A polygonum multiflorum thunb extract, a preparation method and application thereof

Abstract

The application discloses a polygonatum extract, a preparation method and application thereof, and belongs to the technical field of polygonatum extraction; the polygonatum extract is prepared by the following steps: firstly, soaking and grinding the crushed polygonatum to obtain a polygonatum slurry; and then, performing enzyme hydrolysis-fermentation synchronous extraction on the polygonatum slurry, so that the extraction rate of polygonatum polysaccharide and polygonatum saponin in the polygonatum extract can be effectively improved, and remarkable effects can be achieved; specifically, the extraction rate of the polygonatum polysaccharide in the obtained polygonatum extract is above 70.28%, and the extraction rate of the polygonatum saponin is above 22.68%. Moreover, the preparation method of the polygonatum extract is simple in operation and is favorable for actual scale-up production.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of polygonatum extract, and particularly relates to a polygonatum extract, a preparation method and application thereof. BACKGROUND

[0002] Polygonatum is a perennial herb of the genus Polygonatum in the Liliaceae family, and the rhizome is used as medicine, which has the effects of tonifying qi and yin, invigorating the spleen, moistening the lungs, and nourishing the kidney. Modern medical pharmacological studies show that polysaccharides and saponins are the main chemical components of polygonatum, in addition to which flavonoids, alkaloids, vitamins, and various essential amino acids and trace elements are also important components of polygonatum chemical composition, which together determine the wide pharmacological activity of polygonatum, including anti-fatigue, antioxidant, immune regulation, blood glucose and lipid regulation, and anti-tumor.

[0003] Polygonatum was listed in the Chinese medicine list of food and medicine by the National Health Commission in 2002. In recent years, the continuous improvement of national health consciousness has promoted the rapid development of the health industry, and the development and utilization of polygonatum will also be more widely applied. Fresh polygonatum contains calcium oxalate needle crystal bundles, and eating raw polygonatum has a tingling and throat irritation effect, so polygonatum is usually processed by nine steaming and nine processing before eating, but the nine steaming and nine processing process is time-consuming and complicated, and the polysaccharide content of polygonatum is reduced. At present, polygonatum decoction pieces on the market are mostly eaten by directly adding or brewing, which greatly reduces the effective utilization rate of polygonatum. Therefore, it is necessary to efficiently extract polysaccharides and other functional components in polygonatum to improve the effective utilization rate of polygonatum. At present, the extraction of active ingredients in polygonatum is mostly hot water extraction or ultrasonic extraction, which has the limitations of low extraction rate, high energy consumption, high equipment requirements, and cannot be produced on a large scale. In recent years, enzyme extraction and fermentation extraction have been widely studied and applied, but single enzyme hydrolysis or fermentation cannot fully release the functional components in polygonatum. SUMMARY

[0004] The present application aims to overcome the shortcomings of the prior art and provide a polygonatum extract with high extraction rate of polysaccharides and saponins and capable of large-scale production, as well as a preparation method and application thereof.

[0005] To achieve the above-mentioned purpose, in the first aspect of the present application, the present application provides a preparation method of a polygonatum extract, which comprises the following steps:

[0006] (1) The crushed polygonatum is added to water for soaking, and after the soaking is completed, it is ground to obtain a polygonatum slurry;

[0007] (2) Enzyme and microorganism are added to the polygonatum slurry for simultaneous extraction by enzyme hydrolysis and fermentation;

[0008] (3) After the simultaneous extraction by enzyme hydrolysis and fermentation, sterilization and filtration are performed, the filtrate is collected and centrifuged, the supernatant is concentrated to obtain the polygonatum extract.

[0009] The present application can effectively improve the extraction rate of polygonatum sibiricum polysaccharide and polygonatum sibiricum saponin in the polygonatum sibiricum extract by first soaking and grinding the crushed polygonatum sibiricum to obtain a polygonatum sibiricum slurry, and then performing enzyme-fermentation synchronous extraction on the polygonatum sibiricum slurry.

[0010] Specifically, in the first aspect, the present application can reduce the particle size of polygonatum sibiricum by soaking and grinding, so that the contact area between enzymes and microorganisms and polygonatum sibiricum is increased in the subsequent enzyme-fermentation synchronous extraction process, the interface reaction efficiency is improved, and the release of active ingredients in polygonatum sibiricum is more beneficial. In the second aspect, the present application can further improve the extraction rate of polygonatum sibiricum polysaccharide and polygonatum sibiricum saponin by enzyme-fermentation synchronous extraction, and the enzymes and microorganisms can work synergistically.

[0011] As a preferred embodiment of the preparation method of the present application, in step (2), before adding enzymes and microorganisms to the polygonatum sibiricum slurry for enzyme-fermentation synchronous extraction, a step of adding esterase to the polygonatum sibiricum slurry for first enzyme hydrolysis and enzyme inactivation is further included.

[0012] The present application has found that further selecting the addition of esterase to the polygonatum sibiricum slurry for first enzyme hydrolysis in enzyme-fermentation synchronous extraction can effectively achieve the purpose of degreasing of polygonatum sibiricum raw materials, improve the interface reaction efficiency of subsequent enzyme-fermentation synchronous extraction, and further improve the extraction rate of polygonatum sibiricum polysaccharide and polygonatum sibiricum saponin.

[0013] Exemplarily, the esterase includes any one of carboxylic acid ester hydrolase and glycerol ester hydrolase. The esterase can be purchased from Wuhan Xinhua Yang Biological Co., Ltd. Jiulong Branch, etc.

[0014] As a preferred embodiment of the preparation method of the present application, in step (2), the enzyme includes at least one of cellulase, papain, and beta-glucanase.

[0015] Exemplarily, the cellulase, papain, and beta-glucanase can be purchased from Wuhan Xinhua Yang Biological Co., Ltd. Jiulong Branch or Nanning Pangbo Biological Engineering Co., Ltd.

[0016] Preferably, the enzyme is papain.

[0017] As a preferred embodiment of the preparation method of the present application, in step (2), the microorganism includes at least one of yeast and lactic acid bacteria.

[0018] Preferably, the lactic acid bacteria includes at least one of plant lactobacillus, lactobacillus acidophilus, and paracasei lactobacillus.

[0019] Exemplarily, the yeast can be purchased from Angel Yeast Co., Ltd. and the like; the Lactobacillus plantarum, Lactobacillus acidophilus and Lactobacillus paracasei can be purchased from Weike Biotech (Suzhou) Co., Ltd., Shandong Zhongke Jiayi Biological Engineering Co., Ltd. and the like.

[0020] Preferably, the microorganism is yeast.

[0021] As a preferred embodiment of the preparation method of the present application, in step (1), the mass ratio of the crushed Huangjing and water is 1:(5-10).

[0022] Exemplarily, the mass ratio of the crushed Huangjing and water can be any point value or any two-point range value between 1:(5-10), such as 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, etc.

[0023] Preferably, in step (1), the mass ratio of the crushed Huangjing and water is 1:(5-8).

[0024] As a preferred embodiment of the preparation method of the present application, in step (1), the soaking time is 0.5-1.5h.

[0025] Exemplarily, the soaking time can be any point value or any two-point range value between 0.5-1.5h, such as 0.5h, 1h, 1.5h, etc.

[0026] Preferably, in step (1), the soaking time is 1-1.5h.

[0027] As a preferred embodiment of the preparation method of the present application, in step (1), the grinding rate is 5-20kg / min.

[0028] Exemplarily, the grinding rate can be any point value or any two-point range value between 5-20kg / min, such as 5kg / min, 10kg / min, 15kg / min, 20kg / min, etc.

[0029] Preferably, in step (1), the grinding rate is 5-10kg / min.

[0030] As a preferred embodiment of the preparation method of the present application, in step (1), the fineness of the Huangjing slurry is ≤5mm.

[0031] Preferably, in step (1), the fineness of the Huangjing slurry is ≤2mm.

[0032] The present application researches and finds that when the mass ratio of Huangqi and water in the soaking process and the soaking time are further selected within the above range, and the grinding rate and the fineness of the Huangqi slurry after grinding are selected within the above range, especially within the further preferred range, the particle size of Huangqi can be better refined, the interface reaction rate of subsequent enzymolysis and microbial fermentation can be improved, the extraction rate of Huangqi polysaccharide and Huangqi saponin can be increased, and the sensory score of Huangqi extract can be improved.

[0033] As a preferred embodiment of the preparation method of the present application, in the step (2), the addition amount of the first enzymolysis esterase is 0.2-1.5% based on the mass of the crushed Huangqi.

[0034] For example, the addition amount of the first enzymolysis esterase can be any point value or any two-point range value between 0.2-1.5% based on the mass of the crushed Huangqi, such as 0.2%, 0.5%, 0.8%, 1%, 1.2%, 1.5%, etc.

[0035] Preferably, in the step (2), the addition amount of the first enzymolysis esterase is 0.5-0.8% based on the mass of the crushed Huangqi.

[0036] As a preferred embodiment of the preparation method of the present application, the temperature of the first enzymolysis is 40-60℃, and the time is 1-3h.

[0037] For example, the temperature of the first enzymolysis can be any point value or any two-point range value between 40-60℃, such as 40℃, 45℃, 50℃, 55℃, 60℃, etc.; the time can be any point value or any two-point range value between 1-3h, such as 1h, 2h, 3h, etc.

[0038] Preferably, the temperature of the first enzymolysis is 50-60℃, and the time is 2-3h.

[0039] As a preferred embodiment of the preparation method of the present application, during the first enzymolysis, the material-liquid ratio of the Huangqi slurry is 1:(10-40), and the pH value is 4.0-7.0.

[0040] For example, during the first enzymolysis, the material-liquid ratio of the Huangqi slurry can be any point value or any two-point range value between 1:(10-40), such as 1:10, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40, etc.; the pH value can be any point value or any two-point range value between 4.0-7.0, such as 4, 4.5, 5, 5.5, 6, 6.5, 7, etc.

[0041] Preferably, during the first enzymolysis, the material-liquid ratio of the Huangqi slurry is 1:(18-22), and the pH value is 5.0-6.0.

[0042] The present application research found that when the addition amount of esterase in the first esterase enzymolysis process, the temperature and time of the first enzymolysis, and the solid-liquid ratio and pH value of the first enzymolysis of rhizoma polygonati slurry are within the above range, especially within the further preferred range, the lipid components in the rhizoma polygonati raw material can be better removed to achieve the purpose of degreasing, avoiding the conventional use of chemical reagents for degreasing, and at the same time, the interface reaction rate of subsequent enzymolysis-microorganism simultaneous extraction can be more effectively improved to increase the extraction rate of rhizoma polygonati polysaccharide and rhizoma polygonati saponin.

[0043] As a preferred embodiment of the preparation method of the present application, in the step (2), the mass percentage of enzyme is 0.1-1.5% and the mass percentage of microorganism is 1-10% based on the mass of the crushed rhizoma polygonati in the enzymolysis-fermentation simultaneous extraction.

[0044] For example, the mass percentage of enzyme can be any point value or any two-point range value between 0.1-1.5%, such as 0.1%, 0.5%, 1%, 1.5%, etc., and the mass percentage of microorganism can be any point value or any two-point range value between 1-10%, such as 1%, 2%, 4%, 6%, 8%, 10%, etc.

[0045] Preferably, in the step (2), the mass percentage of enzyme is 0.5-1.5% and the mass percentage of microorganism is 2-8% based on the mass of the crushed rhizoma polygonati in the enzymolysis-fermentation simultaneous extraction.

[0046] As a preferred embodiment of the preparation method of the present application, in the step (2), the extraction temperature is 25-40℃ and the extraction time is 12-60h in the enzymolysis-fermentation simultaneous extraction.

[0047] For example, the extraction temperature can be any point value or any two-point range value between 25-40℃, such as 25℃, 30℃, 35℃, 40℃, etc., and the extraction time can be any point value or any two-point range value between 12-60h, such as 12h, 15h, 20h, 25h, 30h, 35h, 40h, 45h, 50h, 55h, 60h, etc.

[0048] Preferably, in the step (2), the extraction temperature is 25-35℃ and the extraction time is 12-36h in the enzymolysis-fermentation simultaneous extraction.

[0049] The present application research found that in the enzymolysis-fermentation simultaneous extraction, the addition amount of enzyme and microorganism and the extraction temperature and time will affect the extraction rate of rhizoma polygonati polysaccharide and rhizoma polygonati saponin, and when the parameters are further selected within the above range, especially within the further preferred range, the extraction rate of rhizoma polygonati polysaccharide and rhizoma polygonati saponin is higher, and the sensory score of rhizoma polygonati extract is also higher.

[0050] As a preferred embodiment of the preparation method of the present application, the temperature for enzyme inactivation is 90-100 DEG C, and the time is 10-20 min.

[0051] As a preferred embodiment of the preparation method of the present application, the temperature for sterilization is 85-115 DEG C, and the time is 10-30 min.

[0052] Preferably, the temperature for sterilization is 90-100 DEG C, and the time is 10-20 min.

[0053] As a preferred embodiment of the preparation method of the present application, the filtration is performed by using a filter screen with a mesh size of 200-500.

[0054] Exemplarily, the mesh size of the filter screen can be any point value or any two-point range value between 200-500, such as 200 mesh, 300 mesh, 400 mesh, 500 mesh, etc.

[0055] As a preferred embodiment of the preparation method of the present application, the centrifugal speed is 3000-5000 rpm, and the time is 10-20 min.

[0056] Exemplarily, the centrifugal speed can be any point value or any two-point range value between 3000-5000 rpm, such as 3000 rpm, 3500 rpm, 4000 rpm, 4500 rpm, 5000 rpm, etc.; and the time can be any point value or any two-point range value between 10-20 min, such as 10 min, 15 min, 20 min, etc.

[0057] As a preferred embodiment of the preparation method of the present application, the temperature for concentration is ≤80 DEG C, and the vacuum degree for concentration is ≤-0.07 MPa.

[0058] In the second aspect of the present application, the present application provides a polygonatum extract prepared by the preparation method of the present application.

[0059] In the third aspect of the present application, the present application provides the use of the polygonatum extract in the preparation of products for improving immunity, regulating blood sugar and blood lipid, resisting tumor, and resisting oxidation.

[0060] As a preferred embodiment of the use of the present application, the product includes food and health care products.

[0061] Compared with the prior art, the present application has the following beneficial effects:

[0062] 1. This invention uses a colloid mill to prepare Polygonatum slurry from raw materials. After being treated with a colloid mill, the particle size of the raw materials becomes smaller, which increases the specific surface area for enzyme and microbial interaction, making it more conducive to the release of functional components in Polygonatum.

[0063] 2. This invention utilizes esterase hydrolysis to degrease Polygonatum odoratum raw materials, avoiding the cumbersome process of degreasing with chemical reagents, improving the interfacial reaction efficiency of simultaneous and synergistic extraction of enzymes and microorganisms, and making it more conducive to the extraction of functional components from Polygonatum odoratum.

[0064] 3. This invention selects enzyme systems and microorganisms suitable for the extraction of Polygonatum polysaccharides and Polygonatum saponins from Polygonatum. By utilizing exogenous enzymes and microbial fermentation for simultaneous and synergistic extraction through enzymatic hydrolysis and fermentation, the extraction rates of Polygonatum polysaccharides and Polygonatum saponins are improved. The extraction rate of Polygonatum polysaccharides reaches over 70.28%, and the extraction rate of Polygonatum saponins is over 22.68%, thereby enhancing the effective utilization rate of Polygonatum.

[0065] 4. The preparation method of Polygonatum extract provided by the present invention is simple to operate and is conducive to large-scale production. Attached Figure Description

[0066] Figure 1 The sensory evaluation results of the Polygonatum extracts prepared in Example 7 and Comparative Example 12 are shown in the figure.

[0067] Figure 2 This is a schematic diagram of the preparation process of the Polygonatum extract of the present invention. Detailed Implementation

[0068] To better illustrate the purpose, technical solution, and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments.

[0069] Unless otherwise specified, the reagents, methods and equipment used in this invention are all conventional reagents, methods and equipment in the field; and unless otherwise specified, the raw materials used in parallel experiments are from the same batch.

[0070] Example 1

[0071] This invention provides a Polygonatum extract, the preparation process of which is shown in the schematic diagram below. Figure 2 As shown, it includes the following steps:

[0072] (1) After cleaning the moldy, insect-infested and impurity parts of Polygonatum, crush it with a pulverizer to obtain crushed Polygonatum. Then, add 1.0 kg of crushed Polygonatum to 5 times the mass of purified water and soak for 1 hour. After soaking, grind it with a colloid mill at a grinding rate of 5 kg / min until the fineness is ≤2 mm to obtain Polygonatum slurry.

[0073] (2) After adjusting the material-liquid ratio of the polygonatum sibiricum preparate to 1:20 and the pH value to 5.0, 0.6wt% glycerol ester hydrolytic enzyme is added to perform first enzymolysis for 2h at 50°C. After the first enzymolysis, the enzyme is inactivated at 95°C for 10min and cooled to 25°C;

[0074] (3) Then, 1wt% papain and 2wt% yeast are added to the system cooled to 25°C to perform enzymolysis-fermentation synchronous extraction for 20h at 30°C. After the enzymolysis-fermentation synchronous extraction, the temperature is raised to 95°C to sterilize for 10min;

[0075] (4) After sterilization, the filter cloth with a mesh of 300 is used to filter, the filtrate is collected, and the filtrate is centrifuged at 4000rpm for 15min. After centrifugation, the supernatant is taken, and then concentrated under the condition that the temperature is ≤80°C and the vacuum degree is ≤-0.07MPa to obtain the polygonatum sibiricum extract.

[0076] Example 2

[0077] The polygonatum sibiricum extract provided by the embodiment of the present application has the only difference from the preparation method of Example 1 that cellulase is used to replace papain.

[0078] Example 3

[0079] The polygonatum sibiricum extract provided by the embodiment of the present application has the only difference from the preparation method of Example 1 that β-glucanase is used to replace papain.

[0080] Example 4

[0081] The polygonatum sibiricum extract provided by the embodiment of the present application has the only difference from the preparation method of Example 1 that the mixture of cellulase, papain and β-glucanase (wherein the mass ratio of cellulase, papain and β-glucanase is 1:8:1) is used to replace papain.

[0082] Example 5

[0083] The polygonatum sibiricum extract provided by the embodiment of the present application has the only difference from the preparation method of Example 1 that plant lactobacillus is used to replace yeast.

[0084] Example 6

[0085] The polygonatum sibiricum extract provided by the embodiment of the present application has the only difference from the preparation method of Example 1 that the first enzymolysis in step (2) is not performed.

[0086] Example 7

[0087] The present application provides a kind of Rhizoma Polygonati extract, the only difference between the preparation method of the Rhizoma Polygonati extract and the embodiment 1 is that the mass of the Rhizoma Polygonati after crushing is 50.0 kg.

[0088] Comparative Example 1

[0089] The present application provides a kind of Rhizoma Polygonati extract, the only difference between the preparation method of the Rhizoma Polygonati extract and the embodiment 1 is that the Rhizoma Polygonati is not ground after soaking.

[0090] Comparative Example 2

[0091] The present application provides a kind of Rhizoma Polygonati extract, the only difference between the preparation method of the Rhizoma Polygonati extract and the embodiment 1 is that the Rhizoma Polygonati is not ground after soaking.

[0092] (1) After removing the moldy, insect-infested parts and impurities in the Rhizoma Polygonati, the Rhizoma Polygonati is crushed with a crusher to obtain crushed Rhizoma Polygonati. Then, 1.0 kg of the crushed Rhizoma Polygonati is added to 5 times the mass of purified water for soaking for 1 h. After soaking, the Rhizoma Polygonati is ground with a colloid mill at a grinding rate of 5 kg / min until the fineness is ≤2 mm to obtain a Rhizoma Polygonati slurry.

[0093] (2) After adjusting the solid-liquid ratio of the Rhizoma Polygonati slurry to 1:20 and the pH value to 5.0, 0.6 wt% glycerol ester hydrolase is added for first enzymolysis at 50°C for 2 h.

[0094] (3) After the first enzymolysis, 1 wt% papain is added for enzymolysis at 50°C for 2 h. After the enzymolysis, the enzyme is inactivated at 95°C for 10 min and cooled to 25°C.

[0095] (4) Then, 2 wt% yeast is added to the system cooled to 25°C for fermentation extraction at 30°C for 20 h. After the fermentation extraction, the temperature is raised to 95°C for sterilization for 10 min.

[0096] (5) After sterilization, the filtrate is collected by filtering with 300 mesh filter cloth. The filtrate is centrifuged at 4000 rpm for 15 min. After centrifugation, the supernatant is collected. Then, the Rhizoma Polygonati extract is obtained by concentrating at a temperature ≤80°C and a vacuum degree ≤-0.07 MPa.

[0097] Comparative Example 3

[0098] The present application provides a kind of Rhizoma Polygonati extract, the only difference between the preparation method of the Rhizoma Polygonati extract and the embodiment 1 is that the Rhizoma Polygonati is not ground after soaking.

[0099] (1) The moldy, insect-transformed parts and impurities in the Rhizoma Polygonati were removed and cleaned, and then ground by a grinder to obtain ground Rhizoma Polygonati. Then, 1.0 kg of the ground Rhizoma Polygonati was added to 5 times the amount of purified water to soak for 1 h. After soaking, the Rhizoma Polygonati was ground by a colloid mill at a grinding rate of 5 kg / min until the fineness was ≤2 mm to obtain a Rhizoma Polygonati slurry;

[0100] (2) After adjusting the material-liquid ratio of the Rhizoma Polygonati slurry to 1:20 and the pH value to 5.0, 0.6 wt% glycerol ester hydrolase was added for first enzymolysis at 50°C for 2 h. After the first enzymolysis, the enzyme was inactivated at 95°C for 10 min and cooled to 25°C;

[0101] (3) Then, 2 wt% yeast was added to the system cooled to 25°C for fermentation extraction at 30°C for 20 h. After the fermentation extraction, the system was sterilized at 95°C for 10 min, and then cooled to 50°C and 1 wt% papain was added for enzymolysis at 50°C for 2 h. After the enzymolysis, the enzyme was inactivated at 95°C for 10 min.

[0102] (5) After the enzyme was inactivated, the filtrate was collected by filtering with a 300-mesh filter cloth, and then centrifuged at 4000 rpm for 15 min. The supernatant was collected, and then concentrated under the conditions of a temperature ≤80°C and a vacuum degree ≤-0.07 MPa to obtain a Rhizoma Polygonati extract.

[0103] Comparative Example 4

[0104] The Rhizoma Polygonati extract provided by the comparative example of the present application has a preparation method that is only different from that of Example 1 in that no yeast is added in step (3), and the enzymolysis temperature of papain is adjusted to 30°C for enzymolysis for 20 h.

[0105] Comparative Example 5

[0106] The Rhizoma Polygonati extract provided by the comparative example of the present application has a preparation method that is only different from that of Example 1 in that no yeast is added in step (3), and the enzymolysis temperature of papain is adjusted to 50°C for enzymolysis for 2 h.

[0107] Comparative Example 6

[0108] The Rhizoma Polygonati extract provided by the comparative example of the present application has a preparation method that is only different from that of Example 1 in that no yeast is added in step (3), and cellulase is used for enzymolysis, and the enzymolysis temperature is adjusted to 50°C for enzymolysis for 2 h.

[0109] Comparative Example 7

[0110] The Rhizoma Polygonati extract provided by the comparative example of the present application has a preparation method that is only different from that of Example 1 in that no yeast is added in step (3), and β-glucanase is used for enzymolysis, and the enzymolysis temperature is adjusted to 50°C for enzymolysis for 2 h.

[0111] Comparative Example 8

[0112] The present comparative example provides a Huangqi extract, the only difference between the preparation method of the Huangqi extract and that of Example 1 is that no papain is added in step (3).

[0113] Comparative Example 9

[0114] The present comparative example provides a Huangqi extract, the only difference between the preparation method of the Huangqi extract and that of Example 1 is that no papain is added in step (3), and the fermentation is carried out with 4 wt% of Lactobacillus plantarum, the fermentation temperature is 37℃, and the fermentation time is 24h.

[0115] Comparative Example 10

[0116] The present comparative example provides a Huangqi extract, the preparation method of the Huangqi extract is hot water extraction, comprising the following steps:

[0117] (1) After removing the moldy, insect-infested parts and impurities of Huangqi, the Huangqi is crushed with a crusher to obtain crushed Huangqi, then 1.0 kg of the crushed Huangqi is added to 5 times the mass of purified water for soaking for 1h, after the soaking is completed, the Huangqi is ground with a colloid mill at a grinding rate of 5 kg / min, and the grinding fineness is ≤2mm to obtain a Huangqi slurry;

[0118] (2) The material-liquid ratio of the Huangqi slurry is adjusted to 1:20 by adding water, and the Huangqi is extracted at 100℃ for 2h, then filtered with 300 mesh filter cloth, and then 8 times the mass of water of the crushed Huangqi is added, and the Huangqi is extracted at 100℃ for 1h, and the filtrates of the two extractions are combined;

[0119] (3) The filtrate is centrifuged at 4000 rpm for 15 min, the supernatant is taken after centrifugation, then concentrated under the condition of temperature ≤80℃ and vacuum degree ≤-0.07 MPa, then sterilized at 110℃ for 20 min to obtain a Huangqi extract.

[0120] Comparative Example 11

[0121] The present comparative example provides a Huangqi extract, the only difference between the preparation method of the Huangqi extract and that of Comparative Example 10 is that no grinding treatment is carried out in step (1).

[0122] Comparative Example 12

[0123] The present comparative example provides a Huangqi extract, the only difference between the preparation method of the Huangqi extract and that of Comparative Example 10 is that no crushing and no grinding treatment is carried out in step (1).

[0124] Effect Example

[0125] The extraction rates of the polysaccharide and the saponin in the rhizoma polygonati extract prepared from the effect example exploration examples 1-7 and the comparative examples 1-12 of the present application were investigated, and the test method included the following contents:

[0126] 1. The content of the polysaccharide was determined according to the determination method in the Chinese Pharmacopoeia 2020 edition, specifically as follows:

[0127] (1) Preparation of the reference solution: about 38 mg of anhydrous glucose reference substance dried at 105 DEG C to a constant weight was accurately weighed, placed in a 100 mL volumetric flask, dissolved and diluted with water to the mark, and shaken to obtain the reference solution;

[0128] (2) Preparation of the standard curve: 0.1 mL, 0.2 mL, 0.3 mL, 0.4 mL and 0.5 mL of the reference solution were accurately measured and placed in 10 mL stoppered test tubes, respectively, and water was added to 2.0 mL, shaken, slowly added with 0.05% anthrone-sulfuric acid solution to the mark in an ice water bath, mixed, placed in a water bath for 10 minutes after cooling, taken out, immediately placed in an ice water bath for 10 minutes, taken out, and determined as a blank with the corresponding reagent. The absorbance was determined at a wavelength of 582 nm, the absorbance was taken as the vertical coordinate, and the concentration was taken as the horizontal coordinate to draw a standard curve;

[0129] (3) Determination method: 5 mL of the sample was accurately measured and placed in a round-bottom flask, 25 mL of water was added, mixed, 120 mL of anhydrous ethanol was slowly added, mixed, placed in a 90 DEG C water bath for heating and refluxing for 0.5 hours, filtered while hot, the residue was washed with 80% hot ethanol for 2 times, 10 mL each time, the residue and filter paper were placed in a flask, 150 mL of water was added, placed in a boiling water bath for heating and refluxing for 1 hour, filtered while hot, the residue and flask were washed with hot water for 4 times, 10 mL each time, the filtrate and washing liquid were combined, cooled, transferred to a 250 mL volumetric flask, added with water to the mark, shaken, accurately measured 2 mL, placed in a 10 mL stoppered test tube, and determined according to the method in the preparation of the standard curve from "slowly added with 0.05% anthrone-sulfuric acid solution to the mark in an ice water bath" to determine the absorbance, and the amount of anhydrous glucose in the sample solution was read from the standard curve to calculate.

[0130] 2. Determination of the content of the saponin

[0131] (1) Preparation of the standard curve: 0.0 mL, 0.4 mL, 0.6 mL, 0.8 mL, 1.0 mL and 1.2 mL of the ginsenoside Re standard solution were taken in 10 mL stoppered colorimetric tubes, placed in a water bath to evaporate the solvent, 0.2 mL of vanillin solution was added, then 0.8 mL of perchloric acid was added, mixed, the residue was completely dissolved, placed in a 60 DEG C water bath for heating for 10 min, taken out, cooled in an ice bath, 5.0 mL of glacial acetic acid was added, shaken, and determined as a blank with the corresponding reagent, and the absorbance was immediately determined at a wavelength of 560 nm;

[0132] (2) Assay: 3 cm activated and eluted macroporous resin was loaded into a glass chromatographic column with an inner diameter of 1.5 cm, 1.0 mL sample was added, eluted with 25 mL water, the eluate was discarded, and then 25 mL 70% ethanol was used to elute ginsenosides at a speed of not more than 3 mL / min until the eluate was colorless, the eluate was collected in an evaporating dish, placed in a 60°C water bath to evaporate, the residue was dissolved with a small amount of methanol and transferred to a 10 mL stoppered colorimetric tube, the absorbance was measured according to the method under the preparation of the standard curve from "placing in a water bath to evaporate the solvent" to the standard solution, and the total saponin content was calculated according to the standard curve.

[0133] 3. Sensory evaluation of the extract of Polygonatum

[0134] The sensory evaluation was performed by 12 people who met the sensory evaluation criteria, with a full score of 100 points, and the evaluation indexes included taste (30 points), smell (30 points), color (20 points), and tissue morphology (20 points), and the specific evaluation criteria were shown in Table 1.

[0135] Table 1. Sensory evaluation criteria

[0136]

[0137] Through analysis of the recorded data, the results of the extraction rates of polysaccharides and saponins of Polygonatum of the examples and the comparative examples were shown in Table 2.

[0138] Table 2. Extraction rates of polysaccharides and saponins of Polygonatum

[0139]

[0140]

[0141] As can be seen from Table 2, when the preparation method provided by the present application was used, the extraction rates of polysaccharides and saponins of Polygonatum in the obtained extract of Polygonatum were high; specifically, the extraction rate of polysaccharides of Polygonatum was above 70.28%, and the extraction rate of saponins of Polygonatum was above 22.68%.

[0142] As can be seen from Example 1 and Comparative Example 1, when no colloid milling and pulping treatment is performed, the extraction rates of Polygonatum polysaccharides and Polygonatum saponins in the obtained Polygonatum extract show a significant decreasing trend. As can be seen from Example 1 and Comparative Examples 2-3, when no simultaneous enzymatic hydrolysis-fermentation extraction is performed, the extraction rates of Polygonatum polysaccharides and Polygonatum saponins in the obtained Polygonatum extract also show a significant decreasing trend. As can be seen from Example 1 and Comparative Examples 4-9, when only enzymatic hydrolysis or fermentation is selected in the simultaneous enzymatic hydrolysis-fermentation extraction, the effect of the present invention cannot be achieved. As can be seen from Example 1 and Comparative Examples 10-12, when conventional hot water extraction is used, the extraction rates of Polygonatum polysaccharides and Polygonatum saponins in the Polygonatum extract show a significant decreasing trend.

[0143] As can be seen from Examples 1 and 2-4, in the simultaneous enzymatic hydrolysis-fermentation extraction, the choice of enzyme also affects the overall performance of the extract. When papain is further selected as the enzyme, the extraction rates of Polygonatum polysaccharides and Polygonatum saponins in the obtained Polygonatum extract are higher. As can be seen from Examples 1 and 5, in the simultaneous enzymatic hydrolysis-fermentation extraction, the choice of microorganism also affects the overall performance of the extract. When yeast is further selected as the microorganism, the extraction rates of Polygonatum polysaccharides and Polygonatum saponins in the obtained Polygonatum extract are higher. As can be seen from Examples 1 and 6, when the first enzymatic hydrolysis of esterase is not introduced, although the extraction rates of Polygonatum polysaccharides and Polygonatum saponins in the Polygonatum extract are better than those in the comparative example, they are still somewhat lower than those in Example 1, indicating that the first enzymatic hydrolysis of esterase can effectively help improve the extraction rate of the active ingredients in Polygonatum.

[0144] In addition, sensory evaluation was performed on the Polygonatum extracts prepared in Example 7 and Comparative Example 12, and the sensory evaluation results are as follows: Figure 1 As shown, from Figure 1 It can be clearly seen that the Polygonatum extract prepared by the preparation method provided by the present invention has a high sensory score.

[0145] Finally, it should be noted that the above embodiments are used to illustrate the technical solutions of the present invention and not to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A method for preparing a polygonatum extract, characterized by, The preparation method comprises the following steps: (1) taking the crushed polygonatum sibiricum to soak in water, grinding after the soaking is finished, and obtaining a polygonatum sibiricum slurry; the fineness of the polygonatum sibiricum slurry is ≤2 mm; (2) adding enzymes and microorganisms to the polygonatum sibiricum slurry for simultaneous extraction by enzymolysis-fermentation; (3) sterilizing, filtering, collecting the filtrate and centrifuging after the simultaneous extraction by enzymolysis-fermentation, taking the supernatant, and concentrating to obtain a polygonatum sibiricum extract; Before the step (2), the method further comprises the steps of adding esterase to the polygonatum sibiricum slurry for first enzymolysis and enzyme inactivation; In the step (2), the enzymes comprise papain; In the step (2), the microorganisms comprise yeast; In the step (2), the extraction temperature in the simultaneous extraction by enzymolysis-fermentation is 25-35℃, and the extraction time is 12-36 h.

2. The production method according to claim 1, characterized by, In the step (1), the mass ratio of the crushed polygonatum sibiricum to water is 1:(5-10); And / or, the soaking time is 0.5-1.5 h; And / or, the grinding rate is 5-20 kg / min.

3. The preparation method according to claim 1, characterized in that, In the step (2), the addition amount of the esterase for the first enzymolysis is 0.2-1.5% based on the mass of the crushed polygonatum sibiricum; And / or, the temperature for the first enzymolysis is 40-60℃, and the time is 1-3 h; And / or, during the first enzymolysis, the material-liquid ratio of the polygonatum sibiricum slurry is 1:(10-40), and the pH value is 4.0-7.

0.

4. The method of claim 1, wherein, In the step (2), the mass percentage of the enzymes is 0.1-1.5% and the mass percentage of the microorganisms is 1-10% based on the mass of the crushed polygonatum sibiricum in the simultaneous extraction by enzymolysis-fermentation.

5. The preparation method according to claim 1, characterized in that, The temperature for the enzyme inactivation is 90-100℃, and the time is 10-20 min; And / or, the temperature for the sterilization is 85-115℃, and the time is 10-30 min.

6. A polygonum multiflorum extract, characterized by, The polygonatum sibiricum extract is prepared by the preparation method in any one of claims 1-5.

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