Passion fruit megastigmane glycoside compound, preparation method and application thereof

Abstract

The present invention belongs to the technical field of extraction, separation and purification of plant components, and specifically relates to passion fruit megastigmane glycoside compounds, their preparation methods, and applications. The passion fruit megastigmane glycoside compounds provided by the present invention are derived from passion fruit juice and are natural plant secondary metabolites. These disaccharide compounds, formed by linking a C13 norsesquiterpene aglycone to glucose and rhamnose via an oxyglycosidic bond, represent a novel class of compounds with broad application prospects in anti-anxiety, sedative-hypnotic, or flavor-enhancing products.

Description

Technical Field

[0001] The invention belongs to the technical field of extraction, separation and purification of plant components, and particularly relates to a passion fruit megastigmane glycoside compound and a preparation method and application thereof. Background Art

[0002] Currently, the anti-anxiety, sedative and hypnotic products on the market mainly contain melatonin health supplements or benzodiazepines such as oxazepam, lorazepam, and alprazolam. These products or drugs can cause dependence or toxic side effects in the human body. γ-Aminobutyric acid (γ-GABA) is an important inhibitory transmitter in the human central nervous system and one of the most important neurotransmitters in brain tissue. It has physiological activities such as anti-anxiety, sedation and hypnosis. In addition, γ-GABA metabolism is regulated by γ-aminobutyric acid transaminase (γ-GABA-T). Therefore, by inhibiting the activity of γ-GABA-T, the level of γ-GABA metabolism can be reduced, and the level of γ-GABA in the body can be increased, thus exerting its physiological functions of anti-anxiety, sedation and hypnosis.

[0003] Molecular docking, a computer-aided drug design technique, uses chemometrics and other disciplines to simulate molecular geometry and intermolecular forces to identify and predict the complex structure of ligands and receptors. Essentially, it involves the mutual recognition of two or more molecules and has become a relatively mature direct drug design method in recent years. Molecular docking not only allows for detailed investigation of interactions between ligands (drug molecules) and receptors (known target proteins or active sites), but also enables the discovery and optimization of the molecular structures of lead compounds. It is a key structure-based drug design method in modern innovative drug research.

[0004] Currently, several compounds have been successfully marketed using computer-aided drug design, including Roche's Saquinavir, an HIV protein inhibitor based on HIV proteolytic enzymes; Biota's Relenza, an anti-influenza drug based on neuraminidase; Novartis' Gleevec, an anti-chronic myeloid leukemia drug based on Abl tyrosine kinase; and Merck Sharp & Dohme's Dorzolamid, a glaucoma drug based on carbonic anhydrase. Therefore, molecular docking and computer-aided drug design can rapidly discover natural products that modulate γ-GABA.

[0005] Megastigmane glycosides are a unique class of natural products, formed by linking C13 norosequiterpenoid aglycones to sugars. These compounds not only possess a wide range of physiological activities, including anxiolytic, sedative, and sleep-inducing properties, but also serve as aroma precursors, releasing numerous aroma components such as ionol, ionone, and megastigmatriene upon thermal decomposition. Furthermore, research has shown that plant aromatics are effective in alleviating various anxiety and sleep disorders. The Compendium of Materia Medica lists 35 species of fragrant woods, 56 species of aromatic herbs, and various aromatherapy methods. Therefore, the aroma produced by thermal decomposition of megastigmane glycosides possesses anxiolytic, sedative, and hypnotic properties, and therefore holds great promise for their application in aromatherapy products and tobacco products.

[0006] Passion fruit contains many plant secondary metabolites that are beneficial to human health and are commonly used to treat symptoms such as anxiety, insomnia, epilepsy, and nervous tension. These uses are similar to sedatives and hypnotic drugs. Its roots, vines, flowers, and fruits can be used as medicine, with good efficacy for ailments such as neuralgia, insomnia, and wind-heat-induced dizziness. However, the material basis for passion fruit's use in treating anxiety and insomnia remains unclear. Therefore, the discovery and development of natural products from passion fruit with anti-anxiety, sedative, and hypnotic properties is an urgent need for high-value utilization of passion fruit. Summary of the Invention

[0007] To address the deficiencies of the prior art, the present invention provides a passion fruit megastigmane glycoside compound, a preparation method, and an application thereof. The megastigmane glycoside compound provided by the present invention is derived from passion fruit juice, a specialty fruit, and is a novel structural compound.

[0008] The technical solution of the present invention to solve the above technical problems is as follows:

[0009] A passion fruit megastigmane glycoside compound, whose specific structural formula is any one of the following structures:

[0010]

[0011] The beneficial effects of the passion fruit megastigmane glycoside compound of the present invention are:

[0012] The passion fruit megastigmane glycoside compound of the present invention is derived from passion fruit juice, is a natural plant secondary metabolite, is a disodium glycoside compound formed by connecting a C13 norosequiterpene aglycone with glucose and rhamnose via an oxygen glycosidic bond, and is a new type of structural compound.

[0013] A second object of the present invention is to provide a method for preparing the passion fruit megastigmane glycoside compound. The technical solution of the present invention to solve the above technical problem is as follows: A method for preparing the passion fruit megastigmane glycoside compound comprises the following steps:

[0014] Step 1: Passion fruit pulp with seeds and pomace removed is passed through a high-speed continuous flow centrifuge for solid-liquid separation to obtain passion fruit clarified juice.

[0015] Step 2: The passion fruit clarified juice obtained in step 1 is first separated by a ceramic membrane with a molecular weight cutoff of 100kD to 200kD, and the obtained permeate is further separated by an ultrafiltration membrane of 2kD to 4kD, and the permeate is collected to obtain a passion fruit juice ultrafiltrate.

[0016] Step 3: After the passion fruit juice ultrafiltrate obtained in step 2 is adsorbed on a non-polar or weakly polar macroporous resin, the column is first rinsed with 4 to 6 times the column volume of pure water, and then gradient eluted with 4 to 6 times the column volume of a 10% low-carbon alcohol solution and a 20% to 70% low-carbon alcohol solution, respectively, and the 20% to 70% low-carbon alcohol eluate is collected, concentrated at 50° C., and dried to obtain a passion fruit juice extract rich in megastigmane glycoside compounds.

[0017] Step 4: The passion fruit juice extract rich in megastigmane glycoside compounds obtained in step 3 was repeatedly injected into a high performance liquid chromatography column, and the chromatographic peaks with retention times of 29.4 min, 37.0 min, 42.6 min, 47.9 min and 49.9 min were accumulated and collected, respectively, and concentrated and dried to obtain compound 1, compound 2, compound 3, compound 4 and compound 5, respectively.

[0018] The beneficial effects of preparing passion fruit megastigmane glycoside compounds according to the present invention are:

[0019] Passion fruit juice has a complex composition and high water content. Current technologies require multiple column chromatography separations, which consumes significant manpower, material resources, and time. Furthermore, there are no reports of combining membrane separation with column chromatography to prepare passion fruit megastigmane glycoside compounds. Therefore, the present invention utilizes membrane separation and macroporous resin separation techniques to not only effectively address the high water content of the juice but also rapidly and extensively enrich the target megastigmane glycoside compound. Subsequently, high-column efficiency and high-sensitivity preparative liquid chromatography separation techniques are used to efficiently prepare high-purity megastigmane glycoside compound monomers.

[0020] Compared with existing separation technologies, the method of the present invention can achieve rapid enrichment of megastigmane glycoside compounds, accurately prepare target components, effectively reduce the separation process, and achieve simultaneous preparation of multiple monomer compounds, providing a convenient way for the rapid preparation of passion fruit megastigmane glycoside compounds.

[0021] On the basis of the above technical solution, the present invention can also be improved as follows.

[0022] Furthermore, in step 1, the high-speed continuous flow centrifugal speed is 8000 r / min to 20000 r / min.

[0023] The above method has the further beneficial effect that high-speed centrifugal force can effectively remove various solid impurities such as tiny plant fibers and suspended particles in the juice to obtain clarified juice, and continuous flow centrifugal separation is more efficient.

[0024] Furthermore, in step 3, the non-polar macroporous resin is any one of D101, HPD-100, X-5, XND-11, H103, ADS-17, HP-20, and D3520; the weakly polar macroporous resin is any one of AB-8, D201, HPD-300, DS-401, and DM-130.

[0025] A further beneficial effect of the above method is that the non-polar and weakly polar macroporous resin of the above specific type can effectively adsorb the megastigmane glycoside compound in passion fruit juice.

[0026] Furthermore, in step 3, the low-carbon alcohol is any one of methanol and ethanol.

[0027] A further beneficial effect of the above method is that methanol and ethanol have a good elution ability for the non-polar and weakly polar macroporous resins adsorbed with the megastigmane glycoside compound.

[0028] Furthermore, in step 3, the concentration step must be carried out at 50° C. in order to avoid structural changes of the target compound caused by long-term high-temperature concentration.

[0029] Furthermore, in step 4, the high performance liquid chromatography preparation parameters are: 10 mm × 250 mm shim-pack GISC18 chromatographic column, the particle size of the filler is 10 μm, the column temperature is 30 ° C, the mobile phase is acetonitrile and water, and the elution mode is gradient elution: 0 min, the volume ratio of acetonitrile to water is 19:81; 55 min, the volume ratio of acetonitrile to water is 19:81; 60 min, the volume ratio of acetonitrile to water is 40:60.

[0030] A further beneficial effect of the above method is that under the chromatographic conditions, five megastigmane glycoside monomer compounds can be prepared simultaneously.

[0031] The third object of the present invention is to provide a use of a passion fruit megastigmane glycoside compound in the preparation of anxiolytic, sedative, hypnotic or flavor-enhancing products.

[0032] Furthermore, the present invention provides the use of the above-mentioned megastigmane glycoside compound in the preparation of medicines, functional foods or tobacco products.

[0033] The present invention conducted molecular docking computer-aided drug design studies on passion fruit megastigmane glycoside compounds and γ-aminobutyric acid transaminase (PDB: 10HW). The results demonstrated that the megastigmane glycoside compounds prepared by the present invention all exhibited strong binding activity to γ-aminobutyric acid transaminase. In particular, the binding energies of Compounds 1 and 2 with γ-aminobutyric acid transaminase were comparable to those of the antianxiety and sedative-hypnotic drug oxazepam. This indicates that the megastigmane glycoside compounds obtained by the present invention have broad application prospects in antianxiety and sedative-hypnotic products.

[0034] The present invention conducted a study on the pyrolysis-gas chromatography / mass spectrometry of passion fruit megastigmane glycoside compounds, which showed that the megastigmane glycoside compounds degraded under heat to release a large amount of flavor components such as ionol, ionone and megastigmatriene, which have a flavoring effect and can be used in products such as cigarettes to improve the flavor and quality of the products. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 This is the HR-ESI-MS spectrum of compound 1 of Example 1 of the present invention.

[0036] Figure 2 This is the 1H-NMR spectrum of Compound 1 of Example 1 of the present invention.

[0037] Figure 3 This is the 13C-NMR spectrum of Compound 1 of Example 1 of the present invention.

[0038] Figure 4 This is the HR-ESI-MS spectrum of compound 2 in Example 1 of the present invention.

[0039] Figure 5 This is the 1H-NMR spectrum of Compound 2 of Example 1 of the present invention.

[0040] Figure 6 This is the 13C-NMR spectrum of Compound 2 of Example 1 of the present invention.

[0041] Figure 7 This is the HR-ESI-MS spectrum of compound 3 in Example 1 of the present invention.

[0042] Figure 8 This is the 1H-NMR spectrum of compound 3 in Example 1 of the present invention.

[0043] Figure 9 This is the 13C-NMR spectrum of compound 3 in Example 1 of the present invention.

[0044] Figure 10 This is the HR-ESI-MS spectrum of compound 4 in Example 1 of the present invention.

[0045] Figure 11 This is the 1H-NMR spectrum of compound 4 in Example 1 of the present invention.

[0046] Figure 12 This is the 13C-NMR spectrum of compound 4 in Example 1 of the present invention.

[0047] Figure 13 This is the HR-ESI-MS spectrum of compound 5 in Example 1 of the present invention.

[0048] Figure 14 This is the 1H-NMR spectrum of compound 5 in Example 1 of the present invention.

[0049] Figure 15 This is the 13C-NMR spectrum of Compound 5 of Example 1 of the present invention.

[0050] Figure 16 Molecular docking diagram of compound 1 and γ-aminobutyrate transaminase.

[0051] Figure 17 Molecular docking diagram of compound 2 and γ-aminobutyrate transaminase.

[0052] Figure 18 Molecular docking diagram of compound 3 and γ-aminobutyrate transaminase.

[0053] Figure 19 Molecular docking diagram of compound 4 and γ-aminobutyrate transaminase.

[0054] Figure 20 Molecular docking diagram of compound 5 and γ-aminobutyrate transaminase. DETAILED DESCRIPTION

[0055] The principles and features of the present invention are described below. The examples given are only used to explain the present invention and are not used to limit the scope of the present invention.

[0056] Example 1

[0057] Step 1: Take 40 kg of passion fruit pulp from which seeds and pomace have been removed, and pass it through a continuous flow high-speed centrifuge at a rotation speed of 15,000 r / min to obtain passion fruit clarified juice.

[0058] Step 2: The passion fruit clarified juice obtained in step 1 is first separated by a ceramic membrane with a molecular weight cutoff of 150kD, and the permeate is then separated by an ultrafiltration membrane with a molecular weight cutoff of 3kD, and the membrane permeate is collected to obtain a passion fruit juice ultrafiltrate.

[0059] Step 3: The passion fruit juice ultrafiltrate obtained in Step 2 is injected into a D101 macroporous resin column for adsorption. After adsorption, the column is first rinsed with 5 column volumes of pure water, then gradient eluted with 5 column volumes of 10% ethanol and then 50% ethanol, respectively. The 50% ethanol eluate is collected, concentrated at 50°C, and dried to obtain a passion fruit juice extract rich in megastigmane glycoside compounds.

[0060] Step 4: The passion fruit juice extract rich in megastigmane glycoside compounds described in Step 3 was repeatedly injected into a high-performance liquid chromatography column for preparation. The high-performance liquid chromatography preparation parameters were: a 10 mm × 250 mm Shim-Pack GIS C18 column, a filler particle size of 10 μm, a column temperature of 30°C, a mobile phase of acetonitrile and water in a volume ratio, and a gradient elution mode: 0 min, a volume ratio of acetonitrile to water of 19:81; 55 min, a volume ratio of acetonitrile to water of 19:81; 60 min, a volume ratio of acetonitrile to water of 40:60. The chromatographic peaks at retention times of 29.4 min, 37.0 min, 42.6 min, 47.9 min, and 49.9 min were accumulated and collected, and the mixture was concentrated and dried to obtain Compound 1, Compound 2, Compound 3, Compound 4, and Compound 5, respectively. It was determined that the purities of Compound 1, Compound 2, Compound 3, Compound 4 and Compound 5 were 92.1%, 90.3%, 93.2%, 92.8% and 90.7%, respectively.

[0061] Structure identification:

[0062] 1. Structural identification of compound 1

[0063] Amorphous powder, m / z 561.2558 [M+HCOO] - , molecular formula is C 25 H 40 O 11 , 1 H-NMR and 13 The C-NMR data are shown in Table 1.

[0064] Table 1 Compound 1 1 H-NMR and13 C-NMR data (deuterated methanol)

[0065]

[0066]

[0067] 2. Structure identification of compound 2

[0068] Amorphous powder, m / z 561.2534 [M+HCOO] - , molecular formula is C 25 H 40 O 11 , 1 H-NMR and 13 The C-NMR data are shown in Table 2.

[0069] Table 2 Compound 2 1 H-NMR and 13 C-NMR data (deuterated methanol)

[0070]

[0071]

[0072] 3. Structural identification of compound 3

[0073] Amorphous powder, m / z 563.2730 [M+HCOO] - , molecular formula is C 25 H 42 O 11 , 1 H-NMR and 13 The C-NMR data are shown in Table 3.

[0074] Table 3 Compound 3 1 H-NMR and 13 C-NMR data (deuterated methanol)

[0075]

[0076]

[0077] 4. Structural identification of compound 4

[0078] Amorphous powder, m / z 563.2696 [M+HCOO] - , molecular formula is C 25 H 42 O 11 , 1 H-NMR and 13 The C-NMR data are shown in Table 4.

[0079] Table 4 Compound 4 1 H-NMR and 13 C-NMR data (deuterated methanol)

[0080]

[0081]

[0082]

[0083] 5. Structure identification of compound 5

[0084] Amorphous powder, m / z 563.2686 [M+HCOO] - , molecular formula is C 25 H 42 O 11 , 1 H-NMR and 13 The C-NMR data are shown in Table 5.

[0085] Table 5 Compound 5 1 H-NMR and 13 C-NMR data (deuterated methanol)

[0086]

[0087]

[0088] Example 2

[0089] The preparation of the passion fruit megastigmane glycoside compound of this embodiment includes the following steps:

[0090] Step 1: Take 40 kg of passion fruit pulp from which seeds and pomace have been removed, and pass it through a continuous flow high-speed centrifuge at a rotation speed of 20,000 r / min to obtain passion fruit clarified juice.

[0091] Step 2: The passion fruit clarified juice obtained in step 1 is first separated by a ceramic membrane with a molecular weight cutoff of 200kD, and the permeate is then separated by an ultrafiltration membrane with a molecular weight cutoff of 4kD, and the membrane permeate is collected to obtain a passion fruit juice ultrafiltrate.

[0092] Step 3: injecting the passion fruit juice ultrafiltrate obtained in step 2 into an AB-8 macroporous resin column for adsorption. After the adsorption is completed, the column is first rinsed with 4 times the column volume of pure water, and then gradient eluted with 4 times the column volume of 10% methanol and 70% methanol, respectively. The 70% methanol eluate is collected, concentrated at 50° C., and dried to obtain a passion fruit juice extract rich in megastigmane glycoside compounds.

[0093] Step 4: The passion fruit juice extract rich in megastigmane glycoside compounds described in Step 3 was repeatedly injected into a high-performance liquid chromatography column for preparation. The high-performance liquid chromatography preparation parameters were: a 10 mm × 250 mm Shim-Pack GIS C18 column, a filler particle size of 10 μm, a column temperature of 30°C, a mobile phase of acetonitrile and water in a volume ratio, and a gradient elution mode: 0 min, a volume ratio of acetonitrile to water of 19:81; 55 min, a volume ratio of acetonitrile to water of 19:81; 60 min, a volume ratio of acetonitrile to water of 40:60. The chromatographic peaks at retention times of 29.4 min, 37.0 min, 42.6 min, 47.9 min, and 49.9 min were collected, concentrated, and dried to obtain Compound 1, Compound 2, Compound 3, Compound 4, and Compound 5, respectively.

[0094] Structural identification is the same as in Example 1.

[0095] Example 3

[0096] The preparation of the passion fruit megastigmane glycoside compound of this embodiment includes the following steps:

[0097] Step 1: Take 40 kg of passion fruit pulp from which seeds and pomace have been removed, and pass it through a continuous flow high-speed centrifuge at a rotation speed of 8000 r / min to obtain passion fruit clarified juice.

[0098] Step 2: The passion fruit clarified juice obtained in step 1 is first separated by a ceramic membrane with a molecular weight cut-off of 100 kD, and the permeate is then separated by an ultrafiltration membrane with a molecular weight cut-off of 2 kD, and the membrane permeate is collected to obtain a passion fruit juice ultrafiltrate.

[0099] Step 3: The passion fruit juice ultrafiltrate of step 2 is injected into an HP-20 macroporous resin column for adsorption. After the adsorption is completed, the column is first rinsed with 6 times the column volume of pure water, and then gradient eluted with 6 times the column volume of 10% ethanol and 30% ethanol, respectively. The 30% ethanol eluate is collected, concentrated at 50° C., and dried to obtain a passion fruit juice extract rich in megastigmane glycoside compounds.

[0100] Step 4: The passion fruit juice extract rich in megastigmane glycoside compounds described in Step 3 was repeatedly injected into a high-performance liquid chromatography column for preparation. The high-performance liquid chromatography preparation parameters were: a 10 mm × 250 mm Shim-Pack GIS C18 column, a filler particle size of 10 μm, a column temperature of 30°C, a mobile phase of acetonitrile and water in a volume ratio, and a gradient elution mode: 0 min, a volume ratio of acetonitrile to water of 19:81; 55 min, a volume ratio of acetonitrile to water of 19:81; 60 min, a volume ratio of acetonitrile to water of 40:60. The chromatographic peaks at retention times of 29.4 min, 37.0 min, 42.6 min, 47.9 min, and 49.9 min were collected, concentrated, and dried to obtain Compound 1, Compound 2, Compound 3, Compound 4, and Compound 5.

[0101] Structural identification is the same as in Example 1.

[0102] Example 4

[0103] (1) Molecular docking and computer-aided drug design research on passion fruit megastigmane glycoside compounds and γ-aminobutyric acid transaminase.

[0104] The docked ligand compounds are Compound 1, Compound 2, Compound 3, Compound 4, and Compound 5 described in the present invention. The docked protein is γ-aminobutyric acid transaminase (GABA-T, PDB: 10HW). AutoDock Vina software was used to optimize the ligands and proteins required for molecular docking, including removing or hydrogenating the crystal structure of the target protein, modifying amino acids, optimizing energy, and adjusting force field parameters. Finally, the optimized target structure and the ligand structure were molecularly docked using pyrx software, and the binding energy (kcal / mol) value represents the binding ability of the two. The lower the binding energy, the more stable the binding between the ligand and the receptor. It was visualized and analyzed using Pymol, and the 2D graph was visualized using Discovery Studio 2020Client. The docking results are shown in Table 6

[0105] Table 6 Molecular docking binding energy of passion fruit megastigmane glycoside compounds and γ-aminobutyric acid transaminase

[0106] Compound Binding energy (kcal / mol) Compound 1 -9.1 Compound 2 -9.1 Compound 3 -8.2 Compound 4 -8.7 Compound 5 -8.5 Oxazepam -9.0

[0107] It is generally believed that a binding energy between a ligand compound and a target protein less than -4.25 kcal / mol indicates a certain binding activity between the two, and a binding energy less than -7.0 kcal / mol indicates a strong binding activity.

[0108] The results in Table 6 show that the binding energy of passion fruit megastigmane glycoside compounds with GABA-T is less than -8.0 kcal / mol, and the binding energy of compound 1 and compound 2 with GABA-T is comparable to that of the sedative hypnotic drug oxazepam, indicating that passion fruit megastigmane glycoside compounds have the ability to regulate GABA-T activity. Further visualization analysis of the binding protein revealed that passion fruit megastigmane glycoside compounds mainly bind to γGABA-T through hydrogen bonds, Pi-Signa, Pi-Alkyl and Alkyl interactions. The results are as follows Figures 16-20 shown.

[0109] Example 5

[0110] Analysis of Pyrolysis Compositions of Glycosides from Passion Fruit Megastigmane

[0111] Given the structural similarity of the passion fruit megastigmane glycoside compounds prepared in the present invention, Compounds 1 and 4 were selected for thermal pyrolysis-gas chromatography / mass spectrometry analysis at a pyrolysis temperature of 500°C. The results are as follows:

[0112] (1) The main pyrolysis products of compound 1 are: α-cyclocitral (5.78%), ionone (17.59%), (2-trans)-1-(2,6,6-trimethyl-2-cyclohexen-1-yl)-2-buten-1-one (5.05%), 2,6,6-trimethyl-1-[(1-cis)-3-methyl-1,3-butadienyl]-1,3-cyclohexadiene (9.16%), 3-oxo-β-ionone (6.25%), dihydro-3-oxo-α-ionone (13.48%), dihydro-3-oxo-α-ionol (27.29%).

[0113] (2) The main thermal decomposition products of compound 4 are: megastigmatrienone (68.3%) and 3-oxo-7,8-dihydro-α-ionol (9.28%).

[0114] Passion fruit megastigmane glycoside compounds degrade upon heating, primarily producing odoriferous derivatives such as ionone, ionol, and megastigmatrienone. This indicates that the passion fruit megastigmane glycoside compounds described herein have a flavoring effect and can be used as flavor enhancers. Cigarette products typically burn at temperatures between 300 and 800°C, within the decomposition temperature range of megastigmane glycoside compounds. Adding megastigmane glycoside compounds to tobacco products releases aromatic components upon heating, improving the quality of tobacco products by enhancing their flavor and neutralizing unpleasant notes. Furthermore, the pleasant aroma can soothe emotions and have a calming effect.

[0115] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A method for preparing passion fruit megastigmane glycoside compound, characterized in that: The following steps are involved: Step 1: Passion fruit pulp with seeds and pomace removed is subjected to solid-liquid separation in a high-speed continuous flow centrifuge to obtain clarified passion fruit juice; Step 2: The passion fruit clarified juice obtained in step 1 is first separated by a ceramic membrane with a molecular weight cutoff of 100kD to 200kD, and the obtained permeate is further separated by an ultrafiltration membrane of 2kD to 4kD, and the permeate is collected to obtain a passion fruit juice ultrafiltrate; Step 3: After the passion fruit juice ultrafiltrate obtained in step 2 is adsorbed on a non-polar or weakly polar macroporous resin, the column is first rinsed with 4 to 6 times the column volume of pure water, and then gradient eluted with 4 to 6 times the column volume of a 10% low-carbon alcohol solution and a 20% to 70% low-carbon alcohol solution, respectively, and the 20% to 70% low-carbon alcohol eluate is collected, concentrated at 50° C., and dried to obtain a passion fruit juice extract rich in megastigmane glycoside compounds; Step 4: The passion fruit juice extract rich in megastigmane glycoside compounds obtained in step 3 was repeatedly injected into a high performance liquid chromatography column, and the chromatographic peaks with retention times of 29.4 min, 37.0 min, 42.6 min, 47.9 min and 49.9 min were accumulated and collected, respectively. The products were concentrated and dried to obtain Compound 1, Compound 2, Compound 3, Compound 4 and Compound 5, respectively, with the structural formulas as follows: ； In step 3, the non-polar macroporous resin is D101 or HP-20; the weakly polar macroporous resin is AB-8; and the low-carbon alcohol is either methanol or ethanol; In step 4, the high performance liquid chromatography preparation parameters are: 10 mm × 250 mm shim-pack GIS C18 chromatographic column, the particle size of the filler is 10 μm, the column temperature is 30 ° C, the mobile phase is a mixed eluent of acetonitrile and water, the elution mode is gradient elution, 0 min, the volume ratio of acetonitrile to water is 19:81; 55 min, the volume ratio of acetonitrile to water is 19:81; 60 min, the volume ratio of acetonitrile to water is 40:

60.

2. The method for preparing the passion fruit megastigmane glycoside compound according to claim 1, wherein: The high-speed continuous flow centrifuge described in step 1 has a rotation speed of 8000 r / min to 20000 r / min.

3. A use of a passion fruit megastigmane glycoside compound in the preparation of a flavoring product, characterized in that: The passion fruit megastigmane glycoside compound is selected from the following compounds: 。 4. The use according to claim 3, characterized in that: The dosage form is liquid preparation, semi-solid preparation or solid preparation.

5. The use according to claim 3, characterized in that: The product type is cigarette products.

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