A method for simultaneously extracting polysaccharide and triterpenes from poria cocos

Abstract

The application provides a method for simultaneously extracting polysaccharide and triterpenes in Poria cocos, and belongs to the technical field of biological extraction. The application uses specific first solvent and second solvent and water as solvents, increases the solubility of triterpenes in the upper phase, and makes the triterpenes enriched in the upper phase. Meanwhile, the alkali salt is used to dissolve the polysaccharide, so that the polysaccharide is enriched in the lower phase. In the process of extracting Poria cocos, the double water phase is formed, the triterpenes in the upper phase and the polysaccharide in the lower phase are simultaneously extracted and separated, the yield of the polysaccharide is close to 5%, the extraction rate is 79.9%, the yield of the triterpenes is 0.44% (4.4 mg / g), and the extraction rate is 63.7%. Moreover, the method of the application realizes the natural separation of the upper phase and the lower phase by using the solvent and the alkali salt, saves time, labor and materials, is simple to operate, and has a mild reaction, and has wide application value.

Description

Technical Field

[0001] This invention relates to the field of bioextraction technology, and in particular to a method for simultaneously extracting polysaccharides and triterpenes from Poria cocos. Background Technology

[0002] Poria cocos, the dried sclerotium of the fungus Poria cocos (family Polyporaceae), is sweet and bland in taste and neutral in nature. It has the effects of promoting diuresis and eliminating dampness, strengthening the spleen and stomach, and calming the mind and soothing the nerves. It is often used for edema with scanty urine, phlegm retention with dizziness and palpitations, and spleen deficiency with poor appetite. Poria cocos has been used as a traditional Chinese medicine and dietary supplement for over two thousand years, and there is a saying that "nine out of ten prescriptions use Poria cocos." Among the classic prescriptions using Poria cocos are Linggui Zhugan Decoction from *Synopsis of Prescriptions of the Golden Chamber*, Huopu Xialing Decoction from *Medical Origins*, and Chushi Weiling Decoction from *Golden Mirror of Medicine*.

[0003] Poria cocos contains effective components such as polysaccharides, triterpenes, and fatty acids. Poria cocos polysaccharides and triterpenes are rich in pharmacological activities, mainly concentrated in anti-inflammatory, anti-tumor, immunomodulatory, and hypoglycemic effects. Poria cocos polysaccharides also have pharmacological activities such as improving learning and memory, inhibiting myocardial hypertrophy in rats, and preventing stone formation. Poria cocos triterpenes also have sedative activities, reducing organ transplant rejection reactions, and improving polycystic ovary syndrome. Poria cocos polysaccharides and triterpenes have wide applications, and research has found that poria cocos polysaccharides may be developed into potential immune adjuvants.

[0004] Besides its medicinal value, Poria cocos also has edible value, making it a food and medicine homology substance. Common food products include Poria cocos cake, Poria cocos porridge, and Poria cocos pancakes. Triterpenes and polysaccharides can be isolated from Poria cocos and used in the development of functional products such as health supplements, food, and pharmaceuticals.

[0005] However, existing methods for extracting the effective components of Poria cocos all extract triterpenes and polysaccharides separately. For example, the existing technology "Research Progress on Extraction, Structure, Activity and Mechanism of Action of Poria cocos Polysaccharides, Liu Xingwen" discloses: 1. Water extraction method (Poria cocos powder → hot water reflux extraction → concentration → alcohol precipitation → drying → crude Poria cocos polysaccharide), which can only extract water-soluble polysaccharides, requires a long time, and has a low yield of 1.3%. 2. Alkaline extraction method: Poria cocos powder → alkaline solution → neutralization of extract with acetic acid → alcohol precipitation → drying → crude Poria cocos polysaccharide, which can only extract alkali-soluble polysaccharides, which are difficult to dissolve in water. 3. Enzymatic hydrolysis method: Poria cocos powder → adding buffer solution → adding enzyme → heating → enzyme inactivation → filtration → alcohol precipitation → crude Poria cocos polysaccharide; yield 3.56%. 4. Fermentation and alcohol precipitation method: intracellular polysaccharide fermentation broth → mycelium → alkaline solution → concentration → intracellular polysaccharide or fermentation broth → supernatant → concentration → alcohol precipitation → extracellular polysaccharide; the total sugar content of the polysaccharide obtained by this method is lower than that of natural Poria cocos polysaccharide. 5. Ultrasonic-assisted extraction: Poria cocos powder → soaking → ultrasonic-assisted extraction → concentration → crude polysaccharide; yield is low at 5.58%. 6. Microwave-assisted extraction: Poria cocos powder → soaking → microwave-assisted extraction → concentration → crude polysaccharide; yield is 2.96%. 7. Supercritical CO2 extraction: Poria cocos powder → supercritical carbon dioxide extraction → crude polysaccharide. Water-soluble Poria cocos polysaccharide extraction has limitations, yielding only 3.104%. For example, existing technologies disclose extraction methods for triterpenes from Poria cocos: ethanol reflux extraction and methanol reflux extraction. In these methods, the yields of triterpenes and polysaccharides from Poria cocos are low, and simultaneous extraction of triterpenes and polysaccharides cannot be achieved. Summary of the Invention

[0006] The purpose of this invention is to provide a method for simultaneously extracting polysaccharides and triterpenes from Poria cocos. The method is simple and can extract polysaccharides and triterpenes from Poria cocos at the same time, with high yields of polysaccharides and triterpenes.

[0007] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0008] This invention provides a method for simultaneously extracting polysaccharides and triterpenes from Poria cocos, comprising the following steps:

[0009] The alkaline salt and water are mixed, and a first solvent and a second solvent are added to the resulting solution to obtain an aqueous two-phase solution.

[0010] Poria cocos powder was added to the aqueous two-phase mixture, and after shaking extraction, the mixture was separated. The upper phase contained triterpenes, and the lower phase contained polysaccharides.

[0011] The first solvent includes ethanol or ethylene glycol; the second solvent includes acetone or acetonitrile.

[0012] Preferably, the alkaline salt includes potassium carbonate or dipotassium hydrogen phosphate.

[0013] Preferably, the volume ratio of the first solvent to the second solvent is 2-5:2-5.

[0014] Preferably, the ratio of the alkaline salt to the second solvent is 3.5–5.5 g: 2–5 mL.

[0015] Preferably, the mass ratio of the Poria cocos powder to the total volume of water, the first solvent, and the second solvent is 1g:15-50mL.

[0016] Preferably, the temperature for the oscillation extraction is 40–60°C.

[0017] Preferably, the oscillation extraction time is 30 to 80 minutes.

[0018] Preferably, the volume ratio of the first solvent to the second solvent is 2:1; the mass ratio of the Poria cocos powder to the total volume of water, the first solvent, and the second solvent is 1g:25mL; and the shaking extraction temperature is 48℃ and the time is 50min.

[0019] This invention provides a method for simultaneously extracting polysaccharides and triterpenes from Poria cocos. The method utilizes a specific first and second solvent, along with water, as solvents to increase the solubility of triterpenes in the upper phase, leading to their enrichment there. Simultaneously, the method leverages the dissolution of polysaccharides by alkali salts, resulting in their enrichment in the lower phase. This forms an aqueous two-phase extraction process, achieving simultaneous extraction and separation of triterpenes in the upper phase and polysaccharides in the lower phase. The polysaccharide yield is close to 5%, with an extraction rate of 79.9%, while the triterpenoid yield is 0.44% (4.4 mg / g), with an extraction rate of 63.7%. Furthermore, this method utilizes solvents and alkali salts to achieve natural separation of the upper and lower phases, saving time, labor, and materials. It is simple to operate, has a mild reaction, and possesses broad application value. Attached Figure Description

[0020] Figure 1 The graph shows the yield (a) and extraction rate (b) of polysaccharides and triterpenes under different conditions in Example 1.

[0021] Figure 2 The graph shows the yield (a) and extraction rate (b) of polysaccharides and triterpenes under different conditions in Example 2.

[0022] Figure 3 The graph shows the yield (a) and extraction rate (b) of polysaccharides and triterpenes under different conditions in Example 3.

[0023] Figure 4 The graph shows the yield (a) and extraction rate (b) of polysaccharides and triterpenes under different conditions in Example 4.

[0024] Figure 5 The graph shows the yield (a) and extraction rate (b) of polysaccharides and triterpenes under different conditions in Example 5.

[0025] Figure 6The graph shows the yield (a) and extraction rate (b) of polysaccharides and triterpenes under different conditions in Example 6.

[0026] Figure 7 The graph shows the yield (a) and extraction rate (b) of polysaccharides and triterpenes under different conditions in Example 7. Detailed Implementation

[0027] This invention provides a method for simultaneously extracting polysaccharides and triterpenes from Poria cocos, comprising the following steps:

[0028] The alkaline salt and water are mixed, and a first solvent and a second solvent are added to the resulting solution to obtain an aqueous two-phase solution.

[0029] Poria cocos powder was added to the aqueous two-phase mixture, and after shaking extraction, the mixture was separated. The upper phase contained triterpenes, and the lower phase contained polysaccharides.

[0030] The first solvent includes ethanol or ethylene glycol; the second solvent includes acetone or acetonitrile.

[0031] In this invention, unless otherwise specified, all raw materials required for preparation are commercially available products well known to those skilled in the art.

[0032] In this invention, the alkaline salt preferably includes potassium carbonate or dipotassium hydrogen phosphate. This invention utilizes the dissolution of polysaccharides by the alkaline salt to achieve polysaccharide extraction.

[0033] In this invention, the volume ratio of the first solvent to the second solvent is preferably 2-5:2-5, more preferably 3-4:2-4, and most preferably 2:1; the amount ratio of the alkali salt to the second solvent is preferably 3.5-5.5g:2-5mL, more preferably 4.0-5.0g:2-4mL, and even more preferably 5g:2mL.

[0034] In this invention, an alkaline salt and water are mixed, and a first solvent and a second solvent are added to the resulting solution. Preferably, the resulting mixture is allowed to stand and separate into two aqueous phases. This invention does not have a special limitation on the standing and separation time, as long as a clear separation interface is formed.

[0035] In this invention, the preferred method for preparing the Poria cocos powder includes: cutting Poria cocos blocks into 1 cm³ pieces. 3 The lumps were dried at 60℃ for 4 hours, then pulverized into fine powder using a pulverizer. The powder was then sieved through a No. 4 sieve with a mesh size of 60, and the sieved material was collected and stored in a dry environment to obtain Poria cocos powder.

[0036] In this invention, the mass ratio of the Poria cocos powder to the total volume of water, the first solvent, and the second solvent is preferably 1g:15-50mL, more preferably 1g:12.5-25mL, and even more preferably 1g:15-20mL.

[0037] In this invention, the temperature of the shaking extraction is preferably 40-60°C, more preferably 45-55°C, and even more preferably 45-48°C, and the shaking extraction time is preferably 30-80 min, more preferably 40-60 min, and even more preferably 45-50 min.

[0038] The present invention does not impose any special limitations on the oscillation conditions for the oscillation extraction; oscillation can be performed according to a process well known in the art.

[0039] As a preferred embodiment of the present invention, the volume ratio of the first solvent and the second solvent is 2:1; the mass ratio of the Poria cocos powder to the total volume of water, the first solvent, and the second solvent is 1g:25mL; and the shaking extraction temperature is 48℃ and the time is 50min.

[0040] After the shaking extraction is completed, the present invention preferably centrifuges or filters the resulting mixture to remove herbaceous fibers and separate them to obtain upper and lower phase materials. The present invention does not have any particular limitation on the filtration process; any process well known in the art can be followed.

[0041] In this invention, the obtained upper phase material mainly includes a first solvent and a second solvent; the lower phase material mainly includes an alkaline salt solution, with triterpenes enriched in the upper phase and polysaccharides enriched in the lower phase.

[0042] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0043] In the following embodiments,

[0044] 1. Polysaccharide content determination: Phenol-sulfuric acid method, refer to "Spectrophotometric Method for Determination of Ginseng Polysaccharides in Ginseng" DB 22 / T1685-2012

[0045] Concentrated sulfuric acid, commercially available, analytical grade, concentration 98wt%; Ethanol solution (80%): Add 80mL of anhydrous ethanol to 20mL of water and mix well.

[0046] Glucose standard (C6H) 12 O6 (CAS No.: 50-99-7): Purity ≥ 99.0%.

[0047] Glucose stock solution (10 mg / mL): Accurately weigh 1.0000 g of glucose standard dried to constant weight at 105℃, place it in a 100 mL volumetric flask, dissolve and dilute to the mark to prepare a 10 mg / mL standard stock solution.

[0048] Glucose working solution (0.1 mg / mL): Take 1 mL of the above (10 mg / mL) stock solution and dilute to 100 mL to prepare a 0.1 mg / mL working solution.

[0049] Phenol solution (50g / L): Take 5.0g of superior grade phenol reagent, dilute it with aqueous solution to 100mL volumetric flask, mix well, and store in a refrigerator at 4℃ for one month.

[0050] Construction of the standard working curve: Accurately transfer 0, 0.1, 0.2, 0.4, 0.6, 0.8, 1.0, 1.2, 1.4, and 1.6 mL of glucose standard solution into 25 mL stoppered graduated test tubes. Add water to 2.0 mL, then add 1.0 mL of 5% phenol solution to each tube and shake well. Quickly add 5 mL of concentrated H2SO4 solution and shake for 5 min. Let stand for 10 min, then heat in a boiling water bath for 20 min. Remove and cool to room temperature. Measure the absorbance at 486 nm, using the reagent blank as a reference.

[0051] Sample determination: The test solution is first diluted 20 times, then 0.2 mL is accurately transferred and 1.8 mL of water is added to a 25 mL stoppered graduated test tube. 1.0 mL of 5 wt% phenol solution is added and shaken well. 5 mL of concentrated sulfuric acid is quickly added, shaken for 5 min, left to stand for 10 min, heated in a boiling water bath for 20 min, removed and cooled to room temperature, and the absorbance is measured at 486 nm with the reagent blank as a reference.

[0052] 2. Determination of triterpenes: Refer to Appendix A of Hunan Provincial Local Food Safety Standard "Food Safety Standard for Poria" DBS43 / 014-2022: All reagents used in this method are of analytical grade, and the water is grade III water as specified in GB / T 6682.

[0053] Principle: Triterpenoids in Poria cocos react with vanillin under acidic conditions to produce a blue-purple product, which has maximum absorption at a wavelength of 550 nm. The absorbance value is directly proportional to the total triterpenoid content.

[0054] 5% Vanillin-Glacial Acetic Acid Solution: Weigh 5g of vanillin, dissolve it in 70mL of glacial acetic acid, and then dilute to 100mL with glacial acetic acid. Oleanolic Acid Standard: CAS No. 508-02-1, purity ≥99%.

[0055] Oleanolic acid standard stock solution: Accurately weigh 20 mg (accurate to 0.1 mg) of oleanolic acid standard dried to constant weight at 105℃, dissolve in methanol and dilute to 100 mL. The mass concentration of oleanolic acid in this standard solution is 200 μg / mL. Store in a sealed container at 4℃. Shelf life is 1 month.

[0056] Analytical balance: sensitivity 0.1 mg. Spectrophotometer: equipped with 1 cm cuvette, wavelength 550 nm.

[0057] Water bath: Temperature control range 60–100℃. Centrifuge: 8000 r / min.

[0058] Standard curve:

[0059] Accurately transfer 0 mL, 0.1 mL, 0.2 mL, 0.3 mL, 0.4 mL, and 0.5 mL of oleanolic acid standard stock solution into 10 mL test tubes, with standard masses of 0 μg, 20 μg, 40 μg, 60 μg, and 100 μg, respectively. Place the test tubes in a water bath at 90℃-100℃ to evaporate the solvent. Add 0.1 mL of 5% vanillin-glacial acetic acid solution and 0.8 mL of perchloric acid, mix well, and incubate in a 60℃ water bath for 20 min to develop the color. Remove the tubes and immediately cool them in an ice-water bath for 5 min to terminate the color development reaction. Add 5.0 mL of glacial acetic acid, mix well, and let stand at room temperature for 10 min. Immediately use a 1 cm cuvette, zeroing with a 0 tube, and measure the absorbance at 550 nm. Plot a standard curve with the mass of the oleanolic acid standard as the ordinate and absorbance as the abscissa.

[0060] Sample Measurement

[0061] Accurately transfer 0.4 mL of sample extract into a 10 mL test tube, place it in a water bath at 100 °C to evaporate the solvent, and simultaneously prepare a reagent blank. Calculate the total triterpenoid content based on the absorbance of the sample extract.

[0062] In the following examples, the yield is calculated as follows:

[0063] The yield (Y1, %) of triterpenes is calculated as shown in Equation 1:

[0064]

[0065] C1: Concentration of triterpenes in the extract, mg / mL;

[0066] V1: Volume of the upper phase, mL;

[0067] m: Mass of Poria cocos powder, in grams.

[0068] The yield (Y2, %) of polysaccharides is calculated as shown in Equation 2:

[0069]

[0070] C2: Concentration of polysaccharides in the extract, mg / mL;

[0071] V2: Volume of the lower phase, mL;

[0072] m: Mass of Poria cocos powder, in grams.

[0073] Calculation of extraction rate y:

[0074]

[0075] Y: Yield of polysaccharides or triterpenes, %;

[0076] m s The content of polysaccharides or triterpenes in Poria cocos, %.

[0077] In the following embodiments:

[0078] Preparation of Poria cocos powder: Cut Poria cocos pieces into 1cm cubes. 3 The lumps were dried at 60℃ for 4 hours, then pulverized into fine powder using a pulverizer. The powder was then sieved through a No. 4 sieve with a mesh size of 60, and the sieved material was collected and stored in a dry environment to obtain Poria cocos powder.

[0079] Example 1

[0080] Take nine 20mL stoppered test tubes and add the following to each:

[0081] 1#: Mix 4g of dipotassium hydrogen phosphate with 13mL of water, then add 5mL of ethanol, shake well, and let stand to separate into two aqueous phases.

[0082] 2#: Mix 4g of dipotassium hydrogen phosphate with 13mL of water, then add 5mL of ethanol and 2mL of acetone, shake well and let stand to separate into two aqueous phases.

[0083] 3#: Mix 4g of potassium carbonate with 13mL of water, then add 7mL of ethanol, shake well, and let stand to separate into two aqueous phases.

[0084] 4#: Mix 4g of potassium carbonate with 13mL of water, then add 5mL of ethanol and 2mL of acetone, shake well and let stand to separate into two aqueous phases.

[0085] 5#: Mix 4g of potassium carbonate with 13mL of water, then add 5mL of ethanol and 2mL of acetonitrile, shake well and let stand to separate into two aqueous phases.

[0086] 6#: Sodium hydroxide solution, 0.15mol / L, 13mL.

[0087] 7#: Pure water, 13mL.

[0088] 8#: Methanol, 13 mL.

[0089] 9#: Ethanol, 13 mL.

[0090] Add 0.5g of Poria cocos powder to each of the nine stoppered test tubes. After extracting the mixture by shaking at 48℃ for 40min, remove the herbaceous fibers by centrifugation from tubes #1 to #5, separate the upper and lower phases, measure the volume of each phase using a syringe, and transfer them to dry vials. Quantify the components of the upper and lower phases separately.

[0091] Products #6 and #7 were used for direct determination of triterpenoid and polysaccharide content; products #8 and #9 were used for determination of triterpenoids only (polysaccharides are insoluble in methanol and ethanol).

[0092] Test results show that the upper phase contains triterpenes and the lower phase contains polysaccharides.

[0093] Test results are available Figure 1 ;from Figure 1 As can be seen, under the same conditions, the aqueous solution of potassium carbonate + ethanol + acetone yielded the highest yield and extraction rate of triterpenes and polysaccharides, which was better than that of 0.15M sodium hydroxide, dipotassium hydrogen phosphate, and potassium carbonate. Moreover, the upper and lower phases naturally separate in this method, which is beneficial for the development of subsequent products.

[0094] Example 2

[0095] Different amounts of potassium carbonate (3.5, 4, 4.5, 5, 5.5, 6 g) and 13 mL of water were added to different 50 mL centrifuge tubes and mixed to form an aqueous solution. Then, 5 mL of ethanol and 2 mL of acetone were added, mixed, and allowed to stand to separate into two aqueous phases. Then, 0.5 g of Poria cocos powder was added, and the resulting mixture was extracted by shaking at 48 °C for 40 min. Herbal fibers were removed by centrifugation, and the upper and lower phases were separated.

[0096] The volumes of the upper and lower phases were measured using a syringe and transferred to dry vials. The triterpenes and polysaccharides in the upper and lower phases were then quantified separately. The results are shown in [Figure number missing]. Figure 2 ;Depend on Figure 2 It can be seen that when the amount of potassium carbonate used is 5g, the yield and extraction rate of polysaccharides and triterpenes are relatively high.

[0097] Example 3

[0098] Table 1 Raw material usage in Example 3

[0099] Potassium carbonate (g) Water volume (mL) Acetone volume (mL) Total volume (mL) 5g 14.5 0.5 20 5g 14 1 20 5g 13.5 1.2 20 5g 13 2 20 5g 12.5 2.5 20 5g 12 3 20 5g 11 4 20 5g 10 5 20

[0100] According to the dosage in Table 1, 5 mL of ethanol and 25% potassium carbonate were added to eight 50 mL centrifuge tubes, respectively. Different volumes of acetone (0.5, 1, 1.5, 2, 2.5, 3, 4, 5 mL) were added to maintain a total volume of 20 mL. The mixture was mixed and allowed to stand to separate into two aqueous phases. Then, 1 g of Poria cocos powder was added. The resulting mixture was extracted by shaking at 48 °C for 40 min. Herbal fibers were removed by filtration, and the upper and lower phases were separated.

[0101] The volumes of the upper and lower phases were measured, and the contents were transferred separately into dry vials using a syringe. The triterpenes in the upper phase and the polysaccharides in the lower phase were then quantitatively determined. The results are shown below. Figure 3 .

[0102] Depend on Figure 3 It can be seen that the optimal amount of acetone is 2 mL.

[0103] Example 4

[0104] Table 2 Raw material usage in Example 4

[0105]

[0106]

[0107] According to the dosage in Table 2, add 2 mL of acetone, 25% potassium carbonate (i.e., 5 g of potassium carbonate) and water to eight 50 mL centrifuge tubes, and add different volumes of ethanol (3, 3.5, 4, 5, 7, 8, 9, 10 mL) to maintain a total volume of 20 mL. Mix well and let stand to separate the layers to form an aqueous two-phase mixture. Then add 1 g of Poria cocos powder. After extracting the mixture by shaking at 48 °C for 40 min, remove the herbal fibers by filtration and separate the upper and lower phases.

[0108] The volumes of the upper and lower phases were measured and transferred separately into dry vials using a syringe. The triterpenes in the upper phase and the polysaccharides in the lower phase were quantitatively determined, and the results are shown below. Figure 4 .

[0109] Depend on Figure 4 It can be seen that the optimal amount of ethanol is 4 mL.

[0110] Example 5

[0111] Add 5g of potassium carbonate and 14mL of water to five 50mL centrifuge tubes respectively, shake well, add 2mL of acetone and 4mL of ethanol to maintain a total volume of 20mL, mix well and let stand to separate into two aqueous phases; add 1g of Poria cocos powder, and then extract the resulting mixture by shaking at 20℃, 30℃, 40℃, 48℃ and 60℃ for 40min respectively. After that, remove the herbal fiber by filtration and separate the upper phase and the lower phase.

[0112] The volumes of the upper and lower phases were measured and transferred separately into dry vials using a syringe. The triterpenes in the upper phase and the polysaccharides in the lower phase were quantitatively determined, and the results are shown below. Figure 5 .

[0113] Depend on Figure 5 It can be seen that the optimal extraction temperature is 48℃.

[0114] Example 6

[0115] Add 5g of potassium carbonate and 14mL of water to six 50mL centrifuge tubes, shake well, add 2mL of acetone and 4mL of ethanol to maintain a total volume of 20mL, mix well and let stand to separate into two aqueous phases, add 1g of Poria cocos powder, and then extract the resulting mixture at 48℃ for different times (20, 30, 40, 50, 60, 80 min). After that, remove the herbal fiber by filtration and separate the upper and lower phases.

[0116] The volumes of the upper and lower phases were measured, and the contents were transferred separately into dry vials using a syringe. The triterpenes in the upper phase and the polysaccharides in the lower phase were then quantitatively determined. The results are shown in [Figure number missing]. Figure 6 .

[0117] Depend on Figure 6 It can be seen that the optimal shaking time is 50 minutes.

[0118] Example 7

[0119] Add 5 g of potassium carbonate and 14 mL of water to six 50 mL centrifuge tubes, shake well, add 2 mL of acetone and 4 mL of ethanol to maintain a total volume of 20 mL, mix well, and allow to stand to separate into aqueous two phases. Then add different masses of Poria cocos powder (0.5, 0.8, 1, 1.3, 1.6, 2 g) to the centrifuge tubes, i.e., material-to-liquid ratios of 1:40, 1:25, 1:20, 1:15, 1:12.5, and 1:10 g:mL, respectively. Extract the resulting mixture at 48 °C with shaking for 50 min, then remove the herbal fibers by filtration, separating the upper and lower phases.

[0120] The volumes of the upper and lower phases were measured, and the contents were transferred separately into dry vials using a syringe. The triterpenes in the upper phase and the polysaccharides in the lower phase were then quantitatively determined. The results are shown in [Figure number missing]. Figure 7 .

[0121] Depend on Figure 7 It can be seen that the optimal material-to-liquid ratio is 1:25.

[0122] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A method for simultaneously extracting polysaccharides and triterpenes from Poria cocos, comprising the following steps: The alkaline salt and water are mixed, and a first solvent and a second solvent are added to the resulting solution to obtain an aqueous two-phase solution. Poria cocos powder was added to the aqueous two-phase mixture, and after shaking extraction, the mixture was separated. The upper phase contained triterpenes, and the lower phase contained polysaccharides. The first solvent is ethanol; the second solvent is acetone; the alkaline salt is potassium carbonate; The volume ratio of the first solvent to the second solvent is 2~5:2~5; The ratio of the alkaline salt to the second solvent is 3.5~5.5g:2~5mL; The mass ratio of the Poria cocos powder to the total volume of water, the first solvent, and the second solvent is 1g:15~50mL.

2. The method according to claim 1, characterized in that, The temperature for the oscillation extraction is 40~60℃.

3. The method according to claim 1 or 2, characterized in that, The oscillation extraction time is 30~80 minutes.

4. The method according to claim 1, characterized in that, The volume ratio of the first solvent to the second solvent is 2:1; the mass ratio of the Poria cocos powder to the total volume of water, the first solvent, and the second solvent is 1g:25mL; the shaking extraction temperature is 48℃ and the time is 50min.

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