CN108285827B - Grape seed oil and preparation method thereof - Google Patents
Grape seed oil and preparation method thereof Download PDFInfo
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- CN108285827B CN108285827B CN201810315870.6A CN201810315870A CN108285827B CN 108285827 B CN108285827 B CN 108285827B CN 201810315870 A CN201810315870 A CN 201810315870A CN 108285827 B CN108285827 B CN 108285827B
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- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/02—Pretreatment
- C11B1/04—Pretreatment of vegetable raw material
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23D—EDIBLE OILS OR FATS, e.g. MARGARINES, SHORTENINGS, COOKING OILS
- A23D9/00—Other edible oils or fats, e.g. shortenings, cooking oils
- A23D9/007—Other edible oils or fats, e.g. shortenings, cooking oils characterised by ingredients other than fatty acid triglycerides
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23D—EDIBLE OILS OR FATS, e.g. MARGARINES, SHORTENINGS, COOKING OILS
- A23D9/00—Other edible oils or fats, e.g. shortenings, cooking oils
- A23D9/02—Other edible oils or fats, e.g. shortenings, cooking oils characterised by the production or working-up
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- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/02—Pretreatment
- C11B1/025—Pretreatment by enzymes or microorganisms, living or dead
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- C11B3/00—Refining fats or fatty oils
- C11B3/02—Refining fats or fatty oils by chemical reaction
- C11B3/06—Refining fats or fatty oils by chemical reaction with bases
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- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
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- C11B3/00—Refining fats or fatty oils
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Abstract
The invention belongs to the field of food processing, and discloses grape seed oil and a preparation method thereof. The invention adopts the composite plant hydrolase and the acid protease to extract the grape seed oil by the stepwise hydrolysis method, greatly reserves the content of active ingredients, particularly polyphenol in the grape seed oil, and simultaneously considers the extraction rate of the grape seed oil. Compared with the grape seed oil prepared by squeezing or leaching, the grape seed oil prepared by the method greatly retains active ingredients in the grape seed oil, and the prepared grape seed oil is clear and transparent and can be used as high-end health-care oil. The preparation method disclosed by the invention is short in time, and can effectively destroy the cell wall of the grape seed kernel cell, so that the oil in the cell is easier to extract, and the extraction rate of the grape seed oil is effectively improved. By adopting the alkali extraction method for demulsification, the solvent residue is avoided, and meanwhile, the acid value of the grape seed crude oil can be effectively reduced by 0.168(KOH)/(mg/g), and the later alkali refining process is not needed. The method is simple to operate, low in cost and suitable for industrial production.
Description
Technical Field
The invention belongs to the field of food processing, and particularly relates to grape seed oil and a preparation method thereof, in particular to grape seed oil with high active ingredients, especially polyphenol content, and a preparation method thereof.
Background
With the continuous improvement of the social living standard, the dietary habits of people also change, and a lot of people suffer from three-high disease, namely hyperglycemia, hypertension and hyperlipidemia, caused by overeating. Among them, patients with hyperglycemia are most common, and need to strictly control diet, have low sugar and low salt, and need to take hypoglycemic drugs for severe patients. This also allows many hyperglycemic patients to choose between a diet with little or no oil release.
Grape seed oil is high-grade edible oil obtained by further processing grape seeds which are byproducts of wine brewing or fruit juice squeezing. The grape seed oil is rich in nutrition, and the unsaturated fatty acid is more than 90%, mainly comprising linoleic acid (more than 70%), oleic acid, palmitic acid and stearic acid. Linoleic acid is essential fatty acid, is one of important components constituting human cell membrane and skin, and can maintain adult blood lipid balance, reduce cholesterol, and improve cardiovascular diseases. Grape seed oil also contains phenolic compounds, such as gallic acid, catechin, epicatechin, procyanidin, etc., and recent studies have reported that these phenolic compounds have a wide range of biological activities: antioxidant, anticancer, free radical scavenging, antiinflammatory, antiviral, and blood sugar lowering effects. In addition, the grape seed oil also contains phytosterol, squalene, various fat-soluble vitamins (A, E, D, K, P) and various trace elements (Ca, Fe, Zn, Mn and Cu).
The antioxidant effect of the grape seed oil on mouse organs is researched by the syringa pubescens and the like, and the content of lipid peroxide in mouse tissues and the activity of glutathione peroxidase and superoxide dismutase are measured. The results show that the activities of the liver and kidney organs glutathione peroxidase and superoxide dismutase of the white mice in the grape seed oil experimental group are higher than those of the control group, the content of malondialdehyde in each organ is obviously lower than that of the control group, and the grape seed oil is proved to have a certain antioxidation effect. Lutterodt et al, studied the radical scavenging ability of cold pressed Chardonnay, Massachan, ruby, and Congder grape seed oils, expressed as Trolox equivalent, to the DPPH radical scavenging ability of the seed oil, and showed DPPH radical scavenging rates ranging from 0.07 to 2.22 mmol/g. The nutritional value and the medical function of the grape seed oil are fully determined by medical circles and nutriologists at home and abroad.
At present, grape seed oil on the market mostly adopts cold pressing and solvent extraction technologies. The oil yield is not high by a squeezing method, and the solvent leaching method has the problem of solvent residue. The aqueous enzymatic method is an environment-friendly oil extraction technology, and degrades plant cell walls through crushing and enzymolysis to release oil in oil cells. Compared with the traditional process, the aqueous enzymatic method has the following advantages: avoids the influence of high temperature on the active ingredients of the materials, has higher free oil yield, light color and small protein denaturation degree, and is beneficial to environmental protection and the sustainable development of the oil industry. In recent years, a great deal of literature is continuously carrying out process optimization aiming at improving the extraction rate of the grape seed oil by the aqueous enzymatic method. The Plumbum preparatium (2017) is subjected to enzymolysis for 9h by using complex enzyme (cellulase, pectinase, xylanase and trypsin), and demulsification is performed by a freeze-thaw method, so that the oil yield is 9.947%, and the acid value is 0.52 (KOH)/(mg/g). The extraction rate of grape seed oil obtained by enzymolysis and organic solvent extraction of homoxylon (2009) and the like reaches 88.9 percent, and the acid value is 0.43 (KOH)/(mg/g); the capene and the like (2015) are subjected to enzymolysis for 2 hours by using cellulase, and then are subjected to enzymolysis for 1.5 hours by using neutral protease, and the extraction rate of the grape seed oil is 77.48%; on the basis, ultrasonic pretreatment is carried out for 15min, and the extraction rate reaches 87.65%; then microwave demulsification is adopted, the extraction rate can reach 93.83 percent, and the acid value is 2.16 (KOH)/(mg/g). Ljjrox was hydrolyzed with steam pretreatment knot and cellulase/protease for 8h, with an oil yield of 13.26% and an acid number of 1.24 (KOH)/(mg/g).
However, the research on the active ingredients of the grape seed oil prepared by the enzymolysis process is relatively less, and the phenolic substances in the grape seed oil are the main antioxidant substances of the grape seed oil and are most easily oxidized and degraded in the enzymolysis process. Therefore, a preparation method of grape seed oil capable of improving the content of active ingredients, particularly polyphenol in the grape seed oil and simultaneously considering the extraction rate of the grape seed oil is urgently needed.
Disclosure of Invention
In view of the above, the present invention aims to provide a grape seed oil with high content of active ingredients, especially polyphenols, and a preparation method thereof, aiming at the defects of the prior art.
One object of the invention is to provide grape seed oil with a total phenol content of more than or equal to 30 mg/kg.
Furthermore, the grape seed oil has unsaturated fatty acid not less than 86.63%, total tocopherol not less than 253.6mg/kg, total sterol not less than 2.588mg/g, and acid value of 0.168 (KOH)/(mg/g).
Another object of the present invention is to provide a method for preparing grape seed oil, comprising the steps of:
(1) crushing grape seeds, and sieving to obtain grape seed powder;
(2) adding water into the grape seed powder obtained in the step (1), uniformly mixing to obtain slurry, adjusting the pH of the slurry to acidity, adding mixed enzyme, and performing enzymolysis reaction to obtain a first mixed solution;
(3) adjusting the pH value of the first mixed solution obtained in the step (2) to acidity, and adding acidic protease for enzymolysis to obtain acidolysis solution;
(4) adding the acidolysis solution obtained in the step (3) into an alkali solution to adjust the pH value to be alkaline, and reacting to obtain a second mixed solution;
(5) and (4) inactivating enzyme of the second mixed solution obtained in the step (4), performing centrifugal separation, and taking upper-layer free oil to obtain the grape seed oil.
The method comprises the step (1) of pretreating grape seeds to obtain grape seed powder, wherein the pretreatment comprises crushing and sieving.
In some embodiments, the sieving in step (1) is through a 20-60 mesh sieve.
In some embodiments of the invention, the grape seed meal in step (1) has a moisture and volatiles content of 7.0% to 9.0%, an oil content of 14 to 20%, a crude protein content of 8 to 10%, and a fiber content of 26 to 30%.
The step (2) of the invention carries out mixed enzyme enzymolysis on the grape seed powder. Firstly, adding water into grape seed powder to mix to prepare slurry, adjusting the pH value of the slurry to acidity, adding mixed enzyme to carry out enzymolysis of the mixed enzyme, and carrying out enzymolysis reaction to obtain a first mixed solution.
In some embodiments, the slurry in step (2) is prepared from water and grape seed meal in a volume-to-mass ratio of 3-6 (mL):1(g) and mixing uniformly.
In some embodiments, the slurry in step (2) is prepared from water and grape seed meal in a volume-to-mass ratio of 5 (mL):1(g) and mixing uniformly. In some embodiments, the slurry in step (2) is prepared from water and grape seed meal in a volume-to-mass ratio of 6(mL):1(g) and mixing uniformly.
In some embodiments, the slurry pH is adjusted to 2-6 in step (2). In some embodiments, the slurry pH is adjusted to 5 in step (2).
In some embodiments, the mixed enzymes of step (2) are cellulase, pectinase, hemicellulase and xylanase (0.6-0.7): (0.1-0.15): (0.1-0.2): (0.1-0.15) in a mass ratio. In some embodiments, the mixed enzymes of step (2) are cellulases, pectinases, hemicellulases and xylanases in a ratio of 0.6:0.1:0.2:0.1 mass ratio.
In some embodiments, the amount of blending enzyme added in step (2) is 1wt% to 2 wt% of the slurry. In some embodiments, the amount of mixed enzyme added in step (2) is 2 wt% of the slurry.
In some embodiments, the enzymatic reaction is at 40-60 ℃ for 1-3 h.
In the step (3), the first mixed solution is subjected to acid protease enzymolysis. Firstly, adjusting the pH value of the first mixed solution in the step (2) to be acidic, and then adding acidic protease for enzymolysis to obtain acidolysis solution.
In some embodiments, the pH is adjusted to 2-6 in step (3). In some embodiments, the pH is adjusted to 2 in step (3).
In some embodiments, the amount of acidic protease added in step (3) is 1wt% to 2 wt%. In some embodiments, the acidic protease is added in an amount of 1wt% in step (3).
In some embodiments, the enzymatic hydrolysis reaction in step (3) is at 40-60 ℃ for 1-3 h. In some embodiments, the enzymatic hydrolysis reaction in step (3) is at 50 ℃ for 3 h.
The step (4) of the invention carries out alkali extraction on the obtained acidolysis solution. And (4) adding an alkali solution into the acidolysis solution obtained in the step (3) to adjust the pH value to be alkaline, and reacting to obtain a second mixed solution.
In some embodiments, the pH in step (4) is adjusted to 8-10. In some embodiments, the pH in step (4) is adjusted to 8.
In one embodiment of the present invention, the alkali solution in step (4) is sodium hydroxide and distilled water at a mass-to-volume ratio of 20-40 (g): 1(L) configuration.
In some embodiments, the reacting in the step (4) is at 40-60 ℃ for 20-40 min. In some embodiments, the reacting in step (4) is at 50 ℃ for 30 min.
The step (5) of the invention is to extract the grape seed oil from the second mixed solution. And (4) inactivating enzyme of the second mixed solution in the step (4), performing centrifugal separation, and taking upper-layer free oil to obtain the grape seed oil.
In some embodiments, the enzyme deactivation in the step (5) is that the slurry is heated to 80-90 ℃ to deactivate the enzyme for 10-20min, and then cooled. In some embodiments, the enzyme deactivation in the step (5) is to heat the slurry to 85 ℃ to deactivate the enzyme for 15min, and then cool the slurry.
In some embodiments, the centrifugation speed in the step (5) is 3000-4000r/min, and the centrifugation is performed for 10-20 min. In some embodiments, the centrifugation speed in step (5) is 3500r/min and the centrifugation is 15 min.
The invention also provides the grape seed oil obtained by the method.
In a specific embodiment, the result of examining the blood sugar reducing effect of the grape seed oil prepared by the invention shows that the grape seed oil prepared by the invention can effectively improve the glucose tolerance of mice, obviously reduce the contents of cholesterol, triglyceride, insulin and HbA1c of the mice and has better effect than corn oil. The contents of interleukin-6, tumor necrosis factor and C-reactive protein in the Grape Seed Oil (GSO) group prepared by the invention are all reduced. The result shows that the grape seed oil prepared by the invention has obvious effect of assisting in reducing blood sugar. Therefore, the invention also provides the application of the grape seed oil in health-care food.
The invention adopts the composite plant hydrolase and the acid protease to extract the grape seed oil by the stepwise hydrolysis method, greatly reserves the content of active ingredients, particularly polyphenol in the grape seed oil, and simultaneously considers the extraction rate of the grape seed oil. Compared with the prior art, the invention has one of the following beneficial effects:
(1) compared with the grape seed oil prepared by squeezing or leaching, the method greatly reserves the active ingredients in the grape seed oil, such as unsaturated fatty acid (86.63%), total phenol (37.46mg/kg), total tocopherol (253.6mg/kg) and total sterol (2.588mg/g), and the content of the active ingredients is obviously higher than that of the grape seed oil prepared by the existing aqueous enzymatic method. The prepared grape seed oil is clear and transparent and can be used as high-end health care oil.
(2) The preparation method disclosed by the invention is short in time, and can effectively destroy the cell wall of the grape seed kernel cell, so that the oil in the cell is easier to extract, and the extraction rate of the grape seed oil is effectively improved.
(3) Compared with other aqueous enzymatic methods which adopt organic solvents to extract oil, the method avoids solvent residues, can effectively reduce the acid value of 0.168(KOH)/(mg/g) of crude grape seed oil, and does not need a later alkali refining process.
(3) The method is simple to operate, low in cost and suitable for industrial production.
Detailed Description
The invention discloses grape seed oil and a preparation method thereof. Those skilled in the art can modify the process parameters appropriately to achieve the desired results with reference to the disclosure herein. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods and products of the present invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications of the methods described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of the present invention without departing from the spirit and scope of the invention.
In order to further understand the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Unless otherwise specified, the raw materials or auxiliary materials (enzyme preparations) used in the examples of the present invention are commercially available. In the following examples, corn oil and lard are commercially available.
And (3) determining the oil content of grape seeds: measured by Soxhlet extraction according to GB/T5009.6-2003.
Determination of acid value of grape seed oil: according to GB/T5530-2005 animal and vegetable oil acid value and acidity determination, a cold solvent method is adopted for determination.
The oil yield determination method comprises the following steps: the grape seed oil extraction rate (%) (free oil mass/grape seed kernel mass) × 100.
The extraction rate measuring method comprises the following steps: the grape seed oil extraction rate (%) (% free oil mass/(grape seed powder mass × grape seed oil content) × 100.
And (3) determining the total phenol content of the grape seed oil: and measuring by adopting a Fulin phenol method spectrophotometer according to GB/T8313-2008 'detection method for the content of tea polyphenol and catechins in tea'.
Total tocopherol assay: sample pretreatment: weighing 1.5g (accurate to 0.001g) of oil sample, placing in a brown volumetric flask, adding a proper amount of n-hexane for dissolution, carrying out ultrasonic treatment for 5min, then carrying out constant volume to 10ml, and carrying out liquid chromatography HPLC analysis after passing through a 0.22um microporous filter membrane.
HPLC conditions: the chromatographic column is
Silica (4.6 mm. times.250 mm, 5 μm); the ultraviolet detection wavelength is 295nm, and the mobile phase is n-hexane: isopropanol 98.5: 1.5; the flow rate is 1.0 ml/min; the column temperature is 25 ℃; the amount of the sample was 10.0. mu.l.
The total sterol determination method comprises the following steps: sample pretreatment: weighing 0.2 + -0.01 g grape seed oil, adding 5 alpha-cholestanol standard solution 0.1mg/mL as internal standard, adding 3mL 2mol/L KOH-CH3CH2Treating OH solution with ultrasonic wave for 5min, saponifying with 85 deg.C water bath for 30min, cooling, adding 2mL water and 5mL N-hexane, oscillating, centrifuging at 4000r/min for 5min, collecting supernatant, extracting the lower layer with 5mL N-hexane twice, mixing organic layers, washing with water to neutrality, and recovering N2Blow-drying, adding 200 μ L derivatization reagent BSTFA + TMCS (99:1), and 75 deg.C waterBathing for 30min, filtering with 0.22 μm microporous membrane, and performing GC-MS analysis.
Gas chromatography conditions: a chromatographic column: thermo TR-5MS (30m × 0.25mm, 0.25 μm), sample size 1.0 μ L, injection port temperature 280 deg.C, helium He, flow rate 1.2 mL/min. Temperature programming: the initial temperature is 200 ℃, the temperature is increased to 250 ℃ at the speed of 10 ℃/min, the temperature is maintained for 1min, the temperature is increased to 300 ℃ at the speed of 5 ℃/min, and the temperature is maintained for 20 min.
The mass spectrum conditions comprise interface temperature of 250 ℃, ionization mode EI, electron energy of 70eV, ion source temperature of 250 ℃ and mass scanning range m/z of 50-550 amu.
Unsaturated fatty acid determination method: sample pretreatment: weighing 0.03 + -0.001 g grape seed oil as oil sample, adding 0.5mol/L NaOH-CH32mL of OH solution, uniformly oscillating, heating in water bath at 65 ℃ until oil drops are completely dissolved, standing, cooling, and adding 25% of BF3-CH32mL of OH solution, esterifying for 20min in water bath at 65 ℃, standing and cooling, adding 2mL of n-hexane, fully shaking, adding 2mL of saturated NaCl solution, centrifuging for 15min, taking the upper organic phase in a dry sample bottle, and adding a small amount of anhydrous Na2SO4Removing a trace amount of water.
Gas chromatography conditions: PEG-20M capillary column (30M × 0.32mm, 0.25 μ M); carrier gas: n with a purity of 99.999%2(ii) a Purging flow rate: 3 mL/min; sample introduction amount: 1 mu L of the solution; the split ratio is 80: 1; the temperature of a sample inlet is 250 ℃; the temperature of the detector is 250 ℃; FID detector. Temperature rising procedure: the initial temperature is 150 deg.C, increased to 190 deg.C at 5 deg.C/min, maintained for 2min, and increased to 240 deg.C at 5 deg.C/min, and maintained for 10 min.
The color and luster measuring method comprises the following steps: the color of the grapeseed oil was determined using a color meter. Use of CIE L in measurements*a*b*And (5) representing a color space. Color parameters and definitions: l is*Is lightness (black-white, 0-100), a*Is red-green hue (green-red, -a)*-+a*)、b*Is a yellow-blue hue (blue-yellow, -b)*-+b*)。
Experimental animals: c57BL/6J mice, SPF grade, body weight (18. + -.2) g, supplied by Shanghai Spiker laboratory animals, Inc.
The main apparatus is as follows: glucometer, ROCHE; microplate reader, BioTek staining, usa; ultra-low temperature freezer refrigerator, Thermo Fisher Scientific, usa; model 7200 spectrophotometer, Shanghai spectral element instruments, Inc.; the kit is built into a bioengineering research institute by Nanjing.
Example 1 Effect of the types and amounts of Mixed enzymes on the extraction of grape seed oil
(1) Grape seed pretreatment: crushing grape seeds by a universal flour mill, and sieving by a 50-mesh sieve to obtain a grape seed powder raw material, wherein the water content and volatile matter content of the grape seed powder raw material are 8.65%, the oil content of the grape seed powder raw material is 17.25%, the crude protein content of the grape seed powder raw material is 9.2%, and the fiber content of the grape seed powder raw material is 30.6%.
(2) Mixed enzyme enzymolysis: mixing distilled water and grape seed powder according to a volume mass ratio of 5 (mL):1(g) uniformly mixing to prepare slurry, adding hydrochloric acid to adjust the pH of the slurry to 5, adding mixed enzyme for enzymolysis, wherein the enzymolysis reaction temperature is 50 ℃, the enzymolysis reaction time is 2 hours, and the adding scheme of the mixed enzyme is shown in table 1;
(3) acid protease enzymolysis: adding hydrochloric acid into the slurry, adjusting pH to 4.5, adding acidic protease for enzymolysis, wherein the addition amount is 0.1%, the enzymolysis reaction temperature is 50 ℃, and the enzymolysis reaction time is 2 h.
(4) Alkali liquor extraction: the mass-volume ratio of sodium hydroxide to distilled water is 20-40 (g): 1(L), adding the prepared sodium hydroxide solution into the slurry, adjusting the pH to 8, and carrying out alkali extraction reaction at 50 ℃ for 30 min.
(5) Extracting oil: heating the serous fluid to 85 deg.C, inactivating enzyme for 15min, centrifuging the mixed solution at 3500r/min for 15min, and collecting upper layer free oil to obtain light yellow grape seed oil.
TABLE 1 influence of the types and amounts of blending enzymes on the extraction yield and total phenol content of grapeseed oil
As can be seen from Table 1, the compound enzyme can improve the polyphenol content of the grape seed oil, and the oil yield and the polyphenol content of the grape seed oil extracted by a single enzyme preparation are not high. The compound enzyme contains cellulase, pectinase, hemicellulase and xylanase, the viscosity of the slurry is increased along with the increase of the content of the compound enzyme, the number of emulsifying layers is increased, and the extraction rate of the grape seed oil and the content of total phenols are reduced. The compound enzyme addition amount is 1 percent, the effect is optimal, and the total phenol content is as high as 37.46 mg/kg.
Example 2 Effect of the type and amount of protease added on the extraction of grape seed oil
(1) Grape seed pretreatment: crushing grape seeds by a universal flour mill, and sieving by a 50-mesh sieve to obtain a grape seed powder raw material, wherein the water content and volatile matter content of the grape seed powder raw material are 8.65%, the oil content of the grape seed powder raw material is 17.25%, the crude protein content of the grape seed powder raw material is 9.2%, and the fiber content of the grape seed powder raw material is 30.6%.
(2) Mixed enzyme enzymolysis: mixing distilled water and grape seed powder according to a volume mass ratio of 5 (mL):1(g) mixing uniformly to prepare slurry, adding hydrochloric acid to adjust the pH of the slurry to 5, and adding 1% mixed enzyme (wherein the ratio of cellulase to pectinase to hemicellulase to xylanase is 0.6:0.1:0.2:0.1), the enzymolysis reaction temperature is 50 ℃, and the enzymolysis reaction time is 2 hours;
(3) and (3) protease enzymolysis: adding protease for enzymolysis at 50 deg.C for 2 hr, wherein the protease addition and pH are shown in Table 2.
(4) Alkali liquor extraction: sodium hydroxide and distilled water according to the mass-volume ratio of 20-40 (g): 1(L), adding the prepared sodium hydroxide solution into the slurry, adjusting the pH to 8, and carrying out alkali extraction reaction at 50 ℃ for 30 min.
(5) Extracting oil: heating the serous fluid to 85 deg.C, inactivating enzyme for 15min, centrifuging the mixed solution at 3500r/min for 15min, and collecting upper layer free oil to obtain light yellow grape seed oil.
TABLE 2 influence of protease type and content on grape seed oil extraction yield and total phenol content
As shown in Table 2, the content of polyphenols in the grape seed oil was increased by the acidic protease compared to the neutral protease and the alkaline protease in the same amount. The best effect is achieved when the adding amount of the acid protease is 1 percent, and the total phenol content reaches 37.46 mg/kg.
Example 3 Effect of acid protease enzymolysis temperature, time and pH on extraction of grape seed oil
(1) Grape seed pretreatment: crushing grape seeds by a universal flour mill, and sieving by a 50-mesh sieve to obtain a grape seed powder raw material, wherein the water content and volatile matter content of the grape seed powder raw material are 8.65%, the oil content of the grape seed powder raw material is 17.25%, the crude protein content of the grape seed powder raw material is 9.2%, and the fiber content of the grape seed powder raw material is 30.6%.
(2) Mixed enzyme enzymolysis: mixing distilled water and grape seed powder according to a volume mass ratio of 5 (mL):1(g) are mixed evenly to prepare serous fluid, hydrochloric acid is added to adjust the pH value of the serous fluid to 5, 1 percent of mixed enzyme (wherein the ratio of cellulase to pectinase to hemicellulase to xylanase is 0.6:0.1:0.2:0.1) is added, the enzymolysis reaction temperature is 50 ℃, and the enzymolysis reaction time is 2 hours.
(3) Acid protease enzymolysis: hydrochloric acid was added to the slurry, and acidic protease was added for enzymatic hydrolysis in an amount of 1%, and the influence of the enzymatic hydrolysis temperature, enzymatic hydrolysis time, and pH of the acidic protease on the enzymatic hydrolysis rate and total phenol content is shown in table 3.
(4) Alkali liquor extraction: sodium hydroxide and distilled water according to the mass-volume ratio of 20-40 (g): 1(L), adding the prepared sodium hydroxide solution into the slurry, adjusting the pH to 8, and carrying out alkali extraction reaction at 50 ℃ for 30 min.
(5) Extracting oil: heating the serous fluid to 85 deg.C, inactivating enzyme for 15min, centrifuging the mixed solution at 3500r/min for 15min, and collecting upper layer free oil to obtain light yellow grape seed oil.
TABLE 3 influence of the temperature, time and pH of the acid protease enzymolysis on the extraction yield and total phenol content of grapeseed oil
| Temperature of enzymolysis (. degree.C.) | Time of enzymolysis (h) | pH | Oil yield (%) | Extraction ratio (%) | Total phenol content (mg/kg) |
| 40 | 2 | 4.5 | 9.23 | 53.51 | 34.27 |
| 50 | 2 | 4.5 | 10.29 | 59.65 | 37.46 |
| 60 | 2 | 4.5 | 10.34 | 59.94 | 35.13 |
| 50 | 1 | 4.5 | 6.78 | 39.30 | 32.11 |
| 50 | 3 | 4.5 | 11.26 | 65.28 | 40.12 |
| 50 | 2 | 2 | 9.45 | 54.78 | 42.34 |
| 50 | 2 | 6 | 10.67 | 61.86 | 30.12 |
As can be seen from Table 3, the highest total phenol content of the grape seed oil can reach 42.34mg/kg when the enzymolysis temperature of the acidic protease is 50 ℃, the enzymolysis time is 2h and the pH value is 2.
EXAMPLE 4 Effect of lye extraction temperature and time on grape seed oil extraction
A preparation method of grape seed oil for assisting in reducing blood sugar mainly comprises the following steps
(1) Grape seed pretreatment: crushing grape seeds by a universal flour mill, and sieving by a 50-mesh sieve to obtain a grape seed powder raw material, wherein the water content and volatile matter content of the grape seed powder raw material are 8.65%, the oil content of the grape seed powder raw material is 17.25%, the crude protein content of the grape seed powder raw material is 9.2%, and the fiber content of the grape seed powder raw material is 30.6%.
(2) Mixed enzyme enzymolysis: mixing distilled water and grape seed powder according to a volume mass ratio of 5 (mL):1(g) mixing uniformly to prepare slurry, adding hydrochloric acid to adjust the pH of the slurry to 5, and adding 1% mixed enzyme (wherein the ratio of cellulase to pectinase to hemicellulase to xylanase is 0.6:0.1:0.2:0.1), the enzymolysis reaction temperature is 50 ℃, and the enzymolysis reaction time is 2 hours;
(3) acid protease enzymolysis: adding hydrochloric acid into the slurry, adjusting pH to 4.5, adding acidic protease for enzymolysis at 50 deg.C for 2 hr, with addition amount of 1%.
(4) Alkali liquor extraction: sodium hydroxide and distilled water according to the mass-volume ratio of 20-40 (g): 1(L), the prepared sodium hydroxide solution was added to the slurry to adjust the pH to 8, and the alkali reaction temperature and time were as shown in Table 4.
(5) Extracting oil: heating the serous fluid to 85 deg.C, inactivating enzyme for 15min, centrifuging the mixed solution at 3500r/min for 15min, and collecting upper layer free oil to obtain light yellow grape seed oil.
TABLE 4 influence of alkali extraction temperature and time on grape seed oil extraction yield and total phenol content
As can be seen from Table 4, the highest total phenol content of the grape seed oil was 37.46mg/kg at the alkali extraction temperature of 50 ℃ and the alkali extraction time of 30 min. In the alkali extraction process, the total phenol content in the grape seed oil can be reduced when the temperature is too high or the extraction time is longer.
Example 5 Effect of non-alkali extraction on grape seed oil extraction
(1) Grape seed pretreatment: crushing grape seeds by a universal flour mill, and sieving by a 50-mesh sieve to obtain a grape seed powder raw material, wherein the water content and volatile matter content of the grape seed powder raw material are 8.65%, the oil content of the grape seed powder raw material is 17.25%, the crude protein content of the grape seed powder raw material is 9.2%, and the fiber content of the grape seed powder raw material is 30.6%.
(2) Mixed enzyme enzymolysis: mixing distilled water and grape seed powder according to a volume mass ratio of 5 (mL):1(g) mixing uniformly to prepare slurry, adding hydrochloric acid to adjust the pH of the slurry to 5, and adding 1% mixed enzyme (wherein the ratio of cellulase to pectinase to hemicellulase to xylanase is 0.6:0.1:0.2:0.1), the enzymolysis reaction temperature is 50 ℃, and the enzymolysis reaction time is 2 hours;
(3) acid protease enzymolysis: adding hydrochloric acid into the slurry, adjusting pH to 4.5, adding acidic protease for enzymolysis, wherein the addition amount is 1%, the enzymolysis reaction temperature is 50 ℃, and the enzymolysis reaction time is 2 h. .
(4) Extracting oil: heating the serous fluid to 85 deg.C, inactivating enzyme for 15min, centrifuging the mixed solution at 3500r/min for 15min, and collecting upper layer free oil to obtain light yellow grape seed oil.
At this time, the oil yield of the grapeseed oil was 3.61%, the extraction rate was 20.93%, and the total phenol content was 40.14 mg/kg.
Example 6 Effect of other demulsification methods on grape seed oil extraction
(1) Grape seed pretreatment: crushing grape seeds by a universal flour mill, and sieving by a 50-mesh sieve to obtain a grape seed powder raw material, wherein the water content and volatile matter content of the grape seed powder raw material are 8.65%, the oil content of the grape seed powder raw material is 17.25%, the crude protein content of the grape seed powder raw material is 9.2%, and the fiber content of the grape seed powder raw material is 30.6%.
(2) Mixed enzyme enzymolysis: mixing distilled water and grape seed powder according to a volume mass ratio of 5 (mL):1(g) mixing uniformly to prepare slurry, adding hydrochloric acid to adjust the pH of the slurry to 5, and adding 1% mixed enzyme (wherein the ratio of cellulase to pectinase to hemicellulase to xylanase is 0.6:0.1:0.2:0.1), the enzymolysis reaction temperature is 50 ℃, and the enzymolysis reaction time is 2 hours;
(3) acid protease enzymolysis: adding hydrochloric acid into the slurry, adjusting pH to 4.5, adding acidic protease for enzymolysis, wherein the addition amount is 1%, the enzymolysis reaction temperature is 50 ℃, and the enzymolysis reaction time is 2 h.
(4) Enzyme deactivation: heating the slurry to 85 deg.C to inactivate enzyme for 15 min.
(5) And (3) extraction: adding 100ml n-hexane, and ultrasonic cleaning in ultrasonic cleaner for 30 min.
(6) Extracting oil: and centrifuging the mixed solution for 15min at 3500r/min, taking an upper organic phase, and performing rotary evaporation at 40 ℃ to obtain the light yellow grape seed oil.
At this time, the oil yield of the grape seed oil was 6.01%, the extraction rate was 34.84%, and the total phenol content was 32.14 mg/kg.
Example 7 Effect of feed-liquid ratio on grape seed oil extraction
(1) Grape seed pretreatment: crushing grape seeds by a universal flour mill, and sieving by a 50-mesh sieve to obtain a grape seed powder raw material, wherein the water content and volatile matter content of the grape seed powder raw material are 8.65%, the oil content of the grape seed powder raw material is 17.25%, the crude protein content of the grape seed powder raw material is 9.2%, and the fiber content of the grape seed powder raw material is 30.6%.
(2) Mixed enzyme enzymolysis: uniformly mixing distilled water and grape seed powder to obtain a slurry, adding hydrochloric acid to adjust the pH of the slurry to 5, adding 1% mixed enzyme (wherein the ratio of cellulase, pectinase, hemicellulase and xylanase is 0.6:0.1:0.2:0.1), carrying out enzymolysis reaction at 50 ℃ for 2 hours;
(3) acid protease enzymolysis: adding hydrochloric acid into the slurry, adjusting pH to 4.5, adding acidic protease for enzymolysis, wherein the addition amount is 1%, the enzymolysis reaction temperature is 50 ℃, and the enzymolysis reaction time is 2 h.
(4) Alkali liquor extraction: sodium hydroxide and distilled water according to the mass-volume ratio of 20-40 (g): 1(L), adding the prepared sodium hydroxide solution into the slurry, adjusting the pH to 8, and carrying out alkali extraction reaction at 50 ℃ for 30 min.
(5) Extracting oil: heating the serous fluid to 85 deg.C, inactivating enzyme for 15min, centrifuging the mixed solution at 3500r/min for 15min, and collecting upper layer free oil to obtain light yellow grape seed oil.
TABLE 5 influence of feed liquid ratio on grapeseed oil extraction rate and total phenol content
| Distilled water (mL): grape seed powder mixture (g) | Oil yield (%) | Extraction ratio (%) | Total phenol content (mg/kg) |
| 3:1 | 9.67 | 56.06 | 35.68 |
| 4:1 | 9.90 | 57.39 | 36.52 |
| 5:1 | 10.29 | 59.65 | 37.46 |
| 6:1 | 10.12 | 58.67 | 38.68 |
As can be seen in Table 5, the total phenol content of the grape seed oil extracted by the distilled water and the grape seed powder according to the volume mass ratio of 6(mL) to 1(g) is higher and can reach 38.68 mg/kg.
Example 8 determination of beneficial ingredients and acid number of grapeseed oil
(1) Grape seed pretreatment: crushing grape seeds by a universal flour mill, and sieving by a 50-mesh sieve to obtain a grape seed powder raw material, wherein the water content and volatile matter content of the grape seed powder raw material are 8.65%, the oil content of the grape seed powder raw material is 17.25%, the crude protein content of the grape seed powder raw material is 9.2%, and the fiber content of the grape seed powder raw material is 30.6%.
(2) Mixed enzyme enzymolysis: mixing distilled water and grape seed powder according to a volume mass ratio of 5 (mL):1(g) mixing uniformly to prepare slurry, adding hydrochloric acid to adjust the pH of the slurry to 5, and adding 1% mixed enzyme (wherein the ratio of cellulase to pectinase to hemicellulase to xylanase is 0.6:0.1:0.2:0.1), the enzymolysis reaction temperature is 50 ℃, and the enzymolysis reaction time is 2 hours;
(3) acid protease enzymolysis: adding hydrochloric acid into the slurry, adjusting pH to 4.5, adding acidic protease for enzymolysis, wherein the addition amount is 1%, the enzymolysis reaction temperature is 50 ℃, and the enzymolysis reaction time is 2 h.
(4) Alkali liquor extraction: sodium hydroxide and distilled water according to the mass-volume ratio of 20-40 (g): 1(L), adding the prepared sodium hydroxide solution into the slurry, adjusting the pH to 8, and carrying out alkali extraction reaction at 50 ℃ for 30 min.
(5) Extracting oil: heating the serous fluid to 85 deg.C, inactivating enzyme for 15min, centrifuging the mixed solution at 3500r/min for 15min, and collecting upper layer free oil to obtain light yellow grape seed oil.
Content of beneficial substances in grape seed oil: unsaturated fatty acid (86.63%), total phenols (37.46mg/kg), total tocopherols (253.6mg/kg), total sterols (2.588 mg/g). The acid value of the obtained grapeseed oil was 0.168 (KOH)/(mg/g).
Comparative example 9 comparison of polyphenols by other methods
Crushing grape seeds, sieving with 40 meshes, regulating pH to 6.5, adding 1% of complex enzyme (mass ratio of cellulase, pectinase, xylanase and trypsin is 2:1:2:1), carrying out enzymolysis at 45 ℃ for 9h, inactivating enzyme for 5min, cooling to room temperature, centrifuging at 3900r/min, sucking supernatant clear oil, demulsifying an emulsion by a freeze-thaw method, repeatedly freezing, thawing and centrifuging enzymolysis liquid until the upper layer has no oil layer, and weighing the oil mass (Plumbum 2017). The total phenol content of the grape seed oil obtained in the experiment is 12.79mg/kg, and the acid value is 0.52 (KOH)/(mg/g). The grapeseed oil-like total phenol content was higher and the acid number was lower in example 8 compared to control 9. The enzymolysis time of the comparative example 9 is too long, which is not beneficial to the active ingredients in the crude oil and promotes the oxidation of the grease.
Comparative example 10 comparison of polyphenols by other methods
Pulverizing grape seed, sieving with 40 mesh sieve, adding citric acid-sodium citrate buffer solution at a ratio of 1:3, pH5.5, water bathing at 50 deg.C for 5min, adding 1% (V/W) cellulase, stirring, performing enzymolysis for 1.5h, adding petroleum ether, continuing to act for 30min, centrifuging at 8000r/min for 30min, performing rotary evaporation of oil phase, and weighing the obtained crude oil (homoxylol, 2009). The total phenol content of the grape seed oil obtained in the experiment is 20.14mg/kg, and the acid value is 0.43 (KOH)/(mg/g). The grapeseed oil-like total phenol content was higher and the acid number was lower in example 8 compared to control 10.
Comparison of polyphenols with other methods of comparative example 11
Crushing grape seeds, sieving the crushed grape seeds with 40 meshes, adding a citric acid buffer solution with the pH value of 4.18 according to the material-liquid ratio of 1:6, pretreating the grape seeds with normal-pressure steam for 20min, cooling, adding cellulase according to the addition amount of 1.0% (V/W), performing enzymolysis in a 45 ℃ constant-temperature water bath for 1h, heating to 55 ℃, continuing the enzymolysis for 8h, heating to 85 ℃, keeping the temperature for 10min for enzyme deactivation, cooling to room temperature, centrifuging for 10min at 4500r/min, extracting grease with anhydrous ether, drying, and removing a solvent remained in the grease to obtain the grape seed oil (SANGSUI, 2010, Lv). The total phenol content of the grape seed oil obtained in the experiment is 10.28mg/kg, and the acid value is 1.24 (KOH)/(mg/g).
The grapeseed oil-like total phenol content was higher and the acid number was lower in example 8 compared to control 11. The enzymolysis time of the comparative example 11 is too long, which is not beneficial to the active ingredients in the crude oil and promotes the oxidation of the grease.
Comparative example 12 comparison of polyphenols by other methods
Drying grape seeds at constant temperature, pulverizing, sieving with 80 mesh sieve, adding distilled water (1:6.6), placing in ultrasonic reactor with ultrasonic power of 225W, and ultrasonic processing for 15min at 50 deg.C. Then adjusting pH to 5.0, adding cellulase 2.0%, performing enzymolysis for 2.0h, adjusting pH to 7.0, adding 1.0% neutral protease, and performing enzymolysis for 1.5 h. And (3) after the enzymolysis is finished, inactivating the enzyme in a boiling water bath for 10min, centrifuging for 20min at 4000r/min, and collecting the upper-layer free oil and emulsion. Putting the emulsion into a beaker, placing the beaker in a microwave reactor, and performing microwave for 7min at the microwave power of 500W. The microwaved emulsion was centrifuged at 4000r/min for 15min, the free oil was separated, and the two free oils were combined and weighed (capene, 2015). The total phenolic content of the grape seed oil obtained in the experiment is 18.56mg/kg, and the acid value is 2.16 (KOH)/(mg/g).
The grapeseed oil-like total phenol content was higher and the acid number was lower in example 8 compared to control 12.
Example 13 Effect of traditional pressing and extraction according to the invention on grape seed oil color
The color of the grape seed oil extracted by the traditional pressing method and the example 8 are respectively measured.
TABLE 6 colour of grape seed oil extracted by conventional pressing and example 8
| Extraction method | L* | a* | b* | ΔE* |
| Traditional squeezing method | 83.05±2.96 | -4.10±0.65 | 47.31±1.66 | 47.82±2.02 |
| Example 8 | 84.99±1.00 | -4.07±0.09 | 38.70±0.97 | 39.02±1.13 |
As can be seen from Table 6, the total color difference Δ E of the grape seed oil obtained in example 8*39.02 +/-1.13, and the color value is less than delta E of grape seed oil extracted by a traditional pressing method*The value 47.82 is less color difference and more uniform color of the grapeseed oil obtained in example 8. Brightness values L of two grape seed oils*: traditional squeezing method (83.05)<Example 8 method (84.99) illustrates the brightness value L of grapeseed oil obtained in example 8*Is more white and has transparent color, and is prepared by traditional squeezing methodThe obtained grape seed oil is more turbid. A of two grape seed oils*Are all negative values, b*Both positive values indicate that the color of the grape seed oil is biased towards green and yellow. Meanwhile, the red-green value a of the grape seed oil obtained by the method of example 8*And a yellow-blue value b*Is more neutral than the traditional pressing method.
Example 14 drug effect experiment of assisting hypoglycemic grapeseed oil in reducing blood sugar
(1) Rat breeding
Rats were housed in the SPF class Barrier animal laboratory, the Experimental animal center of university in south of the Yangtze river. The daily care and experimental conditions of the animals meet the standards of the environmental and facility standards of experimental animals of the Ministry of health of the people's republic of China.
Healthy male C57BL/6J mice (20. + -.2 g) were purchased and acclimatized to a general diet (AIN93G) for 5 days, fasted for 16 hours, and the tail blood was collected and randomly divided into a normal group (ND), corn oil group (CO), Lard model control group (Lard) and grape seed oil Group (GSO) obtained by the method of example 8, 12 mice per group, based on the fasting blood glucose level of the mice.
3.2 administration of group feed formulations
The administration of the feed in a grouped feed formula: the feed formula of the normal control group is prepared according to the AIN-93G experimental animal feed formula recommended by research diet company according to American society for nutrition, the feed formula of the model control group is prepared according to the D12492, the feeding time of each group is 8:30-9:00 in the morning, the group is free diet, and the feeds with different formulas are continuously fed for 3 months.
TABLE 7 feed formulations for different experimental groups
(2) Sample collection and index determination
Fasting glucose and glucose tolerance (blood samples): after feeding the feed for 3 months, the mice are fasted for 16 hours without water prohibition, the limit fasting blood sugar is determined as abdominal blood sugar at 0min, 1g/kg body weight glucose solution is injected into the abdominal cavity, and tail vein blood sugar of the mice at 30min, 60min, 90min and 120min after the glucose solution is injected into the abdominal cavity is determined by a glucometer and is used for determining the sugar tolerance.
Insulin, cholesterol, triglyceride, HbA1c levels (blood sample): after the test is finished, animals of each group are fasted for 12 hours, intraperitoneal injection is carried out on the mice by adopting 1% pentobarbital sodium solution, blood is collected from eyeballs after anesthesia, the blood is kept for about 1 hour, and serum is taken out and stored in a refrigerator at the temperature of minus 80 ℃ for standby after 3000g of centrifugation is carried out for 10 minutes.
Inflammatory factors IL-6, TNF-alpha, CRP (tissue), mice post-mortem dissected, liver, kidney, spleen, pancreas, small intestine were taken. The internal organs were stored at-80 ℃ until use.
And (3) respectively measuring each group of biochemical indexes by using a kit, expressing the result by mean +/-SD, analyzing the result by using SPSS statistical software (23.0), and performing ANOVA analysis on the result.
After the experiment, the biochemical index level changes of each group of mice are shown in tables 8-10.
TABLE 8 comparison of glucose tolerance in different groups of mice
Note: in the table, different letters in the same column indicate significant differences.
The results of the experiment in table 8 show that the blood glucose trend of the GSO group is significantly more stable than that of the Lard model group, indicating that the GSO group has better tolerance to glucose injection, and the blood glucose at 60min and 90min is significantly lower than that of the Lard group (p < 0.05). The insulin sensitivity was best in ND mice compared to 60% high fat diet mice. The lower the area under the broken line (AUC), the better the insulin sensitivity of the mice in this group. The area under the curve (AUC) of the ND group mice is the lowest, the AUC of the 60% high fat feed group mice is obviously increased compared with the normal group mice (p is less than 0.01), and the area under the broken line of the GSO group mice is obviously reduced (p is less than 0.01) compared with the Lard group mice, so that the grape seed oil prepared by the method can effectively improve the glucose tolerance of the mice.
TABLE 9 comparison of Cholesterol, Triglycerides, insulin, HbA1c in different groups of mice
The results in table 9 show that the mice in the Lard model (Lard) group all have very significant differences in cholesterol, triglycerides, insulin, HbA1c compared to the blank group (P <0.01), indicating successful modeling.
Compared with the Lard (Lard) model group, the cholesterol, triglyceride, insulin and HbA1c of the grapeseed oil (GSO) group mice have obvious improvement effect on all indexes. Meanwhile, the contents of insulin and HbA1c in mice in a Grape Seed Oil (GSO) group are remarkably reduced (p is less than 0.05) compared with the contents in a Corn Oil (CO) group, which indicates that the grape seed oil prepared by the invention can remarkably reduce the contents of cholesterol, triglyceride, insulin and HbA1c in the mice and has better effect than the corn oil.
TABLE 10 comparison of IL-6, TNF-alpha, CRP in liver of mice of different groups
Note: different letters in the same column indicate significant differences in the table.
The results in table 10 show that the mouse interleukin-6 (IL-6), tumor necrosis factor (TNF- α), C-reactive protein (CRP) in the Lard model (Lard) group have very significant differences (P <0.01) compared with the blank group, indicating that the modeling is successful.
The contents of interleukin-6, tumor necrosis factor and C-reactive protein in the Grape Seed Oil (GSO) group prepared by the invention are all reduced, and the Grape Seed Oil (GSO) group has significant difference (P is less than 0.01) compared with a lard model group. Compared with the corn oil, the content of C-reactive protein in the grape seed oil prepared by the invention is remarkably reduced (P is less than 0.05).
And (4) conclusion: the result shows that the grape seed oil prepared by the invention has obvious effect of assisting in reducing blood sugar.
The above description is only a preferred embodiment of the present invention, and all the solutions, features, principles and equivalents thereof as well as all the simple changes thereof are included in the protection scope of the present invention. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (5)
1. A preparation method of grape seed oil comprises the following steps:
(1) crushing grape seeds, and sieving to obtain grape seed powder;
(2) adding water into the grape seed powder obtained in the step (1), uniformly mixing to obtain slurry, adjusting the pH of the slurry to acidity, adding mixed enzyme, and performing enzymolysis reaction to obtain a first mixed solution; the slurry in the step (2) is prepared from water and grape seed powder according to the volume mass ratio of 3-6 (mL):1(g) mixing uniformly; adjusting the pH value of the slurry to 2-6; the mixed enzyme is cellulase, pectinase, hemicellulase and xylanase (0.6-0.7): (0.1-0.15): (0.1-0.2): (0.1-0.15), and the addition amount of the mixed enzyme is 1-2 wt% of the slurry; the enzymolysis reaction is carried out for 1-3h at 40-60 ℃;
(3) adjusting the pH value of the first mixed solution obtained in the step (2) to acidity, and adding acidic protease for enzymolysis to obtain acidolysis solution; adjusting the pH value to 2-6 in the step (3); the adding amount of the acid protease is 1 to 2 weight percent; the enzymolysis reaction is carried out for 1-3h at 40-60 ℃;
(4) adding the acidolysis solution obtained in the step (3) into an alkali solution to adjust the pH value to be alkaline, and reacting to obtain a second mixed solution; adjusting the pH value to 8-10 in the step (4); the reaction is carried out for 20-40min at 40-60 ℃;
(5) inactivating enzyme of the second mixed solution in the step (4), performing centrifugal separation, and taking upper-layer free oil to obtain grape seed oil;
the total phenol content of the obtained grape seed oil is more than or equal to 30 mg/kg; the content of unsaturated fatty acid is more than or equal to 86.63 percent, the content of total tocopherol is more than or equal to 253.6mg/kg, and the content of total sterol is more than or equal to 2.588 mg/g.
2. The preparation method according to claim 1, wherein the sieving in the step (1) is 20-60 mesh sieving.
3. The preparation method according to claim 1, wherein the enzyme deactivation in the step (5) is carried out by heating the slurry to 80-90 ℃ for 10-20min, and cooling; the centrifugal rotating speed is 3000-4000r/min, and the centrifugation is carried out for 10-20 min.
4. Grape seed oil obtained by the preparation method of any one of claims 1-3.
5. Use of grape seed oil obtained by the method according to any one of claims 1 to 3 for the preparation of a health food.
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Non-Patent Citations (4)
| Title |
|---|
| 4个葡萄品种葡萄籽冷榨油的性质与体外抗氧化活性;郑亚蕾等;《食品科学》;20160215(第03期);第27页摘要、第29页第1.3.4.2-1.3.4.1节、第30页第2.2节、第31页第2.3节 * |
| 关于植物油碱处理的工艺改进;翟岳明;《涂料工业》;19800629(第03期);第12页第2节 * |
| 水酶法提取葡萄籽油的工艺研究;吕姗姗等;《粮油加工》;20101228(第12期);第7-8页第1.1节、第1.3节，第10页第3节 * |
| 超临界CO_2流体提取含原花青素的葡萄籽油;杨晋等;《食品科技》;20091220(第12期);第240页摘要 * |
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