CN107541332B - Moringa oleifera oil and preparation method thereof - Google Patents
Moringa oleifera oil and preparation method thereof Download PDFInfo
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Abstract
The invention discloses moringa oil and a preparation method thereof. The method comprises the steps of firstly carrying out microwave treatment, high-temperature cooking, high-pressure homogenization and alkaline extraction on moringa seeds, and then combining with plant hydrolase enzymolysis and protease enzymolysis, high-temperature treatment, low-temperature freezing, unfreezing and low-temperature high-speed centrifugation to obtain moringa seed oil rich in moringa seed protein peptide, wherein the extraction rate of the moringa seed oil is more than 90%; and finally, adding a moringa leaf extract which is subjected to plant hydrolase enzymolysis, protease enzymolysis, ethanol extraction and macroporous resin enrichment extraction into the moringa seed oil to enhance the oxidation resistance, so as to obtain the moringa oil. The moringa oil disclosed by the invention is high in nutritive value, good in oxidation stability and long in shelf life, and can be widely applied to the food industry.
Description
Technical Field
The invention belongs to the field of deep processing of moringa seeds, and particularly relates to moringa oil and a preparation method thereof.
Background
Moringa oleifera, a plant of the genus Moringa of the family Moringaceae, is native to the Himalayas mountain in the northern part of India, is widely planted in the tropical and subtropical regions of Asia and Africa, has 14 species, is the traditional Moringa oleifera (Moringa oleifera Lam.) in India, has fast growth and wide distribution, and is the Moringa oleifera species with the largest cultivation area and the most researches. The introduction, planting and development research of moringa oleifera in China mainly focuses on Yunnan provinces, Guangxi provinces, Guangdong provinces, Fujian provinces, Guizhou provinces, Taiwan provinces and the like. In 2012, moringa leaves were approved by the national institutes of health and family planning as a new resource food. The whole body of the moringa oleifera is treasure, and leaves, flowers, seeds, tender shoots, tender stems, roots and the like of the moringa oleifera can be eaten, so that the moringa oleifera is a plant with homology of medicine and food. Moringa oleifera is commonly used in India and African countries to treat high-incidence diseases such as hypertension, diabetes, cardiovascular diseases, rheumatism, arthritis and the like.
The moringa oil contains high-content oleic acid and various sterols, and is a high-quality oil. However, due to lack of research on the deep processing technology, the popularization and the deep development of the moringa oil are severely restricted. The traditional oil extraction method mainly comprises a squeezing method and a leaching method, and along with the development of science and technology, the understanding of people on the concept of healthy oil is deepened and the deep pursuit of a green production processing mode is carried out, the defects of the squeezing method and the organic solvent extraction method are gradually revealed. The aqueous enzymatic method has mild action conditions, no organic solvent residue and high oil extraction rate, and is gradually paid more attention.
Oils and fats without antioxidant are easily oxidized. In recent years, natural antioxidants are becoming more popular in the food industry as the market demands for healthy oils and fats have increased. The moringa leaves are rich in polyphenols, have strong oxidation resistance, and can be developed into a natural oil antioxidant to be applied to vegetable oil.
Therefore, the method for efficiently and greenly extracting and preparing the moringa oil has stronger social and economic benefits.
Disclosure of Invention
The invention aims to provide moringa oil. The moringa oil has oleic acid content of 72% and behenic acid content of 3%, high nutritive value, high oxidation stability and long shelf life, and can be widely used in food industry.
The invention also aims to provide a preparation method of the moringa oil. The method comprises the steps of firstly carrying out microwave treatment, high-temperature cooking, high-pressure homogenization and alkaline extraction on moringa seeds, and then combining with plant hydrolase enzymolysis and protease enzymolysis, high-temperature treatment, low-temperature freezing, unfreezing and low-temperature high-speed centrifugation to obtain moringa seed oil rich in moringa seed protein peptide, wherein the extraction rate of the moringa seed oil is more than 90%; and finally, adding a moringa leaf extract which is subjected to plant hydrolase enzymolysis, protease enzymolysis, ethanol extraction and macroporous resin enrichment extraction into the moringa seed oil to enhance the oxidation resistance, so as to obtain the moringa oil.
The purpose of the invention is realized by the following technical scheme.
A preparation method of moringa oil comprises the steps of adopting pretreatment processes of microwave treatment, high-temperature cooking, high-pressure homogenization and ultrasonic-assisted alkaline extraction, and combining treatment processes of plant hydrolase enzymolysis, protease enzymolysis, high-temperature treatment, low-temperature freezing, unfreezing and low-temperature high-speed centrifugation to obtain moringa oil; finally, the moringa oil is prepared by adding the moringa leaf extract, and the method specifically comprises the following steps:
(1) microwave treatment: drying fresh moringa seeds, removing shells, performing microwave treatment, crushing, sieving and performing superfine crushing to obtain moringa seed powder;
(2) high-temperature cooking: steaming the obtained moringa seed powder at high temperature;
(3) high-pressure homogenization: taking moringa seed powder which is steamed at high temperature, adding deionized water, shearing at high speed, passing through a colloid mill, and homogenizing at high pressure to obtain moringa seed homogenate;
(4) ultrasonic-assisted alkaline extraction: adding sodium hydroxide into the moringa seed powder homogenate to adjust the pH value, performing ultrasonic-assisted extraction, and cooling to room temperature to obtain a suspension;
(5) and (3) carrying out enzymolysis on moringa seeds: adjusting the pH value of the suspension, adding plant hydrolase for enzymolysis, adjusting the pH value, adding protease for enzymolysis, performing high-temperature treatment, cooling to room temperature, performing low-temperature freezing treatment, performing low-temperature thawing, and performing low-temperature high-speed centrifugation to obtain moringa seed oil;
(6) preparing the moringa oil: dissolving the moringa oleifera leaf extract in absolute ethyl alcohol, adding the moringa oleifera leaf extract into the moringa oleifera seed oil, and uniformly stirring to obtain the moringa oleifera oil.
Further, in the step (1), the microwave treatment is heating for 2-4min under the microwave condition of 500-700W.
Further, in the step (1), the sieving is performed by a 100-mesh sieve.
Further, in the step (1), the micronization is carried out for 2-3h in a jet mill, and the average particle size of the moringa seed powder obtained by pulverization is 8-12 μm.
Further, in the step (2), the high-temperature cooking is performed for 20-30min under the water vapor condition of 100-120 ℃.
Further, in the step (3), the material-to-liquid ratio of the moringa seed powder subjected to high-temperature cooking to the deionized water is 1: 6-8 g/mL.
Further, in the step (3), the high-speed shearing is carried out at a speed of 8000-10000 rpm for 10-20 minutes.
Further, in the step (3), the number of times of passing through the colloid mill is 2-3.
Further, in the step (3), the pressure of the high-pressure homogenization is 20-40 MPa, and the number of times of the high-pressure homogenization is 2-3.
Further, in the step (4), the pH value is adjusted to be 7.0-9.0.
Further, in the step (4), the ultrasonic power of the ultrasonic-assisted extraction is 600-800W, the time is 10-20min, and the temperature is 40-60 ℃.
Further, in the step (5), the pH value of the suspension is adjusted to 4.4-5.0.
Further, in the step (5), the addition amount of the plant hydrolase is 2-4% of the mass of the moringa seeds.
Further, in the step (5), the plant hydrolase is a composite plant hydrolase of alpha-amylase and Viscozyme L, wherein the addition amount of the alpha-amylase accounts for 60-80% of the total enzyme amount, and the addition amount of the composite plant hydrolase of Viscozyme L accounts for 20-40% of the total enzyme amount.
Further, in the step (5), the plant hydrolase is subjected to enzymolysis for 4-6 hours at 50-56 ℃.
Further, in the step (5), after the plant hydrolase is subjected to enzymolysis, the pH value is adjusted to 7.0-8.0.
Further, in the step (5), the adding amount of the protease is 4-6% of the mass of the moringa seeds.
Further, in the step (5), the protease is a compound protease and an alkaline protease with the trade mark NS37071, the adding amount of the compound protease accounts for 10-30% of the total enzyme amount, and the adding amount of the alkaline protease with the trade mark NS37071 accounts for 70-90% of the total enzyme amount.
Further, in the step (5), the enzymolysis of the protease is carried out for 12-16 hours at 50-56 ℃.
Further, in the step (5), the high temperature treatment is 100-120 ℃ heat treatment for 20-40 min.
Further, in the step (5), the low-temperature freezing treatment is freezing for 8-12h at-18 to-24 ℃.
Further, in the step (5), the low-temperature unfreezing is carried out at 4-8 ℃ and the mixture is placed for 8-12 h.
Further, in the step (5), the low-temperature centrifugation is carried out at 4-8 ℃ and the centrifugation is carried out for 20-30min at 6000-8000 g.
Further, in the step (5), the extraction yield of the moringa seed oil is more than 90%.
Further, in the step (6), the moringa oleifera leaf extract is obtained by carrying out treatment processes of plant hydrolase enzymolysis, protease enzymolysis, ethanol extraction and macroporous resin enrichment, and specifically comprises the following steps:
taking moringa oleifera dry leaves, crushing and sieving to obtain moringa oleifera leaf powder, adding deionized water, uniformly mixing to obtain a suspension, adjusting the pH value of the suspension, adding plant hydrolase for enzymolysis, adjusting the pH value, adding protease for enzymolysis, adding absolute ethyl alcohol, heating and refluxing for extraction, cooling an extracting solution to room temperature, centrifuging to remove residues, taking a supernatant, concentrating under reduced pressure, freeze-drying, taking freeze-dried powder, dissolving in water, passing through a macroporous resin column, performing gradient elution by adopting an ethanol solution, collecting an eluent, and freeze-drying to obtain the moringa oleifera leaf extract.
Further, the sieving is a 40 mesh sieve.
Furthermore, the feed-liquid ratio of the moringa oleifera leaf powder to the deionized water is 1: 8-10 g/mL.
Further, the pH of the suspension is adjusted to 4.4-5.0.
Furthermore, the addition amount of the plant hydrolase is 2-4% of the mass of the dry leaves of the moringa oleifera.
Further, the plant hydrolase is a composite plant hydrolase of trade name Viscozyme L.
Furthermore, the plant hydrolase is subjected to enzymolysis for 4-6 hours at 50-56 ℃.
Further, after the plant hydrolase is subjected to enzymolysis, the pH value is adjusted to 7.0-8.0.
Furthermore, the addition amount of the protease is 4-6% of the mass of the moringa oleifera dry leaves.
Further, the protease is pancreatin and alkaline protease sold under the trade name NS37071, the adding amount of the pancreatin accounts for 40-60% of the total enzyme amount, and the adding amount of the alkaline protease sold under the trade name NS37071 accounts for 40-60% of the total enzyme amount.
Furthermore, the enzymolysis of the protease is carried out for 6-8 hours at 50-56 ℃.
Furthermore, the addition amount of the absolute ethyl alcohol is that the content of the ethyl alcohol in the system reaches 30-50 wt%.
Further, the heating reflux condition is that the heating reflux is carried out for 1-2h at the temperature of 90-100 ℃.
Furthermore, the centrifugation is 4000-6000 g for 10-20 min.
Further, the concentrated solution is concentrated under reduced pressure until the solid content is 20-30%.
Further, the concentration of the lyophilized powder after dissolving in water was 100-200 mg/mL.
Further, the macroporous resin is mitsubishi chemical macroporous adsorption resin SP 207.
Further, the gradient elution is gradient elution by using 0-50 vol% ethanol solution, and specifically comprises the following steps: eluting with 100vol% water for 1-2 column volumes, eluting with 100-60 vol% water and 0-40 vol% ethanol for 3-5 column volumes, eluting with 50 vol% water and 50 vol% ethanol for 6-7 column volumes, and collecting 50 vol% ethanol eluate.
Further, in the step (6), the dissolving ratio of the moringa leaf extract to the absolute ethyl alcohol is 0.1-0.2:3-5 g/mL.
Further, in the step (6), the mass ratio of the moringa leaf extract to the moringa seed oil is 1-2: 10000.
A moringa oil having an oleic acid content of >72% and a behenic acid content of >3% produced by any one of the above methods.
The prepared moringa oil has high nutritive value, good oxidation stability and long shelf life, and can be widely applied to the food industry.
Compared with the prior art, the invention has the following advantages and beneficial effects:
(1) according to the method, the moringa seed oil is obtained by adopting a pretreatment process of microwave treatment, high-temperature cooking, high-pressure homogenization and ultrasonic-assisted alkaline extraction, and combining plant hydrolase enzymolysis, protease enzymolysis, high-temperature treatment, low-temperature freezing, unfreezing and low-temperature high-speed centrifugation, so that the oil yield of the moringa seed is improved;
(2) according to the method, the moringa oleifera leaf extract obtained by enzymolysis of plant hydrolase and protease, ethanol extraction and macroporous resin enrichment is added into the moringa oleifera seed oil to prepare the moringa oleifera oil, so that the oxidation stability of the moringa oleifera oil is greatly improved, and the quality guarantee period of the moringa oleifera oil is prolonged;
(3) according to the method, the moringa oleifera seed oil is extracted by adopting an efficient and green extraction technology, the extraction rate of the moringa oleifera seed oil is more than 90%, the obtained moringa oleifera oil has the oleic acid content of more than 72% and the behenic acid content of more than 3%, no organic solvent residue, no chemical additive, high nutritional value, good oxidation stability and long shelf life, and can be widely applied to the food industry.
Detailed Description
The technical solution of the present invention is further described in detail with reference to the following specific examples, but the scope of the present invention is not limited to the following examples.
In the following examples, the method for measuring the fatty acid composition in fats and oils was as follows:
weighing 10-20mg of a grease sample, adding 2mL of 14% boron trifluoride-methanol solution, and carrying out water bath at 60 ℃ for 30min to carry out methyl esterification; cooling to room temperature, adding 1mL of water and 1mL of n-hexane, mixing uniformly, standing for layering, taking an upper organic layer, adding anhydrous sodium sulfate for dewatering, drying by using high-purity nitrogen, adding 0.4mL of n-hexane and 0.1mL of an internal standard (methyl heptadecanoate-n-hexane solution), filtering by using a 0.22 micron filter membrane, and carrying out GC-MS analysis.
GC conditions were as follows:
(1) TR-5ms elastic quartz chromatographic column with the split ratio of 100: 1;
(2) column temperature program: the initial temperature is 40 deg.C, and is maintained for 2min, then the temperature is increased to 150 deg.C at 10 deg.C/min, and is maintained for 2min, and then the temperature is increased to 280 deg.C at 10 deg.C/min, and is maintained for 5 min;
(3) the carrier gas was high purity He at a flow rate of 1.0 mL/min.
MS conditions:
(1) electron impact ionization ion source (EI), electron energy 70 eV;
(2) the ion source temperature is 230 ℃, and the interface (transmission line) temperature is 250 ℃;
(3) the scanning speed is 3.0scans/s, and the scanning mass range is 33-350 m/z.
The method for measuring the oxidation stability of the grease adopts a Schaal oven method, and the grease is placed in a constant-temperature oven at 60 ℃ for continuously accelerating the oxidation for 14 days; the POV value and acid value of 16.00g of oil samples were measured at 2d intervals.
According to the national standard, the upper limit of the peroxide value of the grease is 6mmol/kg, and then the shelf life of the product is calculated according to the Arrhenius empirical formula, wherein the storage time of the product is 16 days under the condition that the temperature is 60 ℃ and 1 day of an oven method is equivalent to the storage time of the product at 20 ℃; the acid value is determined by GB/T5009.37-2003, and the peroxide value is determined by GB/T5009.37-2003.
Example 1
The preparation method of the moringa oil specifically comprises the following steps:
(1) microwave treatment: drying fresh moringa seeds, removing shells, heating with microwave power of 500W for 2min, pulverizing, sieving with 100 mesh sieve, and pulverizing in a jet mill for 2h to obtain moringa seed powder with average particle size of 12 μm;
(2) high-temperature cooking: steaming the obtained Moringa seed powder at 100 deg.C for 20 min;
(3) high-pressure homogenization: taking the moringa seed powder after high-temperature cooking, adding deionized water according to the material-liquid ratio of 1:6g/mL, shearing at the speed of 8000rpm for 10 minutes, passing through a colloid mill for 2 times to obtain moringa seed homogenate, and homogenizing at 20MPa and high pressure for 2 times;
(4) ultrasonic-assisted alkaline extraction: adding sodium hydroxide into the moringa seed powder homogenate to adjust the pH value to 7.0, extracting for 10min at 40 ℃ under the ultrasonic power of 600W, and cooling to room temperature to obtain a suspension;
(5) and (3) carrying out enzymolysis on moringa seeds: adjusting the pH value of the suspension to 4.4, adding alpha-amylase accounting for 60% of the total enzyme by mass and a compound plant hydrolase of a commercial brand Viscozyme L accounting for 40% of the total enzyme by mass, and carrying out enzymolysis for 4 hours at 50 ℃; after the enzymolysis of plant hydrolase, adjusting the pH value to 7.0, adding compound protease accounting for 4 percent of the mass of the moringa seeds and alkaline protease of a commercial brand NS37071, wherein the adding amount of the compound protease accounts for 10 percent of the total enzyme amount, the adding amount of the alkaline protease of the commercial brand NS37071 accounts for 90 percent of the total enzyme amount, and carrying out enzymolysis for 12 hours at 50 ℃; heating at 100 deg.C for 20min, cooling to room temperature, and freezing at-18 deg.C for 8 hr; standing at 4 ℃ for 8h for low-temperature thawing, and centrifuging at 6000g for 20min to obtain moringa seed oil 1, wherein the extraction rate of the moringa seed oil 1 is 92%;
(6) preparing a moringa oleifera leaf extract: taking dried moringa leaves, crushing, sieving by a 40-mesh sieve, adding deionized water according to the feed-liquid ratio of 1:8g/mL, uniformly mixing, adjusting the pH value of a suspension to 4.4, adding a compound plant hydrolase of a commercial brand Viscozyme L with the mass of 2% of the mass of the dried moringa leaves, and carrying out enzymolysis for 4 hours at 50 ℃; after enzymolysis by plant hydrolase, adjusting the pH value to 7.0, adding pancreatin accounting for 4% of the mass of the dry leaves of the moringa oleifera and alkaline protease of a commercial brand NS37071, wherein the addition amount of the pancreatin accounts for 40% of the total enzyme amount, the addition amount of the alkaline protease of the commercial brand NS37071 accounts for 60% of the total enzyme amount, and carrying out enzymolysis for 6 hours at 50 ℃; adding absolute ethyl alcohol to make the content of the ethyl alcohol in the system reach 30 wt%, heating and refluxing for 1h at 90 ℃, cooling to room temperature, centrifuging for 10min at 4000g, taking supernatant, concentrating under reduced pressure until the content of solid matters is 20%, freeze-drying, taking freeze-dried powder to prepare 100mg/mL aqueous solution, passing through a Mitsubishi chemical macroporous adsorption resin SP207 column, and performing 7-column volume gradient elution by adopting 0-50 vol% ethyl alcohol solution at the flow rate of 2mL/min, specifically: eluting with 100% water and 0% ethanol for 1-5 column volumes, eluting with 50 vol% water and 50 vol% ethanol for 6-7 column volumes, collecting 50 vol% ethanol eluate, and freeze drying to obtain Moringa oleifera leaf extract; the extracted moringa oleifera leaf extract has a polyphenol content of 22%;
(7) preparing the moringa oil: dissolving 0.1g of moringa oleifera leaf extract in 3mL of absolute ethyl alcohol, adding into 1000g of moringa oleifera seed oil, and uniformly stirring to obtain moringa oleifera oil 1.
The prepared moringa oil 1 has the oleic acid content of 74.2 percent, the behenic acid content of 3.18 percent and high nutritional value, and the shelf life of the moringa oil under the conditions of room temperature and light shielding is estimated to be 72 months according to an Arrhenius empirical formula.
Example 2
The preparation method of the moringa oil specifically comprises the following steps:
(1) microwave treatment: drying fresh moringa seeds, removing shells, heating for 3min at the microwave power of 600W, crushing, sieving with a 100-mesh sieve, and crushing in a jet mill for 2.5h to obtain moringa seed powder with the average particle size of 10 mu m;
(2) high-temperature cooking: steaming the obtained Moringa seed powder at 110 deg.C for 25 min;
(3) high-pressure homogenization: taking the moringa seed powder after high-temperature cooking, adding deionized water according to the material-liquid ratio of 1:7g/mL, shearing for 15 minutes at the speed of 9000rpm, passing through a colloid mill for 3 times to obtain moringa seed homogenate, and homogenizing for 3 times at the high pressure of 30 MPa;
(4) ultrasonic-assisted alkaline extraction: adding sodium hydroxide into the moringa seed powder homogenate to adjust the pH value to 8.0, extracting for 15min at 50 ℃ under the ultrasonic power of 700W, and cooling to room temperature to obtain a suspension;
(5) and (3) carrying out enzymolysis on moringa seeds: adjusting the pH value of the suspension to 4.7, adding alpha-amylase accounting for 70% of the total enzyme by mass and a compound plant hydrolase of a commercial brand Viscozyme L accounting for 30% of the total enzyme by mass, and carrying out enzymolysis for 5 hours at 53 ℃; after the enzymolysis of plant hydrolase, adjusting the pH value to 7.5, adding composite protease accounting for 5 percent of the mass of the moringa seeds and alkaline protease of a product number NS37071, wherein the addition amount of the composite protease accounts for 20 percent of the total enzyme amount, the addition amount of the alkaline protease of the product number NS37071 accounts for 80 percent of the total enzyme amount, and carrying out enzymolysis for 14 hours at the temperature of 53 ℃; heating at 110 deg.C for 30min, cooling to room temperature, and freezing at-21 deg.C for 10 hr; standing at 6 deg.C for 10h for thawing at low temperature, centrifuging at 7000g for 25min to obtain Moringa seed oil 2, with extraction rate of Moringa seed oil 2 of 91%;
(6) preparing a moringa oleifera leaf extract: taking dried moringa leaves, crushing, sieving by a 40-mesh sieve, adding deionized water according to the feed-liquid ratio of 1:9g/mL, uniformly mixing, adjusting the pH value of a suspension to 4.7, adding a compound plant hydrolase of a commercial brand Viscozyme L with the mass of 3% of the mass of the dried moringa leaves, and carrying out enzymolysis for 5 hours at 53 ℃; after the enzymolysis of plant hydrolase, adjusting the pH value to 7.5, adding pancreatin which accounts for 50% of the total enzyme amount and alkaline protease which accounts for 50% of the total enzyme amount and is 5% of the mass of the dry leaves of the moringa oleifera and the alkaline protease which is sold under the trade mark NS37071, and carrying out enzymolysis for 7 hours at 53 ℃; adding absolute ethyl alcohol to make the content of the ethyl alcohol in the system reach 40 wt%, heating and refluxing at 95 ℃ for 1.5h, cooling to room temperature, centrifuging for 10min at 5000g, taking supernatant, concentrating under reduced pressure until the content of solid matters is 25%, freeze-drying, taking freeze-dried powder to prepare 150mg/mL aqueous solution, passing through a Mitsubishi chemical macroporous adsorption resin SP207 column, and performing 7 column volume gradient elution by adopting 0-50 vol% ethyl alcohol solution at the flow rate of 2mL/min, specifically: eluting 1-2 column volumes with 100% water and 0% ethanol, eluting 3-5 column volumes with 80% water and 20% ethanol, eluting 6-7 column volumes with 50% water and 50% ethanol, collecting 50% ethanol eluate, and freeze drying to obtain Moringa oleifera leaf extract; the extracted moringa oleifera leaf extract has a polyphenol content of 23%;
(7) preparing the moringa oil: dissolving 0.15g of moringa oleifera leaf extract in 4mL of absolute ethyl alcohol, adding 1000g of moringa oleifera seed oil, and uniformly stirring to obtain moringa oleifera oil 2.
The prepared moringa oil 2 has the oleic acid content of 73.4 percent, the behenic acid content of 3.11 percent and high nutritional value, and the shelf life of the moringa oil under the conditions of room temperature and light shielding is presumed to be 74 months according to an Arrhenius empirical formula.
Example 3
The preparation method of the moringa oil specifically comprises the following steps:
(1) microwave treatment: drying fresh moringa seeds, removing shells, heating for 4min at the microwave power of 700W, crushing, sieving with a 100-mesh sieve, and crushing in a jet mill for 3h to obtain moringa seed powder with the average particle size of 8 mu m;
(2) high-temperature cooking: steaming the obtained Moringa seed powder at 120 deg.C for 30 min;
(3) high-pressure homogenization: taking the moringa seed powder after high-temperature cooking, adding deionized water according to the feed-liquid ratio of 1:8g/mL, shearing at the speed of 10000rpm for 20 minutes, passing through a colloid mill for 3 times to obtain moringa seed homogenate, and homogenizing at 40MPa for 2 times;
(4) ultrasonic-assisted alkaline extraction: adding sodium hydroxide into the moringa seed powder homogenate to adjust the pH value to 9.0, extracting for 20min at the ultrasonic power of 800W and the temperature of 60 ℃, and cooling to room temperature to obtain a suspension;
(5) and (3) carrying out enzymolysis on moringa seeds: adjusting the pH value of the suspension to 5.0, adding alpha-amylase accounting for 80% of the total enzyme by mass and composite plant hydrolase of a commercial brand Viscozyme L accounting for 20% of the total enzyme by mass, and carrying out enzymolysis for 6 hours at 56 ℃; after the enzymolysis of plant hydrolase, adjusting the pH value to 8.0, adding composite protease with the mass of 6 percent of that of the moringa seeds and alkaline protease with the trade mark NS37071, wherein the addition amount of the composite protease accounts for 30 percent of the total enzyme amount, the addition amount of the alkaline protease with the trade mark NS37071 accounts for 70 percent of the total enzyme amount, and carrying out enzymolysis for 16 hours at 56 ℃; heating at 120 deg.C for 40min, cooling to room temperature, and freezing at-24 deg.C for 12 hr; standing at 8 ℃ for 12h for low-temperature thawing, and centrifuging at 8000g for 30min to obtain Moringa seed oil 3, wherein the extraction rate of the Moringa seed oil 3 is 93%;
(6) preparing a moringa oleifera leaf extract: taking dried moringa leaves, crushing, sieving by a 40-mesh sieve, adding deionized water according to the feed-liquid ratio of 1:10g/mL, uniformly mixing, adjusting the pH value of a suspension to 5.0, adding a compound plant hydrolase of a commercial brand Viscozyme L with the mass of 4% of the mass of the dried moringa leaves, and carrying out enzymolysis for 6 hours at the temperature of 56 ℃; after enzymolysis by plant hydrolase, adjusting the pH value to 8.0, adding pancreatin which accounts for 60% of the total enzyme amount and alkaline protease of a commercial brand NS37071 which accounts for 6% of the mass of the dry leaves of the moringa oleifera, and carrying out enzymolysis for 8 hours at 56 ℃ under the conditions that the addition amount of the pancreatin accounts for 40% of the total enzyme amount and the addition amount of the alkaline protease of the commercial brand NS37071 accounts for 40% of the total enzyme amount; adding absolute ethyl alcohol to enable the content of the ethyl alcohol in the system to reach 50wt%, heating and refluxing for 2h at 100 ℃, cooling to room temperature, centrifuging for 20min at 6000g, taking supernate, concentrating under reduced pressure until the content of solid matters is 30%, freeze-drying, taking freeze-dried powder to prepare 200mg/mL aqueous solution, passing through a Mitsubishi chemical macroporous adsorption resin SP207 column, and performing 7-column volume gradient elution by adopting 0-50 vol% ethyl alcohol solution at the flow rate of 2mL/min, wherein the method specifically comprises the following steps: eluting 1-2 column volumes with 100% water and 0% ethanol, eluting 3-5 column volumes with 60% water and 40% ethanol, eluting 6-7 column volumes with 50% water and 50% ethanol, collecting 50% ethanol eluate, and freeze drying to obtain Moringa oleifera leaf extract; the extracted Moringa oleifera leaf extract contains 24% of polyphenols.
(7) Preparing the moringa oil: dissolving 0.2g of moringa oleifera leaf extract in 5mL of absolute ethyl alcohol, adding 1000g of moringa oleifera seed oil, and uniformly stirring to obtain moringa oleifera oil 3.
The prepared moringa oil 3 has the oleic acid content of 73.1 percent, the behenic acid content of 3.23 percent and high nutritional value, and the shelf life of the moringa oil under the conditions of room temperature and light shielding is estimated to be 78 months according to an Arrhenius empirical formula.
Comparative example 1
The preparation method of the moringa oil specifically comprises the following steps:
(1) microwave treatment: drying fresh moringa seeds, removing shells, performing microwave treatment at a power of 700W for 4min, pulverizing, sieving with a 100-mesh sieve, and pulverizing in a jet mill for 3h to obtain moringa seed powder with an average particle size of 8 μm;
(2) high-temperature cooking: steaming the obtained Moringa seed powder at 120 deg.C for 30 min;
(3) high-pressure homogenization: taking the moringa seed powder after high-temperature cooking, adding deionized water according to the feed-liquid ratio of 1:8g/mL, shearing at the speed of 10000rpm for 20 minutes, passing through a colloid mill for 3 times to obtain moringa seed homogenate, and homogenizing at 40MPa for 2 times;
(4) ultrasonic-assisted alkaline extraction: adding sodium hydroxide into the moringa seed powder homogenate to adjust the pH value to 9.0, extracting for 20min at the ultrasonic power of 800W and the temperature of 60 ℃, and cooling to room temperature to obtain a suspension;
(5) and (3) carrying out enzymolysis on moringa seeds: adjusting the pH value of the suspension to 5.0, adding alpha-amylase accounting for 80% of the total enzyme by mass and composite plant hydrolase of a commercial brand Viscozyme L accounting for 20% of the total enzyme by mass, and carrying out enzymolysis for 6 hours at 56 ℃; after the enzymolysis of plant hydrolase, adjusting the pH value to 8.0, adding composite protease with the mass of 6 percent of that of the moringa seeds and alkaline protease with the trade mark NS37071, wherein the addition amount of the composite protease accounts for 30 percent of the total enzyme amount, the addition amount of the alkaline protease with the trade mark NS37071 accounts for 70 percent of the total enzyme amount, and carrying out enzymolysis for 16 hours at 56 ℃; heating at 120 deg.C for 40min, cooling to room temperature, and freezing at-24 deg.C for 12 hr; standing at 8 ℃ for 12h for low-temperature thawing, and centrifuging at 8000g for 30min to obtain Moringa seed oil 4, wherein the extraction rate of the Moringa seed oil 4 is 93%;
(6) preparing the moringa oil: dissolving 0.2g of tert-butyl hydroquinone (TBHQ) in 5mL of absolute ethanol, adding into 1000g of moringa seed oil, and stirring to obtain moringa oil 4.
The prepared moringa oil 4 has the oleic acid content of 73.3 percent, the behenic acid content of 3.13 percent and high nutritional value, and the shelf life of the moringa oil under the conditions of room temperature and light shielding is estimated to be 68 months according to an Arrhenius empirical formula.
Comparative example 2
The preparation method of the moringa oil specifically comprises the following steps:
(1) microwave treatment: drying fresh moringa seeds, removing shells, heating for 4min at the microwave power of 700W, crushing, sieving with a 100-mesh sieve, and crushing in a jet mill for 3h to obtain moringa seed powder with the average particle size of 8 mu m;
(2) high-temperature cooking: steaming the obtained Moringa seed powder at 120 deg.C for 30 min;
(3) high-pressure homogenization: taking the moringa seed powder after high-temperature cooking, adding deionized water according to the feed-liquid ratio of 1:8g/mL, shearing at the speed of 10000rpm for 20 minutes, passing through a colloid mill for 3 times to obtain moringa seed homogenate, and homogenizing at 40MPa for 2 times;
(4) ultrasonic-assisted alkaline extraction: adding sodium hydroxide into the moringa seed powder homogenate to adjust the pH value to 9.0, extracting for 20min at the ultrasonic power of 800W and the temperature of 60 ℃, and cooling to room temperature to obtain a suspension;
(5) and (3) carrying out enzymolysis on moringa seeds: adjusting the pH value of the suspension to 5.0, adding alpha-amylase accounting for 80% of the total enzyme by mass and composite plant hydrolase of a commercial brand Viscozyme L accounting for 20% of the total enzyme by mass, and carrying out enzymolysis for 6 hours at 56 ℃; after the enzymolysis of plant hydrolase, adjusting the pH value to 8.0, adding composite protease with the mass of 6 percent of that of the moringa seeds and alkaline protease with the trade mark NS37071, wherein the addition amount of the composite protease accounts for 30 percent of the total enzyme amount, the addition amount of the alkaline protease with the trade mark NS37071 accounts for 70 percent of the total enzyme amount, and carrying out enzymolysis for 16 hours at 56 ℃; heating at 120 deg.C for 40min, cooling to room temperature, and freezing at-24 deg.C for 12 hr; standing at 8 deg.C for 12h for thawing at low temperature, and centrifuging at 8000g for 30min to obtain Moringa seed oil 5;
the prepared moringa seed oil 5 has the extraction rate of 93 percent, the oleic acid content of 72.5 percent and the behenic acid content of 3.21 percent, has high nutritional value, and the shelf life of the moringa seed oil under the conditions of room temperature and light shielding is estimated to be 14 months according to an Arrhenius empirical formula.
The total fatty acid composition of the moringa oils prepared in examples 1-3 and comparative examples 1-2 is shown in table 1.
TABLE 1 Moringa oil Total fatty acid composition
As can be seen from Table 1, the fatty acid compositions of the moringa oil prepared in examples 1-3 and comparative examples 1-2 are similar, the oleic acid content is the highest and exceeds 72%, the behenic acid content exceeds 3%, and the oil extraction rate is higher than 90%; in addition, the moringa oleifera seed oil in the embodiments 1-3 is added with the moringa oleifera leaf extract, while TBHQ is added in the comparative example 1, no antioxidant is added in the comparative example 2, and the moringa oleifera leaf extract can obviously improve the oxidation stability of the moringa oleifera seed oil and is superior to the oxidation resistance of TBHQ to the moringa oleifera seed oil by combining the difference of the quality guarantee periods of grease.
The method comprises the steps of adopting a pretreatment process of microwave treatment, high-temperature cooking, high-pressure homogenization and ultrasonic-assisted alkaline extraction, then combining plant hydrolase enzymolysis, protease enzymolysis, high-temperature treatment, low-temperature freezing, unfreezing and low-temperature high-speed centrifugation to obtain moringa seed oil, and adding a moringa leaf extract prepared by plant hydrolase enzymolysis, protease enzymolysis, ethanol extraction and macroporous resin enrichment into the moringa seed oil to prepare the moringa oil. The moringa seed oil prepared by adopting an efficient and green extraction technology has high extraction rate, and the obtained moringa oil has no organic solvent residue, no chemical additive, high nutritive value and long shelf life, and can be widely applied to the food industry.
The above examples are only preferred embodiments of the present invention, which are intended to be illustrative and not limiting, and those skilled in the art should understand that they can make various changes, substitutions and alterations without departing from the spirit and scope of the invention.
Claims (9)
1. A preparation method of moringa oil is characterized in that a pretreatment process of microwave treatment, high-temperature cooking, high-pressure homogenization and ultrasonic-assisted alkaline extraction is adopted, and a treatment process of plant hydrolase enzymolysis, protease enzymolysis, high-temperature treatment, low-temperature freezing, unfreezing and low-temperature high-speed centrifugation is combined to obtain moringa oil; finally, the moringa oil is prepared by adding the moringa leaf extract, and the method specifically comprises the following steps:
(1) microwave treatment: drying fresh moringa seeds, removing shells, performing microwave treatment, crushing, sieving and performing superfine crushing to obtain moringa seed powder;
(2) high-temperature cooking: steaming the obtained moringa seed powder at high temperature;
(3) high-pressure homogenization: taking moringa seed powder which is steamed at high temperature, adding deionized water, shearing at high speed, passing through a colloid mill, and homogenizing at high pressure to obtain moringa seed homogenate;
(4) ultrasonic-assisted alkaline extraction: adding sodium hydroxide into the moringa seed powder homogenate to adjust the pH value, performing ultrasonic-assisted extraction, and cooling to room temperature to obtain a suspension;
(5) and (3) carrying out enzymolysis on moringa seeds: adjusting the pH value of the suspension, adding plant hydrolase for enzymolysis, adjusting the pH value, adding protease for enzymolysis, performing high-temperature treatment, cooling to room temperature, performing low-temperature freezing treatment, performing low-temperature thawing, and performing low-temperature high-speed centrifugation to obtain moringa seed oil;
(6) preparing the moringa oil: dissolving the moringa oleifera leaf extract in absolute ethyl alcohol, adding the moringa oleifera leaf extract into moringa oleifera seed oil, and uniformly stirring to obtain the moringa oleifera oil; the moringa oleifera leaf extract is obtained by extracting the following steps:
taking moringa oleifera dry leaves, crushing and sieving to obtain moringa oleifera leaf powder, adding deionized water, mixing uniformly to obtain a suspension, adjusting the pH value of the suspension, adding plant hydrolase for enzymolysis, adjusting the pH value, adding protease for enzymolysis, adding absolute ethyl alcohol, heating and refluxing for extraction, cooling an extracting solution to room temperature, centrifuging to remove residues, taking a supernatant, concentrating under reduced pressure, freeze-drying, taking freeze-dried powder, dissolving in water, passing through a macroporous resin column, performing gradient elution by adopting an ethanol solution, collecting an eluent, and freeze-drying to obtain a moringa oleifera leaf extract; the feed-liquid ratio of the moringa oleifera leaf powder to the deionized water is 1: 8-10 g/mL; adjusting the pH value of the suspension to 4.4-5.0; the adding amount of the plant hydrolase is 2-4% of the weight of the dry moringa leaves; the plant hydrolase is a composite plant hydrolase of a commodity brand Viscozyme L; the plant hydrolase enzymolysis is carried out for 4-6 hours at 50-56 ℃; after plant hydrolase enzymolysis, adjusting the pH value to 7.0-8.0; the addition amount of the protease is 4-6% of the mass of the moringa leaves; the gradient elution is gradient elution by adopting 0-50 vol% ethanol solution, and specifically comprises the following steps: eluting with 100vol% water for 1-2 column volumes, eluting with 100-60 vol% water and 0-40 vol% ethanol for 3-5 column volumes, eluting with 50 vol% water and 50 vol% ethanol for 6-7 column volumes, and collecting 50 vol% ethanol eluate.
2. The method for preparing Moringa oleifera oil according to claim 1, wherein in step (1), the microwave treatment power is heating for 2-4min under the microwave condition of 500-700W; the sieving is to sieve through a 100-mesh sieve; the ultra-micro crushing is to crush the moringa oleifera seeds for 2 to 3 hours in a jet mill, and the average particle size of the crushed moringa oleifera seeds is 8 to 12 mu m; in the step (2), the high-temperature cooking is performed for 20-30min under the steam condition of 100-120 ℃.
3. The preparation method of moringa oil according to claim 1, wherein in the step (3), the feed-liquid ratio of the moringa seed powder subjected to high-temperature cooking to the deionized water is 1: 6-8 g/mL; the high-speed shearing is carried out at the speed of 8000-10000 rpm for 10-20 minutes; the number of times of passing through the colloid mill is 2-3; the pressure of the high-pressure homogenization is 20-40 MPa, and the number of times of the high-pressure homogenization is 2-3; in the step (4), the pH value is adjusted to be 7.0-9.0; the ultrasonic power of the ultrasonic-assisted extraction is 600-.
4. The method for preparing moringa oil according to claim 1, wherein in the step (5), the pH value of the suspension is adjusted to 4.4-5.0; the plant hydrolase enzymolysis is carried out for 4-6 hours at 50-56 ℃; after plant hydrolase enzymolysis, adjusting the pH value to 7.0-8.0; the protease enzymolysis is carried out for 12-16 hours at 50-56 ℃; the high-temperature treatment is heat treatment at the temperature of 100-120 ℃ for 20-40 min; the low-temperature freezing treatment is freezing at the temperature of-18 to-24 ℃ for 8 to 12 hours; the low-temperature unfreezing is carried out at 4-8 ℃ and the mixture is placed for 8-12 h; the low-temperature centrifugation is carried out for 20-30min at 6000-8000 g under the condition of 4-8 ℃; the extraction yield of the moringa seed oil is more than 90 percent.
5. The preparation method of moringa oil according to claim 1, wherein in the step (5), the addition amount of the plant hydrolase is 2-4% of the mass of the moringa seeds; the plant hydrolase is a composite plant hydrolase of alpha-amylase and a commodity brand Viscozyme L, the addition amount of the alpha-amylase accounts for 60-80% of the total enzyme amount, and the addition amount of the composite plant hydrolase of the commodity brand Viscozyme L accounts for 20-40% of the total enzyme amount; the adding amount of the protease is 4-6% of the mass of the moringa seeds; the protease is compound protease and alkaline protease of a commercial brand NS37071, the adding amount of the compound protease accounts for 10-30% of the total enzyme amount, and the adding amount of the alkaline protease of the commercial brand NS37071 accounts for 70-90% of the total enzyme amount.
6. The method for preparing moringa oil according to claim 1, wherein in the step (6), the screening is performed by a 40-mesh screen; the protease is pancreatin and alkaline protease with a commercial brand NS37071, the adding amount of the pancreatin accounts for 40-60% of the total enzyme amount, and the adding amount of the alkaline protease with the commercial brand NS37071 accounts for 40-60% of the total enzyme amount; the enzymolysis of the protease is carried out for 6-8 hours at 50-56 ℃.
7. The preparation method of moringa oil according to claim 1, wherein in the step (6), the absolute ethyl alcohol is added in an amount which enables the content of the ethyl alcohol in the system to reach 30-50 wt%; the heating reflux is carried out for 1-2h at the temperature of 90-100 ℃; centrifuging at 4000-6000 g for 10-20 min; concentrating the mixture under reduced pressure until the content of solid matters is 20-30%; the concentration of the freeze-dried powder after being dissolved in water is 100-200 mg/mL; the macroporous resin is Mitsubishi chemical macroporous adsorption resin SP 207.
8. The method for preparing moringa oil according to claim 1, wherein in the step (6), the dissolving ratio of the moringa leaf extract to the absolute ethyl alcohol is 0.1-0.2:3-5 g/mL; the mass ratio of the moringa leaf extract to the moringa seed oil is 1-2: 10000.
9. A moringa oil obtained by the method of any one of claims 1-8, wherein the oleic acid content is >72% and the behenic acid content is > 3%.
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