidOrganic acids43.781.9Level 2171.095C7H11NO5189.06359188.05672[M − H]−−0.6791397041.99N-acetyl-dl-glutamic acidAmino acidsNA87.1Level 2181.099C6H10O6178.04763177.04068[M − H]−−0.59177666674.9Δ-Gluconic acidδ-lactoneCarbohydrates and derivativesNA90.6Level 2191.106C9H8O3164.04747182.0813[M + NH4]+0.74246283118.72-Hydroxycinnamic acidPhenylpropanoids77.286.1Level 2201.134C4H6O4118.02667117.01939[M − H]−0.54319647862.4Succinic acidOrganic acids91.991Level 1211.14C10H13N5O4267.09628312.09454[M + FA−H]−−1.76302054711.9AdenosineNucleotides and analogues86.185.2Level 2221.142C7H10O7206.0424205.03548[M − H]−−1.251187152213-Hydroxy-3-(methoxycarbonyl) pentanedioic acidOthers4.887.8Level 2231.143C5H5N5135.05446134.04736[M − H]−−0.25111025841.7AdenineNucleotides and analogues76.281.1Level 1241.15C10H13N5O5283.0917284.09888[M + H]+0.12843230783.7GuanosineNucleotides and analogues84.8NALevel 2251.153C5H5N5O151.04949152.05672[M + H]+0.53291864775.2GuanineNucleotides and analogues8091.6Level 1261.181C10H13N5O3251.10188252.10921[M + H]+0.17597456561.52′-DeoxyadenosineNucleotides and analogues84.593.6Level 2271.187C6H12O6180.06332161.0457[M − H − H2O]−−0.40136214268.5L-SorboseCarbohydrates and derivatives66.289.4Level 1281.47C11H15N5O4281.11214282.11905[M + H]+−0.94592442404.42′-o-MethyladenosineNucleotides and analoguesNA85.2Level 2293.116C11H9NO2187.06332188.07059[M + H]+−0.03184845897.5Indole-3-acrylic acidOthers83.990.6Level 2303.123C11H12N2O2204.08972203.08231[M − H]−−0.7546680672.16L-TryptophanAmino acids9292.9Level 1313.29C11H15N5O3S297.08932298.09692[M + H]+−0.82822526337.55′-s-Methyl-5′-thioadenosineNucleotides and analoguesNA93Level 2324.218C6H6O3126.03166125.02444[M − H]−−0.2723877907.51PyrogallolPhenolic acids85.769.8Level 2334.221C7H6O5170.02147169.01414[M − H]−−0.3371859123.52Gallic acidPhenolic acids85.584.6Level 2344.236C7H12O4160.07347159.06624[M − H]−−0.56770557559.23-Methyladipic acidOrganic acids87.196.9Level 2354.352C12H12N2O2216.08996217.09717[M + H]+0.37179365585.22,3,4,9-Tetrahydro-1h-β-carboline-3-carboxylic acidOrganic acidsNA92.9Level 2364.734C6H12O3132.07867131.07133[M − H]−0.21111408164.42-Hydroxycaproic acidOrganic acidsNA89.2Level 2375.007C8H15NO3173.10533174.11256[M + H]+0.7749085835.4N-Acetyl-l-leucineAmino acids84.288.9Level 2385.273C8H15NO3173.10522172.09795[M − H]−0.17228758811.12-(Acetylamino)hexanoic acidOrganic acidsNA94.9Level 2395.495C9H10O3166.06292165.05562[M − H]−−0.4771202767.79L-(-)-3-Phenyllactic acidOthers79.892.4Level 2405.783C11H18N2O2210.13664211.14392[M + H]+−0.87148811351.4Cyclo(leucylprolyl)Amino acidsNA86.6Level 2415.832C8H14O4174.08912173.08185[M − H]−−0.50361132054.6Suberic acidOrganic acids89.292.8Level 2425.85C11H13NO3207.08931206.08209[M − H]−−1.14138547722N-Acetyl-L-phenylalanineAmino acids89.488.5Level 2436.629C9H14O4186.08906185.0818[M − H]−−0.7868946334.821-(Carboxymethyl)cyclohexanecarboxylic acidOrganic acidsNA89.1Level 2447.097C12H10N4O2242.08046243.08763[M + H]+0.35161472292.4LumichromeOthers89.639.1Level 2457.105C9H16O4188.10468187.09747[M − H]-−0.973465873760Azelaic acidOrganic acids93.296.7Level 1467.755C12H14O2190.09933191.1066[M + H]+−0.2720998200.24LigustilideOthers87.5NALevel 2477.998C12H16O3208.10987209.11719[M + H]+−0.388947399.972Β-AsaroneOthers8285.7Level 2488.296C10H18O4202.12036201.1131[M − H]−−0.74435644695.33-Tert-butyladipic acidOrganic acidsNA86.6Level 2498.504C15H10O6286.04722285.04053[M − H]−−1.8017137849.28LuteolinFlavonoids94.386.7Level 2509.458C15H10O5270.05241269.04538[M − H]−−1.529772253.507ApigeninFlavonoids96.286.7Level 1519.756C16H12O6300.06343301.07077[M + H]+0.1517886631.77DiosmetinFlavonoids7888.5Level 2529.757C16H12O6300.06297299.05612[M − H]−−1.3910020073.46HispidulinFlavonoids9379.8Level 1539.785C15H20O3248.14096249.14859[M + H]+−1.14765330608.2(3ar,8r,8ar,9ar)-8-Hydroxy-8a-methyl-3,5-bis(methylene)decahydronaphtho[2,3-b]furan-2(3 h)-one （Others81.789.9Level 25410.161C12H18O2194.13062195.13806[M + H]+−0.32139186228.7SedanolideOthersNA88.1Level 25510.163C18H34O5330.24027329.23309[M − H]−−1.084975697767(15z)-9,12,13-Trihydroxy-15-octadecenoic acidOrganic acidsNA90.2Level 25610.459C12H22O4230.15155229.1442[M − H]−−1.1318453143.37Dodecanedioic acidOrganic acidsNA90.1Level 25711.508C20H30O4334.21425335.22144[M + H]+−0.48288459427.22-Hydroxy-4,5′,8a′-trimethyl-1′-oxo-4-vinyloctahydro-1′h-spiro[cyclopentane-1,2′-naphthalene]-5′-carboxylic acidOrganic acids68.185.2Level 25811.573C10H20O3188.14116187.13385[M − H]−−0.4661126120.0910-Hydroxydecanoic acidOrganic acidsNA85.5Level 25911.73C22H26O6386.17282387.17999[M + H]+−0.30162320645.1Bis(methylbenzylidene) sorbitolOthers15.491.5Level 26011.791C16H12O5284.06798283.06070[M − H]−−1.7519826019.94AcacetinFlavonoids94.683.2Level 16111.863C15H18O3246.12566247.13295[M + H]+0.2743672909.94ArglabinTerpenoids6586.7Level 26212.006C15H20O3248.14096249.14842[M + H]+−1.1416706391.7Atractylenolide IIITerpenoids92.992.4Level 16312.437C14H26O4258.18284257.17584[M − H]−−1.033550240.872Tetradecanedioic acidOrganic acidsNA88.7Level 26412.818C20H32O3320.23487321.24219[M + H]+−0.87240249419.63-Methyl-5-[(1 s,2 r,4ar)-1,2,4a,5-tetramethyl-7-oxo-1,2,3,4,4a,7,8,8a-octahydro-1-naphthalenyl] pentanoic acidOrganic acids78.190.5Level 26512.858C20H32O2304.23998305.24768[M + H]+−0.836899745.63Arachidonic acidOrganic acids61.989.3Level 26612.934C24H30O6414.20434415.21118[M + H]+0.241507553854Bis(4-ethylbenzylidene) sorbitolOthersNA94.9Level 26712.976C9H10O3166.06299167.07022[M + H]+−0.0292932147.44Ethyl parabenOthers8487.1Level 26812.977C11H14O3194.09427195.1017[M + H]+−0.12215420117.34-Ethoxy ethylbenzoateOthers6990.6Level 26913.247C18H34O4314.24521313.23795[M − H]−−1.601064033527(±)9,10-Dihydroxy-12z-octadecenoic acidOrganic acidsNA94.9Level 27013.379C12H26O4S266.15457265.14728[M − H]−−2.31919909720.2Dodecyl sulfateOthersNA94.3Level 27113.427C20H28O2300.20881301.21606[M + H]+−0.39260138250.1IsotretinoinOrganic acidsNA85.1Level 27213.908C18H35NO2297.26671298.27426[M + H]+−0.2257608549.562-Aminooctadec-4-yne-1,3-diolOthersNA92.4Level 27314.152C18H28O3292.20372293.21088[M + H]+−0.424772792399 s,13r-12-Oxophytodienoic acidOrganic acidsNA92.4Level 27414.574C12H16O2192.11508193.12238[M + H]+0.28134018825.8Senkyunolide AOthers85.9NALevel 27514.763C21H44NO7P453.28511452.27756[M − H]-−0.95115696629.3Glycerophospho-n-palmitoyl ethanolamineOthersNA93.5Level 27614.975C18H30O2278.22423279.23184[M + H]+−1.243057598727Α-Eleostearic acidUnsaturated fatty acids9196.7Level 27715.058C18H30O3S326.19129325.18396[M − H]−−0.84934129316.54-Dodecylbenzenesulfonic acidOthersNA92.4Level 27815.149C20H32O3320.23482321.24207[M + H]+−1.02654199927.6(3 s)-3-Methyl-5-[(1 s,8ar)-2,5,5,8a-tetramethyl-4-oxo-1,4,4a,5,6,7,8,8a-octahydro-1-naphthalenyl] pentanoic acidOrganic acids64.389.1Level 27915.292C18H30O2278.22423279.23172[M + H]+−1.261402325459Pinolenic acidOrganic acids90.295.8Level 28015.332C14H28O3244.20347243.19621[M − H]−−1.5222659032.03(r)-3-Hydroxy myristic acidOrganic acidsNA88.9Level 28115.832C18H30O3294.2195295.22685[M + H]+0.00358460636019-Oxo-10(e),12(e)-octadecadienoic acidOrganic acidsNA97.6Level 28216.357C30H46O4470.33934471.34756[M + H]+−0.5730986480.7418-β-glycyrrhetinic acidTerpenoids87.390.6Level 28317.175C20H32O3320.23482321.24222[M + H]+−1.02129134534.2Ginkgolic acidPhenolic acids6686.1Level 28417.422C19H32O2292.23998293.24725[M + H]+−0.8784449956.049(z),11(e),13(e)-Octadecatrienoic acid methyl esterOthersNA94Level 28517.727C20H34O2306.25582307.26297[M + H]+−0.2159379266.82Linolenic acid ethyl esterOthers91.196.2Level 28617.935C21H38O4354.27685355.28397[M + H]+−0.44469393052.81-Linoleoyl glycerolOthersNA94.4Level 28718.154C16H32O3272.23457271.22726[M − H]-−2.12232231687416-Hydroxyhexadecanoic acidOrganic acidsNA93.3Level 28818.184C22H36O3348.26628349.2735[M + H]+−0.4779521502.693-Methyl-5-(5,5,8a-trimethyl-2-methylene-7-oxodecahydro-1-naphthalenyl) pentyl acetateOthersNA85Level 28918.301C20H30O3318.21935319.22678[M + H]+−0.4687070678.6411-α-hydroxy-17-methyltestosteroneSteroids74.985.5Level 29018.528C16H30O2254.22444255.23161[M + H]+−0.5493625271.65Palmitoleic acidOrganic acids79.897.2Level 29118.532C16H30O2254.22421253.21733[M − H]−−1.4575050753.73Δ2-Trans-hexadecenoic acidUnsaturated fatty acidsNA86.4Level 29218.608C18H37NO2299.28227300.28955[M + H]+−0.5260691604.46Palmitoyl ethanolamideOthersNA95.2Level 29318.618C20H39NO2325.29787326.30499[M + H]+−0.64205900632.2Oleoyl ethanolamideOthersNA94.2Level 29418.94C20H32O3320.23475319.228[M − H]−−1.2211930213.55-[(3z)-5-Hydroxy-3-methyl-3-penten-1-yl]-1,4a-dimethyl-6-methylenedecahydro-1-naphthalenecarboxylic acidOrganic acids80.586.2Level 29518.948C18H32O2280.23944279.23248[M − H]−−2.8119540670539(z),11(e)-Conjugated linoleic acidUnsaturated fatty acids85.887.3Level 29619.741C18H35NO281.27169282.27902[M + H]+−0.61800775254.3OleamideOthersNA97.3Level 29720.138C16H33NO255.25604256.26334[M + H]+−0.68144475650.8HexadecanamideOthersNA90.2Level 29820.821C20H34O2306.25582307.26343[M + H]+−0.216347388.901Linolenic acid ethyl esterOthers90.296.2Level 19920.977C21H42O4358.30821359.31543[M + H]+−0.27268288479.91-StearoylglycerolOthersNA89.1Level 210021.524C18H34O2282.25578283.26306[M + H]+−0.3421237622.49Ethyl palmitoleateOthers64.893.9Level 210122.225C18H37NO283.28734284.29741[M + H]+−0.61133000171.1StearamideOthersNA90.6Level 210222.746C20H38O2310.28707311.29428[M + H]+−0.3722068099.07Ethyl oleateOthers57.896.3Level 2Compound 49, C15H10O6, was easily deprotonated in the negative ion mode to produce ion m/z 285 [M − H]− and then RDA cracking to get 2 fragment ions m/z 133 [C8H6O2 – H]− and m/z 151 [C7H4O4 – H]−. Compound 49 was identified as luteolin compared to the database and references [36]. The MS2 spectrum is shown in Figure 2.Figure 2
The MS2 spectrum of luteolin.Compound 60 produced ionm/z 283 [M − H]−, then lost the methyl group, and got ion 268 [M − H − CH3]−, combined with retention time, MS data, and MS2 data, compound 60 was identified as acacetin by comparin with the database and references [37]. The MS2 spectrum is shown in Figure 3.Figure 3
The MS2 spectrum of acacetin.Compound 62 was easily protonated in the positive ion mode to produce ionm/z 249 [M + H]+, then lost the hydroxyl group, and got ion m/z 231 [M + H – H2O]+, Compound 62 was identified as atractylenolide III by comparing with the database and references [38]. The MS2 spectrum is shown in Figure 4.Figure 4
The MS2 spectrum of acacetin atractylenolide III.


## 4. Discussion and Conclusion

In this study, 102 compounds were preliminarily identified inH. erinaceus, including organic acids, nucleotides and analogues, amino acids, carbohydrates and derivatives, flavonoids, unsaturated fatty acids, terpenoids, phenolic acids, phenylpropanoid, and steroid. It revealed that the compounds in H. erinaceus were diverse. Previous studies showed that the chemical constituents of H. erinaceus include terpenoids, phenolics, steroids, pyranones, fatty acids, and alkaloids; thereinto, reports about terpenoids and phenolics in H. erinaceus were more [3–16]. Terpenoids and phenolics were less in this study; the reasons may be different material parts, different origins, different varieties, different extraction methods, and so on. In addition, nucleotides were also the main constituents in H. erinaceus. Yan et al. [39] studied the content of five nucleosides in H. erinaceus from different habitats by high-performance liquid chromatography; the results showed that different origins of H. erinaceus had the same nucleoside components, such as cytosine, inosine, and adenosine, and the content of inosine and adenosine in H. erinaceus was higher. In this study, 10 nucleotides and analogues were identified, including adenosine, adenine, guanosine, guanine, and uridine.Most of the compounds inH. erinaceus had a good biological activity; researches about polysaccharide were more, which had a wide variety of pharmacological functions such as antimicrobial, antidiabetic, and antihypertension ones [1]. Small molecular compounds isolated from H. erinaceus also had many biological activities. Diterpenoids in H. erinaceus could promote the synthesis of nerve growth factors, such as Erinacines A, Erinacines B, Erinacines C, Erinacines D, Erinacines E, Erinacines F, Erinacines G, Erinacines H, and Erinacines I [4–6]. Phenolics compounds in H. erinaceus could also promote the synthesis of nerve growth factors, such as Hericenone C, Hericenone D, Hericenone E, and Hericenone H [12, 13]. Hericenones A and B showed cytotoxicity against HeLa cells [11]. In addition, Hericenone B was found to be a potential antiplatelet aggregation agent [40]. Steroids in H. erinaceus could activate the transcriptional activity of PPARs, PPARα, and PPARγ, such as erinarols A and erinarols B. In addition, nucleosides and flavonoids may be the main active components of H. erinaceus. Nucleoside components have many biological activities, such as antitumor, antivirus, and gene therapy. Studies have shown that adenosine, inosine, and guanosine have many pharmacological effects such as regulating immunity, neuroprotection, and treatment of cardiovascular diseases [41]. Flavonoids also have many pharmacological effects, such as neuroprotection, antimyocardial ischemia, hypotension, improved learning and memory, antigastric ulcer, protection of reproductive tissue, anti-inflammatory, and antitumor [42].In a word,H. erinaceus has high nutritional value and medicinal value. At present, it has been developed into a variety of functional foods, including health wine, health drinks, healthy yogurt, tea, cans, and health vinegar [2], which was of great significance to study the chemical composition of H. erinaceus. In this study, 102 compounds were preliminarily identified to provide reference for the follow-up study of H. erinaceus.



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*Source: 5560626-2021-03-29.xml*