Gastrodia elata and walnut beverage and preparation method thereof

Abstract

The invention relates to a gastrodia elata walnut beverage and a preparation method thereof, and belongs to the technical field of food. The gastrodia elata and walnut beverage disclosed by the invention is prepared from the following raw materials in parts by weight: 10 to 15 parts of dried gastrodia elata, 100 to 120 parts of walnut kernels, 1 to 1.5 parts of ficin, 25 to 30 parts of white granulated sugar and / or 30 to 35 parts of a sweetening agent, 0.1 to 0.15 part of vitamin E, 1 to 1.5 parts of sucrose ester, 1.5 to 2 parts of sodium caseinate, 0.3 to 0.5 part of sodium erythorbate, 0.6 to 0.8 part of sodium citrate and 0.8 to 1.0 part of sodium bicarbonate. Active substances rich in gastrodia elata mainly comprise gastrodia elata polysaccharide and gastrodin (polyphenol), which can be exactly combined with an active substance walnut polypeptide of walnuts, so that better biological activity is obtained. By means of the method, combination of gastrodia elata polysaccharide, gastrodin and walnut polypeptide can be promoted, the obtained product is higher in release, digestion, absorption and activity, and the effects of improving sleep quality, relieving brain fatigue and supplementing brain nutrition can be achieved.

Description

Technical Field

[0001] The invention belongs to the technical field of food, and in particular relates to a gastrodia elata walnut beverage and a preparation method thereof. Background Art

[0002] Sleep is an active process of the human body and one of the indispensable life activities of the human body. Insomnia is a kind of "sleep disorder", and its main clinical manifestations are difficulty falling asleep, early awakening, and poor sleep quality. Nowadays, factors such as fast-paced social life, invisible work pressure, and bad living habits induce insomnia. The incidence rate is increasing year by year and tends to be younger. It has become an epidemic in modern urban civilization, seriously affecting people's health and daily life. Insomnia, depression, memory degeneration and other symptoms caused by learning and life pressure, chronic stress, environmental factors, etc. can lead to brain lesions, thereby damaging cognitive and memory functions. Therefore, people of different ages have the problem of decreased memory function.

[0003] Gastrodia elata Blume was originally named Red Arrow, which was first recorded in Shennong's Herbal Classic. In the Song Dynasty, Kaibao Materia Medica began to record the name of Gastrodia elata. In Compendium of Materia Medica, Li Shizhen combined the two and called them Gastrodia elata. Gastrodia elata is mainly produced in Zhaotong, Yunnan, and is also distributed in Guizhou, Hubei and other places. Modern pharmacological research shows that Gastrodia elata is rich in a variety of active substances such as gastrodin and p-hydroxybenzyl alcohol, and has good sedative and anticonvulsant effects.

[0004] my country's walnut production accounts for 49.7% of the world's total, making it a veritable walnut producing country. The production of walnuts has increased fivefold in the past decade. With the continuous increase in walnut production, the walnut industry is also developing steadily. Walnuts contain 15%-20% high-quality protein, 18 kinds of amino acids, 60%-70% fat (more than 90% are unsaturated fatty acids), rich in linolenic acid and a variety of vitamins and minerals. The bioactive substances isolated from walnut green peels, walnut kernels and walnut meal have many health benefits in helping to improve brain health, improve metabolic syndrome, enhance immunity and resist fatigue. Walnuts contain a variety of bioactive substances that are beneficial to the human body, mainly by reducing oxidative stress and inflammatory reactions, promoting neurotransmitters, regulating intestinal flora, and promoting brain health.

[0005] Walnuts are rich in protein that can be hydrolyzed into walnut peptides. In recent years, studies on walnut peptides have found that they have good biological activity. In order to improve the solubility and absorbability of polypeptides, they are often modified, such as glycosylation modification, polyphenol non-covalent modification, etc. The existing tonic beverage processing methods mostly use the physical mixing of gastrodia and walnuts, and there are also physical mixing of gastrodin extracted from gastrodia and walnut peptides produced after walnut enzymatic hydrolysis. The gastrodia walnut beverage prepared by physical mixing has only the superposition of the effects of the two raw materials, and the process is more complicated and needs to be pre-treated separately, especially after enzymatic hydrolysis, proteins / peptides of different molecular weights need to be separated and purified, and gastrodin needs to be purified after extraction.

[0006] Based on this, the present invention is proposed. Summary of the invention

[0007] In order to solve the above technical problems, the present invention provides a gastrodia elata walnut beverage and a preparation method thereof. The active substances rich in gastrodia elata are mainly gastrodia elata polysaccharides and gastrodin (polyphenols), which can be combined with walnut polypeptide, an active substance of walnut, to obtain better biological activity. The method of the present invention can promote the combination of gastrodia elata polysaccharides, gastrodin and walnut polypeptide, and the obtained product has stronger release, digestion, absorption and activity, which can improve sleep quality, reduce brain fatigue and supplement brain nutrition.

[0008] The first object of the present invention is to provide a gastrodia elata walnut beverage, the raw materials of which include, by weight: 10-15 parts of dried gastrodia elata, 100-120 parts of walnut kernels, 1-1.5 parts of ficin, 25-30 parts of white sugar and / or 30-35 parts of sweetener, 0.1-0.15 parts of vitamin E, 1-1.5 parts of sucrose ester, 1.5-2 parts of sodium caseinate, 0.3-0.5 parts of sodium isoVC, 0.6-0.8 parts of sodium citrate, and 0.8-1.0 parts of sodium bicarbonate.

[0009] In one embodiment of the present invention, the enzymatic activity of the ficin is 100 U / g-150 U / g.

[0010] In one embodiment of the present invention, one unit of enzyme activity refers to the amount of enzyme required to produce a ΔA280 of 1.0 per minute at a final volume of 10 mL (1 cm optical path), 37° C., and pH 7.0 when measuring the trichloroacetic acid (TCA) soluble product of casein.

[0011] In one embodiment of the present invention, the sweetener is obtained by mixing erythritol, steviol glycoside and mogroside in a mass ratio of 99.78:0.14:0.08.

[0012] In one embodiment of the present invention, the dried gastrodia elata is produced in Guizhou Province, and the gastrodia elata in Dafang County, Bijie City, Guizhou Province is the best. Guizhou Province is one of the production areas of gastrodia elata in China, with unique natural conditions. The gastrodia elata produced there is of good quality, and it is known as "the best gastrodia elata comes from Guizhou". "Chinese Materia Medica" states under the gastrodia elata: "The quality of the gastrodia elata produced in Guizhou is good, sold nationwide, and exported." "Chinese Authentic Medicinal Materials" lists gastrodia elata as the first "expensive medicine". Among the gastrodia elata in Guizhou, Dafang gastrodia elata is the best among the gastrodia elata in Guizhou, and it ranks first in both appearance and internal quality, and there is also a saying that "the best gastrodia elata in Guizhou is Dafang".

[0013] In one embodiment of the present invention, the walnut kernel is produced in Guizhou Province, and preferably in Hezhang County, Bijie City. Hezhang walnuts are rich in protein, unsaturated fatty acids, minerals, vitamins B and E. Regular consumption can prevent cell aging, strengthen the brain, enhance memory and delay aging. It is an ideal skin beauty agent for the human body and has a great effect on preventing and treating premature graying and shedding of hair. The special vitamin ingredients in it can reduce the intestinal absorption of cholesterol, and are suitable for patients with arteriosclerosis, hypertension and coronary heart disease.

[0014] In one embodiment of the present invention, the ficin has the advantages of high temperature resistance, high activity, good stability, insensitivity to pH changes, etc., and can hydrolyze walnut protein into appropriate polypeptides.

[0015] A second object of the present invention is to provide a method for preparing the gastrodia elata walnut beverage, comprising the following steps:

[0016] S1. Fry the walnut kernels, add walnut oil, beat into walnut paste, and pass through a 100-mesh sieve to obtain walnut paste;

[0017] S2, stirring the dried Gastrodia elata and the walnut paste described in S1 in water, adding ficin to perform enzymolysis in stages to obtain an enzymolysis solution;

[0018] S3, cooling the enzymatic hydrolyzate described in S2 to 50° C.-60° C., adding vitamin E, sucrose ester, sodium caseinate and water, stirring evenly, and homogenizing to obtain a mixed solution;

[0019] S4. Cool the mixed solution described in S3 to 30° C.-40° C., add white sugar and / or sweetener, sodium isoVC, sodium citrate and sodium bicarbonate, stir evenly, and obtain the Gastrodia elata walnut beverage through homogenization, filtration and sterilization.

[0020] In one embodiment of the present invention, in S1, the mass volume ratio of the walnut kernel and the walnut oil is (100-120):(10-15) g / mL.

[0021] In one embodiment of the present invention, in S2, the mesh size of the dried Gastrodia elata is 100-120 mesh;

[0022] The mass volume ratio of the dry Gastrodia elata and water is (10-15):(300-400) g / mL.

[0023] In one embodiment of the present invention, in S2, the staged enzymatic hydrolysis includes two stages, the first stage is enzymatic hydrolysis at 40°C-50°C and 100rpm-150rpm for 3h-4h; the second stage is heat preservation at 90°C-100°C for 9min-11min.

[0024] In one embodiment of the present invention, in S3, the volume of the mixed solution is 1000 mL;

[0025] The homogenization pressure is 38Mpa-42Mpa, and the number of times is 2 times.

[0026] In one embodiment of the present invention, in S4, the homogenization pressure is 28Mpa-32Mpa, and the number of times is 1 time.

[0027] In one embodiment of the present invention, in S4, the sterilization temperature is 120°C-122°C, and the time is 9min-11min.

[0028] The technical solution of the present invention has the following advantages over the prior art:

[0029] (1) Glycosylation modification refers to the modification between monosaccharides, oligosaccharides or polysaccharides and proteins / peptides through covalent linkage. The most common one is the Maillard reaction, which is the condensation and rearrangement between carbonyl compounds (reducing sugars) and amino compounds (amino acids, peptides, proteins), and finally generates a complex polymer with better antioxidant properties and free radical scavenging ability. Non-covalent modification of polyphenols has highly reactive groups, which are easy to form stable complexes with proteins / peptides, and can improve antioxidant activity, emulsification activity and stability. The preparation method of the present invention uses ficin for auxiliary processing, which can promote the quantitative and directional hydrolysis of walnut protein into polypeptides that are more easily reactive with gastrodia polysaccharides and gastrodin, and obtain a product with higher activity.

[0030] (2) The Gastrodia elata walnut beverage of the present invention can improve sleep quality, reduce brain fatigue, and supplement brain nutrition, and is suitable for drinking before going to bed. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] In order to make the content of the present invention more clearly understood, the present invention is further described in detail below according to specific embodiments of the present invention in conjunction with the accompanying drawings, wherein:

[0032] Figure 1The polysaccharide content of the gastrodia elata from different origins and different textures of the present invention;

[0033] Figure 2 The yield of walnut peptides from different origins and different processing methods of the present invention;

[0034] Figure 3 This is the SDS-PAGE image of the enzymatic hydrolyzate in Test Example 1 of the present invention;

[0035] Explanation of the accompanying drawings: M-Marker, 1-Comparative Example 9, 2-Comparative Example 2, 3-Comparative Example 3, 4-Comparative Example 7, 5-Comparative Example 4, 6-Comparative Example 5, 7-Example 1, 8-Comparative Example 13, 9-Example 2, 10-Comparative Example 10, 11-Comparative Example 6, 12-Comparative Example 8, 13-Example 3, 14-Comparative Example 11, 15-Example 4, 16-Comparative Example 12. DETAILED DESCRIPTION

[0036] The present invention is further described below in conjunction with the accompanying drawings and specific embodiments so that those skilled in the art can better understand the present invention and implement it. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. It should be understood that the specific embodiments are only used to explain the present invention, but the embodiments are not intended to limit the present invention.

[0037] In the present invention, unless otherwise explained, technical and scientific terms used in the present invention have the same meanings as commonly understood by those skilled in the art to which the present invention belongs.

[0038] In the present invention, unless otherwise stated, the term "and / or" used in the present invention includes any and all combinations of one or more of the associated listed items.

[0039] In the present invention, unless otherwise stated, the experimental methods used in the embodiments of the present invention are conventional methods unless otherwise stated, and the materials, reagents, etc. used are all commercially available unless otherwise stated.

[0040] In the present invention, unless otherwise specified, the sweetener used in the embodiments of the present invention is obtained by mixing erythritol, steviol glycoside and mogroside in a mass ratio of 99.78:0.14:0.08.

[0041] 1. Comparison of different raw materials

[0042] (1) Comparison of Gastrodia elata from different origins and textures

[0043] Fresh and dried Gastrodia elata from Yunnan, Sichuan, and Dafang, Guizhou were selected for the experiment to compare the polysaccharide content of Gastrodia elata. The fresh Gastrodia elata was first washed, the surface moisture was absorbed, the epidermis was scraped off, and an appropriate amount of pure water was added to pulp it and then freeze-dried into powder to obtain fresh Gastrodia elata freeze-dried powder; the dried Gastrodia elata was directly powdered to obtain dry Gastrodia elata powder. 90% ethanol was added to the powders for reflux defatting. After defatting, wait until the ethanol is completely evaporated, and deionized water is added according to the powder mass: deionized water volume ratio of 1:30. Heat and extract at 75°C for 4h. After concentration, Savage reagent (chloroform: n-butanol = 4:1) was added for deproteinization. This was repeated three times. Anhydrous ethanol was added to the aqueous solution until the ethanol volume concentration reached 80%, and the precipitation was allowed to stand at 4°C. The precipitated part was collected, and the ethanol was evaporated and freeze-dried and weighed. The test results of the polysaccharide content are as follows. Figure 1 As shown. Figure 1 It can be seen that the dried Gastrodia elata from Guizhou Dafang contains the highest content of Gastrodia elata polysaccharides, so the subsequent experiments used dried Gastrodia elata slices from Guizhou Dafang.

[0044] (2) Comparison of walnuts from different origins and processed in different ways

[0045] Walnuts from Yunnan, Sichuan, and Hezhang, Guizhou were selected for the experiment to compare the yield of walnut peptides. Take an appropriate amount of walnuts and mix them with deionized water at a solid-liquid ratio of 1:20, beat them, keep them at 45°C for 3h, adjust the pH to 10, centrifuge them at 4000rpm for 5min, discard the upper floating oil and the lower precipitate, take the middle layer clear liquid, adjust the pH to 4.5, centrifuge them at 4000rpm for 5min, wash the precipitate with deionized water until it is neutral; add 25mL of 10g / 100mL TCA solution, mix well, let it stand, take all the supernatant after centrifugation, filter and take a certain amount of filtrate in the digestion tube, and determine its nitrogen content (N2) after digestion. Walnuts from different origins were washed, drained, and stir-fried over low heat until basically cooked. The skin was peeled / rubbed off, 12 mL of walnut oil was added, and the mixture was beaten into walnut paste, which was passed through a 100-mesh sieve to obtain walnut paste. Dried Gastrodia elata was crushed into 100-mesh powder to obtain Gastrodia elata powder. The Gastrodia elata powder and walnut paste were mixed evenly, 350 mL of purified water was added, and the mixture was stirred and mixed using a food processor. Ficin was added, and the mixture was first enzymolyzed at 45 °C and 125 rpm for 3 h, and then heated to 95 °C and maintained for 10 min to obtain an enzymatic solution. 25 mL of 10 g / 100 mL TCA solution was added, mixed evenly, allowed to stand, and all the supernatant was taken after centrifugation. After filtering, a certain amount of filtrate was taken into a digestive tube, and its nitrogen content (N1) was determined after digestion. The walnut peptide yield is calculated as follows: walnut peptide yield = N1 / N2×100%, where N1 is the soluble nitrogen in 10% TCA, mg; N2 is the total nitrogen in the raw material, mg (total nitrogen is determined by the Kjeldahl method of GB / T 5009.5-2016). The test results of walnut peptide yield are as follows: Figure 2 As shown. Figure 2It can be seen that the walnuts from Hezhang, Guizhou can produce the highest yield of soluble walnut peptides, especially after being processed according to the method of the present invention, the yield of soluble walnut peptides has been significantly improved, so the subsequent experiments use walnut kernels from Hezhang, Guizhou.

[0046] 2. Comparison of different embodiments and comparative examples

[0047] Example 1-4 Gastrodia elata walnut beverage with different raw material dosage and enzymolysis time

[0048] The raw material components and their weight parts are as follows: dried gastrodia elata, walnut kernels, ficin, 28g of white sugar and 32g of sweetener, 0.12g of vitamin E, 1.2g of sucrose ester, 1.8g of sodium caseinate, 0.4g of sodium isoVC, 0.7g of sodium citrate, and 0.9g of sodium bicarbonate;

[0049] Among them, the specific dosage (g) and enzymolysis time (h) of dried gastrodia elata, walnut kernel and ficin are shown in Table 1:

[0050] Table 1

[0051] Sample Gastrodia elata Walnut Ficin Enzyme hydrolysis time Example 1 10 100 1 3 Example 2 15 120 1.5 4 Example 3 10 120 1.5 3 Example 4 15 100 1.5 4

[0052] The specific steps include:

[0053] S1, grinding dry Gastrodia elata into 100 mesh powder to obtain Gastrodia elata powder;

[0054] S2. Wash and drain the walnut kernels, stir-fry over low heat until they are basically cooked, peel / rub off the outer skin, add 12 mL of walnut oil, beat into walnut paste, and pass through a 100-mesh sieve to obtain walnut paste;

[0055] S3, stir the gastrodia powder and walnut paste evenly, add 350 mL of purified water, stir and mix with a food processor, add ficin, first enzymolyze at 45°C and 125 rpm for 3h-4h, then heat to 95°C and maintain for 10min to obtain an enzymolysis solution;

[0056] S4. After the enzymatic hydrolyzate is cooled to 55°C, vitamin E powder, sucrose ester and sodium caseinate are added to make up the liquid volume to 1000mL and stir evenly. Use a homogenizer to homogenize twice at a pressure of 40Mpa to obtain a mixed solution;

[0057] S5. Cool the mixed solution to 35° C., add sodium isoVC, sodium citrate, sodium bicarbonate, white sugar and / or sweetener, stir evenly, homogenize once with a homogenizer at a pressure of 30 MPa, filter through 100 mesh, and sterilize at 121° C. for 10 min to obtain the Gastrodia elata walnut beverage.

[0058] Comparative Examples 1-8 Gastrodia elata walnut beverages with different raw material dosages

[0059] The same as Example 1, except that the specific dosage (g) of dried gastrodia elata, walnut kernel and ficin is as shown in Table 2:

[0060] Table 2

[0061] Sample Gastrodia elata Walnut Ficin Comparative Example 1 10 0 1 Comparative Example 2 0 100 1 Comparative Example 3 5 100 1 Comparative Example 4 20 120 1.5 Comparative Example 5 10 80 1 Comparative Example 6 10 140 1.5 Comparative Example 7 10 100 0.5 Comparative Example 8 15 120 2

[0062] Comparative Examples 9-11 Gastrodia elata walnut beverages with different proteases

[0063] The method is basically the same as Example 1, except that the protease is different, as shown in Table 3:

[0064] Table 3

[0065] Sample Protease Comparative Example 9 Papain Comparative Example 10 Alkaline protease Comparative Example 11 Flavor protease

[0066] Comparative Example 12-13 Gastrodia elata walnut beverage with different enzymolysis time

[0067] The method is basically the same as Example 1, except for the enzymolysis time (h), as shown in Table 4:

[0068] Table 4

[0069] Sample Enzyme hydrolysis time Comparative Example 12 2 Comparative Example 13 5

[0070] Test Example 1 Molecular Weight Analysis and Comparison

[0071] Based on Examples 1-4 and Comparative Examples 2-13, the enzymatic hydrolysate was analyzed by SDS-PAGE to compare the molecular weight and distribution of the proteins. Comparative Example 1 was not analyzed because it only used Gastrodia elata as the raw material and had very little protein content. The analysis method is as follows: take 20 μL of the enzymatic hydrolysate and dilute it 20 times with ultrapure water, take 40 μL of the dilution and add 10 μL of 5×LoadingBuffer (protein sample preparation buffer) thereto, heat at 100°C for 10 minutes, and use 12% protein gel to perform SDS-PAGE electrophoresis on the protein sample. Electrophoresis procedure: run at 85V for 20 minutes and then run at 125V for 1 hour. After the electrophoresis, the protein gel was stained with Coomassie Brilliant Blue Rapid Stain for 20 minutes, then decolorized with pure water, and photographed to observe the protein bands after decolorization for 6 hours. The results are as follows Figure 3 As shown. Figure 3It can be seen that the protein bands of Examples 1-4 are the most abundant and the darkest, especially 17kDa, 55kDa, and 72kDa have special bands; the remaining lanes are substantially free of / extremely shallow, indicating that Examples 1-4 contain proteins that are not present in the comparative examples, which are products of the combination of small molecule walnut peptides and Gastrodia polysaccharides and Gastrodin. Comparative Example 13 also has corresponding bands, but the color is extremely shallow, indicating that the enzymolysis time exceeds the appropriate range, resulting in a reduction in active products; Comparative Examples 9-11 have very few bands, indicating that inappropriate enzymes cannot achieve the corresponding hydrolysis effect, or the enzymolysis effect is too much, resulting in the enzymolysis of walnut protein into short peptides of extremely small molecular weight (≤8kDa), and the synthesis of active products is also reduced accordingly; other inappropriate preparation conditions / raw material compositions also fail to achieve corresponding effects.

[0072] Different proteases have very different amino acid sequences. The closest to ficin (molecular weight of about 26,000) is papain (molecular weight of 23,000-23,400). The amino acid sequences near the active centers of the two are also different, resulting in different substrates (ficin has a significantly higher affinity for substrates than papain), different enzymatic cleavage sites, and different reaction products. Ficin is a cysteine ​​protease (sulfhydryl protease), and its active center contains a sulfhydryl group (-SH). Most neutral proteases are metalloproteases, and their active centers contain metal ions such as Zn 2+ The activity of fig protease is stable when treated with pH 5-10 for 0.5h. The optimal pH of neutral protease is 6-7, and it is rapidly inactivated when it exceeds this range.

[0073] Test Example 2 Emulsion Stability Test

[0074] Take 100 μL of each beverage of Examples 1-4 and Comparative Examples 1-13, add them to 4.9 mL of 0.1% sodium dodecyl sulfate (SDS) solution, mix them and measure the absorbance at 500 nm wavelength as A0, let them stand for 30 minutes and measure the absorbance again as A30, and use 0.1% SDS solution as a blank control. Stability = A0 / (A0-A30)×30, compare the stability of the emulsification effect of the beverage, the results are shown in Table 5:

[0075] Table 5

[0076]

[0077]

[0078] As can be seen from Table 5, the scheme of the embodiment can significantly improve the emulsification effect of the beverage, which cannot be achieved when walnuts or one of the Gastrodia raw materials is missing (Comparative Examples 1-2), and cannot be achieved when the enzymatic hydrolysis is insufficient or too much (Comparative Examples 12-13). Other unsuitable raw material ratios (Comparative Examples 3-8) and protease types (Comparative Examples 9-11) will affect the emulsification effect. Only the scheme of the present invention can achieve the optimal ratio of Gastrodia polysaccharide and walnut polypeptide, and the walnut polypeptide in the walnut and the Gastrodia polysaccharide and gastrodin (polyphenol) in the Gastrodia will undergo glycosylation reaction, and the generated glycosylated polypeptide can significantly improve the emulsification performance, because the sugar end of the glycosylated polypeptide can increase the viscosity of the water phase in the oil-water emulsification system, and the steric hindrance of the peptide end can increase the thickness of the protective film formed by emulsification, thereby enhancing the emulsification stability.

[0079] Test Example 3 Anti-lipid oxidation detection

[0080] Dissolve lecithin in 0.01mol / L phosphate buffer solution with a pH of 7.40 to prepare a 10mg / mL solution, labeled as LLS; add 15g TCA, 0.37g TBA, and 2.0mL concentrated hydrochloric acid to distilled water to prepare a 100mL solution, labeled as TCA-TBA-HCl. Add 2.0mL of LLS solution, 2.0mL of 400μmol / L FeCl3 solution, and 2.0mL of the enzymatic hydrolyzate of Examples 1-4 and Comparative Examples 1-13 to a test tube, mix well, bathe at 37°C for 1h, then add 2.0mL of TBA-TCA-HCl solution, bathe in boiling water for 15min, cool rapidly, centrifuge at 4000rpm for 10min, and measure the absorbance of the supernatant at a wavelength of 532nm. Sample anti-lipid oxidation activity = (A blank - A sample) / A blank. Wherein: A blank is the absorbance value obtained by using deionized water instead of enzymatic solution for detection; A sample is the absorbance value of the sample group. The results are shown in Table 6:

[0081] Table 6

[0082] Sample Anti-lipid oxidation / % Example 1 58 Example 2 60 Example 3 54 Example 4 63 Comparative Example 1 35 Comparative Example 2 25 Comparative Example 3 39 Comparative Example 4 27 Comparative Example 5 34 Comparative Example 6 32 Comparative Example 7 29 Comparative Example 8 36 Comparative Example 9 36 Comparative Example 10 41 Comparative Example 11 35 Comparative Example 12 40 Comparative Example 13 42

[0083] It can be seen from Table 6 that the enzymolysis solution of the embodiment has good resistance to lipid oxidation. This is because the associations formed by the polyphenols in Gastrodia elata and the polypeptides in walnut in the relevant reactions have a certain ability to resist lipid oxidation, and this ability to resist lipid oxidation will change with the dosage of Gastrodia elata, walnut, and protease and the enzymolysis time. Only the scheme of the present invention can achieve the best effect of Gastrodia elata polyphenols and walnut polypeptides.

[0084] Test Example 4 Determination of p-anisidine

[0085] The value of p-anisidine characterizes the amount of oxidation products such as aldehydes and ketones during the oxidation of fats and oils. Fats and oxygen react to generate aldehydes and ketones. Such compounds have toxic side effects such as carcinogenicity, increased blood pressure, and damage to the body's absorption of fat-soluble vitamins, which are detrimental to human health. The content of aldehyde compounds in edible oils and oils is generally expressed by the p-anisidine value. The larger the value, the more serious the deterioration of the oil. Take the enzymatic hydrolyzate of Examples 1-4 and Comparative Examples 1-13 and freeze-dry it, then add walnut oil, stir at 50°C for 10min, cool and filter it, place it in an oven, heat it at 180±2°C for 6h every day, and heat it continuously for 4d. After heating and cooling every day, the sample is sealed and stored in a 4°C refrigerator. Weigh 2.0g of oil sample in a 25mL volumetric flask, dissolve it with isooctane and make up to volume. Accurately weigh 5mL of oil sample in a 25mL colorimetric tube, accurately add 1mL of p-anisidine reagent with a pipette, shake it, and let it stand for 10min. First, use isooctane solvent as blank to measure the absorbance of the oil sample at 350nm wavelength Ablank. Then use 5mL isooctane + 1mL p-anisidine reagent (stand for 10min) as blank to measure the absorbance of the oil solution Asample. p-anisidine = 25 × (1.2Asample-Ablank) / W, where W refers to the mass of the oil sample. The results are shown in Table 7:

[0086] Table 7

[0087]

[0088]

[0089] It can be seen from Table 7 that the enzymatic hydrolyzate of the embodiment has a lower p-anisidine value. This is because the associations formed by the polyphenols in Gastrodia elata and the polypeptides in walnuts in the relevant reactions have a certain ability to resist lipid oxidation, and this ability to resist lipid oxidation will change with the dosage of Gastrodia elata, walnuts, and protease and the enzymatic hydrolysis time. Only the parameter range provided in the invention content can achieve the optimal effect of Gastrodia elata polyphenols and walnut polypeptides.

[0090] Test Example 5 DPPH free radical scavenging experiment

[0091] Prepare 1.75 × 10 -4 mol / L DPPH free radical solution, take 2mL of the enzymatic hydrolysate of Examples 1-4 and Comparative Examples 1-13 and add it to 2mL of DPPH free radical ethanol solution, react in the dark for 1h at room temperature, and then detect its absorbance at a wavelength of 517nm. DPPH scavenging rate = [A blank - (A reaction - A sample)] / A blank × 100%, where A blank is the absorbance value of the DPPH free radical solution, A reaction is the absorbance value of the solution after adding the sample to react for 1h, and A sample is the absorbance value of the mixed enzymatic hydrolysate of Gastrodia elata and walnut, and the results are shown in Table 8:

[0092] Table 8

[0093]

[0094]

[0095] As can be seen from Table 8, the enzymolysis solution of the embodiment has a strong antioxidant (DPPH removal) ability. This is because the polyphenols in Gastrodia elata and the polypeptides in walnut form an association in the relevant reaction, and the antioxidant (DPPH removal) ability of this association is much higher than that of the single components of Gastrodia elata or walnut (Comparative Examples 1-2), and the scavenging ability of this antioxidant group will change with the dosage of Gastrodia elata, walnut, protease and enzymolysis time. Only the solution of the present invention can achieve the best effect of Gastrodia elata polyphenols and walnut polypeptides.

[0096] Test Example 6 Detection of dimethyl disulfide in Gastrodia elata

[0097] Take 2 mL of the enzymatic hydrolysate of Examples 1-4 and Comparative Examples 1-13, place it in a sampling bottle of a solid phase microextraction instrument, perform headspace extraction, the extraction temperature is 85°C, the extraction time is 30 min, and after the extraction is completed, quickly remove the extraction head and immediately insert it into the gas chromatograph injection port. The gas chromatography conditions are as follows:

[0098] Chromatographic column: ZB-5MSI 5% Phenyl-95% DiMethylpolysiloxane (30m×0.25mm×0.25μm) flexible quartz capillary column, injection port temperature 250℃, thermal desorption 3min. Start column temperature 45℃, retain 2in, program temperature rise to 220℃ at 4℃ / min, maintain 2min; vaporization chamber temperature 250℃; high-purity He carrier gas; carrier gas flow rate 1.0mL / min, non-split injection; solvent delay time 1.5min;

[0099] Mass spectrometry conditions: EI source ion source, mass range 20-450 amu; ion source temperature 230°C; quadrupole temperature set to 150°C, interface temperature set to 280°C.

[0100] The content of dimethyl disulfide that can be detected in each sample is shown in Table 9:

[0101] Table 9

[0102] Sample Dimethyl disulfide content / % Example 1 0.08 Example 2 0.11 Example 3 0.04 Example 4 0.13 Comparative Example 1 0.45 Comparative Example 2 0 Comparative Example 3 0.39 Comparative Example 4 0.78 Comparative Example 5 0.49 Comparative Example 6 0.42 Comparative Example 7 0.39 Comparative Example 8 0.66 Comparative Example 9 0.37 Comparative Example 10 0.41 Comparative Example 11 0.41 Comparative Example 12 0.33 Comparative Example 13 0.40

[0103] As can be seen from Table 9, no Gastrodia elata was used in the raw materials of Comparative Example 2, so no dimethyl disulfide was detected; Comparative Example 4 used the largest amount of Gastrodia elata, so the content of dimethyl disulfide was also the highest (0.78%); However, Comparative Example 8 used the same amount of Gastrodia elata as Examples 2 and 4, and the content of dimethyl disulfide in Comparative Example 8 was 0.66%, while Examples 2 and 4 only had 0.11% and 0.13%, respectively. It can be seen that after adding appropriate amounts of walnut and fig proteases and carrying out appropriate reactions, the content of dimethyl disulfide, a bad odor substance in the sample, was reduced; Comparative Examples 5-7 and 9-11 all used the same amount of Gastrodia elata as Examples 1 and 3, but the content of dimethyl disulfide was still relatively high; Comparative Examples 12-13 did not achieve a good effect of removing odor due to inappropriate enzymolysis time.

[0104] Test Example 7 Sensory Evaluation

[0105] The sensory evaluation team selected sensory assessors in accordance with the provisions of GB / T 16291.1-2012, and submitted three samples of Tianma Walnut Drink to each assessor, two of which were exactly the same. The assessors observed and tasted each sample from left to right, and selected a different sample to determine whether the assessors could assess the sensory differences between the different samples. After screening, a sensory evaluation team was formed by 10 assessors (5 men and 5 women).

[0106] Before the test, each team member must conduct a sensory evaluation of the reference material used in order to better understand the various sensory indicators. Using the quantitative description sensory analysis method, the evaluation team evaluates the beverage samples in five aspects: appearance, aroma, taste, mouthfeel, and overall acceptance according to their own preferences. The full score is 10 points. The detailed scoring criteria are shown in Table 10:

[0107] Table 10

[0108]

[0109] All sensory tests were conducted in a sensory evaluation room. To prevent the influence of sequence effects, the samples were randomly marked with three markers and randomly delivered to each sensory evaluator. Each sample was evaluated three times, and the average value was used as the final score. The scoring results of each sample are shown in Table 11:

[0110] Table 11

[0111] Sample Appearance fragrance taste stability Overall acceptance Example 1 8 8 9 10 9 Example 2 9 7 8 9 8 Example 3 8 8 8 9 7 Example 4 9 7 7 9 8 Comparative Example 1 4 4 4 3 5 Comparative Example 2 3 3 5 4 4 Comparative Example 3 6 5 6 7 6 Comparative Example 4 6 1 1 6 3 Comparative Example 5 5 4 5 6 5 Comparative Example 6 7 5 4 5 5 Comparative Example 7 5 5 7 7 6 Comparative Example 8 6 2 2 7 3 Comparative Example 9 4 5 5 7 6 Comparative Example 10 4 6 6 6 5 Comparative Example 11 3 7 6 5 6 Comparative Example 12 3 6 5 3 4 Comparative Example 13 4 6 5 3 4

[0112] As can be seen from Table 11, the results of subjective scoring are consistent with the results of GC-MS and emulsification stability. Comparative Examples 4 and 8, which have high odor content, have the lowest aroma and taste scores; after the samples are placed for the same time, the emulsification conditions of Comparative Examples 1, 2, 12, and 13 have deteriorated. It can be seen that the scheme of the present invention can make the polysaccharides and polyphenols in Gastrodia elata react with the polypeptides in walnuts to generate a complex that helps maintain the stability of the emulsion; it can also eliminate the bad odor in the raw materials, make the olfactory and taste experience of the two more integrated and harmonious, and obtain a brain tonic beverage with better flavor.

[0113] Test Example 8: Experimental observation of mice on the improvement of learning and memory ability

[0114] Male ICR mice were selected for the experiment and randomly divided into groups of 5 mice each. The mouse aging model was first induced by subcutaneous injection of 200 mg / kg D-galactose, and then the beverage solution of Examples 1-4 and Comparative Examples 1-13 was given by gavage once a day for 30 consecutive days. Five mice in the blank group were gavaged with sodium carboxymethylcellulose solution and subcutaneously injected with normal saline; five mice in the model group were gavaged with sodium carboxymethylcellulose solution and subcutaneously injected with 200 mg / kg D-galactose to induce a mouse aging model. On the 31st day of the experiment, a mouse jumping platform experiment was conducted, and the mice were placed in a jumping platform box for adaptive training. The electric rod at the bottom of the jumping platform box was powered by alternating current. When the mice were shocked, they would jump onto the platform to avoid harmful stimuli. Before the test, the mice were placed in a training box for 5 minutes to adapt. After 24 hours, the mice were placed on the platform again for the experiment. The time required for the mice to jump off the platform for the first time within 5 minutes (jump platform latency) and the number of times they jumped off the platform within 5 minutes (number of errors) were recorded to compare the learning and memory abilities of the mice: the shorter the jump platform latency and the more errors, the worse the learning and memory abilities of the mice; the longer the jump platform latency and the fewer errors, the better the learning and memory abilities of the mice. The beverage's ability to repair brain aging in mice is shown in Table 12:

[0115] Table 12

[0116]

[0117]

[0118] As can be seen from Table 12, it can be seen from the data of the blank group and the model group that in order to avoid electric shock, normal mice will stay on the platform for a long time (170±12s), and the number of jumps is less (2 times). After inducing aging of mice with D-galactose, their learning and memory functions decline, the platform incubation period decreases to 78±19s, and the number of jumps is 6 times. After gavage with the beverage of Example 1-4, brain function is restored, the average platform time can be restored to 146s, and the average number of jumps is 2.5 times. The comparative examples do not obtain a better brain nourishing effect. Comparative Examples 1-8 use different amounts of Gastrodia elata and / or walnut as raw materials for brain tonic beverages. Although the brain function of mice is improved compared with the model group, it is far less than that of Examples 1-4; even if the Gastrodia elata and walnut selected are consistent with Example 2 as in Comparative Example 8, but because more ficin is added, the effective ingredients are hydrolyzed too much, which reduces the brain tonic effect. It can be seen that the amount of raw materials, the type of protease, and the enzymolysis time are closely related to the brain-tonifying effect, and the scheme of the present invention has achieved remarkable results.

[0119] Test Example 9: Volunteer Experiment to Observe the Improvement of Sleep Quality

[0120] Twenty volunteers with sleep disorders were selected for a population test. The inclusion criteria were as follows: (1) One or more of the following sleep symptoms: ① Difficulty maintaining sleep; ② Difficulty falling asleep; ③ Waking up earlier than expected. (2) One or more daytime symptoms: ① Lack of energy or fatigue; ② Impaired social function (such as school, family or career); ③ Memory loss and lack of concentration; ④ Emotional restlessness or irritability; ⑤ Behavioral problems (such as aggressiveness, hyperactivity or impulsiveness); ⑥ Daytime sleepiness; ⑦ Accidents or mistakes; ⑧ Lack of energy, motivation or work initiative; ⑨ Worry about one's own sleep quality. (3) The above (1) and (2) occur at least 3 times a week. (4) Insomnia is not caused by an inappropriate sleeping environment (quiet, dark, comfortable or safe environment) or opportunity (sufficient sleep time). The control group was given a commercially available soy milk beverage half an hour before going to bed every night, and the observation group was given a Gastrodia elata walnut brain tonic beverage (Example 1, Comparative Example 1-2) half an hour before going to bed every night. The beverage was taken for 3 consecutive weeks, and the scores of the Pittsburgh Sleep Quality Index (PSQI) before and after taking the beverage were compared to evaluate the effect of the Gastrodia elata walnut brain tonic beverage provided by the present invention on subjective sleep time, sleep onset time, sleep time, sleep efficiency, sleep disorders, etc. The PSQI scores (x±s) of volunteers with sleep disorders before and after treatment are shown in Table 13:

[0121] Table 13

[0122] Group n Before treatment After treatment Before and after difference Control group 5 10.32±1.21 9.78±1.80 -0.54 Example 1 5 11.87±2.22 7.09±1.98 -4.78 Comparative Example 1 5 12.23±1.44 10.62±0.87 -1.61 Comparative Example 2 5 10.51±0.99 9.02±2.35 -1.49

[0123] It can be seen from Table 13 that both Example 1 and Comparative Examples 1-2 are helpful in improving sleep quality, and the effect of Example 1 is particularly good, achieving more significant effects than Comparative Example 1 (using only Gastrodia elata) and Comparative Example 2 (using only walnuts).

[0124] Obviously, the above embodiments are merely examples for clear explanation and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived from these are still within the protection scope of the invention.

Claims

1. A gastrodia elata walnut beverage, characterized in that: The raw materials of the gastrodia elata walnut beverage include, by weight: 10-15 parts of dried gastrodia elata, 100-120 parts of walnut kernels, 1-1.5 parts of ficin, 25-30 parts of white sugar and / or 30-35 parts of sweetener, 0.1-0.15 parts of vitamin E, 1-1.5 parts of sucrose ester, 1.5-2 parts of sodium caseinate, 0.3-0.5 parts of sodium isoVC, 0.6-0.8 parts of sodium citrate, and 0.8-1.0 parts of sodium bicarbonate.

2. The gastrodia elata walnut beverage according to claim 1, characterized in that The enzymatic activity of the ficin is 100U / g-150U / g.

3. The gastrodia elata walnut beverage according to claim 1, characterized in that The sweetener is obtained by mixing erythritol, steviol glycoside and mogroside in a mass ratio of 99.78:0.14:0.

08.

4. A method for preparing a gastrodia elata walnut beverage as claimed in any one of claims 1 to 3, characterized in that: The following steps are involved: S1. Fry the walnut kernels, add walnut oil, beat into walnut paste, and pass through a 100-mesh sieve to obtain walnut paste; S2, stirring the dried Gastrodia elata and the walnut paste described in S1 in water, adding ficin to perform enzymolysis in stages to obtain an enzymolysis solution; S3, cooling the enzymatic hydrolyzate described in S2 to 50° C.-60° C., adding vitamin E, sucrose ester, sodium caseinate and water, stirring evenly, and homogenizing to obtain a mixed solution; S4. Cool the mixed solution described in S3 to 30° C.-40° C., add white sugar and / or sweetener, sodium isoVC, sodium citrate and sodium bicarbonate, stir evenly, and obtain the Gastrodia elata walnut beverage through homogenization, filtration and sterilization.

5. The method for preparing the gastrodia elata walnut beverage according to claim 4, characterized in that: In S1, the mass volume ratio of the walnut kernel and the walnut oil is (100-120):(10-15) g / mL.

6. The method for preparing the gastrodia elata walnut beverage according to claim 4, characterized in that: In S2, the mesh size of the dried Gastrodia elata is 100-120 mesh; The mass volume ratio of the dry Gastrodia elata and water is (10-15):(300-400) g / mL.

7. The method for preparing the gastrodia elata walnut beverage according to claim 4, characterized in that: In S2, the staged enzymatic hydrolysis includes two stages. The first stage is enzymatic hydrolysis at 40°C-50°C and 100rpm-150rpm for 3h-4h; the second stage is heat preservation at 90°C-100°C for 9min-11min.

8. The method for preparing the gastrodia elata walnut beverage according to claim 4, characterized in that: In S3, the volume of the mixed solution is 1000 mL; The homogenization pressure is 38Mpa-42Mpa, and the number of times is 2 times.

9. The method for preparing the gastrodia elata walnut beverage according to claim 4, characterized in that: In S4, the homogenization pressure is 28Mpa-32Mpa, and the number of times is 1 time.

10. The method for preparing the gastrodia elata walnut beverage according to claim 4, characterized in that: In S4, the sterilization temperature is 120°C-122°C, and the time is 9min-11min.

Images