Composite extraction process of wolfberry combined polyphenol
By employing a step-by-step process involving pretreatment of wolfberry powder, compound enzymatic hydrolysis, and ultrasonic treatment, the problems of low extraction rate and easy loss of bioactivity of bound polyphenols from wolfberry are solved, achieving efficient and green extraction of bound polyphenols, which is suitable for industrial production.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- JIANGNAN UNIV
- Filing Date
- 2026-03-17
- Publication Date
- 2026-05-19
AI Technical Summary
Existing technologies for extracting polyphenols from wolfberry are difficult, have low extraction rates, are prone to loss of bioactivity, and involve complex processes. They also pose problems such as solvent residue and environmental pollution, and cannot meet the needs of efficient and green industrial production.
After pretreatment of wolfberry powder, the method of extraction by ethanol reflux, compound enzymatic hydrolysis and ultrasonic treatment is combined. The enzymatic hydrolysis and ultrasonic treatment are carried out in steps. First, the cell wall structure is degraded by compound enzymes (cellulase, pectinase and acidic protease), then the bound polyphenols are released by ultrasonic treatment, and finally the bound polyphenols are extracted and concentrated by organic solvents to achieve efficient extraction of bound polyphenols.
It achieves efficient extraction of polyphenols, improves extraction rate and enhances bioactivity, and the process is green and controllable, suitable for large-scale production, providing a high-value utilization pathway for wolfberry resources.
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Abstract
Description
Technical Field
[0001] This invention relates to the field of wolfberry polyphenol extraction technology, specifically to a compound extraction process combining wolfberry with polyphenols. Background Technology
[0002] Polyphenols, as widely distributed natural active ingredients in the plant kingdom, have become a research hotspot and core development target in the food, pharmaceutical, and health product fields due to their excellent core biological functions such as antioxidation, anti-inflammation, regulation of metabolism, and immune regulation. They have irreplaceable value in safeguarding human health and expanding the application of natural products. Polyphenols in plants mainly exist in two forms: free and bound. Bound polyphenols refer to complexes formed with cell wall polysaccharides, proteins, and other macromolecules through ester bonds, glycosidic bonds, or hydrophobic interactions. Compared to free polyphenols, bound polyphenols have more significant biological activity and metabolic stability, and can exert longer-lasting physiological functions during digestion and absorption in the body. They represent a high-value portion of plant polyphenol resources that has not yet been fully explored.
[0003] Goji berries, a traditional Chinese food and medicine, are rich in various active ingredients such as phenols, polysaccharides, and flavonoids. Among these, polyphenols are the core material basis for goji berries' antioxidant and immunomodulatory pharmacological effects, giving them extremely high development and utilization value. However, current research and applications of goji berry polyphenols, both domestically and internationally, mostly focus on the extraction and characterization of free polyphenols. Systematic research on bound polyphenols, which have superior biological activity, is relatively scarce, resulting in insufficient utilization of goji berry resources and a waste of high-value components.
[0004] The tight binding of bound polyphenols to plant cell wall macromolecules makes their extraction significantly more difficult than that of free polyphenols. Current technologies for extracting bound polyphenols largely rely on acid-base regulation. While this can break some ester and glycosidic bonds, a key technological bottleneck remains: precise control of acid-base conditions is difficult, easily leading to polyphenol structural degradation, reduced product bioactivity, and potential solvent residue and environmental pollution. This fails to meet the demands of efficient and green industrial production. Summary of the Invention
[0005] To address the aforementioned problems, the purpose of this invention is to provide a composite extraction process for wolfberry combined with polyphenols, offering a reference and new ideas for the in-depth exploration of the physiologically active substances in wolfberry and the application and product development of medicinal and edible by-products.
[0006] To achieve the above objectives, the present invention first provides a compound extraction process for wolfberry combined with polyphenols, comprising the following steps: S1. Raw material pretreatment: The dried wolfberries are crushed and sieved to obtain wolfberry powder; the wolfberry powder is extracted by reflux with ethanol solution, and the wolfberry residue is collected and dried after solid-liquid separation to obtain pretreated wolfberry residue; S2. Enzymatic hydrolysis-ultrasound combined extraction: The pretreated wolfberry residue obtained in step S1 is mixed with water, the pH value is adjusted, and a compound enzyme is added for enzymatic hydrolysis; after enzymatic hydrolysis, ultrasonic treatment is performed, and the supernatant is collected after solid-liquid separation; the compound enzyme includes cellulase, pectinase and acidic protease. S3. Extraction and Concentration: Adjust the pH of the supernatant obtained in step S2 to acidic, extract with an organic solvent, collect the extract and concentrate it to obtain a wolfberry polyphenol extract.
[0007] In one embodiment of the present invention, in step S1, the sieve mesh size is 15-20 mesh, the volume fraction of the added ethanol solution is 60-80%, the material-to-liquid ratio of wolfberry powder to the added ethanol solution is 1g:10-15mL, the reflux extraction temperature is 70-90℃, the reflux extraction time is 60-90min, and the solid-liquid separation is centrifuged at 4000rpm for 15min.
[0008] In one embodiment of the present invention, in step S1, the above reflux extraction-solid-liquid separation steps are repeated 2 to 4 times until no phenolic substances are detected in the final supernatant; the drying is performed at 50°C for 24-36 hours.
[0009] In one embodiment of the present invention, in step S2, the solid-liquid ratio of the pretreated wolfberry residue to water is 1:15~20 g / mL; the pH value is adjusted to 4~5; the temperature during enzymatic hydrolysis is 50-55℃, and the enzymatic hydrolysis time is 1~3h.
[0010] In one embodiment of the present invention, in step S2, the mass ratio of cellulase, pectinase and acidic protease is 1~2:3:2~3, preferably 1:3:3.
[0011] In one embodiment of the present invention, in step S2, the temperature during ultrasonic treatment is 50~60℃, the ultrasonic power is 240~300W, preferably 270W, and the ultrasonic time is 45~60min.
[0012] In this invention, when combining enzymatic hydrolysis with ultrasonic extraction of polyphenols, a step-by-step process of first enzymatic hydrolysis and then ultrasonic treatment is adopted. The step-by-step process is to prevent the cavitation effect, shock wave and microjets of ultrasound during enzymatic hydrolysis from causing irreversible physical damage to the spatial structure of the protein, which would lead to conformational denaturation of the enzyme, loss of active sites and ultimately a significant reduction in enzyme activity.
[0013] In one embodiment of the present invention, after sonication, the sample is centrifuged at 3000-5000 rpm for 8-15 min, and the supernatant is collected.
[0014] In one embodiment of the present invention, in step S3, the pH of the supernatant is adjusted to 3-4, and the organic solvent is ethyl acetate; the extraction is performed 2-4 times.
[0015] In one embodiment of the present invention, in step S3, the concentration is carried out at 40~50°C until the volume is 1 / 10 to 1 / 12 of the volume of the extract.
[0016] In one embodiment of the present invention, the cellulase has an activity of 100,000 U / g, the pectinase has an activity of 50,000 U / g, and the acidic protease has an activity of 100,000 U / g.
[0017] The present invention also provides a wolfberry polyphenol extract prepared according to the above-described composite extraction process.
[0018] The present invention also provides an application of the above-mentioned wolfberry combined with polyphenol extract in the preparation of antioxidants or functional foods.
[0019] Beneficial effects: 1. This invention addresses the problems of low extraction rate, easy loss of activity, complex process, and poor environmental friendliness of bound polyphenols from wolfberry in existing technologies, and provides a composite extraction process for bound polyphenols from wolfberry. Through a process design that combines pretreatment to remove free polyphenols with enzymatic hydrolysis and ultrasonic-assisted ordered extraction, the extraction efficiency and bioactivity of bound polyphenols are both improved. Furthermore, the process is green, controllable, and suitable for large-scale production, providing a new approach for the high-value utilization of wolfberry resources.
[0020] 2. This invention first preferentially removes a large amount of free polyphenols through ethanol reflux, eliminating the competitive interference of free polyphenols on subsequent extraction sites. This allows subsequent steps to precisely target components bound to cell wall macromolecules, creating a pure substrate environment for efficient extraction. Then, a complex enzyme is used to directionally degrade the cell wall structure, breaking the covalent or non-covalent bonds between the bound polyphenols and polysaccharides and proteins, achieving the chemical dissociation of polyphenols. Subsequent ultrasonic treatment utilizes cavitation to break up the dissociated fragments and enhance mass transfer efficiency, promoting the rapid and complete release of bound polyphenols into the extraction solution, thus achieving efficient extraction of bound polyphenols. The bound polyphenol content extracted by this method is as high as 6.02 mg / 100g, far exceeding the bound polyphenol content extracted by existing methods. Detailed Implementation
[0021] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the invention.
[0022] The cellulase involved in the embodiments and comparative examples of this invention was purchased from Shandong Longkete Enzyme Preparation Co., Ltd., with an enzyme activity of 100,000 U / g; the pectinase was purchased from the official website of Yuanye Biotechnology, with an enzyme activity of 50,000 U / g; the acidic protease was purchased from the official website of Phygne, with an enzyme activity of 100,000 U / g; and the neutral protease was purchased from the official website of Phygne, with an enzyme activity of 100,000 U / g.
[0023] Detection of polyphenols A colorimetric method was used to prepare a standard curve using gallic acid: 20.00 mg of gallic acid standard was accurately weighed, dissolved in distilled water, and diluted to a 100 mL amber volumetric flask to obtain a 200 μg / mL standard stock solution. This stock solution was then diluted to prepare standard solutions of 6.25, 12.5, 25, 50, 100, and 200 μg / mL. 0.5 mL of the standard solution was taken, 1 mL of 100% Folin-Ciocalteu reagent was added, and the mixture was stirred and allowed to stand for 5 min. Then, 5 mL of 6% Na₂CO₃ solution was added, and the volume was diluted to 10 mL with distilled water. After stirring thoroughly, the mixture was allowed to stand at room temperature in the dark for 30 min, and the absorbance was measured at 765 nm. The corresponding untreated solution was used as a blank for calibration. A standard curve was plotted with the gallic acid standard solution concentration (μg / mL) on the x-axis and absorbance (A) on the y-axis. Accurately pipette 0.5 mL of the test solution and perform the determination as described above. Substitute the obtained absorbance value into the above regression equation to calculate the polyphenol content in wolfberry.
[0024] The bound polyphenols and total polyphenols were detected using the bound polyphenols and total polyphenol extracts as test solutions, respectively.
[0025] Example 1: A method for extracting polyphenols from wolfberry (1) Raw material pretreatment: Take dried wolfberries, crush them with a pulverizer, and pass them through an 18-mesh sieve to collect wolfberry powder. Weigh 100g of wolfberry powder, add 70% ethanol at a material-to-liquid ratio of 1:12, extract at 80℃ for 60min, transfer the extract to a centrifuge, centrifuge at 4000rpm for 15min, and separate the supernatant from the wolfberry residue. Repeat the above steps 3 times, and take the last supernatant. After confirming that no phenolic substances are detected by the Folin-Ciocalteu method, place the wolfberry residue in a 50℃ drying oven and dry for 24h, then seal and store for later use.
[0026] (2) Extraction of bound polyphenols: Weigh 5g of dried wolfberry residue obtained in step (1), add distilled water at a solid-liquid ratio of 1:20 (g:mL), adjust the pH of the system to 4.40 with 0.1mol / L sodium hydroxide solution, add a compound enzyme (cellulase, pectinase and acidic protease in a mass ratio of 1:3:3), the amount of compound enzyme added is 0.25% of the mass of dried wolfberry residue, control the enzymatic hydrolysis temperature at 55℃ and the enzymatic hydrolysis time at 2h, after the enzymatic hydrolysis is completed, sonicate the hydrolysate with an ultrasonic power of 270W, an ultrasonic time of 45min, and a temperature of 50℃, centrifuge the treated sample at 4000 rpm for 10min, take the supernatant, adjust the pH to 3.40 with 0.1mol / L hydrochloric acid, extract 3 times with an equal volume of ethyl acetate, combine the extracts, rotary evaporate at 45℃, and dilute to 10mL volumetric flask with anhydrous methanol to obtain bound polyphenols.
[0027] The extraction method for total polyphenols is as follows: (1) Raw material pretreatment: Take dried wolfberries, crush them with a pulverizer, pass them through an 18-mesh sieve, and collect the wolfberry powder; (2) Extraction of total polyphenols: Weigh 5g of wolfberry powder obtained in step (1), add distilled water at a solid-liquid ratio of 1:20 (g:mL), adjust the pH of the system to 4.40 with 0.1mol / L sodium hydroxide solution, add a compound enzyme (cellulase, pectinase and acidic protease in a mass ratio of 1:3:3), the amount of compound enzyme added is 0.25% of the mass of dry wolfberry residue, control the enzymatic hydrolysis temperature at 55℃ and the enzymatic hydrolysis time at 2h, after the enzymatic hydrolysis is completed, sonicate the hydrolysate with an ultrasonic power of 270W, an ultrasonic time of 45min, and a temperature of 50℃, centrifuge the treated sample at 4000 rpm for 10min, take the supernatant, adjust the pH to 3.40 with 0.1mol / L hydrochloric acid, extract 3 times with an equal volume of ethyl acetate, combine the extracts, rotary evaporate at 45℃ to 1 / 10 of the extract volume, and dilute to 10mL in a volumetric flask with anhydrous methanol to obtain total polyphenols.
[0028] Example 2 The difference between Example 2 and Example 1 is that the mass ratio of cellulase, pectinase and acidic protease is changed to 2:3:2.
[0029] Comparative Example 1 The difference between Comparative Example 1 and Example 1 is that the mass ratio of cellulase, pectinase and acidic protease was changed to 1:2:2.
[0030] Comparative Example 2 The difference between Comparative Example 2 and Example 1 is that the mass ratio of cellulase, pectinase and acidic protease was changed to 1:1:1.
[0031] Comparative Example 3 The difference between Comparative Example 3 and Example 1 is that the mass ratio of cellulase, pectinase and acid protease was changed to 2:4:1.
[0032] Comparative Example 4 The difference between Comparative Example 4 and Example 1 is that the acidic protease was replaced with a neutral protease.
[0033] Table 1. Content of conjugated polyphenols prepared in Examples 1-2 and Comparative Examples 1-4
[0034] Table 1 shows the content of bound polyphenols prepared in Examples 1-2 and Comparative Examples 1-4. The experimental results show that the content of bound polyphenols prepared in Examples 1-2 is higher than 5.5 mg / 100g, indicating that the method of the present invention helps to obtain a higher content of bound polyphenols. In Comparative Example 4, after replacing the acidic protease in Example 1 with a neutral protease, the extraction rate of bound polyphenols was only 2.36 ± 0.14 mg / 100g, far lower than the extraction rate of 6.02 ± 0.51 mg / 100g in Example 1. There are two main reasons for this phenomenon: First, the proteins in wolfberries that bind to polyphenols may be rich in specific amino acid sequences that acidic proteases prefer. This means that only acidic proteases can effectively hydrolyze these specific protein-polyphenol linkages. If neutral proteases are used, their hydrolysis efficiency will decrease significantly. Second, the optimal pH for neutral proteases is 6.0-8.0, while wolfberries themselves are in an acidic environment. Cellulase and pectinase also have an optimal pH in acidic conditions. When hydrolyzing under acidic conditions, the degree of hydrolysis by neutral proteases is weaker, which further affects the extraction efficiency of bound polyphenols.
[0035] Example 2 and Comparative Examples 1-3 further investigated the effect of the compound ratio of cellulase, pectinase and acidic protease. The results showed that the content of bound polyphenols was the highest when the mass ratio of the three was 1:3:3. When the ratio of the three was in the range of 1-2:3:2-3, a relatively high content of bound polyphenols could be obtained.
[0036] Example 3 The difference between Example 3 and Example 2 is that the ultrasonic power is changed to 240W, 270W, and 300W.
[0037] Table 2. Effect of ultrasonic power on the binding of polyphenols in wolfberry.
[0038] Table 2 shows that the ultrasonic power has a significant impact on the content of bound polyphenols, with the highest extraction rate observed at an ultrasonic power of 270W. Powers below or above 270W further reduced the content of bound polyphenols. The experiment also investigated the total polyphenol content under different ultrasonic powers. In Example 1 (270W), the total polyphenol content was 42.93 ± 1.09 mg / 100g. As the ultrasonic power increased to 300W, the content of bound polyphenols decreased significantly, while the total polyphenol content remained essentially unchanged at 42.87 ± 2.13 mg / 100g. This indicates that ultrasonic power only affects the content of bound polyphenols and does not affect the total polyphenol content.
[0039] Example 4 The difference between Example 4 and Example 2 is that the ultrasound time was changed to 30 min, 45 min, and 60 min.
[0040] Table 3. Effect of ultrasound time on bound polyphenols and total polyphenols in wolfberry.
[0041] Table 3 shows the effects of different ultrasound times on the content of bound polyphenols and total polyphenols in wolfberry. The results show that the extraction rate of bound polyphenols was the highest at 45 min, while the content of total polyphenols was less affected by ultrasound time.
[0042] Example 5 The difference between Example 5 and Example 2 is that the ultrasonic temperature was changed to 40°C, 50°C, and 60°C.
[0043] Table 4. Effect of ultrasonic temperature on bound polyphenols and total polyphenols in wolfberry.
[0044] Table 4 shows the effects of different ultrasonic temperatures on the content of bound polyphenols and total polyphenols in wolfberry. The results show that the extraction rate of bound polyphenols is the highest at 50℃, while the content of total polyphenols is less affected by ultrasonic temperature.
[0045] Example 6 The difference between Example 6 and Example 1 is that the amount of compound enzyme added is changed to 0.2%, 0.25%, and 0.3%.
[0046] Table 5. Effects of compound enzyme dosage on the levels of bound polyphenols and total polyphenols in wolfberry.
[0047] Table 5 shows the effect of the amount of compound enzyme added on the content of bound polyphenols and total polyphenols in wolfberry. The results show that the extraction rate of bound polyphenols is highest when the enzyme addition amount is 0.25%. The experiment also compared the total polyphenol content when the compound enzyme addition amount was 0.25% and 0.3%. The results showed that when the compound enzyme addition amount was 0.25%, the total polyphenol content in wolfberry was 42.93±1.09 mg / 100g, while as the compound enzyme addition amount increased to 0.3%, the total polyphenol content increased to 42.98±1.65 mg / 100g. This indicates that the total polyphenol content is less affected by the amount of enzyme added.
[0048] Antioxidant activity evaluation ABTS free radical scavenging rate: (1) Preparation of ABTS working solution: Prepare 7 mmol / L ABTS aqueous solution and 2.45 mmol / L potassium persulfate aqueous solution respectively. Mix the two solutions thoroughly at a volume ratio of 1:1 and incubate at room temperature of 25°C in the dark for 16 hours to obtain ABTS working stock solution. Dilute ABTS working stock solution with ultrapure water until the absorbance at a wavelength of 734 nm is 1.4, which is the ABTS working solution. Prepare and use immediately.
[0049] (2) Sample addition and reaction: A 96-well plate was used for detection. The ABTS working solution and the test sample were added at a volume ratio of 40:1. 200 μL of ABTS working solution and 5 μL of test sample were added to each well and mixed thoroughly. A blank control group was set up, which replaced the test sample with 5 μL of ultrapure water. The other sample addition and treatment methods were completely the same as those of the experimental group. The sample was incubated at room temperature of 25℃ and in the dark for 6 min.
[0050] (3) Absorbance measurement: After incubation, the absorbance value of each well was measured immediately at a wavelength of 734 nm; three parallel samples were set up for each group of experiments, and the measurement was repeated three times, and the average value was taken.
[0051] (4) Calculation of free radical scavenging rate:
[0052] DPPH free radical scavenging rate: (1) Sample pretreatment: Take the sample to be tested and add anhydrous ethanol at a ratio of 1:9 (sample volume: anhydrous ethanol volume). After mixing thoroughly, shake at room temperature of 25°C for 30 min (shaking frequency 150 rpm), centrifuge at 1000 rpm and room temperature for 10 min, and take the supernatant as the test solution for later use.
[0053] (2) Preparation of DPPH working solution: Weigh the DPPH reagent, dissolve it in anhydrous ethanol and make up to volume to prepare a DPPH working solution with a concentration of 0.2 mmol / L. Store in the dark and refrigerate. Prepare and use immediately.
[0054] (3) Sample addition and reaction: Take 0.05 mL of the supernatant to be tested, add 0.225 mL of anhydrous ethanol and 0.225 mL of DPPH working solution, and vortex thoroughly to mix; Two control groups were set up simultaneously. The blank control group was replaced with 0.05 mL of anhydrous ethanol instead of the supernatant to be tested, and 0.225 mL of anhydrous ethanol and 0.225 mL of DPPH working solution were added. The rest of the treatment was the same. The sample control group was replaced with 0.225 mL of anhydrous ethanol instead of DPPH working solution. The rest of the sample volume and treatment were completely the same as the experimental group (used to subtract the background absorbance interference of the sample, corresponding to the control group in the formula). The samples were incubated at room temperature of 25°C in the dark for 30 min.
[0055] (4) Absorbance measurement: After incubation, the absorbance value of each group was measured immediately at a wavelength of 517 nm; three parallel samples were set up for each group of experiments, and the measurement was repeated three times, and the average value was taken.
[0056] (5) Calculation of free radical scavenging rate:
[0057] The results showed that, compared with the control group (which omitted the steps of adjusting pH and adding compound enzymes in Example 1), the experimental group (Example 1) had a DPPH free radical scavenging rate increased by 34.56% and an ABTS free radical scavenging rate increased by 54.30%, which verified the superiority of the process of the present invention.
[0058] Table 5. Results and Analysis of Antioxidant Activity
[0059] The embodiments provided above are not intended to limit the scope of the invention, nor are the described steps intended to limit the order of execution. Any obvious modifications made to the invention by those skilled in the art based on existing common knowledge also fall within the scope of protection defined by the claims.
Claims
1. A compound extraction process for wolfberry combined with polyphenols, characterized in that, Includes the following steps: S1. Raw material pretreatment: The dried wolfberries are crushed and sieved to obtain wolfberry powder; the wolfberry powder is extracted by reflux with ethanol solution, and the wolfberry residue is collected and dried after solid-liquid separation to obtain pretreated wolfberry residue; S2. Enzymatic hydrolysis-ultrasound combined extraction: The pretreated wolfberry residue obtained in step S1 is mixed with water, the pH value is adjusted, and a compound enzyme is added for enzymatic hydrolysis; after enzymatic hydrolysis, ultrasonic treatment is performed, and the supernatant is collected after solid-liquid separation; the compound enzyme includes cellulase, pectinase and acidic protease. S3. Extraction and Concentration: Adjust the pH of the supernatant obtained in step S2 to acidic, extract with an organic solvent, collect the extract and concentrate it to obtain a wolfberry polyphenol extract.
2. The compound extraction process of wolfberry combined with polyphenols according to claim 1, characterized in that, In step S1, the sieve mesh size is 15-20 mesh, the volume fraction of the added ethanol solution is 60-80%, the material-to-liquid ratio of wolfberry powder to added ethanol solution is 1g:10-15mL, the reflux extraction temperature is 70-90℃, the reflux extraction time is 60-90min, and the solid-liquid separation is centrifuged at 4000rpm for 15min.
3. The compound extraction process of wolfberry combined with polyphenols according to claim 1, characterized in that, In step S2, the solid-liquid ratio of the pretreated wolfberry residue to water is 1:15~20 g / mL; the pH value is adjusted to 4~5; the temperature during enzymatic hydrolysis is 50~55℃, and the enzymatic hydrolysis time is 1~3h.
4. The compound extraction process of wolfberry combined with polyphenols according to claim 1, characterized in that, In step S2, the mass ratio of cellulase, pectinase and acidic protease is 1~2:3:2~3.
5. The compound extraction process of wolfberry combined with polyphenols according to claim 1, characterized in that, In step S2, the temperature during ultrasonic treatment is 50~60℃, the ultrasonic power is 240~300W, and the ultrasonic time is 45~60min.
6. The compound extraction process of wolfberry combined with polyphenols according to claim 1, characterized in that, After sonication, centrifuge at 3000-5000 rpm for 8-15 min, collect the supernatant, and adjust the pH of the supernatant to 3-4. The organic solvent is ethyl acetate. The extraction is performed 2-4 times.
7. The compound extraction process of wolfberry combined with polyphenols according to claim 1, characterized in that, In step S3, the concentration is carried out at 40~50℃ until the volume is 1 / 10~1 / 12 of the volume of the extract.
8. The compound extraction process of wolfberry combined with polyphenols according to claim 1, characterized in that, The cellulase has an activity of 100,000 U / g, the pectinase has an activity of 50,000 U / g, and the acidic protease has an activity of 100,000 U / g.
9. The wolfberry polyphenol extract obtained by the compound extraction process according to any one of claims 1 to 8.
10. The use of the wolfberry polyphenol extract according to claim 9 in the preparation of antioxidants or functional foods.