CN108991471B - Natural food additive with strong antioxidant activity - Google Patents
Natural food additive with strong antioxidant activity Download PDFInfo
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- CN108991471B CN108991471B CN201810678957.XA CN201810678957A CN108991471B CN 108991471 B CN108991471 B CN 108991471B CN 201810678957 A CN201810678957 A CN 201810678957A CN 108991471 B CN108991471 B CN 108991471B
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- anthocyanin
- extract
- purified product
- ethanol
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
- A23L27/10—Natural spices, flavouring agents or condiments; Extracts thereof
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
- A23L27/10—Natural spices, flavouring agents or condiments; Extracts thereof
- A23L27/105—Natural spices, flavouring agents or condiments; Extracts thereof obtained from liliaceae, e.g. onions, garlic
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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Abstract
A natural food additive with strong antioxidant activity belongs to the technical field of food additives. The purple onion skin tannin extract and the seaweed plant pigment extract are prepared by mixing and proportioning a black wolfberry anthocyanin extract and purified product, a black pearl tomato anthocyanin extract and purified product, a purple onion anthocyanin extract and purified product, a pomegranate bark tannin extract and purified product and a seaweed plant pigment extract and according to the weight ratio of 30-40:30-40:10-20:2-10: 2-10. The natural food additive with powerful antioxidant activity has the functions of super oxidation resistance, radiation resistance, aging resistance, fatigue resistance, liver and kidney nourishing, sleep improvement, immunity enhancement, antibiosis and corrosion prevention, is high in health care nutritive value, stable in performance, easy to store and transport for a long time, overcomes the defects of the prior art, and greatly improves the added value of food.
Description
Technical Field
The invention belongs to the technical field of food additives, and particularly relates to a natural food additive with strong antioxidant activity.
Background
The food additive is added into food for improving the quality, color, fragrance and taste of food and prolonging the shelf life of food. With the rapid development of food industry and the current of 'Chongshang green safety and natural color return', a plurality of food additives have the functions of health care and nutrition. For example, the food additive added with vitamin C, seaweed, dendrobium officinale, ganoderma lucidum spore powder, trace elements, amino acid and the like beneficial to human bodies has good functions of health care, oxidation resistance, corrosion prevention and the like, and the food additive refers to a chemical synthetic substance or a natural substance added into food for effectively improving the quality, color, aroma and taste of the food and prolonging the shelf life of the food. Under certain environmental conditions, food loses its original nutritive value and is deteriorated due to the action of microorganisms, mainly food deterioration caused by the propagation of microorganisms. The food preservative can prevent food from putrefaction and deterioration caused by microorganism propagation, and can make food have long shelf life in common natural environment.
With the continuous improvement of the diet level of people, more and more food additives enter the daily life of people and are widely accepted by people. Consumers in today's society are more concerned about the hygiene and health of their own diets. Therefore, various problems related to food additives have naturally become a focus of general attention in recent years also in the diet. More and more food varieties are eaten by people, and more attention is paid to whether additives exist in the food, what additives exist in the food, the health-care nutritive value of the food, no side effect of the food and the like. Therefore, consumers are continuously pursuing natural food additives with high nutrition and health care functions, pure natural, safe eating and no toxic or side effect at the present of pursuing higher and higher color, aroma, taste, shape, sensory quality and health care value.
Anthocyanin is one of the most effective antioxidants from plant sources discovered so far, has super-strong antioxidant and free radical scavenging activities, and has the super-strong effect of improving the immunity of human bodies. Unlike most antioxidants, anthocyanins are responsible for aging and many diseases due to free radicals, and it is estimated that about 90% of senile and degenerative diseases are related to free radicals, so anthocyanins have strong effects of resisting oxidation and enhancing immunity. Anthocyanins are natural sun covers, and can block ultraviolet rays from invading skin, protect human skin from invading by free radicals, and repair injured collagen and elastic fibers. Anthocyanins also improve microcirculation of brain and heart, improve microvascular fragility and permeability, and slow aging and degenerative diseases.
Disclosure of Invention
Aiming at the problems in the prior art, the invention aims to design and provide a technical scheme of a natural food additive with strong antioxidant activity, which has the functions of super oxidation resistance, radiation resistance, aging resistance, fatigue resistance, liver and kidney nourishing, sleep improvement, immunity enhancement, antibiosis and corrosion prevention, high health care nutritive value, stable performance, easy long-term storage and transportation, overcomes the defects of the prior art and greatly improves the added value of food.
The natural food additive with strong antioxidant activity is characterized by being prepared by mixing and proportioning extracted and purified lycium ruthenicum anthocyanin, extracted and purified black pearl tomato anthocyanin, extracted and purified purple onion anthocyanin, extracted and purified pomegranate peel tannin and extracted and purified seaweed plant pigment according to the weight ratio of 30-40:30-40:10-20:2-10: 2-10.
The natural food additive with strong antioxidant activity is characterized in that the extracted and purified lycium ruthenicum anthocyanin is prepared by the following method: placing the lycium ruthenicum murr dried at low temperature into a tissue crushing extractor, adding 50% -70% ethanol solution with the solid-liquid ratio of 1: 2-1: 5 and the pH value of 3.0-4.0, and crushing and extracting for 1-3 min at room temperature. Performing suction filtration, adding 50-65% ethanol solution with pH of 3.0-pH 4.0 and solid-liquid ratio of 1: 2-1: 5 into the obtained filter residue, performing ultrasonic extraction for 1-2 times at room temperature for 20-40 min each time by using an ultrasonic extraction method, and performing suction filtration; mixing the tissue disruption extract and the ultrasonic extract, and concentrating by flash evaporation at 50-60 deg.C to 1/2-1/5 of original volume to obtain concentrated solution; enriching and purifying the total anthocyanin by macroporous adsorption resin Diaion HP 2MGL column chromatography; eluting with distilled water, adjusting the flow rate to 0.5-1.5 mL/min until the eluent is colorless and transparent, eluting with 55-70% ethanol solution with pH of 2.0-4.0, and collecting mauve pigment eluent, wherein the flow rate of the eluent is controlled to 0.5-1.5 mL/min; and (3) carrying out vacuum concentration drying or freeze drying on the collected mauve eluent in a rotary evaporator to obtain the lycium ruthenicum anthocyanin extraction and purification dry powder.
The natural food additive with strong antioxidant activity is characterized in that the extracted and purified product of the anthocyanin of the black pearl tomato is prepared by the following method: placing black pearl tomatoes into a tissue crushing extractor, adding 55-65% ethanol solution with the pH value of 2.0-pH value of 4.0 and the volume ratio of 1: 3-1: 6, crushing and extracting for 1-3 min at room temperature; squeezing with coarse cloth to obtain extractive solution; concentrating the extractive solution in 50-60 deg.C water bath with flash evaporation concentrating device under reduced pressure to 1/3-1/6 of original volume to obtain concentrated solution; enriching and purifying the total anthocyanin by macroporous adsorption resin Diaion HP 2MGL column chromatography; eluting with distilled water, adjusting the flow rate to 0.5-1.5 mL/min until the eluent is colorless and transparent, eluting with 55-70% ethanol solution with pH of 3.0-4.0, and collecting mauve pigment eluent, wherein the flow rate of the eluent is controlled to 0.5-1.5 mL/min; and (3) concentrating and drying the collected mauve eluent in vacuum or freeze drying in a rotary evaporator to obtain the dry powder of the extracted and purified product of the anthocyanin of the black pearl tomato.
The natural food additive with strong antioxidant activity is characterized in that the purple onion anthocyanin extracted and purified product is prepared by the following method: placing purple onions into a tissue crushing extractor, adding 50% -60% ethanol solution with the pH value of 3.0-4.0 and the volume ratio of 1: 3-1: 6, crushing and extracting for 1-3 min at room temperature; performing suction filtration, adding 50-70% ethanol solution with pH of 3.0-pH of 4.0 and volume ratio of 1: 3-1: 6 into the obtained filter residue, performing ultrasonic extraction for 20-40 minutes each time for 1 time at room temperature by adopting an ultrasonic extraction method, and performing suction filtration; mixing the tissue disruption extract and the ultrasonic extract, and concentrating by flash evaporation at 50-60 deg.C to 1/2-1/6 of original volume to obtain concentrated solution; concentrating and purifying the concentrated solution with macroporous adsorbent resin Diaion HP 2MGL column chromatography to obtain purple onion pigment; eluting with distilled water, adjusting the flow rate to 0.5-1.5 mL/min until the eluent is colorless and transparent, eluting with 55-70% ethanol solution with pH of 3.0-4.0, and collecting pigment eluent, wherein the flow rate of the eluent is controlled at 0.5-1.5 mL/min; and (3) concentrating and drying the collected pigment eluent in vacuum or freeze drying in a rotary evaporator to obtain the dry powder of the extracted and purified product of the anthocyanin of the black pearl tomato.
The natural food additive with strong antioxidant activity is characterized in that the pomegranate bark tannin extraction and purification substance is prepared by the following method: placing the pomegranate rind dried at low temperature into a tissue crushing extractor, adding 60% -70% acetone solution with a solid-to-liquid ratio of 1: 4-1: 8, crushing and extracting for 1-3 min at room temperature; suction filtering, flash evaporating and concentrating the extracting solution to 1/4-1/8 of the original volume by adopting a flash evaporation concentration device under the condition of 50-60 ℃, subjecting the obtained concentrated solution to Diaion HP 20 macroporous adsorption resin column chromatography, performing gradient elution by respectively using deionized water, 10% ethanol, 20% ethanol, 40% ethanol, 60% ethanol and 80% ethanol with the liquid-material ratio of 1:20-50 times, according to the flow rate of 2.5-10.0 mL/min, collecting each eluent, combining the 20% ethanol, 40% ethanol and 60% ethanol eluents, performing flash evaporation concentration in a water bath at 50-60 ℃ to obtain 1/10-1/20 of the original volume, and performing rotary evaporation concentration on the obtained concentrated solution by using a rotary evaporator to dry to obtain the purified dry powder of the pomegranate rintannin extract.
The natural food additive with strong antioxidant activity is characterized in that the seaweed phytochrome extraction and purification product is prepared by the following method: placing the dried seaweed in a tissue crushing extractor, adding 70-80% of acetone with a solid-liquid ratio of 1: 3-1: 8, crushing and extracting for 1-3 min at room temperature; suction filtering, and flash evaporating and concentrating the extract at 50-60 deg.C until the concentrated solution does not contain acetone; then extracting with diethyl ether at a volume ratio of 1:1-1:3 for 2-6 times; mixing the ether extracts, steaming at 40 deg.C in water bath under normal pressure to remove ether, and freeze drying the obtained concentrated extract to obtain purified product of seaweed plant pigment.
The natural food additive with strong antioxidant activity is characterized in that the weight ratio of the extracted and purified product of lycium ruthenicum anthocyanin, the extracted and purified product of black pearl tomato anthocyanin, the extracted and purified product of purple onion anthocyanin, the extracted and purified product of pomegranate peel tannin and the extracted and purified product of seaweed plant pigment is 32-38:32-38:12-18:3-8:3-8, preferably 34-35:34-35:15-16:4-5: 4-5.
Lycium ruthenicum Murr, is Lycium ruthenicum Murr, and belongs to Lycium of Solanaceae. Is a multi-thorn shrub with multi branches, the branches are hard and are bent in a zigzag shape, and the branches are white. Lycium ruthenicum Murr is mainly distributed in Qinghai, Ningxia, Xinjiang, Gansu, Shaanxi and other places in China and Europe. The wine is distributed sporadically in Gansu spring, West part of Qinghai, Ningxia, Xinjiang, east part of birch county in Heilongjiang province, west part of inner Mongolia and Tibet. It grows on barren mountains and mountains where people cannot live, and has strong vitality. Lycium ruthenicum Murr has various biological activities, mainly comprising: 1. anti-aging and sleep improvement: a large amount of anthocyanin contained in the lycium ruthenicum has good effects of resisting oxidation, eliminating free radicals of a human body and having remarkable anti-aging effect. The anthocyanidin can cross blood brain barrier, protect cranial nerve from oxidation, stabilize brain tissue function, and protect brain from harm of harmful chemical substances and toxin. The sleep quality can be obviously improved after the anthocyanin in the lycium ruthenicum is eaten. 2. Radiation resistance, improvement of skin: the anthocyanin in the lycium ruthenicum murr is a natural sunlight covering material, can prevent ultraviolet rays from damaging skin, can prevent skin wrinkles from being generated in advance, and can supplement nutrition and eliminate harmful free radicals in vivo. 3. Nourishing liver and kidney and resisting eye fatigue: the anthocyanidin in Lycium ruthenicum Murr can promote regeneration of rhodopsin in retinal cells, prevent severe myopia and retinal detachment, and has effects of resisting asthenopia and improving vision. 4. Improving circulation and enhancing immunity: the anthocyanin in the lycium ruthenicum murr can improve blood circulation, recover lost microvascular efficacy, strengthen vascular elasticity and provide immunity of an organism. In addition, the anthocyanin in the lycium ruthenicum has the effects of reducing blood sugar, lowering blood pressure, regulating lipid metabolism and protecting a cardiovascular system.
The black pearl tomato is originally produced in south America, is a treasure of tomato families, and is named because the color of the fruit and the color of the fruit are black. The fruit can be used as both medicine and food, has strong fruit flavor, moderate sour and sweet taste, high nutritive value, and is easy to be absorbed by human body, and is especially suitable for fresh food. The black pearl tomato is a first-filial generation excellent seed introduced from abroad, and has the advantages of good taste, excellent coloring, high sugar content, good harvest uniformity, strong disease resistance and the like. The black pearl tomato contains lycopene 10 times higher than that of red tomato, and also contains a large amount of antioxidant such as anthocyanin and vitamin C. The anthocyanin in the black pearl tomato can generate strong biological effect in human body, has the effects of protecting kidney, strengthening yang, treating prostatitis, and preventing breast cancer, cervical cancer, prostatic cancer, colon cancer and heart disease, and the eating of the black tomato can reduce the probability of suffering from the prostatic cancer and the heart disease.
The onion is 2-year-old herbaceous plant belonging to genus Allium of family Liliaceae. The onion is a multifunctional and multipurpose food integrating vegetables, medicines, seasonings, sterilization and health care. The purple onions are purple red in appearance, are rich in anthocyanin, contribute to providing colors for fruits and vegetables, belong to flavonoids in fruits and vegetables, cannot generate the anthocyanin by the human body, and are mainly supplemented by taking dark food materials. Purple onions are most effective at killing human cancer cells because the variety contains more of the antioxidant substances anthocyanin and quercetin. Purple onions activate pathways that encourage cancer cell death, they promote the formation of hostile environments in cancer cells, disrupt communication between cancer cells, and thereby inhibit tumor growth. Research shows that the purple onion contains a large amount of anthocyanin, quercetin and other bioactive substances, so that the purple onion has the effects of effectively preventing skin aging, strengthening brain, protecting eyes, protecting liver and kidney, removing in-vivo garbage, purifying blood, preventing hypertension, hyperlipidemia, hyperglycemia, cardiovascular and cerebrovascular diseases, eliminating carcinogenic substances, preventing arthritis, liver and kidney diseases, resisting cancer and sterilizing.
Pomegranate is fragrant, sweet and delicious, is one of the most popular fruits in summer, is rich in nutrition and is good for the whole body. The pericarpium Granati is pericarp of Granati of Punicaceae. The pericarpium Granati contains malic acid and tannin. Tannins, also called tannins, are a class of structurally complex polyphenolic compounds present in plants. Researches show that the pomegranate rind has obvious functions of bacteriostasis and convergence, can cause the intestinal mucosa to be converged, and reduces the secretion of intestinal adhesive wax, so the diarrhea can be effectively treated; pomegranate rind is rich in substances, and has good effects of resisting oxidation, resisting fatigue, scavenging free radicals and the like, and also has the effects of stopping bleeding and astringing.
Algal pigments mainly contain carotenoids, including astaxanthin, which and lycopene, etc., are intermediates in carotenoid synthesis. Widely exists in the biological world, such as shrimp, crab, fish, algae, and plant leaves, flowers, and fruits, and is one of the main carotenoids in marine organisms. Astaxanthin pigment in algae is a carotenoid colorant, is one of the strongest natural antioxidants in the world, can effectively remove oxygen free radicals in cells, enhances the regeneration capacity of the cells, and has the effects of resisting aging, relieving sports fatigue, enhancing vitality, preventing the occurrence and development of chronic diseases such as tumors, cardiovascular diseases, diabetes and the like, delaying aging and the like. Astaxanthin can cross the barrier from blood to brain, so astaxanthin can protect brain, central nervous system and eyes, relieve asthenopia symptom, and improve visual sensitivity; has effects in preventing ultraviolet radiation and relieving pigmentation; regulating cytokines, inhibiting inflammatory cytokine development; anti-inflammatory and anti-infection. The algae plant is a living body which is rich in astaxanthin in nature and is known as a concentrated product of natural astaxanthin. The extraction of astaxanthin from marine and freshwater algae by using algae plants has a wide development prospect and becomes a research hotspot for the international production of natural astaxanthin.
The natural food additive with strong antioxidant activity has the following beneficial effects:
1. according to the instability of anthocyanin structures such as anthocyanins, tannins and carotenoids under the conditions of heat, humidity, light and air, the influence of factors such as heating, humidity, air and light and the characteristic of being easily oxidized, degraded and deteriorated in a solution state, when the anthocyanin of the lycium ruthenicum murr and the pigment of the purple onion are extracted, a room-temperature tissue crushing extraction method and a room-temperature ultrasonic extraction method are combined, effective components in plant tissue cells are quickly and efficiently dissolved in a solvent in a short time through room-temperature tissue crushing extraction, and then the room-temperature ultrasonic extraction is combined, so that the effective components are extracted in a short time; when the anthocyanin of the black pearl tomato, the pomegranate peel tannin and the seaweed plant pigment are extracted, a room-temperature tissue crushing extraction method is adopted to combine the crushing process and the extraction process into one in a few minutes, the processes of crushing, mixing, stirring, homogenizing and extracting are completed simultaneously, the components in the tissue cells are quickly transferred to a solvent in a short time, the quick and complete efficient extraction is realized, the structure of the effective components is furthest ensured not to be damaged, and the high preservation rate and transfer rate of the effective components are ensured.
2. The existing synthetic antioxidant has the advantages of low production cost and low price, but the toxic and side effects of the synthetic antioxidant bring hidden troubles to the application of the food additives. Compared with the prior art, the natural food additive has the advantages of environmental protection, no toxic or side effect and safe use. Most of the existing natural antioxidants are tea polyphenol, vitamin E, flavonoid and polyphenol. As a food additive, it has the disadvantages of high cost and insufficient ability of resisting oxidation and scavenging free radicals. The food additive provided by the invention has the advantages of pure natural raw materials, greenness, safety and no toxic or side effect. The tea polyphenol antioxidant has the advantages of having super-strong activities of resisting oxidation, aging and fatigue and removing free radicals, the activities of resisting oxidation and removing free radicals of the tea polyphenol antioxidant are far greater than that of strong antioxidants such as tea polyphenol and vitamin E, and the antioxidant activity of the tea polyphenol antioxidant is tens of times that of tea polyphenol and VE antioxidant.
3. Anthocyanins, tannins and carotenoid pigments in various fruits, vegetables and plants are all ingredients with strong antioxidant activity, and the fruits, vegetables and plants are not easy to preserve after being mature, are easy to rot and deteriorate, and are quickly oxidized when contacting air. And the fresh fruits and vegetables are used in large quantity, and because the fruits and vegetables contain more sugar, the fruits and vegetables are not suitable for being used by the public for a long time, and the fruits and vegetables are required to be processed in time, such as juicing, disinfecting, canning and sealing, and making into fruit wine, jam and the like, but the fruits and vegetables have the defects of high sugar content, and are not suitable for being used by most people. The invention carries out low-temperature rapid extraction and flash evaporation concentration by a special process and technology, and then carries out enrichment and purification by macroporous resin, thereby removing a large amount of sugar, protein and other impurities, obtaining an extracted and purified substance with high content of effective components, and overcoming the adverse effect brought by directly eating fruits and vegetables.
4. The five natural plants are from different fruit and vegetable plants, and contain different types of antioxidant active ingredients, including anthocyanins, tannins and carotenoids, and effective combinations of three functional ingredients with different structural types play a role in mutual coordination and synergism, so that the activities of resisting oxidation and removing free radicals of the five natural plants are greatly enhanced. A large number of tests and clinical applications show that the natural food additive has the super activities of resisting oxidation, aging and fatigue and removing free radicals, and also has the health-care effect.
5. The natural food additive has the functions of super-strong antioxidation, radioresistance, anti-aging, anti-fatigue, liver and kidney nourishing, sleep improving, immunity enhancing, antibiosis and corrosion prevention, has rich health care and nutrient value and stable performance, is easy to store and transport for a long time, and overcomes the defects of the prior art. Due to the fact that the packaging is carried out in two steps, air and moisture are isolated, the influence of light is avoided, and the stability and the quality guarantee period of the functional components in the product are prolonged. The product can be added into various foods and cakes as food additive, and has bright and beautiful color, and remarkable antioxidant and health-care effects.
Detailed Description
The present invention is further illustrated by the following specific examples. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1: the preparation method comprises the steps of extracting and purifying lycium ruthenicum anthocyanin, extracting and purifying black pearl tomato anthocyanin, extracting and purifying purple onion anthocyanin, extracting and purifying pomegranate peel tannin and extracting and purifying seaweed plant pigment, mixing the materials according to the weight ratio of 40:30:15:5:10, sealing and bagging every 10g in a dark place, and putting the bags into a brown sealed plastic bottle to obtain the purple onion anthocyanin.
The preparation method of the lycium ruthenicum anthocyanin extraction and purification compound comprises the following steps: placing the low temperature air dried Lycium ruthenicum Murr in a tissue crushing extractor, adding 60% ethanol solution with pH 3.0 and solid-to-liquid ratio of 1:4, crushing and extracting at room temperature for 1.5 min. Filtering, adding 60% ethanol solution with pH of 3.0 and solid-to-liquid ratio of 1:4, ultrasonic extracting at room temperature for 30min for 1 time by ultrasonic extraction, and filtering. Mixing the tissue disruption extract and the ultrasonic extract, and concentrating by flash evaporation at 55 deg.C to 1/5 of original volume to obtain concentrated solution. And (3) enriching and purifying the total anthocyanin by macroporous adsorption resin Diaion HP 2MGL column chromatography. Eluting with distilled water, adjusting flow rate to 1.5 mL/min until the eluate is colorless and transparent, eluting with 65% ethanol solution with pH of 4.0, and collecting mauve pigment eluate with flow rate of eluate controlled at 1.5 mL/min. And (3) concentrating and drying the collected mauve eluent in a rotary evaporator in vacuum or freeze drying to obtain lycium ruthenicum anthocyanin extract purified dry powder, weighing and calculating the yield, and drying and storing the sample in a dark place. The yield of the extracted and purified lycium ruthenicum anthocyanin is 14.5%, and the content of total anthocyanidin in the extracted and purified lycium ruthenicum anthocyanin reaches 46.6%.
The preparation method of the black pearl tomato extract purified substance comprises the following steps: placing black pearl tomato in a tissue crushing extractor, adding 55% ethanol solution with pH of 3.0 at a volume ratio of 1:4, crushing and extracting at room temperature for 1 min. Squeezing with coarse cloth to obtain extractive solution. Concentrating the extractive solution in 60 deg.C water bath under reduced pressure to 1/5 of original volume by flash evaporation concentration device to obtain concentrated solution. And (3) enriching and purifying the total anthocyanin by macroporous adsorption resin Diaion HP 2MGL column chromatography. Eluting with distilled water, adjusting flow rate to 1.0 mL/min until the eluate is colorless and transparent, eluting with 55% ethanol solution with pH of 3.0, and collecting mauve pigment eluate with flow rate of eluate controlled at 1.5 mL/min. And (3) concentrating and drying the collected mauve eluent in a rotary evaporator in vacuum or freeze drying to obtain the dry powder of the extracted and purified anthocyanin of the black pearl tomato, weighing and calculating the yield, and drying and storing the sample in a dark place. The yield of the obtained anthocyanin extracted and purified from the black pearl tomatoes is 6.5 percent, and the content of the total anthocyanidin in the extracted and purified product reaches 25.1 percent.
The preparation method of the purple onion pigment extraction and purification product comprises the following steps: placing purple onion in tissue disruption extractor, adding 60% ethanol solution with pH of 4.0 at a volume ratio of 1:4, and disrupting and extracting at room temperature for 1 min. Filtering, adding 60% ethanol solution with pH of 4.0 and volume ratio of 1:5 into the obtained filter residue, performing ultrasonic extraction at room temperature for 1 time for 20 min by ultrasonic extraction method, and filtering. Mixing the tissue disruption extract and the ultrasonic extract, and concentrating by flash evaporation at 50 deg.C to 1/4 of original volume to obtain concentrated solution. And (4) enriching and purifying the purple onion pigment by macroporous adsorption resin Diaion HP 2MGL column chromatography. Eluting with distilled water, adjusting flow rate to 1.5 mL/min until the eluate is colorless and transparent, eluting with 65% ethanol solution with pH of 4.0, and collecting pigment eluate with flow rate of eluate of 1.5 mL/min. And (3) concentrating and drying the collected pigment eluent in a rotary evaporator in vacuum or freeze drying to obtain the dry powder of the extracted and purified product of the anthocyanin of the black pearl tomato, weighing and calculating the yield, and drying and storing the sample in a dark place. The yield of the obtained purple onion pigment extraction and purification product is 10.5%, and the content of total pigments in the extraction and purification product reaches 18.4%.
The preparation method of the pomegranate bark tannin extraction and purification compound comprises the following steps: placing the dried pericarpium Granati in a tissue disruption extractor, adding 70% acetone solution with solid-to-liquid ratio of 1:8, and disrupting and extracting at room temperature for 3 min. Suction filtering, flash evaporating and concentrating the extracting solution to 1/8 of the original volume by adopting a flash evaporation concentration device at the temperature of 60 ℃, passing the obtained concentrated solution through a Diaion HP 20 macroporous adsorption resin column chromatography, performing gradient elution by respectively using deionized water, 10% ethanol, 20% ethanol, 40% ethanol, 60% ethanol and 80% ethanol with the liquid-material ratio of 1: 30 times, collecting the eluates, combining the eluates of 20% ethanol, 40% ethanol and 60% ethanol, performing flash evaporation concentration in a water bath at the temperature of 50-60 ℃ to 1/20 of the original volume, performing rotary evaporation concentration on the obtained concentrated solution by using a rotary evaporator to dry, and obtaining the purified dry powder of the pomegranate rind tannin extract. Weighing and calculating the yield, and keeping the sample dry in the dark. The yield of the purified pomegranate rind tannin extract was 16.3%.
The preparation method of the seaweed plant pigment extraction and purification product comprises the following steps: placing dried Sargassum in tissue crushing extractor, adding 70% acetone at solid-to-liquid ratio of 1:8, and crushing and extracting at room temperature for 1-3 min. Suction filtering, and flash evaporating the extractive solution at 50-60 deg.C to concentrate until the concentrated solution contains no acetone. Then, the mixture was extracted 4 times with diethyl ether at a volume ratio of 1: 2. Mixing the ether extracts, steaming at 40 deg.C in water bath under normal pressure to remove ether, and freeze drying the obtained concentrated extract to obtain purified product of seaweed plant pigment. Weighing and calculating the yield, and keeping the sample dry in the dark. The yield of the purified product of the seaweed phytochrome extraction was 9.4%.
Example 2: the preparation method comprises the steps of extracting and purifying lycium ruthenicum anthocyanin, extracting and purifying black pearl tomato anthocyanin, extracting and purifying purple onion anthocyanin, extracting and purifying pomegranate peel tannin and extracting and purifying seaweed plant pigment, mixing the materials according to the weight ratio of 35: 40:10:5:10, sealing and bagging every 10g in a dark place, and putting the bags into a brown sealed plastic bottle to obtain the purple onion anthocyanin.
The preparation method of the lycium ruthenicum anthocyanin extraction and purification compound comprises the following steps: placing the low temperature air dried Lycium ruthenicum Murr in a tissue crushing extractor, adding 50% ethanol solution with pH 3.0 and solid-to-liquid ratio of 1:5, crushing and extracting at room temperature for 2 min. Filtering, adding 50% ethanol solution with pH of 3.00 and solid-to-liquid ratio of 1:5 into the obtained residue, performing ultrasonic extraction at room temperature for 20 min for 1 time by ultrasonic extraction method, and filtering. Mixing the tissue disruption extract and the ultrasonic extract, and performing flash evaporation concentration at 60 ℃ by using a flash evaporation concentration device to 1/5 of the original volume to obtain a concentrated solution. And (3) enriching and purifying the total anthocyanin by macroporous adsorption resin Diaion HP 2MGL column chromatography. Eluting with distilled water, adjusting flow rate to 1.5 mL/min until the eluate is colorless and transparent, eluting with 60% ethanol solution with pH of 4.0, and collecting mauve pigment eluate with flow rate of eluate controlled at 1.0 mL/min. And (3) concentrating and drying the collected mauve eluent in a rotary evaporator in vacuum or freeze drying to obtain lycium ruthenicum anthocyanin extract purified dry powder, weighing and calculating the yield, and drying and storing the sample in a dark place. The yield of the extracted and purified lycium ruthenicum anthocyanin is 14.4%, and the content of total anthocyanidin in the extracted and purified lycium ruthenicum anthocyanin reaches 51.2%.
The preparation method of the black pearl tomato extract purified substance comprises the following steps: placing black pearl tomato in a tissue crushing extractor, adding 55% ethanol solution with pH of 4.0 at a volume ratio of 1:6, crushing and extracting at room temperature for 2 min. Squeezing with coarse cloth to obtain extractive solution. Concentrating the extractive solution in 50-60 deg.C water bath with flash evaporation concentrating device under reduced pressure to 1/6 of original volume to obtain concentrated solution. And (3) enriching and purifying the total anthocyanin by macroporous adsorption resin Diaion HP 2MGL column chromatography. Eluting with distilled water, adjusting flow rate to 5 mL/min until the eluate is colorless and transparent, eluting with 65% ethanol solution of pH4.0, and collecting mauve pigment eluate with flow rate of eluate controlled at 1.0 mL/min. And (3) concentrating and drying the collected mauve eluent in a rotary evaporator in vacuum or freeze drying to obtain the dry powder of the extracted and purified anthocyanin of the black pearl tomato, weighing and calculating the yield, and drying and storing the sample in a dark place. The yield of the obtained anthocyanin extracted and purified from the black pearl tomatoes is 8.2 percent, and the content of the total anthocyanidin in the extracted and purified product reaches 29.7 percent.
The preparation method of the purple onion pigment extraction and purification product comprises the following steps: placing purple onion in tissue disruption extractor, adding 60% ethanol solution with pH of 3.0 at a volume ratio of 1:5, and disrupting and extracting at room temperature for 1 min. Filtering, adding 60% ethanol solution with pH of 3.0 at a volume ratio of 1:4 into the obtained residue, performing ultrasonic extraction at room temperature for 25 min for 1 time by ultrasonic extraction method, and filtering. Mixing the tissue disruption extract and the ultrasonic extract, and concentrating by flash evaporation at 50-60 deg.C to 1/5 of original volume to obtain concentrated solution. And (4) enriching and purifying the purple onion pigment by macroporous adsorption resin Diaion HP 2MGL column chromatography. Eluting with distilled water, adjusting flow rate to 1.5 mL/min until the eluate is colorless and transparent, eluting with 70% ethanol solution with pH of 4.0, and collecting pigment eluate with flow rate of eluate of 1.5 mL/min. And (3) concentrating and drying the collected pigment eluent in a rotary evaporator in vacuum or freeze drying to obtain the dry powder of the extracted and purified product of the anthocyanin of the black pearl tomato, weighing and calculating the yield, and drying and storing the sample in a dark place. The yield of the obtained purple onion pigment extraction and purification product is 11.2%, and the content of total pigments in the extraction and purification product reaches 19.6%.
The preparation method of the pomegranate bark tannin extraction and purification compound comprises the following steps: placing the dried pericarpium Granati in a tissue disruption extractor, adding 70% acetone solution with solid-to-liquid ratio of 1:6, and disrupting and extracting at room temperature for 3 min. Suction filtering, flash evaporating and concentrating the extracting solution to 1/6 of the original volume by adopting a flash evaporation concentration device at the temperature of 60 ℃, passing the obtained concentrated solution through a Diaion HP 20 macroporous adsorption resin column chromatography, performing gradient elution by respectively using deionized water, 10% ethanol, 20% ethanol, 40% ethanol, 60% ethanol and 80% ethanol with the liquid-material ratio of 1:25 times, collecting the eluates, combining the eluates of 20% ethanol, 40% ethanol and 60% ethanol, performing flash evaporation concentration in a water bath at the temperature of 60 ℃ until reaching 1/20 of the original volume, and performing rotary evaporation concentration on the obtained concentrated solution by using a rotary evaporator until the concentrated solution is dry to obtain the purified dry powder of the pomegranate rind tannin extract. Weighing and calculating the yield, and keeping the sample dry in the dark. The yield of the purified pomegranate rind tannin extract was 16.1%.
The preparation method of the seaweed plant pigment extraction and purification product comprises the following steps: placing dried Sargassum in tissue disruption extractor, adding 75% acetone at solid-to-liquid ratio of 1:6, and disrupting and extracting at room temperature for 3 min. Suction filtering, and flash evaporating the extractive solution at 50-60 deg.C to concentrate until the concentrated solution contains no acetone. Then, the mixture was extracted 3 times with diethyl ether at a volume ratio of 1: 2. Mixing the ether extracts, steaming at 40 deg.C in water bath under normal pressure to remove ether, and freeze drying the obtained concentrated extract to obtain purified product of seaweed plant pigment. Weighing and calculating the yield, and keeping the sample dry in the dark. The yield of the purified seaweed phytochrome extract was 11.8%.
Example 3: the preparation method comprises the steps of extracting and purifying lycium ruthenicum anthocyanin, extracting and purifying black pearl tomato anthocyanin, extracting and purifying purple onion anthocyanin, extracting and purifying pomegranate peel tannin and extracting and purifying seaweed plant pigment, mixing the materials according to the weight ratio of 40:37:10:5:8, sealing and bagging every 10g in a dark place, and putting the bags into a brown sealed plastic bottle to obtain the purple onion anthocyanin.
The preparation method of the lycium ruthenicum anthocyanin extraction and purification compound comprises the following steps: placing the low temperature air dried Lycium ruthenicum Murr in a tissue crushing extractor, adding 55% ethanol solution with pH of 3.0 and solid-to-liquid ratio of 1:5, crushing and extracting at room temperature for 1.5 min. Filtering, adding 55% ethanol solution with pH of 3.0 and solid-to-liquid ratio of 1:2 into the obtained residue, performing ultrasonic extraction at room temperature for 20 min for 2 times, and filtering. Mixing the tissue disruption extract and the ultrasonic extract, and concentrating by flash evaporation at 55 deg.C to 1/4 of original volume to obtain concentrated solution. And (3) enriching and purifying the total anthocyanin by macroporous adsorption resin Diaion HP 2MGL column chromatography. Eluting with distilled water, adjusting flow rate to 1.5 mL/min until the eluate is colorless and transparent, eluting with 65% ethanol solution with pH of 4.0, and collecting mauve pigment eluate with flow rate of eluate controlled at 1.5 mL/min. And (3) concentrating and drying the collected mauve eluent in a rotary evaporator in vacuum or freeze drying to obtain lycium ruthenicum anthocyanin extract purified dry powder, weighing and calculating the yield, and drying and storing the sample in a dark place. The yield of the extracted and purified lycium ruthenicum anthocyanin is 14.6%, and the content of total anthocyanidin in the extracted and purified lycium ruthenicum anthocyanin reaches 55.2%.
The preparation method of the black pearl tomato extract purified substance comprises the following steps: placing black pearl tomato in a tissue crushing extractor, adding 60% ethanol solution with volume ratio of 1:5 and pH of 4.0, crushing and extracting at room temperature for 2 min. Squeezing with coarse cloth to obtain extractive solution. Concentrating the extractive solution in 60 deg.C water bath under reduced pressure to 1/5 of original volume by flash evaporation concentration device to obtain concentrated solution. And (3) enriching and purifying the total anthocyanin by macroporous adsorption resin Diaion HP 2MGL column chromatography. Eluting with distilled water, adjusting flow rate to 1.0 mL/min until the eluate is colorless and transparent, eluting with 60% ethanol solution with pH of 4.0, and collecting mauve pigment eluate with flow rate of eluate controlled at 1.5 mL/min. And (3) concentrating and drying the collected mauve eluent in a rotary evaporator in vacuum or freeze drying to obtain the dry powder of the extracted and purified anthocyanin of the black pearl tomato, weighing and calculating the yield, and drying and storing the sample in a dark place. The yield of the obtained anthocyanin extracted and purified from the black pearl tomatoes is 6.9 percent, and the content of the total anthocyanidin in the extracted and purified product reaches 31.3 percent.
The preparation method of the purple onion pigment extraction and purification product comprises the following steps: placing purple onion in tissue disruption extractor, adding 50% ethanol solution with pH of 4.0 at a volume ratio of 1:4, and disrupting and extracting at room temperature for 1 min. Filtering, adding 60% ethanol solution with pH4.0 at a volume ratio of 1:5 into the obtained residue, performing ultrasonic extraction at room temperature for 30min for 1 time by ultrasonic extraction method, and filtering. Mixing the tissue disruption extract and the ultrasonic extract, and concentrating by flash evaporation at 55 deg.C to 1/5 of original volume to obtain concentrated solution. And (4) enriching and purifying the purple onion pigment by macroporous adsorption resin Diaion HP 2MGL column chromatography. Eluting with distilled water, adjusting flow rate to 1.5 mL/min until the eluate is colorless and transparent, eluting with 60% ethanol solution of pH4.0, collecting pigment eluate, and controlling flow rate of eluate at 1.5 mL/min. And (3) concentrating and drying the collected pigment eluent in a rotary evaporator in vacuum or freeze drying to obtain the dry powder of the extracted and purified product of the anthocyanin of the black pearl tomato, weighing and calculating the yield, and drying and storing the sample in a dark place. The yield of the obtained purple onion pigment extraction and purification product is 9.8 percent, and the content of the total pigment in the extraction and purification product reaches 20.3 percent.
The preparation method of the pomegranate bark tannin extraction and purification compound comprises the following steps: placing the dried pericarpium Granati in a tissue disruption extractor, adding 66% acetone solution with solid-to-liquid ratio of 1:7, and disrupting and extracting at room temperature for 1-3 min. Performing suction filtration, performing flash evaporation concentration on the extracting solution to 1/7 of the original volume by adopting a flash evaporation concentration device at the temperature of 60 ℃, performing gradient elution on the obtained concentrated solution through a Diaion HP 20 macroporous adsorption resin column chromatography by respectively using deionized water, 10% ethanol, 20% ethanol, 40% ethanol, 60% ethanol and 80% ethanol with the liquid-material ratio of 1: 30 times, collecting each eluent, combining 20% ethanol, 40% ethanol and 60% ethanol eluents, performing flash evaporation concentration in a water bath at the temperature of 60 ℃ to 1/20 of the original volume, performing rotary evaporation concentration on the obtained concentrated solution by using a rotary evaporator until the concentrated solution is dry, and obtaining the purified dry powder of the pomegranate rind tannin extract. Weighing and calculating the yield, and keeping the sample dry in the dark. The yield of the purified pomegranate rind tannin extract was 16.7%.
The preparation method of the seaweed plant pigment extraction and purification product comprises the following steps: placing dried Sargassum in tissue disruption extractor, adding 75% acetone at solid-to-liquid ratio of 1:7, and disrupting and extracting at room temperature for 2.5 min. Suction filtering, and flash evaporating and concentrating the extracting solution at 55 ℃ until the concentrated solution does not contain acetone. Then, the mixture was extracted 4 times with diethyl ether at a volume ratio of 1: 2. Mixing the ether extracts, steaming at 40 deg.C in water bath under normal pressure to remove ether, and freeze drying the obtained concentrated extract to obtain purified product of seaweed plant pigment. Weighing and calculating the yield, and keeping the sample dry in the dark. The yield of the purified seaweed phytochrome extract was 17.4%.
The beneficial effects of the present invention are further demonstrated by experimental data below.
1. Examples 1-3 Oxidation and fatigue resistance test
1) Antioxidant and radical scavenging activity assay
Determination of DPPH radical scavenging Capacity: the DPPH method is one of the common methods for detecting the scavenging capacity of free radicals, Trolox is used as a positive control in the sample determination process, and the total antioxidant capacity of the detected sample is converted according to the positive control. The results are expressed as the concentration of Trolox required to achieve a certain concentration of test substance equivalent antioxidant capacity. This process is referred to as the TEAC process. The TEAC value represents the amount of Trolox equivalent to the sample tested at half clearance. A larger TEAC value indicates a stronger radical scavenging ability. IC (integrated circuit)50The value is a parameter often used to evaluate antioxidant capacity and refers to the concentration of antioxidant required to scavenge 50% of the DPPH radicals. The smaller the value, the more 50% clearance is achieved, the free radical scavenging usedThe smaller the concentration dose of the remover, the better the free radical scavenging effect of the remover, and the stronger the antioxidant activity of the corresponding test sample. The determination method comprises the following steps: accurately weighing a certain amount of DPPH reagent, dissolving with 95% ethanol, quantitatively transferring into a 100 mL measuring flask, fixing the volume with 95% ethanol, shaking to obtain DPPH stock solution with a certain concentration, and storing in a refrigerator for later use. Respectively adding sample solutions with different concentrations and DPPH test solution into a 96-hole enzyme label plate, after shaking, keeping the temperature at 24 ℃ for 30min, measuring the light absorption value (Ap) of the enzyme label plate at the wavelength of 517 nm, and simultaneously measuring the blank light absorption value (Ac) of a sample without DPPH and the light absorption value (Amax) of the sample without DPPH. The concentration of each sample is used as the abscissa, the measured radical clearance (Y) is used as the ordinate to draw a quasi-curve, and the IC is calculated according to the regression equation50. The antioxidant ability was evaluated by converting the radical clearance = 1- (Ap-Ac)/Amax × 100% to the TEAC value of the sample.
Determination of oxidation resistance of FRAB: respectively taking 10 mmol. L-1TPTZ (dissolved in 40 mmol. multidot.L)-1Hydrochloric acid), 20 mmol. L-1And 0. lmol. L-1The obtained acetic acid buffer solution (pH 3.6) was mixed in a volume ratio of 1:1:10 to obtain a FRAP reagent. Precisely measuring the sample solution and the FRAP reagent, reacting for 10 minutes, and measuring the absorbance value at 593 nm. And (3) taking the same amount of absolute ethyl alcohol to replace the solution of the sample to be detected, and simultaneously adding a proper amount of FRAP working solution to serve as a blank control. And finally, calculating the antioxidant activity strength of the sample according to the absorbance value. The absorbance value is in direct proportion to the strength of the antioxidant capacity, namely the greater the absorbance value is, the stronger the antioxidant activity is, in other words, the greater the FRAP value is, the stronger the antioxidant activity is.
Determination of total reducing power of TRP: the sample solution and 5 Trolox control solutions with concentration gradient are respectively added into phosphate buffer solution, and then 1% K is added3Fe(CN)6After heating for 30 minutes, the mixture was left at room temperature, 10% trichloroacetic acid was added, and the mixture was centrifuged for 10 minutes after being mixed uniformly. 2.5 mL of the supernatant was taken, distilled water and 0.1% ferric chloride were rapidly added thereto and mixed uniformly, and absorbance measurement was carried out at a wavelength of 700 nm. And drawing a standard curve by taking the concentration of Trolox as an abscissa and the absorbance as an ordinate, wherein the method is as above. ResultsAntioxidant capacity was evaluated by Trolox equivalent TEAC. See table 1.
TABLE 1 results of antioxidative Activity tests (TEAC value, unit: mg TE/g DW) of examples 1-3
| Sample (I) | DPPH method | TRP method | FRAP method |
| Example 1 | 798.4 | 123.3 | 68.3 |
| Example 2 | 823.6 | 134.5 | 72.6 |
| Example 3 | 812.9 | 129.8 | 72.1 |
Test results show that the examples 1-3 have super-strong antioxidant capacity, the antioxidant and free radical scavenging activities of the examples are far greater than those of antioxidants such as tea polyphenol, vitamin E and the like, and the antioxidant activity is tens of times of that of the tea polyphenol and VE.
2) Anti-fatigue test for mouse weight swimming
A clean male Kunming mouse with the weight of 18-22g is taken and injected with 0.5mg of dexamethasone sodium phosphate injection per mouse in an intramuscular way for 7 days continuously to prepare a kidney-yang deficiency animal model. Mice were randomly divided into 6 groups, namely a normal control group, a model group, a positive group, a high dose group (3.0 g/kg), a medium dose group (1.5 g/kg) and a low dose group (0.75 g/kg) as drugs in example 1, 10 mice in each group were orally administered with 0.1mL/10g of test drugs, the normal control group and the model group were injected with normal saline at equal doses during administration, and the positive group was injected with red cow drink at equal doses during administration. The administration is carried out 1 time daily for 15 days. Each group of mice is randomly divided into two groups, one group is used for a weight swimming test, and the other group is used for measuring the content of liver glycogen, serum urea and serum lactic acid of the mice.
Anti-fatigue test of mouse weight swimming: the animals were trained 3 times first, and all mice were placed in a swimming box for forced swimming tests 30min after the last administration. The tail root of the mouse was loaded with 10% weight lead skin. The time from the start of swimming until the mouse head is fully submerged and cannot float out of the water is recorded as the final exhaustive swimming time(s), see table 2. Compared with the normal control group, the mice in the kidney-yang deficiency model group have obviously reduced time for carrying heavy load and swimming. And compared with the model group, the mouse swimming time under load is gradually prolonged along with the increase of the administration dose. The high-dose groups of examples 1-3 prolong the swimming time of the mice with kidney-yang deficiency, which shows that examples 1-3 have good effects of resisting fatigue, tonifying kidney and strengthening yang.
Determination of liver glycogen content: liver glycogen storage directly affects the motor ability of the body. The glycogen content thus indicates how quickly or how much fatigue has occurred. Examples 1-3 As the last administration of the drug for 30min, the mice were placed in a swimming tank with water temperature of 25 ℃ and swim for 90min without load, and after swimming, the mice were taken out, cervical vertebrae were removed, and the liver was taken out. After the liver was rinsed with physiological saline, the content of hepatic glycogen was measured using a glycogen kit and a microplate reader. The results are shown in Table 3. As can be seen from Table 3, liver glycogen content was significantly reduced in the model group as compared with that in the normal group of mice. The liver glycogen content of mice of each medicine group is obviously increased. With the increase of the drug dosage, the content of hepatic glycogen is also obviously increased. Even the high-dose group of the drug group had a higher content of hepatic glycogen than the positive group, and the medium-dose group had a content comparable to that of the positive group. It is shown that the different dose groups of examples 1-3 all show strong liver glycogen storage function, so that examples 1-3 have strong antioxidant and anti-fatigue effects.
Content determination of blood lactic acid: the blood lactic acid measurement is the most common biochemical index for evaluating the degree of physical fatigue during strenuous exercise. Collecting blood intravenously 30min after the last administration, swimming in 30 deg.C water for 10min, collecting blood 0min before and 0min after swimming, centrifuging to obtain serum, and measuring serum lactic acid content with lactic acid test kit and enzyme-labeling instrument. Compared with a normal control group, the blood serum lactic acid content of the mice in the kidney-yang deficiency model group is obviously increased after swimming, and the blood lactic acid concentration is slowly reduced after 30min of rest. The serum lactic acid concentration of the mice in the drug group is obviously reduced after swimming and resting for 30min, and particularly the serum lactic acid content of the mice in the high and medium dosage groups can be most obviously reduced after exercise of the mice with yang deficiency, which shows that the examples 1-3 have obvious fatigue relieving effect. See table 4.
2. Examples 1-3 anti-fatigue and anti-oxidant test effects on people of different age groups
The results of comparing the effects of relieving fatigue using examples 1-3 for young people who have eyestrain caused by long-term use of computer and mobile phones are shown in Table 5.
The improvement was observed after a period of time using examples 1-3 for a 50-60 year old population with prolonged excessive work, excessive stress, poor concentration, poor sleep, physical and mental fatigue, and dull complexion, and the results are shown in Table 6.
Examples 1-3 experimental observations of 60-70 years old aged people with pale complexion, physical weakness, hypoqi, memory deterioration, and facial age spots show improved effects in oxidation resistance, aging resistance, and immunity improvement after three months using examples 1-3. The results are shown in Table 7.
Claims (5)
1. A natural food additive with antioxidant activity is characterized by being prepared by mixing and proportioning a black wolfberry anthocyanin extract purified substance, a black pearl tomato anthocyanin extract purified substance, a purple onion anthocyanin extract purified substance, a pomegranate peel tannin extract purified substance and a seaweed plant pigment extract purified substance according to the weight ratio of 30-40:30-40:10-20:2-10: 2-10;
the lycium ruthenicum anthocyanin extraction and purification compound is prepared by the following method: placing the lycium ruthenicum murr dried at low temperature into a tissue crushing extractor, adding 50% -70% ethanol solution with the solid-liquid ratio of 1: 2-1: 5 and the pH value of 3.0-4.0, and crushing and extracting for 1-3 min at room temperature; performing suction filtration, adding 50-65% ethanol solution with pH of 3.0-pH 4.0 and solid-liquid ratio of 1: 2-1: 5 into the obtained filter residue, performing ultrasonic extraction for 1-2 times at room temperature for 20-40 min each time by using an ultrasonic extraction method, and performing suction filtration; mixing the tissue disruption extract and the ultrasonic extract, and concentrating by flash evaporation at 50-60 deg.C to 1/2-1/5 of original volume to obtain concentrated solution; enriching and purifying the total anthocyanin by macroporous adsorption resin Diaion HP 2MGL column chromatography; eluting with distilled water, adjusting the flow rate to 0.5-1.5 mL/min until the eluent is colorless and transparent, eluting with 55-70% ethanol solution with pH of 2.0-4.0, and collecting mauve pigment eluent, wherein the flow rate of the eluent is controlled to 0.5-1.5 mL/min; vacuum concentrating or freeze drying the collected mauve eluate in a rotary evaporator to obtain dry powder of anthocyanin extract and purified product of Lycium ruthenicum Murr;
the black pearl tomato anthocyanin extract purified product is prepared by the following method: placing black pearl tomatoes into a tissue crushing extractor, adding 55-65% ethanol solution with the pH value of 2.0-pH value of 4.0 and the volume ratio of 1: 3-1: 6, crushing and extracting for 1-3 min at room temperature; squeezing with coarse cloth to obtain extractive solution; concentrating the extractive solution in 50-60 deg.C water bath with flash evaporation concentrating device under reduced pressure to 1/3-1/6 of original volume to obtain concentrated solution; enriching and purifying the total anthocyanin by macroporous adsorption resin Diaion HP 2MGL column chromatography; eluting with distilled water, adjusting the flow rate to 0.5-1.5 mL/min until the eluent is colorless and transparent, eluting with 55-70% ethanol solution with pH of 3.0-4.0, and collecting mauve pigment eluent, wherein the flow rate of the eluent is controlled to 0.5-1.5 mL/min; concentrating and drying the collected mauve eluent in a rotary evaporator in vacuum or freeze drying to obtain dry powder of anthocyanin extract and purified product of black pearl tomato;
the purple onion anthocyanin extracted and purified product is prepared by the following method: placing purple onions into a tissue crushing extractor, adding 50% -60% ethanol solution with the pH value of 3.0-4.0 and the volume ratio of 1: 3-1: 6, crushing and extracting for 1-3 min at room temperature; performing suction filtration, adding 50-70% ethanol solution with pH of 3.0-pH of 4.0 and volume ratio of 1: 3-1: 6 into the obtained filter residue, performing ultrasonic extraction for 20-40 minutes each time for 1 time at room temperature by adopting an ultrasonic extraction method, and performing suction filtration; mixing the tissue disruption extract and the ultrasonic extract, and concentrating by flash evaporation at 50-60 deg.C to 1/2-1/6 of original volume to obtain concentrated solution; concentrating and purifying the concentrated solution with macroporous adsorbent resin Diaion HP 2MGL column chromatography to obtain purple onion pigment; eluting with distilled water, adjusting the flow rate to 0.5-1.5 mL/min until the eluent is colorless and transparent, eluting with 55-70% ethanol solution with pH of 3.0-4.0, and collecting pigment eluent, wherein the flow rate of the eluent is controlled at 0.5-1.5 mL/min; and (3) concentrating and drying the collected pigment eluent in vacuum or freeze drying in a rotary evaporator to obtain the dry powder of the extracted and purified product of the anthocyanin of the black pearl tomato.
2. The natural food additive with antioxidant activity as claimed in claim 1, wherein the purified extract of pomegranate rind tannin is prepared by the following method: placing the pomegranate rind dried at low temperature into a tissue crushing extractor, adding 60% -70% acetone solution with a solid-to-liquid ratio of 1: 4-1: 8, crushing and extracting for 1-3 min at room temperature; suction filtering, flash evaporating and concentrating the extracting solution to 1/4-1/8 of the original volume by adopting a flash evaporation concentration device under the condition of 50-60 ℃, subjecting the obtained concentrated solution to Diaion HP 20 macroporous adsorption resin column chromatography, performing gradient elution by respectively using deionized water, 10% ethanol, 20% ethanol, 40% ethanol, 60% ethanol and 80% ethanol with the liquid-material ratio of 1:20-50 times, according to the flow rate of 2.5-10.0 mL/min, collecting each eluent, combining the 20% ethanol, 40% ethanol and 60% ethanol eluents, performing flash evaporation concentration in a water bath at 50-60 ℃ to obtain 1/10-1/20 of the original volume, and performing rotary evaporation concentration on the obtained concentrated solution by using a rotary evaporator to dry to obtain the purified dry powder of the pomegranate rintannin extract.
3. The natural food additive with antioxidant activity as claimed in claim 1, wherein the seaweed phytochrome extract purified product is prepared by the following method: placing the dried seaweed in a tissue crushing extractor, adding 70-80% of acetone with a solid-liquid ratio of 1: 3-1: 8, crushing and extracting for 1-3 min at room temperature; suction filtering, and flash evaporating and concentrating the extract at 50-60 deg.C until the concentrated solution does not contain acetone; then extracting with diethyl ether at a volume ratio of 1:1-1:3 for 2-6 times; mixing the ether extracts, steaming at 40 deg.C in water bath under normal pressure to remove ether, and freeze drying the obtained concentrated extract to obtain purified product of seaweed plant pigment.
4. The natural food additive with antioxidant activity as claimed in claim 1, wherein the weight ratio of the extracted and purified product of lycium ruthenicum anthocyanin, the extracted and purified product of black pearl tomato anthocyanin, the extracted and purified product of purple onion anthocyanin, the extracted and purified product of pomegranate peel tannin and the extracted and purified product of seaweed plant pigment is 32-38:32-38:12-18:3-8: 3-8.
5. The natural food additive with antioxidant activity as claimed in claim 4, wherein the weight ratio of the extracted and purified product of lycium ruthenicum anthocyanin, the extracted and purified product of black pearl tomato anthocyanin, the extracted and purified product of purple onion anthocyanin, the extracted and purified product of pomegranate peel tannin and the extracted and purified product of seaweed plant pigment is 34-35:34-35:15-16:4-5: 4-5.
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| CN110793963B (en) * | 2019-11-12 | 2022-04-22 | 光明乳业股份有限公司 | Acid indicator based on lycium ruthenicum anthocyanin extract and application thereof |
| CN111035693A (en) * | 2020-01-10 | 2020-04-21 | 中国科学院西北高原生物研究所 | Anthocyanin composition with good stability |
| CN111513147A (en) * | 2020-04-24 | 2020-08-11 | 上海应用技术大学 | Natural lard antioxidant and preparation method thereof |
| CN113729169A (en) * | 2021-08-12 | 2021-12-03 | 四川省食品发酵工业研究设计院有限公司 | Meat colorant with antibacterial effect and preparation method thereof |
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