CN112675084A

CN112675084A - Moringa oleifera extract for promoting collagen secretion and application thereof

Info

Publication number: CN112675084A
Application number: CN202011535487.5A
Authority: CN
Inventors: 熊莉娟; 黄友玉; 王小付; 向通
Original assignee: Kunming Yanzhenqing Biotechnology Co ltd
Current assignee: Honghe Wanniantang Biotechnology Co ltd
Priority date: 2020-12-23
Filing date: 2020-12-23
Publication date: 2021-04-20
Anticipated expiration: 2040-12-23
Also published as: CN112675084B

Abstract

A moringa extract for promoting collagen secretion and application thereof belong to the technical field of plant extracts and application thereof. Pulverizing Moringa oleifera raw material, adding deionized water, adding at least one protease and at least one cellulase, adding an organic solvent, performing reflux extraction and water enzymolysis under stirring, separating an upper organic phase under the condition that the vacuum degree is not lower than 0.04MP, and performing sealed anaerobic fermentation on the rest material system. The fermentation strain is composed of at least one kind of Saccharomyces cerevisiae and at least one kind of organic acid producing strain. Then carrying out solid-liquid separation of the system, and carrying out ultrafiltration membrane filtration on the liquid phase to obtain an extracting solution. The obtained moringa oleifera extract improves the content of necessary components for generating collagen, has good antioxidant activity and has the condition of promoting collagen secretion. Experiments show that the product of the invention can obviously promote the secretion of collagen. The application is as raw material for preparing cosmetics, health products, medicines or feeds.

Description

Moringa oleifera extract for promoting collagen secretion and application thereof

Technical Field

The invention belongs to the technical field of plant extracts and application thereof, and particularly relates to a moringa oleifera extract for promoting collagen secretion and application thereof.

Technical Field

Collagen, also known as Collagen, known by the english collegen name, is a protein with the highest human body content, accounting for about 25% to 33% of the total human body protein, and is distributed throughout various tissues and organs of the whole body, such as skin, tendons, bones, ligaments, cornea, cartilage, bone tissues and the like. The collagen molecule is formed into a three-strand helical structure by mutual winding of three peptide chains, which is called as collagen triple helix, and exists in a collagen fiber form in vivo through multi-stage polymerization to form a reticular structure, so that the skin, tendon, bone, cartilage, bone tissue and the like have mechanical strength. With the increase of people's age, collagen will gradually run off, the structure of collagen fiber will also change, covalent cross-linking more and more, collagen fiber will become more and more brittle and hard, cause tendon, ligament, cartilage mechanical strength to change. For example, when the skin contains 70% collagen, especially the dermis, and the collagen in the dermis is oxidized and broken, the supporting effect on the epidermis is lost, and wrinkles are generated. For example, in bone, collagen accounts for 80% of the bone protein, and loss of collagen can lead to bone fragility and, in severe cases, osteoporosis. For example, articular cartilage mainly comprises collagen, proteoglycan, chondrocytes and water, the collagen forms a net structure, and other components such as proteoglycan and the like are filled in the articular cartilage, so that the articular wear is avoided; when collagen is lost, the net structure formed by the collagen is damaged, the loss of the content is accelerated, the joint is accelerated to be worn under stress, and arthritis appears in severe cases. For example, the major internal organs and tissues of the human body contain collagen, which plays a role in protecting and strengthening the internal organs and preventing prolapse. In addition, the collagen can also improve immunity and prevent degeneration such as intervertebral disc protrusion, blood vessel elasticity deterioration, etc. However, such important collagen is lost year by year, and the secretion level of collagen itself begins to decrease with age, causing various problems to be gradually presented to the human body. It has been sought to effectively supplement or promote collagen production. At present, animal collagen is basically used as a raw material to be decomposed into amino acid or small molecular collagen peptide for eating or being added into cosmetics, but the components are difficult to be combined into collagen after entering the digestive tract or the skin and cannot be realized necessarily. The exogenous collagen is directly supplemented to regenerate endogenous collagen, so that the conversion rate is extremely low and the effect is not obvious. In order to increase collagen, various ways to promote the self-secretion of collagen have been sought.

Moringa oleifera (Moringa oleifera Lam) is a plant of Moringa genus of Moringaceae family, belonging to small arbor plant, native to India, and distributed in dry and hot valley of tropical and subtropical regions. The moringa oleifera is rich in nutrition and rich in protein, mineral substances, vitamins and other components. The moringa oleifera contains 17 amino acids, and is a natural plant which can provide the highest amino acid content and the most complete variety at present. Meanwhile, the content of natural vitamin C in the moringa oleifera is very high and is far higher than that of fruits rich in vitamins such as oranges and the like. Although the article "protective effect of moringa leaf polyphenol extract on skin of photoaged mice" (food and fermentation industry, 2020, 46(18), 67-71) reports that moringa leaf polyphenol extract can increase the content of collagen in skin, it is clearly indicated that the component having this function is polyphenol extract, and does not relate to a technique for promoting collagen secretion by other components than polyphenol in moringa leaf extract. In chinese patent application No. 201780018771.5: 【003】 It is described that "moringa extract alone shows no activity or even a reduced activity in stimulating collagen synthesis in skin cells"; 【0065】 It is stated that "the moringa extract itself causes a reduction in collagen synthesis in the fibroblasts to a large extent".

Disclosure of Invention

The invention aims to provide a moringa oleifera extract for promoting collagen secretion and application thereof.

The invention is characterized in that the moringa extract for promoting collagen secretion of the invention product is prepared by the following steps:

1. crushing the moringa oleifera raw material to obtain the moringa oleifera material. The moringa raw material can be seeds, fruit pods, flowers, leaves and the like of the moringa, and preferably is a moringa fresh fruit pod, a moringa fresh leaf or a moringa seed in the same year.

2. Adding deionized water into the moringa oleifera material obtained in the previous step, wherein the mass ratio of the deionized water to the moringa oleifera material is (2-10): 1.

3. and (2) weighing the amount of each enzyme according to 0.003-0.005 of the mass of the moringa oleifera material obtained in the step 1, wherein the enzyme activity value of each enzyme is 5-20 ten thousand units. Mixing the enzymes, adding deionized water, and mixing to obtain mixed enzyme water solution. The mass ratio of the deionized water to the total amount of the enzyme used for enzymolysis is 5-20: 1. the enzymes used for enzymatic hydrolysis must include at least one protease and at least one cellulase. The protease is preferably bromelain and papain.

4. And (3) dropwise adding the mixed enzyme water solution obtained in the step (3) into the material obtained in the step (2), preferably finishing dropwise adding within 8-10 minutes.

5. Adding an organic solvent into the substance obtained in the step 4, wherein the volume mass ratio of the organic solvent to the moringa oleifera material in the substance obtained in the step 4 is 4-8L: 1 KG. The organic solvent can be petroleum ether, n-hexane, ethyl acetate, dichloromethane, etc., preferably petroleum ether.

6. And (3) performing reflux extraction and water enzymolysis on the product obtained in the step (5) under the stirring condition, and stirring the water for enzymolysis for more than 3.5 hours, preferably 4 hours at constant temperature when the temperature of the system is increased to 52-58 ℃, preferably 55 ℃. The stirring conditions are preferably 58 to 62 revolutions per minute.

7. And (3) separating an upper organic phase in the product obtained in the step (6) under the condition that the vacuum degree is not lower than 0.04MP after the enzymolysis is finished, wherein the organic phase is used as the organic phase, and the rest material system is moringa oleifera material fermentation base liquid.

8. And (4) carrying out sealed anaerobic fermentation on the moringa oleifera material fermentation base liquid obtained in the step 7. The fermentation strain consists of at least one kind of saccharomyces cerevisiae and at least one kind of strain capable of producing organic acid, preferably saccharomycete with envelope covering and lactobacillus plantarum, the addition amount of each strain is 0.05-0.4% of the mass of the moringa oleifera material in the step 1, the fermentation temperature is 28-35 ℃, when the pH value is reduced to 3.5-4.5, the fermentation is stopped, and the fermentation time is preferably 7-15 days. Then, solid-liquid separation of the system was carried out. And collecting the liquid phase, discarding filter residues, and filtering the liquid phase by using an ultrafiltration membrane, wherein the filter pores are 0.28-0.32 mu m, so as to obtain a finished product of the extracting solution.

The detection shows that the obtained extract contains various amino acids, monosaccharide, polysaccharide, vitamin C, etc. Wherein the amino acids include 17 hydrolyzed amino acids and free amino acids such as aspartic acid, glutamic acid, cystine, serine, histidine, glycine, arginine, threonine, alanine, tyrosine, proline, valine, methionine, isoleucine, leucine, phenylalanine, lysine, etc. The monosaccharide includes mannose, ribose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose, xylose, arabinose, and fucose.

Experiments show that the moringa extract has strong antioxidation.

Experiments show that the moringa extract can obviously promote the secretion of collagen. The extract is diluted to 1%, and has DHfa collagen secretion promoting activity, and can increase collagen amount by 13%.

The specific detection conditions of the moringa oleifera pod extract finished product obtained in the embodiment 1 of the invention are as follows:

table I, various amino acids and vitamin C contents in the extract

Name (R)	Test results
		Aspartic acid	0.019％
Threonine	0.011％
		Serine	0.0080％
Glutamic acid	0.023％
		Proline	0.043％
Glycine	0.016％
		Alanine	0.0047％
Cystine	0.0036％
		Valine	0.0097％
Methionine	0.0023％
		Isoleucine	0.022％
Leucine	0.027％
		Tyrosine	0.016％
Phenylalanine	0.016％
		Lysine	0.018％
Histidine	0.0097％
		Arginine	0.068％
Vitamin C	17.0mg/100g

The first table is a related quality detection report form of a finished product quality supervision and inspection research institute in Yunnan province. The quality report sheet shows that a large amount of various amino acids, vitamin C and the like are enriched in the extracting solution, wherein the content of glycine and proline which are necessary components for synthesizing the collagen is obvious.

TABLE II, sample in vitro antioxidation

Sample (I)	Concentration of	Oxidation resistance (%)
			Trolox	25μg/mL	96.315±0.061
Extract liquid	10％	92.282±0.447

The second table shows the data of the natural drug activity screening center of Kunming plant research institute of Chinese academy of sciences, and the experimental results show that the extracting solution has strong DPPH free radical scavenging activity. Even when diluted to 10% concentration, 92.282 + -0.447 (%) clearance was achieved.

Experiment and result of collagen secretion promotion by Kunming plant institute of Chinese academy of sciences

The batch of experimental samples, the reagent summary and the experimental method are as follows:

TABLE III, summary of experimental samples/reagents

Table three is the relevant information for the test samples and reagents used in the batch. The specific experimental method is as follows:

on a 12-well cell culture plate, HDFa cells were mixed with 1% of the sample solution to be tested, and a blank control containing no drug and an HGF- β positive control were set. Culturing at 37 deg.C with 5% CO2 for three days, collecting cell culture supernatant, and storing at-80 deg.C; adding MTS, and detecting the OD value of 490nm by adopting an MTS colorimetric method; and (3) detecting collagen secretion according to a method provided in the collagen ELISA kit, diluting the supernatant by 1:100 times, and measuring an OD value by an enzyme-labeling instrument, wherein the detection wavelength is 450 nm. Calculating to obtain the collagen secretion increase value.

Cell viability (%) ═ experimental well OD490 nm/blank well OD490 nm% 100%

Collagen secretion increase (%) - (experimental well OD450 nm/cell viability/blank well OD450nm-1) × 100%

According to the above detection method, the experimental data are as follows:

TABLE IV sample promotion and cytotoxic effects on HDfa collagen secretion

The experimental result shows that the moringa extract has the activity of promoting the secretion of HDfa collagen. Even when diluted to 1% concentration, the collagen secretion increase of 13.531 + -6.515 (%) was achieved.

Therefore, the single extract has the effect of remarkably promoting the secretion of the collagen, the extracting solution not only provides necessary basic components for synthesizing the collagen, but also has the activity of promoting the secretion of the collagen due to rich nutrient components, and the cytotoxicity experiment shows that the extracting solution is nontoxic, can be used for external application of skin and can be used as health-care food.

The invention adopts a specific water enzymolysis two-phase extraction method, and the obtained moringa extract has the function of promoting collagen secretion. In other words, the extraction process removes substances such as isothiocyanate, thiocarbamate, moringa alkaloid and the like through the processes of water enzymolysis, two-phase extraction and post-fermentation, not only improves the content of necessary components for generating collagen, but also has better antioxidant activity and conditions for promoting collagen secretion.

According to the collagen secretion promoting and antioxidant properties of the extract product, the extract product can be widely applied to the fields of cosmetics, health care products, medicines and the like, can be applied to the skin care fields of facial masks, facial cleansers, face creams, hand creams, eye creams, repair water, body milk, makeup removing water, makeup removing wet tissues and the like in the cosmetics, and can be applied to calcium supplementing, anti-aging and beautifying foods or beverages and the like in the health care product field.

The moringa oleifera extract for promoting collagen secretion is used as a raw material for preparing cosmetics, health products, medicines or feeds.

The invention has the beneficial effects that: the moringa extract has the effect of promoting collagen secretion, and can be widely used as a raw material for preparing cosmetics, health products and medicines.

Detailed Description

Example 1. The method comprises the following steps of preparing a collagen-promoting extracting solution from fresh moringa oleifera pods.

1.5 Kg of organic fresh moringa oleifera pods of the red river valley moringa oleifera industrial company are selected and crushed to obtain moringa oleifera pod pulp.

2. Adding 20Kg of deionized water into the moringa oleifera pod pulp obtained in the previous step, wherein the mass ratio of the deionized water to the moringa oleifera pod pulp is 4: 1.

3. adding various enzymes used for enzymolysis according to the weight of 0.003 proportion of the moringa fruit pod pulp obtained in the step 1, specifically 15g of pectinase, 15g of cellulase, 15g of neutral protease, 15g of bromelain and 15g of papain with the enzyme activity value of 20 ten thousand units, and then adding 750g of deionized water to fully mix to obtain a mixed enzyme aqueous solution. The mass ratio of the deionized water to the enzyme used for enzymolysis is 10: 1.

4. and (4) dropwise adding the mixed enzyme aqueous solution obtained in the step (3) into the material obtained in the step (2), and finishing dropwise adding within 10 minutes.

5. 20L of petroleum ether were added to the product obtained in step 4. The volume mass ratio of the petroleum ether to 5Kg of moringa oleifera pod pulp contained in the product obtained in the step 4 is 4L: 1 Kg.

6. And (3) carrying out reflux extraction and water enzymolysis on the product obtained in the step (5) under the condition of stirring at 62 revolutions per minute, and stirring water at constant temperature for enzymolysis for 4 hours when the temperature of the system is raised to 55 ℃.

7. And (3) after the enzymolysis is finished, separating a petroleum ether phase under the condition that the vacuum degree of a vacuum storage tank is 0.08MP, wherein the petroleum ether phase is used as the petroleum ether phase, and the rest material is the moringa oleifera material fermentation base liquid.

8. And (3) carrying out sealed anaerobic fermentation on the moringa oleifera material fermentation base liquid obtained in the step (7), wherein fermentation strains are 10g of saccharomycete with coated membranes and 20g of lactobacillus plantarum (the addition amounts of the fermentation strains are 0.2% and 0.4% of the mass of the moringa oleifera pod pulp obtained in the step (1)). The fermentation temperature was 32 ℃ and when the pH was reduced to 3.6, the fermentation was stopped for about 8 days. Then solid-liquid separation and filtration are carried out. Filtering with ultrafiltration membrane with filter pores of 0.3 μm to obtain 18Kg of extractive solution.

Example 2. The method comprises the following steps of preparing a collagen-promoting extract from fresh moringa leaves.

1.5 Kg of organic fresh moringa leaves of the moringa industry company of the red river valley are selected and crushed to obtain moringa leaf pulp.

2. Adding 40Kg of deionized water into the moringa leaf pulp obtained in the previous step, wherein the mass ratio of the deionized water to the moringa fruit pod pulp is 8: 1.

3. Adding various enzymes used for enzymolysis according to the weight of 0.005 proportion of the moringa leaf pulp obtained in the step 1, specifically 25g of pectinase, 25g of cellulase, 25g of hemicellulase, 25g of neutral protease and 25g of papain with the enzyme activity value of 10 ten thousand units, and adding 750g of deionized water for fully mixing to obtain a mixed enzyme aqueous solution. The mass ratio of the deionized water to the enzyme used for enzymolysis is 6: 1.

4. and (4) dropwise adding the mixed enzyme aqueous solution obtained in the step (3) into the material obtained in the step (2), and finishing dropwise adding within 8 minutes.

5. 20L of n-hexane was added to the product obtained in step 4. The volume mass ratio of the normal hexane to 5Kg of moringa oleifera leaf pulp contained in the product obtained in the step 4 is 4L: 1 Kg.

7. And (3) after the enzymolysis is finished, separating a normal hexane phase under the condition that the vacuum degree of a vacuum storage tank is 0.08MP, wherein the normal hexane phase is used as the normal hexane phase, and the rest material is the moringa oleifera material fermentation base liquid.

8. And (3) adding fermentation strains into the material from which the normal hexane phase is removed in the step (7) for anaerobic fermentation, wherein the fermentation strains comprise 10g of sacculus-covering saccharomycete, 10g of pichia pastoris, 10g of bifidobacterium and 20g of lactobacillus plantarum. The fermentation temperature was 32 ℃ and when the pH was lowered to 4.1, the fermentation was stopped for about 10 days. Then solid-liquid separation and filtration are carried out. The filtration is carried out by an ultrafiltration membrane with a filter hole of 0.3 mu m to obtain 35Kg of extracting solution. The cell experiment is repeated after the extracting solution is concentrated to 18Kg, and the experimental result shows that the moringa oleifera leaf extracting solution obtained by the process still has the effect of remarkably promoting the secretion of collagen.

Example 3. The method comprises the following steps of preparing collagen-promoting extract from moringa seeds.

1.5 Kg of organic moringa seed kernels of the same year of the red river grain moringa industry company is selected and crushed to about 10 meshes.

2. Adding 20Kg of deionized water into the moringa seed kernel powder obtained in the previous step, wherein the mass ratio of the deionized water to the moringa seed kernel powder is 4: 1.

3. adding various enzymes used for enzymolysis according to the weight of 0.005 proportion of the moringa leaf pulp obtained in the step 1, specifically 25g of pectinase, 25g of cellulase, 25g of hemicellulase, 25g of neutral protease and 25g of papain with the enzyme activity value of 20 ten thousand units, and then adding 750g of deionized water for fully mixing to obtain a mixed enzyme aqueous solution. The mass ratio of the deionized water to the enzyme used for enzymolysis is 6: 1.

5. 20L of ethyl acetate were added to the product obtained in step 4. The volume mass ratio of the ethyl acetate to 5Kg of moringa seed powder contained in the product obtained in the step 4 is 4L: 1 Kg.

7. And after the enzymolysis is finished, separating an ethyl acetate phase under the condition that the vacuum degree of a vacuum storage tank is 0.08MP, wherein the ethyl acetate phase is used as the ethyl acetate phase, and the rest material is the moringa oleifera material fermentation base liquid.

8. And (3) adding fermentation strains into the material from which the ethyl acetate phase is removed in the step (7) for anaerobic fermentation, wherein the fermentation strains comprise 10g of sacculus-covering saccharomycetes, 10g of pichia pastoris, 10g of bifidobacterium, 10g of acetobacter and 20g of lactobacillus plantarum. The fermentation temperature was 32 ℃ and when the pH was reduced to 3.8, the fermentation was stopped for about 11 days. Then solid-liquid separation and filtration are carried out. The filtration is carried out by an ultrafiltration membrane with a filter hole of 0.3 mu m to obtain 18Kg of extracting solution. The cell experiment of the first embodiment is repeated on the extracting solution, and the experimental result shows that the moringa seed kernel extracting solution obtained by the process still has the obvious function of promoting the collagen secretion.

Example 4. And (3) performing a control experiment on feeding of the moringa oleifera extract to livestock and poultry.

Preparing a control feed: common feed: mixing with 35% of bran, 45% of corn flour and 20% of water. The moringa feed comprises the following steps: mixing with 35% of bran, 45% of corn flour, 17% of water, and 3% of Moringa oleifera extract obtained in example 1.

Control experiment of chicken: selecting free-range chicken of the red river valley moringa company as a control: selecting 40 iron-rod partridge chickens of the same variety and the same batch, randomly dividing into two groups, feeding 20 iron-rod partridge chickens in each group for 45 days in independent cages at the same time and in the same amount, randomly sampling 6 iron-rod partridge chickens in each group, and observing the appearance after slaughtering: the moringa oleifera group chicken skin is thick and elastic, the tendon is firm, and the bone is hard, dense and deep. And is independent of individual weight. The experimental result adopts fluorescence spectrum to quantitatively detect the collagen protein, the moringa oleifera group is obviously higher than that of a common feeding group, and specific detection data are shown in the following table.

Table five, data processing of detection results of chicken half-leg collagen fluorescence spectrometers of control group and moringa group

The data in table five show that the feed formulation group using the moringa extract of example 1 significantly increased the collagen content of the chicken.

Control experiment in pigs: selecting pigs of Chili Korea for comparison experiment: selecting 8 commercial pigs of 10 months old in the same nest, randomly dividing into two groups, wherein each group comprises 4 pigs, breeding in groups, feeding the two feeds for 60 days at the same time and the same amount every day, and slaughtering completely, wherein the appearance after slaughtering is more visible: the pigskin of the moringa group is obviously thicker and elastic, the connective tissue connecting the joint head and the joint pit is obviously thicker, and the tendon is firm. And independent of individual weight, the skin of the same part of each pig is unhaired, degreased, washed, cut into fragments and homogenized, and then collagen protein is quantitatively detected by adopting fluorescence spectrum, and the specific detection result is shown in the following table:

sixth, data processing of detection results of pigskin collagen fluorescence spectrometer of control group and moringa group

The data in table six show that the feed formula group using the moringa extract of the example has obvious addition effect on the generation of collagen substances in pigskin.

The specific detection method comprises the following steps: taking the peeled chicken half-leg meat (pigskin) of the same part of each chicken (pig), degreasing, washing with water, and cutting into fragments and homogenizing; 10g of homogenate was weighed, 300ml of 0.5mol/L acetic acid and 0.1g of pepsin (activity value 1:10000) were added thereto, and after 48 hours at 4 ℃, centrifugation was carried out at 10000rpm for 15 minutes. The supernatant was salted out with NaCl to a concentration of 3 mol/L. The precipitated sample was dissolved again in 0.5mol/L acetic acid solution, and salting out was performed with 0.7mol/L NaCl. The precipitate was redissolved in 0.5mol/L acetic acid and dialyzed against 0.1mol/L acetic acid solution for 3 d. Finally, the samples were frozen for 48h using a freeze vacuum dryer (-55 ℃) and stored at 4 ℃ until use.

Dissolving the prepared collagen in 0.1mol/L acetic acid water solution to obtain 0.2-0.6mg/mL collagen solution, and standing at 4 deg.C for 24 hr; 8-Anilinonaphthalene-1-sulfonic Acid (ANS) was dissolved in 0.1mol/L phosphate buffer (pH7.0) to give an ANS solution having a concentration of 8 mmol/L. 2mL of collagen solution was mixed with 20. mu.L of ANS solution, and after standing for 10min, the fluorescence spectrum at 20 ℃ was recorded with a fluorescence spectrophotometer. The excitation wavelength is 380nm, and the emission wavelength is 400-650 nm. The slit widths for both excitation and emission wavelengths were 5 nm.

Example 5. A preparation method of anti-aging health promotion oral liquid for promoting collagen secretion is provided.

The composition consists of the following components: selecting 40-95% of the moringa oleifera stock solution extracted in the example 1 as a main material, and selecting the rest acceptable auxiliary materials from one or a mixture of more than two of fruit powder, edible essence, a sweetening agent, a filling agent, a lubricating agent, a preservative, a suspending agent, an edible pigment, a diluting agent, an emulsifying agent, a disintegrating agent or a plasticizer. Preferably, the fruit powder is one or more of orange powder, lemon powder, cherry powder, apple powder and coconut powder. However, those skilled in the art will recognize that the fruit powder is within the scope of the present invention, and the type of fruit powder is not limited thereto, and the present invention is not limited thereto; preferably, the edible essence is one or a mixture of more than two of orange essence, lemon essence, cherry essence, menthol, apple essence or coconut essence. However, those skilled in the art will recognize that the possible flavors are within the scope of the present invention, and the types of flavors are not limited thereto, and the present invention is not limited thereto; to enhance the mouthfeel of the composition, sweeteners may also be added to increase sweetness. Preferably, the sweetener is one or more of sucralose, AK sugar, aspartame or mogroside. However, those skilled in the art will recognize that any suitable sweetener is within the scope of the present invention, and that the type of sweetener is not limited thereto, and the present invention is not limited thereto.

Example 6. A preparation method of oral liquid for promoting collagen secretion and supplementing calcium and locking calcium is provided.

The method comprises the steps of crushing organic moringa oleifera new leaves of the moringa oleifera industrial company to pulp, weighing 5Kg, transferring the pulp into a vessel containing 20Kg of deionized water, accurately weighing 15g of pectinase, 15g of cellulase, 15g of neutral protease, 15g of bromelain and 15g of papain, mixing the weighed enzymes, fully mixing the enzyme with 750g of deionized water to obtain a mixed enzyme aqueous solution, dropwise adding the mixed enzyme aqueous solution into the vessel containing the moringa oleifera pulp, injecting 20L of petroleum ether solvent, performing reflux extraction and water enzymolysis under the stirring condition, and stirring the mixture at constant temperature for enzymolysis for 4 hours at constant time when the temperature of a system rises to 55 ℃. After the water enzymolysis process is finished, separating a petroleum ether phase for other purposes, continuously adding 20g of capsule-covering membrane-covering saccharomycete and 40g of lactobacillus plantarum into the rest system, transferring the mixture into a closed container for anaerobic fermentation after the addition is finished, controlling the fermentation end point to be PH 3.6, controlling the fermentation period to be T7-10D and the fermentation temperature to be T28-35 ℃, performing solid-liquid separation on the system after the fermentation period is finished, collecting a water phase, discarding filter residues, and filtering the separated water phase by using an ultrafiltration membrane to obtain 15Kg of a finished moringa oleifera new leaf extract.

40-95% of the original liquid of Moringa oleifera is used as main material, 0.1-3% of calcium source (preferably calcium citrate) can be added, and the rest acceptable adjuvants are selected from one or more of fruit powder, edible essence, sweetener, bulking agent, lubricant, antiseptic, suspending agent, edible pigment, diluent, emulsifier, disintegrating agent or plasticizer. Preferably, the fruit powder is one or more of orange powder, lemon powder, cherry powder, apple powder and coconut powder. However, those skilled in the art will recognize that the fruit powder is within the scope of the present invention, and the type of fruit powder is not limited thereto, and the present invention is not limited thereto; preferably, the edible essence is one or a mixture of more than two of orange essence, lemon essence, cherry essence, menthol, apple essence or coconut essence. However, those skilled in the art will recognize that the possible flavors are within the scope of the present invention, and the types of flavors are not limited thereto, and the present invention is not limited thereto; to enhance the mouthfeel of the composition, sweeteners may also be added to increase sweetness. Preferably, the sweetener is one or more of sucralose, AK sugar, aspartame or mogroside. However, those skilled in the art will recognize that any suitable sweetener is within the scope of the present invention, and that the type of sweetener is not limited thereto, and the present invention is not limited thereto.

Although moringa leaves are mentioned in many patents and publications as being useful for calcium supplement, they are proposed in the form of calcium chelate, in which the calcium supplement mechanism is different, and a calcium source can be additionally added, so that the calcium content can be controlled and the shortage or overhigh calcium content can be avoided.

Example 7. A preparation method of Moringa oleifera facial mask essence for promoting collagen secretion is provided.

The mask essence for promoting collagen secretion by moringa oleifera consists of the following components: 15 parts of moringa product raw material, 0.3 part of propylene glycol, 0.6 part of trehalose, 0.5 part of pantothenic acid, 0.2 part of allantoin, 0.2 part of PVM/MA decadiene cross-linked polymer, 0.4 part of xanthan gum, 0.3 part of hydroxyethyl cellulose, 0.3 part of disodium EDTA, 0.05 part of triethanolamine, 0.2 part of sodium hyaluronate, 0.01 part of methyl hydroxybenzoate, 0.01 part of propyl hydroxybenzoate, 0.4 part of PEG-60 anthracene sesame oil, 0.3 part of caprylyl hydroximic acid, 0.02 part of p-hydroxyacetophenone, 0.02 part of rose essence and 100 parts of deionized water in the first embodiment are selected. The facial mask has antioxidant and collagen secretion promoting effects, and has effects of keeping moisture, whitening skin, removing speckle, and resisting wrinkle.

Preparation method

2/3 deionized water is added into a main pot, water bath is started to stir and heat to 85 ℃, 0.01 part of p-hydroxymethyl, 0.01 part of p-hydroxypropyl and 0.02 part of p-hydroxyphenyl acetone are added, the mixture is continuously heated to be completely dissolved, the mixture is cooled to room temperature, then 0.3 part of propylene glycol, 0.6 part of trehalose, 0.5 part of pantothenic acid, 0.2 part of allantoin, 0.2 part of PVM/MA decadiene cross-linked polymer, 0.4 part of xanthan gum, 0.3 part of hydroxyethyl cellulose, 0.3 part of EDTA disodium, 0.05 part of triethanolamine, 0.2 part of sodium hyaluronate, 0.01 part of methyl hydroxybenzoate, 0.01 part of propyl hydroxybenzoate, 0.4 part of PEG-60 anthracene sesame oil, 0.3 part of caprylyl hydroximic acid, 0.02 part of p-hydroxyacetophenone, 0.02 part of rose essence and 1/3 deionized water are respectively added, the mixture is started to stir for 30 minutes, and the mixture is fully and uniformly mixed.

Adjusting pH of Moringa oleifera product raw material to 6.40 with 0.01mol/L sodium hydroxide, adding into main pot, heating and stirring, heating when temperature reaches 85 deg.C, and stirring to cool to room temperature. The obtained product is the moringa oleifera mask essence for promoting collagen secretion.

Example 8. A preparation method of Moringa oleifera repair water is provided.

A preparation method of moringa oleifera repair water comprises the following components: 50-60 parts of deionized water, 30-40 parts of moringa product raw materials, 0.5-1 part of butanediol, 261-2 parts of glyceryl polyether, 5-8 parts of trehalose, 5-8 parts of pantothenic acid, 1-3 parts of allantoin, 10-15 parts of a secondary cracking yeast fermentation product filtrate, 2-5 parts of PEG/PPG-17/6 copolymer, 2-5 parts of beta-glucan, 0.5-1 part of EDTA disodium, 0.5-1 part of sodium hyaluronate, 2-5 parts of nicotinamide, 5-10 parts of yeast amino acids, 2-5 parts of PEG-60 hydrogenated anthracene sesame oil, 0.03-0.05 part of phenoxyethanol, 0.5-1 part of propylene glycol, 0.5-1 part of essence, 0.01-0.03 part of methyl hydroxybenzoate and 0.01-0.03 part of propyl hydroxybenzoate. The repairing water can achieve the effects of moistening skin, caring skin, and making skin tender and elastic.

Preparation method

And (3) stirring and heating 40 parts of prepared deionized water in a water bath kettle to 80-90 ℃, adding 0.03 part of p-hydroxymethyl and 0.01 part of p-hydroxypropyl, and continuously heating until the p-hydroxymethyl and the p-hydroxypropyl are completely dissolved. Sequentially adding 1-2 parts of oil, 0.5-1 part of butanediol, 261-2 parts of glycerol polyether, 5-8 parts of trehalose, 5-8 parts of pantothenic acid, 1-3 parts of allantoin, 1-3 parts of glycerol polymethacrylate, 10-15 parts of a secondary fission yeast fermentation product filtrate, 2-5 parts of PEG/PPG-17/6 copolymer, 2-5 parts of beta-glucan, 0.5-1 part of EDTA disodium, 0.5-1 part of sodium hyaluronate, 2-5 parts of nicotinamide, 5-10 parts of yeast amino acids, 2-5 parts of PEG-60 hydrogenated anthracene sesame oil, 0.03-0.05 part of phenoxyethanol, 0.5-1 part of propylene glycol, 1-3 parts of ethylhexylglycerin and 0.5-1 part of essence.

Adjusting the pH value of the raw material of the moringa product to 6.40 by using 0.01mol/L sodium hydroxide, adding the raw material into a water bath kettle, continuously stirring and heating to 80 ℃, keeping the temperature for 1 hour, cooling and filling to obtain the final product, namely the moringa repairing water.

Example 9. Preparation of moringa oleifera hair conditioner for repairing and relieving itching

The preparation method of the moringa oleifera hair conditioner for repairing and relieving itching comprises the following raw materials: 60% of moringa product raw material, 5% of lanolin, 10% of olive oil, 5% of triethanolamine oleate, 5% of polyoxyethylene castor oil, 5% of lauric acid, 5% of potassium stearate, 1% of EDTA disodium, 1% of sodium sorbate and 1% of essence. The Moringa oleifera hair conditioner has effects of repairing scalp, relieving itching, and smoothing hair. 0032 the materials with the percentage are uniformly mixed, fully stirred for more than 30 minutes, and filled after the materials are fully mixed, and the finally obtained product is the moringa oleifera hair conditioner which has the effects of repairing hair, relieving itching and protecting scalp.

Example 10. A preparation method of Moringa oleifera eye cream is provided.

A moringa oleifera eye cream comprises the following components: 45-55 parts of deionized water, 20-35 parts of moringa product stock solution, 2-5 parts of xanthan gum, 2-5 parts of sodium alginate, 1-5 parts of sodium hyaluronate, 2-6 parts of sorbitan laurate, 2-5 parts of sodium acrylate, 804-8 parts of polysorbate, 1-2 parts of ethylhexylglycerin and 0.5-1 part of phenoxyethanol.

Preparation method

The following raw materials are provided according to parts by weight: 40 parts of deionized water, 25 parts of moringa product stock solution, 5 parts of xanthan gum, 4 parts of sodium alginate, 3 parts of sodium hyaluronate, 6 parts of sorbitan laurate, 4 parts of sodium acrylate, 805 parts of polysorbate-805, 2 parts of ethylhexyl glycerol and 0.5 part of phenoxyethanol.

Adjusting pH of the prepared moringa product stock solution to 6.5-6.8 for later use.

Adding 40 parts of prepared deionized water and 25 parts of moringa oleifera product stock solution into a reaction kettle, stirring and heating in a water bath at the temperature of 80 ℃ for 30min, controlling the rotating speed to be 40-60r/min, and sequentially adding 5 parts of xanthan gum, 4 parts of sodium alginate, 3 parts of sodium hyaluronate, 6 parts of sorbitan laurate, 4 parts of sodium acrylate, 805 parts of polysorbate-805, 2 parts of ethylhexylglycerin and 0.5 part of phenoxyethanol after 30 min. And continuously stirring and heating for 30min, stopping heating, naturally stirring, cooling to room temperature, and filling.

Example 11. Preparation of moringa oleifera hand cream with repairing function

A moringa oleifera hand cream with a repairing function comprises the following components: 10 parts of moringa product raw material, 2 parts of glycerol, 0.3 part of ethylhexyl palmitate, 0.2 part of polyisobutene, 0.5 part of spermaceti stearic acid, 0.2 part of lanolin, 200.3 parts of steareth, 1000.1 parts of PEG (polyethylene glycol) -stearate, 0.4 part of stearate, 210.1 parts of steareth, 0.3 part of tocopheryl acetate, 0.4 part of xanthan gum, 200.3 parts of polysorbitol ester, 0.1 part of allantoin, 130.3 parts of polyacrylate, 0.1 part of phenoxyethanol, 0.4 part of ethylhexyl glycerin, 0.4 part of rose essence, 0.01 part of methylparaben, 0.01 part of propyl hydroxybenzoate and 15 parts of deionized water.

Preparation method

Adding 15 parts of prepared ionized water into a water bath kettle, stirring and heating the ionized water in the water bath kettle to 80-90 ℃, adding 0.01 part of p-hydroxymethyl, 0.01 part of p-hydroxypropyl and 0.02 part of p-hydroxyacetophenone, continuously heating the ionized water until the ionized water is completely dissolved, adding 2 parts of glycerol, 0.3 part of ethylhexyl palmitate, 0.2 part of polyisobutene, 0.5 part of spermaceti stearate, 0.2 part of wool fat, 200.3 parts of steareth, 1000.1 parts of PEG, 0.4 part of stearate, 210.1 parts of steareth, 0.3 part of tocopheryl acetate, 0.4 part of xanthan gum, 200.3 parts of polyglicotriol, 0.1 part of allantoin, 130.3 parts of polyacrylate, 0.1 part of phenoxyethanol, 0.4 part of ethylhexyl glycerol, 0.01 part of methyl hydroxybenzoate and 0.01 part of propyl hydroxybenzoate, sequentially adding the ionized water, continuously stirring, adjusting 10 parts of moringa product raw material to 6.40 parts by using 0.01mol/l of sodium hydroxide, adding the phwood product raw material to mix the mixture to obtain a mixed solution, and (3) fully and uniformly mixing, stopping heating, stirring, cooling to room temperature, adding 0.4 part of rose essence, and continuously stirring for 20 minutes to obtain a final product, namely the moringa oleifera hand cream with the repairing function.

Example 12. A preparation method of Moringa oleifera facial cream with effects of keeping moisture and supplementing water is provided.

A moringa oleifera facial cream capable of preserving moisture and replenishing water comprises the following components: 20 parts of moringa product raw material, 2 parts of glycerol, 2 parts of trehalose, 0.2 part of butanediol, 0.3 part of isononyl isononanoate, 1 part of spermaceti stearic acid, 0.3 part of polydimethylsiloxane, 2 parts of shea butter, 2 parts of glycerol stearate, 20.4 parts of steareth-20.4 parts, PEG-1001 parts, 1 part of stearate, 210.1 parts of steareth-210, 0.2 part of tocopheryl acetate, 2 parts of xanthan gum, 3 parts of sodium hyaluronate, 201 parts of polysorbate-201, 0.5 part of allantoin, 0.5 part of acryloyl dimethyl tauryl/VP copolymer, 0.2 part of polyisobutylene, 0.2 part of dimethiconol, 0.2 part of phenoxyethanol, 2 parts of ethylhexyl glycerol, 1 part of rose essence, 0.01 part of methyl hydroxybenzoate, 0.03 part of propylhydroxybenzoate and 60 parts of deionized water.

Preparation method

60 parts of prepared ionized water is put into a water bath kettle to be stirred and heated to 80-90 ℃, then 0.01 part of p-hydroxymethyl, 0.03 part of p-hydroxypropyl and 0.02 part of p-hydroxyacetophenone are added, the mixture is continuously heated to be completely dissolved, 2 parts of glycerin, 2 parts of trehalose, 0.2 part of butanediol, 0.3 part of isononyl isononanoate, 1 part of spermaceti stearate, 0.3 part of polydimethylsiloxane, 2 parts of shea butter, 2 parts of glycerol stearate, 20.4 parts of steareth-20, 1001 parts of PEG-1001, 1 part of stearate, 210.1 parts of steareth-210, 0.2 part of tocopheryl acetate, 2 parts of xanthan gum, 3 parts of sodium hyaluronate, 201 parts of polysorbate-201, 0.5 part of allantoin, 0.5 part of acryloyl dimethyl taurocarb/VP copolymer, 0.2 part of polyisobutylene, 0.2 part of dimethiconol, 0.2 part of phenoxyethanol and 2 parts of ethylhexyl glycerin are sequentially added, and (3) continuously stirring, adjusting the ph of 20 parts of the raw materials of the moringa oleifera product to 6.40 by using 0.01mol/l of sodium hydroxide, adding the mixed solution, fully and uniformly mixing, stopping heating, stirring and cooling to room temperature, adding 1 part of rose essence, and continuously stirring for 20 minutes to obtain a final product, namely the moringa oleifera cream with the functions of moisturizing and supplementing water.

Example 13. Preparation of moringa oleifera facial cleanser with sun protection and repair functions

A moringa oleifera facial cleanser with sun protection and repair functions comprises the following components: 20 parts of deionized water, 25 parts of a moringa product raw material, 1 part of cocoyl glycine potassium, 0.5 part of lauroyl sarcosine, 1 part of potassium laureth phosphate, 2 parts of decyl glucoside, 1 part of cocamidopropyl betaine, 0.5 part of butanediol, 2 parts of PEG-50 shea butter, 0.5 part of C12-13 alcohol lactate, 0.5 part of betaine, 2 parts of acrylic acid (ester) copolymer, 2 parts of mica, 0.5 part of titanium dioxide, 1 part of PEG-160 sorbitan triisostearate, 81 parts of polyethylene glycol, 1 part of sodium isostearoyl lactate, 0.3 part of phenoxyethanol, 2 parts of ethylhexyl glycerol, 0.5 part of rose essence, 0.03 part of methyl hydroxybenzoate and 0.01 part of propyl hydroxybenzoate. Can effectively inhibit tyrosinase activity and reduce melanin generation, thereby achieving the purpose of whitening and sunscreen, and simultaneously promoting the secretion and synthesis of collagen, and can firstly repair damaged cells.

Preparation method

20 parts of prepared ionized water is put into a water bath kettle to be stirred and heated to 80-90 ℃, then 0.01 part of p-hydroxymethyl and 0.03 part of p-hydroxypropyl are added to be continuously heated to be completely dissolved, 1 part of cocoyl glycine potassium, 0.5 part of lauroyl sarcosine, 1 part of potassium lauryl polyether phosphate, 2 parts of decyl glucoside, 1 part of cocamidopropyl betaine, 0.5 part of butanediol, 2 parts of PEG-50 shea butter resin, 0.5 part of C12-13 alcohol lactate, 0.5 part of betaine, 2 parts of acrylic acid (ester) copolymer, 2 parts of mica, 0.5 part of titanium dioxide, 1 part of PEG-160 sorbitan triisostearate, 81 parts of polyethylene glycol, 1 part of sodium isostearate lactate, 0.3 part of phenoxyethanol and 2 parts of ethylhexyl glycerol are sequentially added to be continuously stirred, 25 parts of raw materials of a moringa product are adjusted to 6.40 by using 0.01mol/l of sodium hydroxide, and then mixed solution is added, and (3) fully and uniformly mixing, stopping heating, stirring and cooling to room temperature, adding 0.5 part of rose essence, and continuously stirring for 20 minutes to obtain a final product, namely the moringa oleifera facial cleanser with the repairing function.

The examples are not intended to limit the scope of the present application, and all equivalent changes or modifications made within the technical spirit of the present invention shall fall within the scope of the present invention, but all products made from the moringa extract of the present invention as a raw material shall fall within the scope of the present invention.

Claims

1. A moringa oleifera extract for promoting collagen secretion is characterized by being prepared by the following steps:

1) crushing a moringa oleifera raw material to obtain a moringa oleifera material;

2) adding deionized water into the moringa oleifera material obtained in the previous step, wherein the mass ratio of the deionized water to the moringa oleifera material is (2-10): 1;

3) weighing the amount of each enzyme used for enzymolysis according to 0.003-0.005 of the mass of the moringa oleifera material obtained in the step 1), wherein the enzyme activity value of each enzyme is 5-20 ten thousand units, combining the enzymes used for enzymolysis, adding deionized water, and fully mixing to obtain a mixed enzyme aqueous solution, wherein the mass ratio of the deionized water to the total amount of all the enzymes is 5-20: 1, the enzymes used for enzymolysis must comprise at least one protease and at least one cellulase;

4) dripping the mixed enzyme aqueous solution obtained in the step 3) into the material obtained in the step 2;

5) adding an organic solvent into the substance obtained in the step 4), wherein the volume mass ratio of the organic solvent to the moringa oleifera material in the substance obtained in the step 4 is 4-8L: 1 KG;

6) carrying out reflux extraction and water enzymolysis on the product obtained in the step 5) under the stirring condition, and stirring the water for enzymolysis for more than 3.5 hours at constant temperature when the temperature of the system is increased to 52-58 ℃;

7) separating the upper organic phase in the product obtained in the step 6) under the condition that the vacuum degree is not lower than 0.04MP after the enzymolysis is finished, wherein the organic phase is used as the organic phase, and the rest material system is moringa oleifera material fermentation base liquid;

8) and (2) carrying out sealed anaerobic fermentation on the moringa oleifera material fermentation base liquid obtained in the step 7), wherein a fermentation strain consists of at least one saccharomyces cerevisiae and at least one strain capable of generating organic acid, the addition amount of each strain is 0.05-0.4% of the mass of the moringa oleifera material obtained in the step 1), the fermentation temperature is 28-35 ℃, when the pH value is reduced to 3.5-4.5, the fermentation is stopped, then, the solid-liquid separation of a system is carried out, the liquid phase is collected, the filter residue is discarded, the liquid phase is subjected to ultrafiltration membrane filtration, and the filtration pore is 0.28-0.32 mu m, so that an extract finished product is obtained.

2. The moringa oleifera extract for promoting collagen secretion according to claim 1, wherein the moringa oleifera raw material in the step 1) is moringa oleifera fresh fruit pod, moringa oleifera fresh leaf or moringa oleifera seed in the same year.

3. The extractive solution of Moringa oleifera capable of promoting collagen secretion according to claim 1, wherein the protease of step 3) is bromelain or papain.

4. The moringa oleifera extract for promoting collagen secretion according to claim 1, wherein the dropwise addition in the step 4) is completed within 8-10 minutes.

5. The collagen secretion promoting moringa oleifera extract as claimed in claim 1, wherein the organic solvent in step 5) is petroleum ether.

6. The moringa oleifera extract for promoting collagen secretion according to claim 1, wherein the timing constant-temperature stirring water enzymolysis temperature in the step 6) is 55 ℃, the time is 4 hours, and the stirring condition is 58-62 revolutions per minute.

7. The moringa oleifera extract for promoting collagen secretion according to claim 1, wherein the fermentation strain of the step 8) is saccharomycete and lactobacillus plantarum, and the fermentation time is 7-15 days.

8. The use of the moringa oleifera extract solution for promoting collagen secretion according to claim 1 as a raw material for preparing cosmetics, health products, drugs or feeds.