A method for detecting chemical components of Tung oil tree

Abstract

The present invention provides a detection method and a separation method for the chemical components of Tung oil. The present invention uses ultra-high performance liquid chromatography combined with a specific mobile phase and elution program to separate 52 compounds from Tung oil leaves. It further uses an analysis method of ultra-high performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry to quickly identify 50 compounds from Tung oil leaves. The results show that Tung oil is rich in flavonoids and phenolic acid components, and also contains phenylpropanoid components, as well as iridoid glycosides and other components. These chemical substances have extremely strong biological activity, indicating that Tung oil has good medicinal potential, and its research and development and utilization can be increased in the future. The present invention enriches the chemical components of Tung oil, and lays the foundation for its pharmacological activity and development and utilization research. At the same time, it provides a new preparation method for 50 compounds, enriching the source of the compounds.

Description

Technical Field

[0001] The invention relates to the technical field of chemical component analysis, and in particular to a method for detecting chemical components of Tung oil tree. Background Art

[0002] Pisonia grandis L., also known as white frost-avoiding tung, maple tung, or thornless vine, is a member of the genus Adenocarpus in the family Mirabilis. It is an evergreen, thornless tree native to India, Sri Lanka, the Maldives, Madagascar, Malaysia, Indonesia, northeastern Australia, and the Pacific Islands. Pisonia grandis is the dominant species in the natural forest communities of China's Xisha Islands, distributed across many islands, with the most abundant growth on Dongdao and Yongxing Island. Pisonia grandis has a loose wood structure and poor quality. Due to wind influence, it has few branches and often grows in clusters. Its leaves are used locally as pig feed. Pisonia grandis grows rapidly, can regenerate broken branches, and is tolerant of salinity and drought. It plays an important role in coastal sand fixation, regulating island climate, and restoring island vegetation. Research on the chemical composition and pharmacological activities of Tung-Tung has revealed that it contains alkaloids, flavonoids, phenols, tannins, oxalic acid, and allantoin, among other chemical components. These compounds possess anti-diabetic, anti-inflammatory, analgesic, anti-inflammatory, wound healing, and ulcer treatment properties, demonstrating their medicinal value. Natural products from higher plants have long been an important source of medicine for humans, and their role in safeguarding human health is increasingly recognized. Traditional Chinese medicine resources are fundamental to the development of Chinese medicine. The use of innovative theories and methods to identify and discover important new resources is a hot topic and a key focus of research on their sustainable utilization.

[0003] In order to further study the chemical components of Tung-Kang-Tung and fully explore the medicinal value of various parts of Tung-Kang-Tung, this study used UPLC-Q-TOF-MS technology to analyze the components of Tung-Kang-Tung, in order to enrich the chemical components of Tung-Kang-Tung, provide a reference for the elucidation of the material basis of the efficacy of Tung-Kang-Tung, and provide a scientific reference basis for its further extraction, separation and pharmacological action research. Summary of the Invention

[0004] The purpose of the present invention is to overcome the deficiencies in the prior art and provide a method for detecting the chemical components of wind-resistant tung, which can effectively separate and identify the chemical components in wind-resistant tung.

[0005] The first aspect of the present invention is to provide a method for detecting the chemical components of wind-resistant tung, using wind-resistant tung leaves as the detection object and using ultra-high performance liquid chromatography-tandem quadrupole time-of-flight mass spectrometry for determination;

[0006] The conditions for ultra-high performance liquid chromatography were:

[0007] The chromatographic column was a 100 mm × 2.1 mm, 1.8 μm Waters ACQUITY UPLC HSS T3 column;

[0008] Column temperature: 38-42°C;

[0009] Volume flow rate: 0.2-0.4mL / min;

[0010] Mobile phase: Mobile phase A is 0.05-0.15% formic acid in water; Mobile phase B is 0.005-0.015% formic acid in acetonitrile;

[0011] The elution gradient is as follows:

[0012]

[0013] Wherein, before the ultra-high performance liquid chromatography-tandem quadrupole time-of-flight mass spectrometry is used for determination, the detection object is extracted with 60%-80% ethanol, preferably with 70% chromatographic ethanol.

[0014] Preferably, the column temperature is 40°C.

[0015] Preferably, the volume flow rate is 0.25-0.35 mL / min, more preferably 0.3 mL / min.

[0016] Preferably, the mobile phase A is a formic acid aqueous solution with a volume concentration of 0.08-0.12%, more preferably a formic acid aqueous solution with a volume concentration of 0.1%.

[0017] Preferably, mobile phase B is a formic acid acetonitrile solution with a volume concentration of 0.008-0.012%, more preferably a formic acid acetonitrile solution with a volume concentration of 0.01%.

[0018] The mass spectrometry conditions of the mass spectrometer are as follows: using an electrospray ionization source (ESI) to collect MS in continuous mode under positive and negative ion conditions. E data.

[0019] Preferably, the mass spectrometry conditions of the mass spectrometer also include: scanning range m / z 50-1500Da, low energy collision voltage (CE) of 6V, high energy collision voltage of 10-45V; capillary voltage of positive ion mode is 3.0kV, capillary voltage of negative ion mode is 2.0kV, cone voltage is 40V, and cone gas flow rate is 50L / hr.

[0020] Preferably, the mass spectrometry conditions of the mass spectrometer also include: scanning time 0.3s, detection time 22min; ion source temperature is 90-110°C, preferably 100°C; auxiliary spray ionization and desolvation gas is high-purity N2, desolvation temperature is 450°C, and desolvation gas flow rate is 600L / hr.

[0021] The detection method of the present invention can identify 50 compounds from the wind-resistant tung tree, including 20 flavonoid components, 10 phenolic acid components, 9 long-chain fatty acid components, 6 phenylpropanoid components, 4 iridoid glycoside components, and 1 lignin component.

[0022] The second aspect of the present invention is to provide a method for separating the chemical components of wind-resistant tung, using wind-resistant tung leaves as the separation object and using ultra-high performance liquid chromatography for separation; the conditions of ultra-high performance liquid chromatography are:

[0023] The conditions for ultra-high performance liquid chromatography were:

[0024] The chromatographic column was a 100 mm × 2.1 mm, 1.8 μm Waters ACQUITY UPLC HSS T3 column;

[0025] Column temperature: 38-42°C;

[0026] Volume flow rate: 0.2-0.4mL / min;

[0027] Mobile phase: Mobile phase A is 0.05-0.15% formic acid in water; Mobile phase B is 0.005-0.015% formic acid in acetonitrile;

[0028] The elution gradient is as follows:

[0029]

[0030] Wherein, before the ultra-high performance liquid chromatography-tandem quadrupole time-of-flight mass spectrometry is used for determination, the detection object is extracted with 60%-80% ethanol, preferably with 70% chromatographic ethanol.

[0031] Preferably, the column temperature is 40°C.

[0032] Preferably, the volume flow rate is 0.25-0.35 mL / min, more preferably 0.3 mL / min.

[0033] Preferably, the mobile phase A is a formic acid aqueous solution with a volume concentration of 0.08-0.12%, more preferably a formic acid aqueous solution with a volume concentration of 0.1%.

[0034] Preferably, mobile phase B is a formic acid acetonitrile solution with a volume concentration of 0.008-0.012%, more preferably a formic acid acetonitrile solution with a volume concentration of 0.01%.

[0035] By adopting the separation method of the present invention, 52 compounds can be separated from the roots of the Auricularia aculeatus.

[0036] The third aspect of the present invention is to provide a method for preparing a compound, wherein the compound is 2-Hydroxyhenicosanoic acid, and / or 2,6-Anhydro-1-O-(3,4,5-trihydroxybenzoyl)-D-mannitol, and / or 3-Hydroxy-4-{[(5xi)-4-O-methyl-D-xylo-hexopyranosyl]oxy}benzoic acid, and / or Ulmoside, and / or Hexopyranoside, 1a, 1b, 2,5a, 6,6a-hexahydro-1a-(hydroxymethyl)-6-methoxyoxireno[4,5]cyclopenta[1,2-c]pyran-2-yl6-O-pentopyranosyl-, and / or 4-Formyl-2-methoxyphenyl hexopyranoside, and / or Isorhamnetin-3-O-glucoside, and / or 13-Oxo-3,6,9,12-tetraoxatetradec-1-ylacetate, and / or Benzoic acid,2-hydroxy-5-[[2-O-[(2S,3R,4R)-tetrahydro-3,4-dihydroxy-4-(hydroxymethyl)-2-furanyl]-beta-D-xylopyranosyl]oxy]-, and / or 3,4,5-Trimethoxyphenyl 6-O-beta-L-glucopyranosyl-beta-L-glucopyranoside, and / or epicatechin, and / or β-Hydroxyacteoside, and / or [(2S,3S,4S,5S)-2,5-Bis(4-hydroxy-3,5-dimethoxyphenyl)-4-(hydroxymethyl)tetrahydro-3-furanyl]methyl beta-D-glucopyranoside, and / or 2H-1-Benzopyran-2-one,4-methyl-7-(beta-D-xylopyranosyloxy)-, and / or Methyl(1S,4aR,7aR )-1-{[(2xi)-6-O-(cyclopropylcarbonyl)-alpha-D-arabino-hexopyranosyl]oxy}-4a-hydroxy-7-(hydroxymethyl)-1,4a,5,7a-tetrahydrocyclopenta[c]pyran-4-carboxylate, and / or (1S,4aR,5S,7aS)-5-({(4xi)-6-Deoxy-4-O-[(2E)-3-(3,4-dimethoxyphenyl)-2-propenoyl]-beta-D-ribo-hexopyranosyl}oxy)-7-(hydroxymethyl)-1,4a,5,7a-tetrahydrocyclopenta[c]pyran-1-yl D-glucopyranoside, and / or Pectolinarigenin, and / or 2-(3,4-Dihydroxyphenyl)ethyl (4xi)-2-O-acetyl-3-O-(6-deoxy-D-idopyranosyl)-4-O-[(2Z)-3-(4-hydroxyphenyl)-2-propenoyl]-beta-D-ribo-hexopyranoside, and / or 2-(3,4-Dihydroxyphenyl)-5,7-dihydroxy-4-oxo-4H-chromen-3-yl 6-O-(6-deoxy-alpha-L-mannopyranosyl)-D-glucopyranoside, and / or Apiin, and / or Acteoside, and / or Isoacteoside, and / or (4aS,5R,6S)-1-Oxo-5-vinyl-4,4a,5,6-tetrahydro-1H,3H-pyrano[3,4-c]pyran-6-yl 4-O-acetyl-6-O-[3-(D-glucopyranosyloxy)-2-hydroxybenzoyl]-D-glucopyranoside, and / or 5,7-Dihydroxy-2-(4-methoxyphenyl)-4-oxo-4H-chromen-3-yl 4-O-(3-O-acetyl-6-deoxy-alpha-D-mannopyranosyl)-beta-L-glucopyranoside, and / or Cirsilineol, and / or 6-Hydroxy-10-(6-hydroxy-1,3-benzodioxol-5-yl)furo[3',4':6,7]naphtho[1,2-d][1,3]dioxol-9(7H)-one, and / or 5,7-Dihydroxy-2-(4-hydroxyphenyl)-4-oxo-4H-chromen-3-yl(5xi)-6-O-(4-O-acetyl-6-deoxy-L-mannopyran osyl)-L-xylo-hexopyranoside, and / or 5-Hydroxy-2-(4-hydroxyphenyl)-6,7-dimethoxy-4-oxo-4H-chromen-3-yl L-galactopyranoside, and / or Limocitrin, and / or Isorhamnetin, and / or 5-Acetamido-1,3-phenylene di(2-furoate), and / or (3,4,5-Trihydroxytetrahydro-2-furanyl)methyl(2Z)-3-(4-hydroxy-3-methoxyphenyl)-2-(2-methoxy-4-{(1E)-3-oxo-3-[(3,4,5-trihydroxytetrahydro-2-furanyl)methoxy]-1-propen-1-yl}phenoxy)acrylate(non-preferred name), and / or Eupatilin, and / or (+)-tanikolide, and / or Farresol, and / or eupatorin, and / or Quercetin 3,4'-dimethyl Ether, and / or Casticin, and / or 3,5-Dihydroxy-6,7,8-trimethoxyflavone, and / or Irisflavone B, and / or 5,7-Dihydroxy-3,6,4'-trimethoxyflavone, and / or Benzoic acid, 2-(acetyloxy)-, anhydride, and / or (12E)-9-Hydroxy-10-oxo-12-octadecenoic acid, and / or Quercetintetramethyl ether, and / or 13-hydroxy-10-oxooctadecenoic acid, and / or beta-dimorphecolic acid, and / or 3,5,7-Trimethoxy-3',4'-methylenedioxyflavone, and / or 10-ketostearic acid, and / or 8-(5-Hexyl-2-furyl)octanoic acid, and / or 12-hydroxy-9E-octadecenoic acid were separated using ultra-high performance liquid chromatography using Tung oil leaves as the separation object.

[0037] The conditions for ultra-high performance liquid chromatography were:

[0038] The chromatographic column was a 100 mm × 2.1 mm, 1.8 μm Waters ACQUITY UPLC HSS T3 column;

[0039] Column temperature: 38-42°C;

[0040] Volume flow rate: 0.2-0.4mL / min;

[0041] Mobile phase: Mobile phase A is 0.05-0.15% formic acid in water; Mobile phase B is 0.005-0.015% formic acid in acetonitrile;

[0042] The elution gradient is as follows:

[0043]

[0044] Wherein, before the ultra-high performance liquid chromatography-tandem quadrupole time-of-flight mass spectrometry is used for determination, the detection object is extracted with 60%-80% ethanol, preferably with 70% chromatographic ethanol.

[0045] Preferably, the column temperature is 40°C.

[0046] Preferably, the volume flow rate is 0.25-0.35 mL / min, more preferably 0.3 mL / min.

[0047] Preferably, the mobile phase A is a formic acid aqueous solution with a volume concentration of 0.08-0.12%, more preferably a formic acid aqueous solution with a volume concentration of 0.1%.

[0048] Preferably, mobile phase B is a formic acid acetonitrile solution with a volume concentration of 0.008-0.012%, more preferably a formic acid acetonitrile solution with a volume concentration of 0.01%.

[0049] The present invention uses ultra-high performance liquid chromatography combined with a specific mobile phase and elution program to separate 52 compounds from the leaves of the wind-resistant tung tree. Further, an analysis method using ultra-high performance liquid chromatography tandem with a quadrupole time-of-flight mass spectrometer is used to quickly identify 50 compounds (including 20 flavonoid components, 10 phenolic acid components, 9 long-chain fatty acid components, 6 phenylpropanoid components, 4 iridoid glycoside components, and 1 lignin component) from the leaves of the wind-resistant tung tree. The results show that the wind-resistant tung tree is rich in flavonoids and phenolic acid components, and also contains phenylpropanoid components, as well as iridoid glycosides and other components. These chemical substances have extremely strong biological activity, indicating that the wind-resistant tung tree has good medicinal potential and can be increased in its research and development and utilization in the future. The present invention enriches the chemical components of the wind-resistant tung tree and lays a foundation for its pharmacological activity and development and utilization research. At the same time, a new preparation method for the 50 compounds is provided, enriching the source of the compounds. BRIEF DESCRIPTION OF THE DRAWINGS

[0050] Figure 1 The total ion current (TIC) and base peak ion current (BPI) graphs of the UPLC-Q-TOF-MS negative ion mode of the wind-resistant tung leaf extract.

[0051] Figure 2 To isolate and identify the structure of flavonoid compounds from Tung oil tree.

[0052] Figure 3 To isolate and identify the chemical structures of other components from Tung oil tree. DETAILED DESCRIPTION

[0053] The present invention will be further described below with reference to the accompanying drawings and in conjunction with specific embodiments to better understand the present invention. Where specific techniques or conditions are not specified in the examples, the methods are based on those described in the literature in this field or on the product specifications. Where the manufacturer of the reagents or instruments is not specified, they are all commercially available conventional products.

[0054] 1. Test materials

[0055] The test sample was collected from the Xisha Islands and identified by Wang Zhunian, a researcher at the Institute of Tropical Crops Germplasm Resources, Chinese Academy of Tropical Agricultural Sciences, as Pisonia grandis R.Br., a wind-resistant tung tree of the genus Pisonia in the family Mirabilis.

[0056] 2 Instruments and reagents

[0057] Instrument: Xevo G2-XS QTof LC / MS: including ACQUITY The equipment used was an I-Class system, a Xevo G2-XS QTof mass spectrometer (Waters, USA); a Waters ACQUITY UPLC HSS T3 (2.1 mm × 100 mm, 1.8 μm) chromatographic column (Waters, USA); a Secura 513-1CN precision balance (Sartorius, Germany); a KQ3200DE CNC ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.); an N1200BV-W mini-concentration system (EYELA Tokyo Rikagaki Co., Ltd., Japan); an Eyela CA-1115B cooling water circulation device (EYELA Tokyo Rikagaki Co., Ltd., Japan); an Eyela FDU-2110 freeze dryer (EYELA Tokyo Rikagaki Co., Ltd., Japan); a Milli-Q ultrapure water system (Merck Millipore, Germany); and a 5810R benchtop high-speed refrigerated centrifuge (Eppendorf Centrifuge, Germany).

[0058] Reagents: Methanol, acetonitrile (mass spectrometry grade, for LC ≥ 99.99%, Fisher Scientific, USA), formic acid (mass spectrometry grade, for LC / MS ≥ 98%-100%, Merck Millipore, Germany), sodium hydroxide (ACS grade, ACS reagent ≥ 97%, Sigma, USA), leucine enkephalin (Standards Kit for Tof G2-S, Waters, USA). Distilled water (Guangzhou Watsons Food and Beverage Co., Ltd.) was obtained; ultrapure water was prepared using a Milli-Q ultrapure water instrument. All other reagents were of analytical grade.

[0059] Material: Pall Syringe filter membrane (0.2 μm, 13 mm, Pall Corporation, USA), disposable sterile syringe with needle (1 mL, Fenglin Medical Instrument Co., Ltd.), Labmed pipette tips 1000 μL, 200 μL, 20 μL (Labmed Biotech, USA), KG2211W 1.5 mL centrifuge tube (KiRGEN, USA), 2 mL transparent screw cap sample bottle (Waters, USA).

[0060] Software: Masslynx V4.1 workstation (Waters, USA), Waters Scientific Information Systems (Waters Corporation, USA).

[0061] 3 Test methods

[0062] 3.1 Preparation of test solution

[0063] Take an appropriate amount of raw material and air-dry it, crush it, and pass it through an 80-mesh sieve. Accurately weigh 10g of sample and extract it with 200mL of 70% chromatographic ethanol under reflux for 1 hour at 70°C. Take 1mL of the extract and perform refrigerated centrifugation (12000rpm, 10min, 10°C). Let it stand for 1-2min. Take 300μL of the supernatant and pass it through a Waters HLB solid-phase microextraction column. Elute it first with 600μL of 5% chromatographic methanol, then with 500μL of 95% chromatographic methanol. Collect the 95% methanol eluate, filter it through a 0.2μm microporous membrane, store it in a 2mL transparent sample bottle, label it TD, and store it in a refrigerator at 4°C until use. All samples should be prepared and used immediately, and the shelf life should not exceed 24h.

[0064] 3.2 UPLC-Q-TOF-MS detection conditions

[0065] Chromatographic analysis conditions: Waters ACQUITY UPLC HSS T3 column (2.1 mm × 100 mm, 1.8 μm), flow rate 0.3 mL min -1 , injection volume 1 μL, column temperature 40°C; mobile phase 0.1% formic acid aqueous solution (A)-0.01% formic acid acetonitrile solution (B), gradient elution program see Table 1.

[0066] Table 1 Gradient elution program

[0067]

[0068] Mass spectrometry detection conditions: electrospray ionization (ESI) source under positive (+) and negative ion (-) conditions, Continuum mode, collecting MS E Data. Calibration solution is 200pg·μL -1 Leucine-enkephalin, 0.5 mM sodium-formate. Scan range (m / z) 50-1500 Da, scan time 0.3 s, detection time 22 min. Low-energy collision voltage (CE) 6 V, high-energy collision voltage 10-45 V; capillary voltage 3.0 kV in positive ion mode, 2.0 kV in negative ion mode, cone voltage 40 V, ion source temperature 100°C, auxiliary spray ionization and desolvation gas high-purity N2, desolvation temperature 450°C, cone gas flow 50 L / hr, desolvation gas flow 600 L / hr.

[0069] 3.3 Data processing

[0070] Masslynx V4.1 software is used to acquire, manage and easily process UPLC / Q Tof MS EThe UNIFI scientific information system is used for data browsing, storage, and comprehensive analysis. Compound identification is performed based on literature data, combined with TCM Chiese [UNIFI 1.7] and ChemSpider online databases.

[0071] 4 Results and Analysis

[0072] The chemical components of the leaves of Tung oil tree were qualitatively analyzed based on UPLC-Q-TOF-MS. The (-)ESI-MS mass spectrometry total ion chromatogram (TIC) had good separation and high response, such as Figure 1 Therefore, negative ion data was imported into UNIFI, and relevant method parameters were set. Based on the accurate relative molecular mass and mass spectral fragments given by the mass spectrometer, the error range was about ±6ppm, and the results of literature reports and online database matching were combined to infer the identification results shown in Table 2. The results showed that 52 compounds were obtained from the leaves of the wind-resistant tung tree, and the structures of 50 of them were identified ( Figure 2 and Figure 3 ), including 20 flavonoid components, 10 phenolic acid components, 9 long-chain fatty acid components, 6 phenylpropanoid components, 4 iridoid glycosides components, and 1 lignin component.

[0073] Table 2 UPLC-Q-TOF-MS identification results of chemical components of wind-resistant tung leaves

[0074]

[0075]

[0076]

[0077]

[0078] The above detailed description of the specific embodiments of the present invention is intended only as an example, and the present invention is not limited to the specific embodiments described above. For those skilled in the art, any equivalent modifications and substitutions made to the practical embodiments are also within the scope of the present invention. Therefore, any equivalent changes and modifications made without departing from the spirit and scope of the present invention should be included within the scope of the present invention.

Claims

1. A method for detecting the chemical components of Tung oil tree, characterized in that: The leaves of Tung oil plant were used as the test object, extracted with 60%-80% ethanol, and determined by ultra-performance liquid chromatography-tandem quadrupole time-of-flight mass spectrometry. The conditions for ultra-high performance liquid chromatography were: The chromatographic column was a 100 mm × 2.1 mm, 1.8 μm Waters ACQUITY UPLC HSS T3 column; Column temperature: 38-42°C; Volume flow rate: 0.2-0.4 mL / min; Mobile phase: Mobile phase A is 0.05-0.15% formic acid in water; Mobile phase B is 0.005-0.015% formic acid in acetonitrile; The elution gradient is as follows: The detected chemical components are 2-Hydroxyhenicosanoic acid, 2,6-Anhydro-1-O-(3,4,5-trihydroxybenzoyl)-D-mannitol, Ulmoside, Hexopyranoside, 1a,1b,2,5a,6,6a-hexahydro-1a-(hydroxymethyl)-6-methoxyoxireno[4,5]cyclopenta[1,2-c]pyran-2-yl 6-O-pentopyranosyl-, 4-Formyl-2-methoxyphenyl hexopyranoside, Isorhamnetin-3-O-glucoside, 13-Oxo-3,6,9,12-tetraoxatetradec-1-yl acetate, Benzoic acid, 2-hydroxy-5-[[2-O-[(2S,3R,4R)-tetrahydro-3,4-dihydroxy-4-(hydroxymethyl)-2-furanyl]-beta-D-xylopyranosyl]oxy]-, 3,4,5-Trimethoxyphenyl 6-O-beta-L-glucopyranosyl-beta-L-glucopyranoside, epicatechin, β-Hydroxyacteoside, [(2S,3S,4S,5S)-2,5-Bis(4-hydroxy-3,5-dimethoxyphenyl)-4-(hydroxymethyl)tetrahydro-3-furanyl]methyl beta-D-glucopyranoside, 2H-1-Benzopyran-2-one, 4-methyl-7-(beta-D-xylopyranosyloxy)-, Pectolinarigenin, 2-(3,4-Dihydroxyphenyl)-5,7-dihydroxy-4-oxo-4H-chromen-3-yl 6-O-(6-deoxy-alpha-L-mannopyranosyl)-D-glucopyranoside, Acteoside, Isoacteoside, 5,7-Dihydroxy-2-(4-methoxyphenyl)-4-oxo-4H-chromen-3-yl 4-O-(3-O-acetyl-6-deoxy-alpha-D-mannopyranosyl)-beta-L-glucopyranoside, Cirsilineol, 6-Hydroxy-10-(6-hydroxy-1,3-benzodioxol-5-yl)furo[3',4':6,7]naphtho[1,2-d][1,3]dioxol-9(7H)-one, Limocitrin, Isorhamnetin, 5-Acetamido-1,3-phenylene di(2-furoate), (3,4,5-Trihydroxytetrahydro-2-furanyl)methyl (2Z)-3-(4-hydroxy-3-methoxyphenyl)-2-(2-methoxy-4-{(1E)-3-oxo-3-[(3,4,5-trihydroxytetrahydro-2-furanyl)methoxy]-1-propen-1-yl}phenoxy)acrylate (non-preferred name), Eupatilin, (+)-tanikolide, Farresol, eupatorin, Quercetin 3,4'-dimethyl Ether, Casticin, 3,5-Dihydroxy-6,7,8-trimethoxyflavone, IrisflavoneB, 5,7-Dihydroxy-3,6,4'-trimethoxyflavone, Benzoic acid, 2-(acetyloxy)-, anhydride, (12E)-9-Hydroxy-10-oxo-12-octadecenoic acid, Quercetin tetramethylether, 13-hydroxy-10-oxooctadecenoic acid, beta-dimorphecolic acid, 3,5,7-Trimethoxy-3',4'-methylenedioxyflavone, 10-ketostearic acid, 8-(5-Hexyl-2-furyl)octanoic acid, and 12-hydroxy-9E-octadecenoic acid., 2. The chemical composition detection method according to claim 1, characterized in that: Extraction was performed using 70% ethanol.

3. The chemical composition detection method according to claim 1, characterized in that: The column temperature is 40° C.; the volume flow rate is 0.25-0.35 mL / min; the mobile phase A is a formic acid aqueous solution with a volume concentration of 0.08-0.12%; and the mobile phase B is a formic acid acetonitrile solution with a volume concentration of 0.008-0.012%.

4. The chemical composition detection method according to claim 3, characterized in that: The volume flow rate was 0.3 mL / min; mobile phase A was a 0.1% by volume formic acid aqueous solution; and mobile phase B was a 0.01% by volume formic acid acetonitrile solution.

5. The chemical composition detection method according to claim 1, characterized in that: The mass spectrometry conditions of the mass spectrometer were as follows: an electrospray ion source was used to collect MS in continuous mode under positive and negative ion conditions. E data.

6. The chemical composition detection method according to claim 3, characterized in that: The mass spectrometry conditions of the mass spectrometer also include: scanning range m / z 50-1500Da, low energy collision voltage 6 V, high energy collision voltage 10-45 V; capillary voltage 3.0 kV for positive ion mode, 2.0 kV for negative ion mode, cone voltage 40 V, cone gas flow 50 L / hr; The scan time was 0.3 s, and the detection time was 22 min. The ion source temperature was 90-110°C. The auxiliary spray ionization and desolvation gases were high-purity N2, the desolvation temperature was 450 °C, and the desolvation gas flow rate was 600 L / hr.

7. The chemical composition detection method according to claim 6, characterized in that The ion source temperature was 100 °C.

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