CN111820314A - A kind of anti-fatigue health care product prepared by using deer blood polypeptide and wolfberry polysaccharide - Google Patents

Abstract

The invention provides a deer blood polypeptide. The prepared deer blood polypeptide and wolfberry polysaccharide are used as effective ingredients to prepare a composition with anti-fatigue effect; the prepared composition has a synergistic effect, and can be used for food and functional products for relieving fatigue. Foodstuffs, Foodstuffs for Special Medical Purposes and Medicines. In the invention, the deer blood polypeptide prepared by enzymatic hydrolysis of deer blood with collagenase and bromelain is used in combination with lycium barbarum polysaccharide. active.

Description

A kind of anti-fatigue health care product prepared by using deer blood polypeptide and wolfberry polysaccharide

technical field

The invention belongs to the technical field of active substance extraction and application, in particular to a deer blood polypeptide and an anti-fatigue health care product prepared by using the deer blood polypeptide and wolfberry polysaccharide jointly.

Background technique

Fatigue refers to the inability of the body to initiate or sustain a certain intensity of activity during physical activity. In today's society, due to the accelerated pace of work and life, irregular work and rest and other factors, many people have been under greater psychological pressure for a long time or are in a state of overworked overload. Dizziness, headache, loss of appetite, poor energy, memory loss, etc. According to a survey by the World Health Organization, more than 35% of people in the world are in a state of fatigue, especially among middle-aged men who are in a state of fatigue as high as 60%-75%. Fatigue can cause the accumulation of oxygen free radicals, immune dysfunction, etc., which can lead to the occurrence and development of other diseases, and seriously affect people's work and life. Therefore, products that can relieve human fatigue have attracted more and more attention.

Deer blood is also widely circulated among the people and is regarded as the top quality of traditional Chinese medicine. "Compendium of Materia Medica" is recorded as a great tonic for deficiency, nourishing essence and blood, detoxification of pox, drug poison and so on. In modern times, animal experiments and clinical studies have proved that deer blood has many physiological activities such as beauty, treatment of anemia, immune regulation, anti-aging, improvement of memory, anti-fatigue, and improvement of sexual function. However, if the deer blood dry powder is directly consumed as an anti-fatigue product, it will have a heavy bloody smell and poor palatability; in addition, it is not easy to be absorbed and digested by the human body, resulting in poor anti-fatigue effect, and the consumption needs to be increased; If the amount exceeds 20 grams, it will cause the user to experience discomfort such as getting angry.

Lycium barbarum polysaccharide is the most important active ingredient in Lycium barbarum. Studies have shown that Lycium barbarum polysaccharide has significant anti-fatigue, anti-aging and immune-enhancing effects. The prepared deer blood polypeptide and Lycium barbarum polysaccharide are used as active ingredients to study their anti-fatigue function, aiming to develop food, functional food, food for special medical purpose and medicine for relieving fatigue.

SUMMARY OF THE INVENTION

The purpose of the present invention is to provide a deer blood polypeptide. The prepared deer blood polypeptide and Lycium barbarum polysaccharide are used as active ingredients to prepare a composition with anti-fatigue effect; the prepared composition has a synergistic effect and can be used for relieving fatigue. Food, functional food, food for special medical purpose and medicine.

The present invention first provides a deer blood polypeptide, and its preparation method is as follows:

1) Homogenization: homogenize the deer blood to be extracted to prepare a homogenate;

The conditions of homogenization are as follows: the rotating speed of the high-speed shearing machine is 8000-12000rpm, and the shearing time is 30-50min;

2) Enzymatic hydrolysis: adding collagenase and bromelain to the homogenate in step 1) for enzymatic hydrolysis to obtain an enzymatic hydrolysis solution, and centrifuging to get the supernatant;

Preferably, the enzyme activity ratio of collagenase and bromelain is 1:10;

The enzymolysis conditions are 3-5h at 35-55°C;

As a specific record of the embodiment, enzymatic hydrolysis was performed at 50°C for 5h;

The rotating speed of described centrifugation is >6000rpm;

3) Membrane filtration: the supernatant prepared in step 2) is first filtered with a membrane with a molecular weight of 3500Da, and then with a 160Da membrane, and the filtrate is the deer blood polypeptide solution;

4) Drying: the deer blood polypeptide concentrate in step 3) is spray-dried to obtain deer blood polypeptide powder.

The deer blood polypeptide provided by the present invention is used for preparing food or health care product with anti-fatigue effect;

Another aspect of the present invention provides an anti-fatigue health care product, which is a deer blood polypeptide prepared by the above method;

Lycium barbarum polysaccharide is also added to the above anti-fatigue health care products.

The deer blood polypeptide prepared by the invention and the wolfberry polysaccharide are used in combination, and the anti-fatigue activity of the combination of the deer blood polypeptide and the wolfberry polysaccharide is obviously higher than that of the single deer blood polypeptide and the wolfberry polysaccharide.

Description of drawings

Figure 1: Diagram of the exhaustive swimming experiment in mice.

Detailed ways

Deer blood polypeptide is a polypeptide with a molecular weight of 500-2000 Da, which is hydrolyzed by deer blood protease, and is the main active substance in deer blood that exerts the above functions. Deer blood protease hydrolysis is not only beneficial to the body's absorption, but also can improve the physiological activity. In existing studies, papain and alkaline protease were used to hydrolyze camel blood, and the anti-fatigue activity of the prepared polypeptide was not good. Therefore, the present invention takes red deer blood as the research object, systematically investigates the appropriate protease and proportion for enzymatic hydrolysis of deer blood protein, and deer blood is efficiently and reasonably decomposed into active peptides with the highest anti-fatigue activity.

The present invention will be described in detail below with reference to the embodiments.

Example 1 Screening of Deer Blood Polypeptide Preparation Method

10kg of deer blood was taken, homogenized by high-speed shearing, added with 3% of the weight of deer blood with different enzymes, and enzymatically hydrolyzed at 50 °C for 3 hours. The swimming time of the mice was significantly increased (Table 1), indicating that the above two enzymatic hydrolysates have anti-fatigue effects.

Table 1 The exhaustive swimming experiment of rats with different enzymolysis products

*p<0.05, the difference is significant

In order to further improve the yield and anti-fatigue effect of enzymolysis products, collagenase and bromelain were compounded, and the anti-fatigue effect was screened by changing the ratio of enzyme activity.

The results showed that when the ratio of collagenase and bromelain enzyme activities was 1:10, the anti-fatigue activity of the composition was the best (as shown in Table 2). , and then separated with a 160Da membrane to obtain three components of enzymatic hydrolysis products, by screening the activities of the three groups of products

Table 2: Rat exhaustive swimming experiment of enzymatic hydrolysis products with different enzyme activity ratios

*p<0.05, the difference is significant

It was found that the products with molecular weights between 160Da-3500Da had the best anti-fatigue effect (as shown in Table 3). Finally, the enzymatic hydrolysis product of deer blood polypeptide was obtained, and 0.85 kg of deer blood polypeptide powder was obtained by spray drying.

Table 3: Rat exhaustive swimming experiment with different molecular weight polypeptides

*p<0.05, the difference is significant

Finally, the specific steps of the preparation method of the deer blood polypeptide with anti-fatigue effect determined in the present invention are as follows:

1) Homogenization: homogenize the deer blood to be extracted to prepare a homogenate;

A specific condition of homogenization is as follows: the rotating speed of the high-speed shearing machine is 8000-12000rpm, and the shearing time is 30-50min;

2) enzymolysis: in the homogenate of step 1), add collagenase and bromelain to carry out enzymolysis, obtain enzymolysis solution, and centrifuge to get supernatant; wherein the ratio of the addition of collagenase and bromelain in homogenate is 1: 50-5:1, the enzymolysis conditions are 3-5h at 35-55°C; the speed of the centrifugation is >6000rpm;

3) Membrane filtration: the supernatant liquid prepared in step 2) is first filtered with a membrane with a molecular weight of 3500Da, and then with a membrane of 160Da, and the filtrate is the deer blood polypeptide solution;

4) Drying: the deer blood polypeptide concentrate in step 3) is spray-dried to obtain deer blood polypeptide powder.

Example 2:

Using the method determined in Example 1, the deer blood polypeptide was prepared under the above conditions, and the content of the polypeptide was 87.65% as determined by Kjeldahl method, and the polypeptide contained glycine, glutamic acid, arginine, lysine, tyrosine amino acid, the total amino acid content (TAA) reached 75.58%, of which the essential amino acid (EAA) content was 36.51%, the EAA/TAA value was 49.13%, and the EAA/NEAA value was 98%; the first limiting amino acid was tryptophan.

Table 4: Deer blood polypeptide amino acid content (unit: %)

The results of exhaustive swimming experiment (Fig. 1) showed that the swimming time of mice given deer blood polypeptide was significantly enhanced (P<0.05), and physical fatigue could be better relieved at the three dose levels. In particular, at the same concentration and dose, the effect of using deer blood meal is significantly lower than that of deer blood polypeptide. Therefore, by the method determined in this Example 1, the obtained deer blood polypeptide significantly enhanced its effect of relieving physical fatigue.

Table 5: Peptide and Deer Blood Meal Rats Exhausted Swimming Experiment

*p<0.05, the difference is significant

Example 3: Preparation of Lycium barbarum polysaccharide

Take 5 kilograms of wolfberry fruit, wash with tap water and purified water in turn, pulverize after drying, and pass through a 15-mesh sieve. Add the crushed and sieved wolfberry fruit into the subcritical extraction device, and then add 25m ³ of butane. After 10 min of nitrogen gas, subcritical extraction was performed. The extraction pressure was 5MPa, and the extraction time was 30min. The solvent was removed, and the fat-soluble components of wolfberry and the residue of wolfberry were collected. Add 5 times of deionized water to the above-mentioned wolfberry slag, and shear it with a high-speed shearing machine for 30 minutes to obtain a wolfberry soaking solution, and the shearing speed is 10,000 rpm. Transfer the sheared medlar soaking solution to the extraction tank, and then add 7 times the volume of deionized water to extract for 2 hours, and the extraction temperature is 60 ° C; for the second extraction, add 10 times the amount of deionized water. After the extraction, centrifuge with a tube centrifuge at 16,000 rpm. After 2 extractions, the extraction solution I was obtained by combining. The above-mentioned extract I was passed through a microfiltration membrane system with a molecular weight of 1 million at a temperature of 50° C. and a pressure of 2 bar. Collect the extract that passes through the microfiltration membrane to obtain the extract II. The above-mentioned extract II was concentrated through an ultrafiltration membrane system with a molecular weight of 1000, the filtration temperature was 30 °C, and the pressure was 10 bar. Collect the extraction solution retained by the ultrafiltration membrane to obtain extraction solution III. Continue to concentrate the extract III under reduced pressure, and the temperature is less than 60°C. When concentrated to a volume of 2.5 times the mass of the raw material of wolfberry, it is placed in an alcohol precipitation tank. While stirring, ethanol was added to make the ethanol concentration reach 75%, and then the stirring was continued for 30 min. Still 8h. The supernatant was separated and recovered, and the precipitate was air-dried for 12 h at a drying temperature of 65°C. Obtain brown wolfberry polysaccharide. The above-mentioned products are determined by the phenol-sulfuric acid method, and the content of Lycium barbarum polysaccharide is 53.21%.

Example 4 Preparation of anti-fatigue chewable tablets

Mix 750g deer blood polypeptide, 750g wolfberry polysaccharide, 500g mannitol, 500g xylitol, 500g lactose, 8.5g citric acid, add 2.5% povidone and 3% carboxymethyl cellulose calcium to granulate, 60 ℃ Dry in vacuo to give dry granules. The dry granules are sorted through a 12-mesh nylon sieve, then the granules are fully mixed with 10.5g of magnesium stearate and 10.5g of talc, and then passed through a 12-mesh nylon sieve. bacteria and packaging to obtain chewable tablets.

The quality inspection of the prepared chewable tablets is as follows:

1. Sensory indicators

Table 6: Chewable Tablets Sensory Quality Analysis Results

2. Physical and chemical indicators:

Chewable tablets had an average tablet weight of 1 g/tablet, and the difference in tablet weight was <3%. The heavy metal content of the chewable tablet was determined according to the provisions of the Chinese Pharmacopoeia, and the results were as follows: lead < 0.2 mg/kg, arsenic < 0.1 mg/kg.

According to the microbial limit test method specified in the Chinese Pharmacopoeia, the chewable tablets were tested for microorganisms, and the test results were as follows: the total number of bacteria <1000cfu/g, the total number of molds <20, and no coliform bacteria, Staphylococcus aureus and Salmonella were detected.

3. Hardness index:

The hardness of the chewable tablet is tested, and the test result is about 4kg, which fully complies with the tablet regulations.

Example 5: Effect of anti-fatigue chewable tablet

The chewable tablets prepared in Example 4 were taken and dissolved in distilled water; the total concentrations of deer blood polypeptide and wolfberry polysaccharide were adjusted to be 200 mg/ml, 400 mg/ml and 800 mg/ml, respectively. Samples are available now. Seventy-five rats were randomly divided into 5 groups. The blank control group was not given any operation. The other groups were given the corresponding drug 1ml/100g by gavage every day, once/d, for 30 consecutive days. 24 hours after the last administration of the test samples, the rats with tails bearing 3% body weight lead skin were used to swim in the swimming box. The water depth is not less than 50cm, and the water temperature is 31℃±1.0℃. When the rat swims to the bottom of the water for 3 consecutive times for more than 10 seconds each time, it is regarded as exhausted.

Table 7: Deer blood polypeptide and Lycium barbarum polysaccharide composition of rat exhaustive swimming test table

*p<0.05, the difference is significant

The data in Table 5 shows that the exhaustive swimming time of the rats in the high-dose group was significantly higher than that in the blank control group, indicating that the high-dose deer blood polypeptide and Lycium barbarum polysaccharide have anti-fatigue effects.

Claims (8)

1. a deer blood polypeptide, is characterized in that, the preparation method of described deer blood polypeptide is as follows: 1) Homogenization: homogenize the deer blood to be extracted to prepare a homogenate; 2) Enzymatic hydrolysis: adding collagenase and bromelain to the homogenate in step 1) for enzymatic hydrolysis to obtain an enzymatic hydrolysis solution, and centrifuging to get the supernatant; 3) Membrane filtration: the supernatant prepared in step 2) is first filtered with a membrane with a molecular weight of 3500Da, and then with a 160Da membrane, and the filtrate is the deer blood polypeptide solution; 4) Drying: the deer blood polypeptide concentrate in step 3) is spray-dried to obtain deer blood polypeptide powder. 2. deer blood polypeptide as claimed in claim 1, is characterized in that, the condition of homogenization in the step 1) of described preparation method is as follows: high-speed shearing machine rotating speed is 8000-12000rpm, and shearing time is 30-50min . 3. deer blood polypeptide as claimed in claim 1, is characterized in that, in the step 2) of described preparation method, the enzyme activity ratio of collagenase and bromelain is 1:10. 4 . The deer blood polypeptide according to claim 1 , wherein the enzymatic hydrolysis condition in step 2) of the preparation method is enzymatic hydrolysis at 50° C. for 5 h. 5 . 5. deer blood polypeptide as claimed in claim 1, is characterized in that, in step 2) of described preparation method, the rotating speed of centrifugation is >6000rpm. 6. The application of the deer blood polypeptide according to any one of claims 1-5 in the preparation of food or health care product with anti-fatigue effect. 7. An anti-fatigue health product, characterized in that, the anti-fatigue health product comprises the deer blood polypeptide according to any one of claims 1-5. 8. The anti-fatigue health-care product according to claim 7, wherein the anti-fatigue health-care product is further added with Lycium barbarum polysaccharide.

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