CN105777926A - Method for comprehensively extracting effective components from walnuts - Google Patents

Abstract

The invention discloses a method for comprehensively extracting active ingredients from walnuts. The steps are as follows: 1) adding water to walnut kernels and grinding them into walnut pulp, inactivating enzymes, adding compound enzymes, reacting, and centrifuging to obtain three parts: clear oil, hydrolyzate and residue , collect the clear oil to obtain the fatty acid; 2) inactivate the hydrolyzate, add ethanol to precipitate, centrifuge, collect the supernatant and the first precipitate, and freeze-dry the supernatant to obtain the active peptide; 3) to step 1) to obtain Add deionized water to the residue, ultrasonic-vacuum extraction, and suction filtration to obtain polysaccharide extract; evaporate and concentrate, add ethanol for precipitation, and centrifuge to obtain the second precipitate, which is combined with the first precipitate, decolorized, and centrifuged , dried to obtain polysaccharides. The method of the invention can obtain fatty acids, active peptides and polysaccharides in walnut kernels, has simple operation, less time consumption, low energy consumption, environment-friendly solvent, high purity and good quality of active peptides, and high yield of fatty acids, active peptides and polysaccharides.

Description

A method for comprehensively extracting active ingredients from walnuts

technical field

The invention relates to a method for comprehensively obtaining active ingredients from walnuts.

Background technique

Walnut (Juglansregia L.) has another name called Juglans regia L., belongs to Juglandaceae Juglans plant. In my country, walnut cultivation area ranks first in the world. Walnut is an excellent dried fruit food that integrates protein, fat, cellulose, vitamins and other nutrients. It is rich in nutrition, high in economic value, warm in nature, sweet in taste, non-toxic, and has the functions of invigorating the stomach, nourishing blood, moistening the lungs, and nourishing the mind. and other effects.

There are many pharmacological activities of walnuts, the most important ones are: anti-tumor activity, anti-oxidation, antibacterial effect and hypotensive effect.

According to reports, every 100g of walnut kernel contains 63.00-70.00g of high-quality fat, 14.60-19.00g of protein, 5.40-10.00g of carbohydrate, 280.00mg of phosphorus, 85.00mg of calcium, 2.60mg of iron, 3.00mg of potassium, 0.36mg of vitamin A, vitamin A B10.26mg, vitamin B20.15mg, niacin 1.00mg, riboflavin 0.11mg, niacin 1.00mg, thiamine 0.17mg, also contains a small amount of selenium, manganese, chromium and other minerals and vitamin E, vitamin K, etc. , rich in lecithin.

The composition with the highest content in the walnut kernel is fat, and the oil content of the walnut kernel is as high as 60% to 70%, and the content of unsaturated fatty acid in the walnut oil is also as high as 90%. In particular, walnut kernels contain relatively high levels of linoleic acid and linolenic acid, which are beneficial to human health. Palmitic acid and stearic acid are saturated fatty acids, and excessive use can cause cardiovascular disease. Compared with other edible oil fatty acids, walnut oil Contains relatively low levels of palmitic and stearic acids.

The content of protein in walnuts accounts for about 13-16%. There are 4 kinds of proteins in total. The highest content is gluten, which is an alkali-soluble protein, accounting for more than 70% of the total walnut protein. The other three are globulins in order of content. , albumin and prolamins. After walnut protein is hydrolyzed by protease, walnut polypeptide can be obtained, which belongs to biologically active peptides and has various pharmacological activities such as antioxidant activity, antibacterial activity and blood pressure lowering activity.

Walnuts contain a high content of polysaccharides, polysaccharides and their complexes are one of the most abundant natural macromolecules in nature, and play a vital role in maintaining life activities. It has shown good application prospects in immune regulation, anti-tumor, anti-virus, anti-oxidation and hypoglycemic.

At present, the method of extracting fatty acids mainly adopts the organic phase leaching method. The extraction process is easy to cause pollution, and the obtained fatty acids often contain organic reagent residues, which reduces the quality. The methods for extracting active peptides and polysaccharides are mainly protease hydrolysis and simple hot water extraction, which take a long time and have a low yield.

Contents of the invention

The purpose of the invention is to overcome the deficiencies of the prior art, a method for comprehensively extracting active ingredients from walnuts.

Technical method of the present invention is summarized as follows:

A method for comprehensively extracting active ingredients from walnuts is characterized in that it comprises the following steps:

1) Add water to walnut kernels and grind them into walnut pulp, inactivate enzymes, add compound enzymes, the compound enzymes are 2% to 5% of the mass of walnut kernels, and react for 1.5 hours at 30°C-60°C and pH 7-9 ~4h, centrifuge to obtain three parts: clear oil, hydrolyzate and residue, collect clear oil to obtain fatty acid;

2) heating the hydrolyzate to 80-100°C to inactivate the enzyme, adding ethanol to ethanol volume concentration above 70% for precipitation, centrifuging, collecting the supernatant and the first precipitate, and freeze-drying the supernatant to obtain the active peptide;

3) Add deionized water to the residue obtained in step 1) according to the ratio of mass ratio of 1:20 to 1:40, and add deionized water under ultrasonic power of 400W to 700W, vacuum degree of 0.02MPa to 0.08MPa, Under the conditions, extract for 10min-50min, filter with suction, repeat the extraction of the filter residue twice, combine the filtrate to obtain the extract of polysaccharide; evaporate and concentrate the extract of polysaccharide at 40-60°C and 0.06-0.1MPa to obtain a concentrate , adding ethanol to the concentrated solution to ethanol volume concentration above 70% for precipitation, centrifugal separation, collecting the second precipitate, merging the second precipitate and the first precipitate into a total precipitate, and using Wash with water and ethanol for decolorization, centrifuge and dry to obtain polysaccharide.

The complex enzyme is composed of pectinase, cellulase and protease at a ratio of 1:1:1~3.

The protease is at least one of alkaline protease, trypsin, neutral protease and compound plant hydrolyzing protease.

The method of the present invention can obtain fatty acids, active peptides and polysaccharides in walnut kernels, has simple operation, less time consumption, low energy consumption, environmentally friendly solvent, high purity of active peptides, good quality, high yield of fatty acids, active peptides and polysaccharides, Respectively above 47.5%, 17.6% and 9.3%, can reach 61.6%, 24.2% and 11.3%.

Description of drawings

Fig. 1 is the comparison of the average yields of fatty acids, active peptides and polysaccharides obtained by walnuts using traditional methods and the method (comprehensive) of the present invention.

Fig. 2 is a process flow chart of the method of the present invention.

detailed description

The compound plant proteolytic enzyme was produced by Tianjin Nuoao Enzyme Preparation Co., Ltd.

A method for comprehensively extracting active ingredients from walnuts provided by the present invention will be described in detail below in conjunction with specific examples.

Example 1

A method for comprehensively extracting active ingredients from walnuts, comprising the steps of:

1) Soak walnut kernels in 10 times the mass of water at 4°C for 24 hours, grind them into walnut pulp, inactivate enzymes at 100°C for 5 minutes, add compound enzyme, the compound enzyme is 2% of the mass of walnut kernels, and the pH is 7.5 at 40°C Under certain conditions, react for 4 hours, centrifuge to obtain clear oil, hydrolyzate and residue three parts, collect clear oil, obtain fatty acid, yield rate is 52.9%; The compound enzyme is formed by the mass ratio of 1:1:1:1 pectinase, fiber Sulfase, trypsin and neutral protease;

2) heating the hydrolyzate described in step 1) to 100°C to inactivate enzymes, adding ethanol to ethanol volume concentration of 70% for precipitation, centrifuging, collecting the supernatant and the first precipitate, and freeze-drying the supernatant to obtain Active peptide, the yield is 18.3%;

3) According to the mass ratio of 1:20, add deionized water to the residue obtained in step 1), place it in an ultrasonic-vacuum device, and extract it for 10 minutes under the conditions of ultrasonic power 600W, vacuum degree 0.02MPa, and 80°C , suction filtration, and the filter residue was repeatedly extracted twice, and the filtrates were combined to obtain a polysaccharide extract; the polysaccharide extract was evaporated and concentrated at 40°C and 0.1MPa to obtain a concentrated solution, and ethanol was added to the concentrated solution to reach the ethanol volume concentration 70% precipitated, centrifuged, collected the second precipitate, combined the second precipitate and the first precipitate into a total precipitate, washed and decolorized with absolute ethanol 2 times the mass of the total precipitate, A total of 5 times, centrifuged, and the precipitate was dried to obtain polysaccharide with a yield of 9.7%.

Example 2

A method for comprehensively extracting active ingredients from walnuts, comprising the steps of:

1) Soak walnut kernels in 10 times the mass of water at 4°C for 24 hours, grind them into walnut pulp, inactivate enzymes at 100°C for 5 minutes, add compound enzyme, the compound enzyme is 3% of the mass of walnut kernels, at 45°C, the pH is 8 Under certain conditions, react for 2 hours and centrifuge to obtain three parts: clear oil, hydrolyzate and residue, collect the clear oil, and obtain fatty acid with a yield of 52.3%; and compound plant hydrolyzate;

2) Warm up the hydrolyzate described in step 1) to 90°C to inactivate the enzyme, add ethanol to ethanol volume concentration of 75% for precipitation, centrifuge, collect the supernatant and the first precipitate, freeze-dry the supernatant to obtain Active peptide, the yield is 17.6%;

3) According to the mass ratio of 1:30, add deionized water to the residue obtained in step 1), place it in an ultrasonic-vacuum device, and extract it for 20 minutes under the conditions of ultrasonic power 500W, vacuum degree 0.05MPa, and 70°C , suction filtration, and the filter residue was repeatedly extracted twice, and the filtrates were combined to obtain a polysaccharide extract; the polysaccharide extract was evaporated and concentrated at 50°C and 0.08MPa to obtain a concentrated solution, and ethanol was added to the concentrated solution to reach the ethanol volume concentration 80% precipitated, centrifuged, collected the second precipitate, combined the second precipitate and the first precipitate into a total precipitate, washed and decolorized with absolute ethanol 2 times the mass of the total precipitate, A total of 5 times, centrifuged, and the precipitate was dried to obtain polysaccharide with a yield of 9.3%.

Example 3

A method for comprehensively extracting active ingredients from walnuts, comprising the steps of:

1) Soak walnut kernels in 10 times the mass of water at 4°C for 24 hours, grind them into walnut pulp, inactivate enzymes at 100°C for 5 minutes, add compound enzyme, the compound enzyme is 5% of the mass of walnut kernels, and the pH is 7 at 30°C Under certain conditions, react for 3 hours, and centrifuge to obtain clear oil, hydrolyzate and residue three parts, collect clear oil, and obtain fatty acid with a yield of 47.5%; , cellulase, alkaline protease, neutral protease and compound plant hydrolytic protease;

2) Warm up the hydrolyzate described in step 1) to 80°C to inactivate the enzyme, add ethanol to ethanol volume concentration of 80% for precipitation, centrifuge, collect the supernatant and the first precipitate, freeze-dry the supernatant to obtain Active peptide, the yield is 19.2%;

3) According to the mass ratio of 1:40, add deionized water to the residue obtained in step 1), place it in an ultrasonic-vacuum device, and extract it for 30 minutes under the conditions of ultrasonic power 400W, vacuum degree 0.07MPa, and 75°C , suction filtration, and the filter residue was repeatedly extracted twice, and the filtrates were combined to obtain a polysaccharide extract; the polysaccharide extract was evaporated and concentrated at 60°C and 0.06MPa to obtain a concentrated solution, and ethanol was added to the concentrated solution to reach the ethanol volume concentration 85% precipitated, centrifuged, collected the second precipitate, combined the second precipitate and the first precipitate into a total precipitate, washed and decolorized with absolute ethanol 2 times the mass of the total precipitate, A total of 5 times, centrifuged, and the precipitate was dried to obtain polysaccharide with a yield of 10.4%.

Example 4

A method for comprehensively extracting active ingredients from walnuts, comprising the steps of:

1) Soak walnut kernels in 10 times the mass of water at 4°C for 24 hours, grind them into walnut pulp, inactivate enzymes at 100°C for 5 minutes, add compound enzyme, the compound enzyme is 4% of the mass of walnut kernels, and the pH is 9 at 60°C Under the condition of 1.5h, centrifuge, get clear oil, hydrolyzate and residue three parts, collect clear oil, obtain fatty acid, yield rate is 61.6%; Composite enzyme is composed of pectinase, Composition of cellulase, trypsin and alkaline protease;

2) Warm up the hydrolyzate described in step 1) to 95°C to inactivate the enzyme, add ethanol to ethanol volume concentration of 90% for precipitation, centrifuge, collect the supernatant and the first precipitate, freeze-dry the supernatant to obtain Active peptide, the yield is 24.2%;

3) According to the mass ratio of 1:35, add deionized water to the residue obtained in step 1), place it in an ultrasonic-vacuum device, and extract it for 50 minutes under the conditions of ultrasonic power 700W, vacuum degree 0.08MPa, and 60°C , suction filtration, and the filter residue was repeatedly extracted twice, and the filtrates were combined to obtain a polysaccharide extract; the polysaccharide extract was evaporated and concentrated at 45°C and 0.1MPa to obtain a concentrated solution, and ethanol was added to the concentrated solution to reach the ethanol volume concentration 90% precipitated, centrifuged, collected the second precipitate, combined the second precipitate and the first precipitate into a total precipitate, washed and decolorized with absolute ethanol 2 times the mass of the total precipitate, A total of 5 times, centrifuged, and the precipitate was dried to obtain polysaccharide with a yield of 11.3%.

Example 5

The traditional method extracts the method for active ingredient respectively from walnut, comprises the following steps:

1) Grind the walnut kernels with a medicinal material grinder to obtain broken walnut kernels, add petroleum ether 8 times the mass of the walnut kernels, react for 60 minutes under the condition of ultrasonic power of 300W, and filter with suction to collect the supernatant and the first residue , the supernatant was evaporated and concentrated under the conditions of 40°C and 0.1MPa to remove petroleum ether to obtain fatty acid with a yield of 34.0%;

2) After the first residue degreased in step 1), add deionized water at a mass ratio of 1:35, extract at 100°C for 2.5 hours, filter with suction, and repeat the extraction of the filter residue twice, and combine and collect The filtrate and the second residue, the filtrate, evaporate and concentrate at 40°C and 0.1MPa to obtain a concentrated solution, add ethanol to the concentrated solution to ethanol volume concentration of 70% for precipitation, centrifuge, collect the precipitate, and use the precipitate Wash and decolorize with 2 times the mass of absolute ethanol, 5 times in total, centrifuge, precipitate and dry to obtain polysaccharide with a yield of 6.7%;

3) After the second middle residue of step 2), add deionized water in a ratio of 1:20 by mass ratio, and then add alkaline protease which is 6% of the second residue quality, at 50 ° C, pH is 9 Under the conditions, react for 2.5 hours, centrifuge, collect the hydrolyzate, and freeze-dry to obtain the active peptide with a yield of 19.6%.

Example 6

The traditional method extracts the method for active ingredient respectively from walnut, comprises the following steps:

1) Grind the walnut kernels with a medicinal material grinder to obtain broken walnut kernels, add petroleum ether 10 times the mass of the walnut kernels, react for 40 minutes under the condition of ultrasonic power of 450W, filter with suction, and collect the supernatant and the first residue , the supernatant was evaporated and concentrated under the conditions of 45°C and 0.08MPa to remove petroleum ether to obtain fatty acid with a yield of 35.1%;

2) After the first residue degreased in step 1), add deionized water at a mass ratio of 1:40, extract at 90°C for 2 hours, filter with suction, repeat the extraction of the filter residue twice, and collect the filtrate And the second kind of residue, the filtrate is evaporated and concentrated under the conditions of 45°C and 0.08MPa to obtain a concentrated solution, and ethanol is added to the concentrated solution until the volume concentration of ethanol is 80% for precipitation, centrifuged, and the precipitate is collected. Wash and decolorize with 100% absolute ethanol, 5 times in total, centrifuge, precipitate and dry to obtain polysaccharide with a yield of 6.1%;

3) After the second residue of step 2), add deionized water at a mass ratio of 1:25, and then add trypsin that is 6% of the mass of the second residue, at 40 ° C, pH is 8 conditions , reacted for 3.5 hours, centrifuged, collected the hydrolyzate, and freeze-dried to obtain the active peptide with a yield of 17.3%.

Traditional method references:

1. Ning Yu, Lin Lijie, Wang Fengjun. Ultrasonic-assisted extraction of walnut catalpa oil and its fatty acid analysis[J]. Food Science and Technology, 2014,39(10):184-188.

2. Zhao Kentian, Yan Xiujuan. Research on hot water extraction technology of soluble polysaccharides from the kernels of walnut catalpa [J]. China Forestry By-products, 2008(2):17-18.

3. Zhang Wencui, Zhang Peizheng. Process Optimization of Enzymatic Preparation of Peptides from Walnut[J]. Chinese Food and Nutrition, 2010(4):52-55.

Claims (3)

1. a method for comprehensively extracting active ingredient from walnut, is characterized in that comprising the steps: 1) Add water to walnut kernels and grind them into walnut pulp, inactivate enzymes, add compound enzymes, the compound enzymes are 2% to 5% of the mass of walnut kernels, and react for 1.5 hours at 30°C-60°C and pH 7-9 ~4h, centrifuge to obtain three parts: clear oil, hydrolyzate and residue, collect clear oil to obtain fatty acid; 2) heating the hydrolyzate to 80-100°C to inactivate the enzyme, adding ethanol to ethanol volume concentration above 70% for precipitation, centrifuging, collecting the supernatant and the first precipitate, and freeze-drying the supernatant to obtain the active peptide; 3) Add deionized water to the residue obtained in step 1) according to the ratio of mass ratio of 1:20 to 1:40, and add deionized water under ultrasonic power of 400W to 700W, vacuum degree of 0.02MPa to 0.08MPa, Under the conditions, extract for 10min-50min, filter with suction, repeat the extraction of the filter residue twice, combine the filtrate to obtain the polysaccharide extract; evaporate and concentrate the polysaccharide extract at 40-60°C and 0.06-0.1Mpa to obtain a concentrate , adding ethanol to the concentrated solution to ethanol volume concentration above 70% for precipitation, centrifugal separation, collecting the second precipitate, merging the second precipitate and the first precipitate into a total precipitate, and using Wash with water and ethanol for decolorization, centrifuge and dry to obtain polysaccharide. 2. The method according to claim 1, characterized in that the compound enzyme is composed of pectinase, cellulase and protease in a ratio of 1:1:1 to 3. 3. The method according to claim 2, characterized in that the protease is at least one of alkaline protease, trypsin, neutral protease and compound plant hydrolytic protease.