Method for extracting Taiwanin E and Diphyllin from barks of Taiwanin

Abstract

The invention belongs to the technical field of substance extraction, and provides a method for extracting Taiwanin E and Diphyllin from barks of Taiwanin. According to the method disclosed by the invention, the taiwania flousiana bark powder is extracted by utilizing the ethanol solution, so that lignans in the taiwania flousiana bark powder can be preliminarily extracted; and the obtained extract liquid is subjected to reflux extraction under the condition that the volume concentration of ethanol is 60-90%, so that Taiwanin E and Diphyllin can be extracted more efficiently. The method comprises the following steps: extracting Taiwanin E and Diphyllin into an ethyl acetate layer through ethyl acetate extraction, and removing part of impurities; and then the ethyl acetate layer is subjected to vacuum concentration and recrystallization, so that the purity of Taiwanin E and Diphyllin can be further improved. According to the preparation method provided by the invention, the extraction rate of Taiwanin E and Diphyllin is high.

Description

Technical Field

[0001] The invention relates to the technical field of material extraction, in particular to a method for extracting Taiwanin E and Diphyllin from the stem bark of Taiwania baldwinii. Background Art

[0002] Taiwanin E is an important biologically active substance with a molecular formula of C 20 H 12 O 7 , molecular weight is 364.06, and the structural formula is shown in Formula 1. It has insecticidal activity against the second, third, and fourth instar larvae of Aedes aegypti, herbicidal activity, and certain cancer stem cell inhibitory activity.

[0003]

[0004] Diphyllin is a kind of aromatic naphthyl lignan lactone that can be isolated from many traditional medicinal plants. Its molecular formula is C 21 H 16 O 7 , molecular weight is 380.09, and the structural formula is shown in Formula 2. Studies have found that it has rich biological activities, and Diphyllin and its derivatives have V-ATPase inhibition, anti-tumor, anti-viral, anti-inflammatory and antioxidant activities.

[0005]

[0006] Currently, there is a problem of low extraction rate in extracting Taiwanin E and Diphyllin from medicinal plants. Summary of the invention

[0007] In view of this, the object of the present invention is to provide a method for extracting Taiwanin E and Diphyllin from the stem bark of Taiwania baldness. The method provided by the present invention has a high extraction rate for Taiwanin E and Diphyllin.

[0008] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0009] The present invention provides a method for extracting Taiwanin E and Diphyllin from the stem bark of Taiwania truncatula, comprising the following steps:

[0010] Extracting the Taiwania truncatula bark powder with an ethanol solution to obtain an extract;

[0011] The extract is subjected to reflux extraction under the condition that the volume concentration of ethanol is 60 to 90% to obtain an extract;

[0012] Concentrating the extract under reduced pressure to obtain an extract concentrate;

[0013] After the extract is dissolved in water, it is extracted with ethyl acetate, and the ethyl acetate layer is collected;

[0014] The ethyl acetate layer is sequentially subjected to vacuum concentration and recrystallization to obtain an extract containing Taiwanin E and Diphyllin.

[0015] Preferably, the particle size of the Taiwania baldnessii stem bark powder is 0.02-20 mm, and the volume concentration of the ethanol solution is 90-95%.

[0016] Preferably, the usage ratio of the Taiwania baldnessii stem bark powder and the ethanol solution is 30-75 g: 631.6-1578.9 mL.

[0017] Preferably, the leaching temperature is 65-70° C. and the leaching time is 18-36 hours.

[0018] Preferably, the reflux extraction is performed 2 to 4 times, and the time of each reflux extraction is greater than or equal to 1 hour.

[0019] Preferably, the reduced pressure concentration is performed until there is no alcohol smell.

[0020] Preferably, the ethyl acetate extraction is an equal volume extraction, and the number of ethyl acetate extractions is 4 to 6 times.

[0021] Preferably, the temperature of the vacuum concentration is 45-50°C.

[0022] Preferably, the recrystallization reagent is a mixed solvent of dichloromethane and methanol, and the volume ratio of dichloromethane to methanol in the mixed solvent of dichloromethane and methanol is 1:1;

[0023] The dissolution temperature of the recrystallization is 45 to 55°C;

[0024] The crystallization temperature of the recrystallization is room temperature;

[0025] The number of times of the recrystallization is 2 to 4 times.

[0026] The present invention also provides an extract containing Taiwanin E and Diphyllin obtained by the method described in the above technical scheme, wherein the mass content of Taiwanin E in the extract containing Taiwanin E and Diphyllin is 47.32-47.36%, and the mass content of Diphyllin is 46.91-46.95%.

[0027] The invention provides a method for extracting Taiwanin E and Diphyllin from the stem bark of Taiwania truncatula.

[0028] The present invention uses an ethanol solution to extract the Taiwania baldness bark powder, and the extraction can preliminarily extract the lignans in the Taiwania baldness bark powder; then the extract is subjected to reflux extraction under the condition that the ethanol volume concentration is 60-90%, and Taiwanin E and Diphyllin can be extracted more efficiently. Ethyl acetate extraction can extract Taiwanin E and Diphyllin into an ethyl acetate layer and remove some impurities; and the ethyl acetate layer is then vacuum concentrated and recrystallized to further improve the purity of Taiwanin E and Diphyllin. The preparation method provided by the present invention has a high extraction rate for Taiwanin E and Diphyllin. At the same time, the method of the present invention is simple, easy to operate, and suitable for industrial production. Furthermore, before the ethyl acetate extraction, the present invention performs reduced pressure concentration and then performs equal volume extraction, thereby saving the use of ethyl acetate. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 The HPLC detection spectrum of the obtained extract is shown in FIG.

[0030] Figure 2 is the standard curve of Taiwanin E;

[0031] Figure 3 This is the standard curve of Diphyllin;

[0032] Figure 4 This is the HPLC detection spectrum of TaiwaninE obtained in Example 1;

[0033] Figure 5 This is the HPLC detection spectrum of Diphyllin obtained in Example 1;

[0034] Figure 6 HR-ESI-MS high-resolution mass spectrum of the extract Taiwanin E of Example 1;

[0035] Figure 7 The extract of Taiwanin E in Example 1 1 H-NMR spectrum;

[0036] Figure 8 The extract of Taiwanin E in Example 1 13 C-NMR spectrum;

[0037] Fig. 9HSQC spectrum of the extract Taiwanin E of Example 1;

[0038] Fig.10 HR-ESI-MS high resolution mass spectrum of the extract Diphyllin of Example 1;

[0039] Fig.11 The extract of Diphyllin in Example 1 1 H-NMR spectrum;

[0040] Fig.12 The extract of Diphyllin in Example 1 13 C-NMR spectrum;

[0041] Fig.13 This is the HSQC spectrum of the extract Diphyllin of Example 1. DETAILED DESCRIPTION

[0042] The present invention provides a method for extracting Taiwanin E and Diphyllin from the stem bark of Taiwania truncatula, comprising the following steps:

[0043] Extracting the Taiwania truncatula bark powder with an ethanol solution to obtain an extract;

[0044] The extract is subjected to reflux extraction under the condition that the volume concentration of ethanol is 60 to 90% to obtain an extract;

[0045] Concentrating the extract under reduced pressure to obtain an extract concentrate;

[0046] After the extract is dissolved in water, it is extracted with ethyl acetate, and the ethyl acetate layer is collected;

[0047] The ethyl acetate layer is sequentially subjected to vacuum concentration and recrystallization to obtain an extract containing Taiwanin E and Diphyllin.

[0048] Unless otherwise specified, the raw materials used in the present invention are preferably commercially available products.

[0049] The invention extracts the Taiwania baldnessii stem bark powder with an ethanol solution to obtain an extract.

[0050] In the present invention, the particle size of the Taiwania baldukki bark powder is preferably 0.02 to 20 mm, more preferably 0.1 to 10 mm, and particularly preferably 2 to 5 mm. In the present invention, the method for preparing the Taiwania baldukki bark powder preferably comprises the following steps: crushing the raw Taiwania baldukki bark to obtain the Taiwania baldukki bark powder. The present invention does not specifically limit the parameters of the crushing, as long as the Taiwania baldukki bark powder with a particle size of 0.02 to 20 mm can be obtained. In the present invention, the volume concentration of the ethanol solution is preferably 90 to 95%, particularly preferably 95%.

[0051] In the present invention, the usage ratio of the Taiwania baldnessii stem bark powder and the ethanol solution is preferably 30-75 g:631.6-1578.9 mL, specifically preferably 30 g:631.6 mL or 75 g:1578.9 mL.

[0052] In the present invention, the leaching temperature is preferably 65-70° C., and the leaching time is preferably 18-36 h, specifically preferably 18 h, 24 h, 30 h or 36 h.

[0053] In the present invention, the leaching is preferably carried out in an extraction tank.

[0054] After the leaching, the present invention preferably does not perform any operation to obtain the leaching solution.

[0055] In the present invention, the extraction can extract the lignan components with important pharmacological activities from the stem bark of Taiwania truncatula, and Taiwanin E and Diphyllin are representative components of this type of compound.

[0056] After obtaining the leaching solution, the present invention performs reflux extraction on the leaching solution under the condition that the ethanol volume concentration is 60-90% to obtain the extract.

[0057] In the present invention, the extract itself contains ethanol of a certain volume concentration; based on the requirement for the volume concentration of ethanol in the reflux extraction process, the extract can be treated, and the treatment preferably includes dilution with water, addition of ethanol to increase concentration, or direct reflux extraction without any treatment.

[0058] In the present invention, during the reflux extraction process, the volume concentration of ethanol is 60-90%, and is preferably 60%, 70%, 80% or 90%.

[0059] In the present invention, the number of reflux extractions is preferably 2 to 4 times, more preferably 3 times; the time for each reflux extraction is preferably greater than or equal to 1 hour, more preferably 1 hour.

[0060] After the reflux extraction, the present invention preferably further comprises: cooling the obtained reflux extraction liquid to 30-40°C and filtering to obtain the extract. In the present invention, the filtration equipment preferably comprises a vacuum pump and a Buchner funnel.

[0061] In the present invention, the reflux extraction can significantly improve the extraction efficiency of the target compound. This method is more efficient than a single immersion or percolation method because it allows the solvent to pass through the raw material multiple times, thereby increasing the extraction opportunity and completely extracting the main components contained in the stem bark of Taiwania truncatula.

[0062] After obtaining the extract, the present invention concentrates the extract under reduced pressure to obtain an extract concentrate.

[0063] In the present invention, the vacuum concentration is preferably performed until there is no alcohol taste. The parameters of the vacuum distillation are not specifically limited in the present invention, as long as there is no alcohol taste.

[0064] After obtaining the extract, the present invention dissolves the extract in water, extracts it with ethyl acetate, and collects the ethyl acetate layer.

[0065] In the present invention, the water-soluble water is preferably pure water.

[0066] In the present invention, the ethyl acetate extraction is preferably an equal volume extraction. In the present invention, the volume ratio of the water-soluble water to the ethyl acetate extracted with ethyl acetate is preferably 1:1. In the present invention, the number of ethyl acetate extractions is preferably 4 to 6 times, more preferably 5 times.

[0067] After collecting the ethyl acetate layer, the present invention sequentially performs vacuum concentration and recrystallization on the ethyl acetate layer to obtain an extract containing Taiwanin E and Diphyllin.

[0068] In the present invention, the temperature of the vacuum concentration is preferably 45-50° C., and the time of the vacuum concentration is not specifically limited as long as the solvent can be evaporated.

[0069] In the present invention, the reagent for recrystallization is preferably a mixed solvent of dichloromethane and methanol, and the volume ratio of dichloromethane and methanol in the mixed solvent of dichloromethane and methanol is preferably 1:1; the present invention does not specifically limit the amount of the reagent for recrystallization, as long as the extract extract obtained by vacuum concentration can be completely dissolved. In the present invention, the dissolution temperature of the recrystallization is preferably 45 to 55°C, and specifically preferably 45°C, 50°C or 55°C. In the present invention, the crystallization temperature of the recrystallization is preferably room temperature. In the present invention, the number of recrystallizations is preferably 2 to 4 times, and more preferably 3 times. In the present invention, the crystallization time of each recrystallization is preferably 6 to 18 hours, and more preferably 12 hours. In the present invention, the crystallization method of the recrystallization is preferably standing.

[0070] In the present invention, the recrystallization process is preferably as follows: the extract obtained by vacuum concentration is completely dissolved with a recrystallization reagent at a dissolution temperature; then crystallization is performed at a crystallization temperature; after the crystallization is completed, filtering is performed to obtain a powder and a filtrate; the filtrate is heated to a dissolution temperature, then crystallized at the crystallization temperature, and after the crystallization is completed, filtering is performed to obtain a powder and a filtrate; the process of heating the filtrate is repeated, and the powder collected is the extract containing TaiwaninE and Diphyllin.

[0071] The method provided by the invention has high extraction rate for Taiwanin E and Diphyllin.

[0072] After obtaining the extract containing Taiwanin E and Diphyllin, the present invention preferably further comprises separating and purifying the extract containing Taiwanin E and Diphyllin, and the separation and purification preferably comprises the following steps:

[0073] Dissolve the extract containing Taiwanin E and Diphyllin to obtain a sample for the machine;

[0074] The sample was purified by high performance liquid chromatography to obtain Taiwanin E and Diphyllin, respectively.

[0075] In the present invention, the dissolving reagent is preferably a mixed solvent of methanol and acetonitrile, and the volume ratio of methanol to acetonitrile in the mixed solvent of methanol and acetonitrile is 2:1.

[0076] In the present invention, the equipment for high performance liquid chromatography purification is preferably an Agilent 1260 high performance liquid chromatograph.

[0077] In the present invention, the parameters of the HPLC purification include: the mobile phase includes mobile phase A and mobile phase B, the mobile phase A is preferably methanol, and the mobile phase B is preferably pure water; the gradient elution program is preferably: 0 to 40 minutes, the volume fraction of mobile phase A changes from 45% to 100% at a constant rate; the flow rate is preferably 1.0 mL / min; the detection wavelength is preferably 290 nm; the column temperature is preferably 30°C; and the injection volume is preferably 10 μL.

[0078] The present invention also provides an extract containing Taiwanin E and Diphyllin obtained by the method described in the above technical scheme, wherein the mass content of Taiwanin E in the extract containing Taiwanin E and Diphyllin is 47.32-47.36%, and the mass content of Diphyllin is 46.91-46.95%.

[0079] The extract containing Taiwanin E and Diphyllin provided by the invention has high mass content of Taiwanin E and Diphyllin.

[0080] The following is a detailed description of the method for extracting Taiwanin E and Diphyllin from the stem bark of Taiwania baldnesswood provided by the present invention in conjunction with the examples, but they should not be construed as limiting the scope of protection of the present invention.

[0081] Example 1

[0082] A method for extracting Taiwanin E and Diphyllin from the stem bark of Taiwania truncatula, comprising the following steps:

[0083] (1) Grinding: Take 30.0 g of the raw material of Taiwania truncatum stem bark and grind it into the size of soybeans to obtain Taiwania truncatum stem bark powder with a particle size of 2 to 5 mm.

[0084] (2) Extraction: Add 30.0 g of Taiwania truncatula bark powder and 631.6 mL of 95% ethanol solution into an extraction tank, heat to 65° C. and soak for 24 h to obtain an extract.

[0085] (3) Extraction: Add 368.4 mL of pure water into the extraction tank, dilute the ethanol to a volume concentration of 60%, reflux extract at 90° C. for 3 times, each extraction time is 1 h, and combine the reflux extraction liquid.

[0086] (4) Filtration: Cool the refluxed extract to 40°C, and then use a vacuum pump and a Buchner funnel to filter to obtain an extract.

[0087] (5) Concentration under reduced pressure: The extract was concentrated under reduced pressure until there was no alcohol taste, and 2.9 g of extract concentrate and 2.6 L of recovered ethanol were obtained.

[0088] (6) Extraction: 2.9 g of the extract obtained in step (5) was completely dissolved in 200 mL of pure water, extracted five times with 200 mL of ethyl acetate, and 1 L of the ethyl acetate layer was collected.

[0089] (7) Vacuum concentration: 1 L of the ethyl acetate layer from step (6) was vacuum concentrated at 50°C, and the solvent was evaporated to dryness to obtain 0.9956 g of extract. The obtained extract was subjected to HPLC detection, and the results were as follows: Figure 1 As shown, from Figure 1 It can be seen that the extract of the ethyl acetate part is rich in components, and the components are separated in the 290nm band, with a strong absorption peak near 19.6min and 23.5min respectively.

[0090] (8) Recrystallization: At 50°C, use 20 mL of a mixed solvent of dichloromethane and methanol in a volume ratio of 1:1 to completely dissolve the extract of step (7); then let it stand at room temperature for 12 h to crystallize, filter out the precipitated powder, and repeat 3 times until precipitation is complete to obtain 49.2 mg of a white powdery solid.

[0091] Taiwanin standard was dissolved in a mixed solvent of methanol and acetonitrile with a volume ratio of 2:1 to prepare Taiwanin standard solutions with concentrations of 0.2 mg / mL, 0.4 mg / mL, 0.6 mg / mL, 0.8 mg / mL and 1.0 mg / mL. The Taiwanin standard solutions of various concentrations were determined by high performance liquid chromatography detection, and a standard curve was established with the concentration of the Taiwanin standard solution as the abscissa and the peak area corresponding to the Taiwanin standard solution as the ordinate. The results are shown in FIG. Figure 2 As shown, Figure 2 is the standard curve of Taiwanin E.

[0092] The Diphyllin standard was dissolved in a mixed solvent of methanol and acetonitrile in a volume ratio of 2:1 to prepare Diphyllin standard solutions with concentrations of 0.2 mg / mL, 0.4 mg / mL, 0.6 mg / mL, 0.8 mg / mL and 1.0 mg / mL. The Diphyllin standard solutions of various concentrations were determined by high performance liquid chromatography detection, and a standard curve was established with the concentration of the Diphyllin standard solution as the horizontal axis and the peak area of ​​the corresponding Diphyllin standard solution as the vertical axis. The results are as follows: Figure 3 As shown, Figure 3 This is the standard curve of Diphyllin.

[0093] The obtained 49.2 mg white powder solid was subjected to high performance liquid chromatography (HPLC) and then Figure 2 and Figure 3 The standard curve was used for quantification, and the results were as follows: the white powder solid contained 23.3 mg of Taiwanin E, with a content of 47.36%; and 23.1 mg of Diphyllin, with a content of 46.95%. Among them, the extraction rate of Taiwanin E = 23.3 mg / 30.0 g = 0.078%, and the extraction rate of Diphyllin = 23.1 mg / 30.0 g = 0.077%.

[0094] (9) Purification: The white powdery solid obtained in (8) was purified. The purification parameters included: the white powder obtained in (8) was dissolved in a mixed solvent of methanol and acetonitrile in a volume ratio of 2:1, and then gradient elution was performed using an Agilent 1260 high performance liquid chromatograph. The mobile phases included mobile phase A and mobile phase B, wherein mobile phase A was methanol and mobile phase B was pure water. The gradient elution program was: 0 to 40 min, the volume fraction of mobile phase A was uniformly changed from 45% to 100%, the flow rate was 1.0 mL / min; the detection wavelength was 290 nm; the column temperature was 30° C.; the injection volume was 10 μL, and Taiwanin E and Diphyllin were obtained.

[0095] The obtained Taiwanin E and Diphyllin were tested, and the results were as follows: Figures 4 to 13 As shown, Figure 4 The HPLC detection spectrum of the obtained Taiwanin E; Figure 5 is the HPLC detection spectrum of the obtained Diphyllin; Figure 6 is the HR-ESI-MS high-resolution mass spectrum of the obtained Taiwanin E; Figure 7 For the obtained Taiwanin E 1 H-NMR spectrum; Figure 8 For the obtained TaiwaninE 13 C-NMR spectrum; Fig. 9 is the HSQC spectrum of Taiwanin E obtained; Fig.10 is the HR-ESI-MS high-resolution mass spectrum of the obtained Diphyllin; Fig.11 For the obtained Diphyllin 1 H-NMR spectrum; Fig.12 For the obtained Diphyllin 13 C-NMR spectrum; Fig.13 The HSQC spectrum of Diphyllin is obtained from Figures 4 to 13It can be seen that the retention time of pure TaiwaninE is 23.449 min, and its molecular formula is C 20 H 12 O 7 , the molecular weight is 364.06, and the structural formula is shown in Formula 1; the retention time of pure Diphyllin is 19.592min, and its molecular formula is C 21 H 16 O 7 , molecular weight is 380.09, and the structural formula is as shown in Formula 2.

[0096] Example 2

[0097] A method for extracting Taiwanin E and Diphyllin from the stem bark of Taiwania truncatula, comprising the following steps:

[0098] (1) Grinding: Take 75.0 g of the raw material of Taiwania truncatum stem bark and grind it into the size of soybeans to obtain Taiwania truncatum stem bark powder with a particle size of 2 to 5 mm.

[0099] (2) Soaking: Add 75.0 g of Taiwania truncatula bark powder and 1578.9 mL of 95% ethanol solution into an extraction tank, heat to 65° C. and soak for 24 hours to obtain an extract.

[0100] (3) Extraction: Add 921.1 mL of pure water into the extraction tank, dilute the ethanol to a volume concentration of 60%, perform reflux extraction at 90° C. for 3 times, each extraction time is 1 h, and combine the reflux extraction liquid.

[0101] (4) Filtration: Cool the refluxed extract to 40°C, and then use a vacuum pump and a Buchner funnel to filter to obtain an extract.

[0102] (5) Concentration under reduced pressure: The extract was concentrated under reduced pressure until there was no alcohol taste, and 3.9 g of extract concentrate and 6.6 L of recovered ethanol were obtained.

[0103] (6) Extraction: 3.9 g of the extract obtained in step (5) was completely dissolved in 300 mL of pure water, extracted five times with 300 mL of ethyl acetate, and 1.5 L of the ethyl acetate layer was collected.

[0104] (7) Vacuum concentration: 1.5 L of the ethyl acetate layer from step (6) was concentrated under vacuum at 50° C., and the solvent was evaporated to dryness to obtain 1.6573 g of extract.

[0105] (8) Recrystallization: At 50°C, use 50 mL of a mixed solvent of dichloromethane and methanol in a volume ratio of 1:1 to completely dissolve the extract of step (7); then let it stand at room temperature for 12 h to crystallize, filter out the precipitated powder, and repeat 3 times until precipitation is complete to obtain 97.2 mg of a white powdery solid.

[0106] The obtained white powdery solid contained 46.0 mg of Taiwanin E, with a content of 47.32%; and 45.6 mg of Diphyllin, with a content of 46.91%.

[0107] Among them, the extraction rate of Taiwanin E = 46 mg / 75 g = 0.061%, and the extraction rate of Diphyllin = 45.6 mg / 75 g = 0.0608%.

[0108] Comparative Example 1

[0109] After the collection of the stem bark of Taiwania baldness, it was first dried in the shade (92kg), then placed in an oven and heated at 60°C to dry, and then crushed to pass through a 40-mesh sieve. The plant dry powder was extracted with 5 times the volume (V / m) of methanol, each extraction was 48 hours, repeated 3 times, and the filtrates were combined and concentrated to obtain a methanol extract. The extract was dissolved in a small amount of methanol, suspended in 4 times the volume of deionized water, and then extracted with petroleum ether and ethyl acetate 3 times each, and the petroleum ether layer (110g) and ethyl acetate layer (650g) were obtained by combining and concentrating.

[0110] The ethyl acetate layer extract was separated into 9 samples Fr.1 to Fr.9 by silica gel column chromatography (chloroform / methanol = 1:0 → 0:1). Fr.3 (165 g) was further subjected to normal phase silica gel column chromatography (petroleum ether / acetone = 99:1 → 50:50), and after TLC detection and merging, 10 samples Fr.3.1 to Fr.3.10 were obtained. Among them, Fr.3.2 (6.6 g) was subjected to reverse phase medium pressure chromatography (methanol / water = 40% → 100%) to obtain 4 components Fr.3.2.1 to Fr.3.2.4, of which the second component (Fr.3.2.2) was purified by dextran gel, eluted with chloroform: methanol = 1:1, and obtained compound 1 (Taiwanin E, 34 mg) and compound 2 (Taiwanin H, 23 mg). After calculation, the extraction rate of compound 1Taiwanin E was 34mg / 92kg=3.70×10 -5 %.

[0111] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A method for extracting Taiwanin E and Diphyllin from the stem bark of Taiwania truncatula, comprising the following steps: Extracting the Taiwania truncatula bark powder with an ethanol solution to obtain an extract; The extract is subjected to reflux extraction under the condition that the volume concentration of ethanol is 60 to 90% to obtain an extract; Concentrating the extract under reduced pressure to obtain an extract concentrate; After the extract is dissolved in water, it is extracted with ethyl acetate, and the ethyl acetate layer is collected; The ethyl acetate layer is sequentially subjected to vacuum concentration and recrystallization to obtain an extract containing Taiwanin E and Diphyllin.

2. The method according to claim 1, characterized in that The particle size of the Taiwania baldnessii stem bark powder is 0.02-20 mm, and the volume concentration of the ethanol solution is 90-95%.

3. The method according to claim 1 or 2, characterized in that: The usage ratio of the Taiwania baldnessii stem bark powder and the ethanol solution is 30-75 g: 631.6-1578.9 mL.

4. The method according to claim 1, characterized in that: The extraction temperature is 65-70° C. and the extraction time is 18-36 hours.

5. The method according to claim 1, characterized in that The number of reflux extractions is 2 to 4 times, and the time of each reflux extraction is greater than or equal to 1 hour.

6. The method according to claim 1, characterized in that The reduced pressure concentration is performed until there is no alcohol smell.

7. The method according to claim 1, characterized in that The ethyl acetate extraction is an equal volume extraction, and the number of ethyl acetate extractions is 4 to 6 times.

8. The method according to claim 1, characterized in that The temperature of the vacuum concentration is 45-50°C.

9. The method according to claim 1, characterized in that: The recrystallization reagent is a mixed solvent of dichloromethane and methanol, wherein the volume ratio of dichloromethane to methanol in the mixed solvent is 1:1; The dissolution temperature of the recrystallization is 45 to 55°C; The crystallization temperature of the recrystallization is room temperature; The number of times of the recrystallization is 2 to 4 times.

10. An extract containing Taiwanin E and Diphyllin obtained by the method according to any one of claims 1 to 9, characterized in that: In the extract containing Taiwanin E and Diphyllin, the mass content of Taiwanin E is 47.32-47.36%, and the mass content of Diphyllin is 46.91-46.95%.

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