Camel placenta hydrolysate as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of preparation of peptides, and particularly relates to camel placenta hydrolysate as well as a preparation method and application thereof. According to the preparation method of the camel placenta hydrolysate and the preparation method of the camel placenta hydrolysate provided by the invention, the camel placenta hydrolysate has remarkable scavenging activity on DPPH, ABTS, hydroxyl free radicals and iron ions, so that the antioxidant activity is exerted; in a fruit fly acute oxidative damage model, the camel placenta hydrolysate can prolong the survival time of fruit flies in an oxidative stress environment; in a normal fruit fly model, the camel placenta hydrolysate can prolong the life of normal aging fruit flies, improve the action ability of the aging fruit flies and play an anti-aging role by improving the activity of in-vivo antioxidant enzymes such as total superoxide dismutase, copper-zinc superoxide dismutase and catalase and reducing the malondialdehyde level on the premise of not reducing the food intake of the fruit flies. The camel placenta hydrolysate can be used as an ideal auxiliary antioxidant and anti-aging medicine and health food raw material.

Description

Technical Field

[0001] The present invention belongs to the technical field of peptide preparation, and particularly relates to a camel placenta hydrolyzate, a preparation method and an application thereof. Background Art

[0002] Aging is an inevitable law in the biological life process. With the development of science and the improvement of living standards, the research on anti-aging drugs has gradually become very important. Anti-aging drugs can be divided into natural anti-aging drugs and synthetic anti-aging drugs. Synthetic anti-aging drugs are widely used because of their low price, but their safety has been questioned. The research on natural anti-aging drugs has gradually become a new research hotspot. Among them, animal-derived natural anti-aging drugs include carnosine, chitosan, antioxidant enzymes, etc., all of which show good anti-aging effects.

[0003] The placenta is the afterbirth during the delivery of mammals. It is a transitional organ for the exchange of substances between the mother and the fetus. It is produced by the connection between the allantoic chorion and the uterine mucosa, and has functions such as gas exchange, providing nutrition, metabolism, defense, immunity and endocrine. With the continuous expansion of the camel breeding scale, the camel placenta resources have become more and more abundant. The research on camel placenta hydrolyzate in the prior art takes the peptide yield as an index, and there is no research on camel placenta hydrolyzate with both anti-aging and antioxidant effects. Summary of the Invention

[0004] The purpose of the present invention is to provide a camel placenta hydrolyzate, a preparation method and an application thereof. The prepared camel placenta hydrolyzate has significant auxiliary antioxidant and anti-aging activities.

[0005] In order to solve the above technical problems, the following technical solutions are proposed:

[0006] The present invention provides a preparation method of a camel placenta hydrolyzate, comprising: hydrolyzing a camel placenta with alkaline protease to obtain a camel placenta extract; the mass ratio of the alkaline protease to the camel placenta is (0.025 - 0.1):100.

[0007] Preferably, the temperature of the hydrolysis is 40°C, and the time of the hydrolysis is 2 - 4 h.

[0008] Preferably, after the hydrolysis, it further includes inactivating the enzyme and centrifuging, and the obtained supernatant is the camel placenta hydrolyzate; the temperature of the inactivation treatment is 85 - 100°C, and the time is 15 min;

[0009] Before the camel placenta is hydrolyzed, it further includes homogenizing; the rotation speed of the homogenization is 3000 - 8000 rpm, and the time is 5 - 15 min.

[0010] The present invention provides a method for preparing camel placenta hydrolyzate, comprising: concentrating and drying the camel placenta hydrolyzate successively to obtain camel placenta hydrolyzate; the camel placenta hydrolyzate is obtained by the preparation method described in the above technical solution.

[0011] Preferably, the concentration method includes vacuum concentration.

[0012] Preferably, the drying method includes freeze-drying; the temperature of the freeze-drying is -55 to -45 °C, and the vacuum degree is 10 to 30 Pa.

[0013] The present invention provides camel placenta hydrolyzate prepared by the preparation method described in the above technical solution, and the protein content of the camel placenta hydrolyzate is 8% to 12%. 。

[0014] The present invention provides the application of the camel placenta hydrolyzate prepared by the preparation method described in the above technical solution or the camel placenta hydrolyzate prepared by the preparation method described in the above technical solution in the preparation of antioxidant products and / or the preparation of anti-aging products.

[0015] The present invention provides the application of the preparation method of the camel placenta hydrolyzate described in the above technical solution or the preparation method of the camel placenta hydrolyzate described in the above technical solution in any one or more of the following 1) to 3):

[0016] 1) Improving the antioxidant property of camel placenta hydrolyzate;

[0017] 2) Improving the anti-aging performance of camel placenta hydrolyzate;

[0018] 3) Improving the hydrolysis degree of camel placenta hydrolyzate.

[0019] The present invention provides a product for assisting antioxidant and / or anti-aging, comprising the camel placenta hydrolyzate described in the above technical solution.

[0020] The beneficial effects of the present invention: The present invention provides a method for preparing camel placenta hydrolyzate, and the obtained camel placenta extract has the effects of antioxidant and anti-aging.

[0021] The present invention also provides a method for preparing camel placenta hydrolysate. Through experimental research, it is found that in vitro, the obtained camel placenta hydrolysate has significant scavenging activities against DPPH, ABTS, hydroxyl radicals and ferric ions, thus exerting antioxidant activity; in the in vivo Drosophila acute oxidative damage model, the prepared camel placenta hydrolysate can extend the survival time of Drosophila in an oxidative stress environment; in the in vivo normal Drosophila model, the prepared camel placenta hydrolysate can, without reducing the food intake of Drosophila, extend the lifespan of normal aging Drosophila, improve the locomotor ability of aging Drosophila, and exert anti-aging effects by increasing the activities of in vivo antioxidant enzymes such as total superoxide dismutase (T-SOD), copper-zinc superoxide dismutase (Cu-Zn SOD), catalase (CAT), etc. and reducing the malondialdehyde (MDA) level. The camel placenta hydrolysate can be used as an ideal raw material for auxiliary antioxidant, anti-aging drugs and health foods. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 It is a graph showing the effects of different addition amounts of alkaline protease on the antioxidant capacity and yield of camel placenta extract;

[0023] Figure 2 It is a graph showing the effect of camel placenta hydrolysate on the crawling ability of Drosophila; among them, a is female Drosophila and b is male Drosophila;

[0024] Figure 3 It is a graph showing the survival curve of Drosophila treated with camel placenta hydrolysate;

[0025] Figure 4 It is a graph showing the effect of camel placenta hydrolysate on the T-SOD activity in Drosophila; on the left is after feeding for 25 days, and on the right is after feeding for 45 days;

[0026] Figure 5 It is a graph showing the effect of camel placenta hydrolysate on the Cu-Zn SOD activity in Drosophila; on the left is after feeding for 25 days, and on the right is after feeding for 45 days;

[0027] Figure 6 It is a graph showing the effect of camel placenta hydrolysate on the CAT activity in Drosophila; on the left is after feeding for 25 days, and on the right is after feeding for 45 days;

[0028] Figure 7 It is a graph showing the effect of camel placenta hydrolysate on the MDA level in Drosophila; on the left is after feeding for 25 days, and on the right is after feeding for 45 days;

[0029] Figure 8 It is a graph showing the effect of camel placenta hydrolysate on the lifespan of Drosophila with acute hydrogen peroxide injury; on the left is female Drosophila (Female), and on the right is male Drosophila (Male);

[0030] Figure 9Degree of hydrolysis of camel placenta extract by different proteases;

[0031] Figure 10 Influence diagram of ABTS radical scavenging activity of camel placenta extract by different proteases. Specific implementation mode

[0032] The present invention provides a preparation method of a camel placenta hydrolysate, including: hydrolyzing camel placenta with alkaline protease to obtain a camel placenta extract. The alkaline protease of the present invention improves the antioxidant activity of the camel placenta extract.

[0033] As an alternative implementation mode, the camel placenta in the present invention is a fresh camel placenta, and the fresh camel placenta is a dehydrated-free, odor-free, clean and healthy camel placenta. The present invention has no special limitation on the source of the camel placenta, and conventional commercially available products can be used.

[0034] As an alternative implementation mode, before hydrolysis, the present invention also includes homogenizing the camel placenta to obtain a homogenate. As an alternative implementation mode, the rotation speed of the homogenization in the present invention is 3000-8000 rpm, and can also be 4500-7000 rpm, more preferably 6000 rpm; the time of the homogenization is 5-15 min, and can also be 8-12 min, more preferably 10 min. The homogenization mode in the present invention is high-speed homogenization, and the high-speed homogenization is beneficial to the hydrolysis of camel placenta and improves the hydrolysis efficiency.

[0035] As an alternative implementation mode, after adjusting the pH of the homogenate, the present invention mixes the alkaline protease with the homogenate and performs hydrolysis to obtain a hydrolysate. The pH of the homogenate in the present invention is adjusted to 10.0, and the reagent used for the adjustment includes sodium hydroxide solution. In the initial hydrolysis in the present invention, the mass ratio of the alkaline protease to the camel placenta is (0.025-0.1):100, and can also be (0.05-0.8):100. The enzyme activity of the alkaline protease added in the present invention is 200 U / mg; the hydrolysis time is 2-4 h, more preferably 3 h, and the hydrolysis temperature is 40°C. The selection of the hydrolysis temperature and time can improve the activity of the alkaline protease, and the hydrolysis degree and antioxidant and anti-aging properties of the camel placenta can be improved under the combined action of the alkaline protease, hydrolysis temperature and time.

[0036] As an alternative embodiment, the enzymes in the hydrolyzate are inactivated and centrifuged, and the obtained supernatant is the camel placenta hydrolyzate. The temperature of the inactivation treatment in the present invention is 85 - 100 °C, or can also be 90 - 98 °C; the time of the inactivation treatment is 15 min. Performing the inactivation treatment can eliminate enzyme activity and prevent the enzymes from affecting subsequent processes. The rotation speed of the centrifugation in the present invention is 3000 - 8000 rpm, or can also be 4500 - 7000 rpm, more preferably 6000 rpm; the time of the centrifugation is 5 - 15 min, or can also be 8 - 12 min, more preferably 10 min.

[0037] The camel placenta hydrolyzate prepared by the present invention has good antioxidant properties and also has the effect of anti - aging.

[0038] The present invention provides a preparation method of camel placenta hydrolyzate, including: successively concentrating and drying the camel placenta hydrolyzate to obtain camel placenta hydrolyzate; the camel placenta hydrolyzate is obtained by the preparation method described in the above technical solution.

[0039] As an alternative embodiment, the concentration method in the present invention includes vacuum concentration. The temperature of the vacuum concentration is not higher than 70 °C, and the vacuum concentration is to 1 / 2 of the original volume. The concentration in the present invention can reduce the material processing amount in the subsequent drying process. The content of protein in the concentrated hydrolyzate obtained in the present invention is high. The protein content in the concentrated hydrolyzate is measured to be 10% by the BCA method, and the BCA protein quantification kit is used for the BCA method measurement.

[0040] As an alternative embodiment, the concentrated product in the present invention is dried to obtain a dried product. The drying method in the present invention includes freeze - drying. The temperature of the freeze - drying is - 55 - - 45 °C, or can also be - 53 - - 48 °C, more preferably - 50 °C; the vacuum degree of the freeze - drying is 10 - 30 Pa, or can also be 18 - 22 Pa. The drying can improve the stability and activity of the camel placenta hydrolyzate, and is easy to store and transport.

[0041] As an alternative embodiment, the dried product in the present invention is sieved to obtain camel placenta hydrolyzate. There are no special requirements for the mesh number of the sieving in the present invention, and it can be selected according to actual needs. There are no special limitations on the sieving method in the present invention, and conventional methods can be used. As an alternative embodiment, the camel placenta hydrolyzate can also be packaged and stored in the present invention.

[0042] The present invention prepares camel placenta hydrolyzate through crushing, alkaline protease hydrolysis, centrifugation, concentration, freeze - drying and sieving. The camel placenta hydrolyzate of the present invention is a mixture containing various camel placenta peptides, and the prepared camel placenta hydrolyzate has significant auxiliary antioxidant and anti - aging activities.

[0043] The present invention provides a camel placenta hydrolyzate prepared by the preparation method described in the above technical solution. The protein mass content of the camel placenta hydrolyzate is 8% - 12%, more preferably 10%. The camel placenta hydrolyzate prepared by the present invention contains a variety of camel placenta peptides and has good auxiliary antioxidant and anti-aging activities, and can be used as an ideal raw material for auxiliary antioxidant and anti-aging drugs and health foods.

[0044] The present invention provides the use of the camel placenta hydrolyzate prepared by the preparation method described in the above technical solution or the camel placenta hydrolyzate prepared by the preparation method described in the above technical solution in the preparation of antioxidant products. As an alternative embodiment, the products of the present invention include drugs and / or health products; the present invention has no special limitation on the preparation method of the products, and conventional methods can be used. The product dosage forms of the present invention include but are not limited to tablets, granules, oral liquids and sprays. The antioxidant of the present invention includes increasing the activity of oxidase and / or reducing the level of malondialdehyde; the oxidase includes one or more of superoxide dismutase, copper-zinc superoxide dismutase and catalase.

[0045] The present invention provides the use of the camel placenta hydrolyzate prepared by the preparation method described in the above technical solution or the camel placenta hydrolyzate prepared by the preparation method described in the above technical solution in the preparation of anti-aging products. As an alternative embodiment, the products include drugs and / or health products; the present invention has no special limitation on the preparation method of the products, and conventional methods can be used. The product dosage forms of the present invention include but are not limited to tablets, granules, oral liquids and sprays. In the normal Drosophila model, the camel placenta hydrolyzate of the present invention can, without reducing the food intake of Drosophila, prolong the lifespan of normally aging Drosophila, improve the locomotor ability of aging Drosophila, and play an anti-aging role by increasing the activities of antioxidant enzymes in vivo such as total superoxide dismutase, copper-zinc superoxide dismutase and catalase and reducing the level of malondialdehyde.

[0046] The present invention provides the use of the preparation method of the camel placenta hydrolyzate described in the above technical solution or the preparation method of the camel placenta hydrolyzate described in the above technical solution in improving the antioxidant property of the camel placenta hydrolyzate. The antioxidant of the present invention includes increasing the activity of oxidase and / or reducing the level of malondialdehyde; the oxidase includes one or more of superoxide dismutase, copper-zinc superoxide dismutase and catalase.

[0047] The present invention provides the use of the preparation method of the camel placenta hydrolyzate described in the above technical solution or the preparation method of the camel placenta hydrolyzate described in the above technical solution in improving the degree of hydrolysis of the camel placenta hydrolyzate.

[0048] The present invention provides the use of the preparation method of the camel placenta hydrolyzate described in the above technical solution or the preparation method of the camel placenta hydrolyzate described in the above technical solution in improving the anti-aging performance of the camel placenta hydrolyzate.

[0049] The present invention provides a product for assisting antioxidant and / or anti-aging, which comprises the camel placenta hydrolyzate described in the above technical solution. As an optional embodiment, the product comprises a drug and / or a health product; the present invention has no special limitation on the preparation method of the product, and a conventional method can be adopted. The dosage forms of the product of the present invention include but are not limited to tablets, granules, oral liquids and sprays.

[0050] In vitro studies have found that the camel placenta hydrolyzate prepared by the present invention has significant scavenging activities against DPPH, ABTS, hydroxyl radicals and ferric ions, thereby exerting antioxidant activity. In the in vivo Drosophila acute oxidative damage model, the camel placenta hydrolyzate prepared by the present invention can prolong the survival time of Drosophila in an oxidative stress environment; in the in vivo normal Drosophila model, the prepared camel placenta hydrolyzate can, without reducing the food intake of Drosophila, prolong the lifespan of normal aging Drosophila by increasing the activities of in vivo antioxidant enzymes such as total superoxide dismutase (T-SOD), copper-zinc superoxide dismutase (Cu-Zn SOD), catalase (CAT), etc. and reducing the level of malondialdehyde (MDA), and improve the locomotor ability of aging Drosophila, thereby exerting an anti-aging effect.

[0051] According to the "Compendium of Materia Medica" records, "it tastes sweet, salty and warm in nature, enters the lung, liver and kidney meridians, and has the effects of nourishing qi and blood, and strengthening the kidney and replenishing essence", and is listed as a top-grade medicinal material. Existing studies have shown that the placenta contains many active ingredients, which have functions such as promoting growth, anti-fatigue, anti-aging, enhancing immunity, anti-inflammatory, and promoting wound healing. It also contains rich nutritional ingredients, mainly regulatory peptides, hormones, growth factors, cytokines, enzymes and enzyme inhibitors, amino acids, trace elements, vitamins, collagen, etc. And modern nutritional research has found that after humans ingest proteins, they need to be enzymatically hydrolyzed into low peptide forms by various enzymes in the digestive tract for absorption, and only a very small amount is absorbed in the form of free amino acids. Moreover, low peptides are digested faster and absorbed more than free amino acids, and have biological potency and nutritional value. Therefore, the present invention processes and prepares camel placenta into placenta peptides and conducts in vitro and in vivo experiments, and finds that the camel placenta hydrolyzate has the effects of assisting antioxidant and anti-aging.

[0052] In order to further illustrate the present invention, the technical solutions provided by the present invention will be described in detail below with reference to the accompanying drawings and embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0053] Example 1-1

[0054] The preparation method of the camel placenta hydrolyzate is as follows:

[0055] (1) Crushing: crushing fresh camel placenta by high-speed homogenization to prepare camel placenta homogenate; the speed of high-speed homogenization is 6000 rpm, and the time is 10 min;

[0056] (2) Protease hydrolysis: NaOH solution was added to the camel placenta homogenate to adjust the pH to 10.0, and then 0.025% alkaline protease by weight of the camel placenta homogenate was added, followed by hydrolysis at 40°C for 3.268 hours, and then heated to 90°C for 15 minutes for inactivation; the enzymatic activity of the added alkaline protease was 200 U / mg;

[0057] (3) Centrifugation: The hydrolyzate obtained in step (2) is centrifuged at high speed to retain the supernatant; the centrifugation speed is 6000 rpm, the time is 10 min, and the supernatant obtained is camel placenta hydrolyzate. The protein content in the concentrated hydrolyzate is determined by the BCA method, and the BCA method is determined using a BCA protein quantitative kit.

[0058] Example 1-2

[0059] (1) Concentration: The supernatant obtained in Example 1-1 was concentrated under reduced pressure at 60° C.; the temperature of the concentrated under reduced pressure was not higher than 70° C., and the supernatant was concentrated under reduced pressure to 1 / 2 of the original volume;

[0060] (2) Drying: freeze-dry the concentrated hydrolyzate using a freeze dryer; freeze-drying parameters are -50±5°C and 10-30Pa;

[0061] (3) Sieving: After the dry powder is sieved, it is packaged and stored to obtain camel placenta hydrolyzate.

[0062] Examples 1-3

[0063] Same as Example 1-1, the only difference is that the mass ratio of alkaline protease to camel placenta is 0.05:100.

[0064] The obtained supernatant is camel placenta hydrolyzate, and the camel placenta hydrolyzate is concentrated, dried and sieved according to the scheme of Example 1-2 to obtain camel placenta hydrolyzate.

[0065] Examples 1-4

[0066] Same as Example 1-1, the only difference is that the mass ratio of alkaline protease to camel placenta is 0.1:100.

[0067] The obtained supernatant is camel placenta hydrolyzate, and the camel placenta hydrolyzate is concentrated, dried and sieved according to the scheme of Example 1-2 to obtain camel placenta hydrolyzate.

[0068] Examples 1-5

[0069] Same as Example 1-1, the only difference is that the mass ratio of the alkaline protease to the camel placenta is 0.25:100.

[0070] The obtained supernatant is the camel placenta hydrolyzate, and the camel placenta hydrolyzate is concentrated, dried and sieved according to the scheme of Example 1-2 to obtain the camel placenta hydrolyzate.

[0071] Comparative Example 1-1

[0072] Same as Example 1-1, the only difference is that the mass ratio of the alkaline protease to the camel placenta is 0.0125:100.

[0073] The obtained supernatant is the camel placenta hydrolyzate, and the camel placenta hydrolyzate is concentrated, dried and sieved according to the scheme of Example 1-2 to obtain the camel placenta hydrolyzate.

[0074] The ABTS scavenging rate and yield of the camel placenta hydrolyzates obtained in Example 1-2, Example 1-3, Example 1-4, Example 1-5 and Comparative Example 1-1 were measured. When measuring, the concentration of the camel placenta hydrolyzate was 1 mg / mL, and the results are shown in Figure 1 . Yield (%) = mass of freeze-dried powder of placenta hydrolyzate / mass of fresh placenta × 100.

[0075] According to Figure 1 It can be seen that the yield and antioxidant ability of the camel placenta extract first increase and then decrease with the increase of the enzyme addition amount. To a certain extent, increasing the enzyme dosage will accelerate the reaction rate, but after adding too much enzyme, competitive inhibition will occur, hindering the progress of the reaction. When the addition amount of alkaline protease is 0.05% of the mass of the camel placenta, the ABTS radical scavenging rate reaches the maximum, which is 69.63%, significantly higher than other groups (P < 0.05). At this time, the yield is 1.63%. After continuing to increase the addition amount of alkaline protease, although the yield increases to a certain extent, the antioxidant activity significantly decreases. Therefore, the enzyme addition amount range of 0.025% - 0.1% is selected for subsequent experiments.

[0076] Example 1-6

[0077] The preparation method of the camel placenta hydrolyzate is as follows:

[0078] (1) Crushing: The fresh camel placenta is broken by high-speed homogenization to make a camel placenta homogenate; the rotation speed of high-speed homogenization is 6000 rpm, and the time is 10 min;

[0079] (2) Protease hydrolysis: Add NaOH solution to the camel placenta homogenate to adjust the pH to 10.0, then add alkaline protease at 0.076% of the mass of the camel placenta homogenate. After that, hydrolyze at 40 °C for 3.268 h, and then heat to 90 °C and maintain for 15 min for inactivation treatment; the enzyme activity of the added alkaline protease is 200 U / mg;

[0080] (3) Centrifugation: Centrifuge the hydrolyzate obtained in step (2) at high speed and retain the supernatant; the centrifugation speed is 6000 rpm and the time is 10 min. The obtained supernatant is the camel placenta hydrolyzate. The protein content in the concentrated hydrolyzate is determined to be 10% by the BCA method, and the BCA protein quantification kit is used for the BCA method determination.

[0081] Examples 1 - 7

[0082] The supernatants obtained in Examples 1 - 6 are camel placenta hydrolyzates. The camel placenta hydrolyzates are concentrated, dried and sieved according to the schemes of Examples 1 - 2 to obtain camel placenta hydrolysates.

[0083] Example 2

[0084] Evaluate the in vitro antioxidant activity of the camel placenta hydrolysates prepared in Examples 1 - 7:

[0085] Prepare sample solutions with the camel placenta hydrolysates prepared in Examples 1 - 7, and the concentrations of the camel placenta hydrolysates in the sample solutions are 1, 2, 3, 4, and 5 mg / mL respectively.

[0086] DPPH free radicals exist in the form of single electrons and can pair electrons with antioxidant substances, causing the system to change from purple to yellow. Therefore, the free radical scavenging ability of DPPH is usually used to evaluate the activity of antioxidant substances. Mix 1 mL of sample solutions with different concentrations with 1 mL of DPPH ethanol solution, react in the dark for 30 min, centrifuge at 4000 r / min for 10 min, and measure the absorbance at 517 nm.

[0087] Use distilled water to replace the sample solution as a blank control, and calculate the DPPH free radical scavenging rate according to the following formula.

[0088] DPPH free radical scavenging rate = (A0 - Ai) / A0 × 100%;

[0089] In the formula, A0: absorbance of the blank control group; Ai: absorbance of the sample group.

[0090] The results are shown in Table 1. It can be seen that the prepared camel placenta hydrolysates have significant scavenging activity against DPPH free radicals and show a dose - dependence. The IC 50 value (i.e., the concentration required for the sample to scavenge 50% of DPPH free radicals) is 2.282 mg / mL.

[0091] Table 1 DPPH radical scavenging rate of camel placenta hydrolysates at different concentrations (%)

[0092]

[0093]

[0094] Note: Different lowercase letters represent significant differences at the P < 0.01 level.

[0095] Example 3

[0096] In vitro antioxidant activity evaluation of the camel placenta hydrolysates prepared in Examples 1 - 7:

[0097] The camel placenta hydrolysates prepared in Examples 1 - 7 were formulated into sample solutions, and the concentrations of the camel placenta hydrolysates in the sample solutions were 0.2, 0.4, 0.6, 0.8, and 1.0 mg / mL, respectively.

[0098] Preparation of ABTS test solution: ABTS stock solution (7.4 mmol / L, 0.4 mL): Take 0.0045 g of ABTS and 1.1025 mL of distilled water; K2S2O8 stock solution (2.6 mmol / L, 1.43 mL): Take 0.0025 g of K2S2O8 and add 3.575 mL of distilled water; Mix the ABTS stock solution and the K2S2O8 stock solution, and let it stand at room temperature in the dark for 12 h, then dilute it 50 times with absolute ethanol.

[0099] Detection of A0 value: Take 1.6 mL of this ABTS solution and mix it thoroughly with 0.6 mL of absolute ethanol, then detect the absorbance at 734 nm.

[0100] Detection of A value: Take 1.6 mL of ABTS test solution, mix it thoroughly with 0.6 mL of camel placenta hydrolysates at different gradient concentrations respectively, and then measure the absorbance at 734 nm.

[0101] ABTS radical scavenging rate = (A0 - A) / A0 × 100%;

[0102] The results are shown in Table 2. It can be seen that the prepared camel placenta hydrolysates have significant scavenging activity against ABTS radicals and show a dose - dependence.

[0103] Table 2 ABTS radical scavenging rate of camel placenta hydrolysates at different concentrations (%)

[0104] Concentration (mg / mL) 0.2 0.4 0.6 0.8 1 Clearance rate (%) <![CDATA[31.89±1.34 e > <![CDATA[50.57±3.75 d > <![CDATA[65.98±2.77 c > <![CDATA[75.24±1.97 b > <![CDATA[81.87±1.09 a >

[0105] Note: Different lowercase letters represent significant differences at the P < 0.01 level.

[0106] Example 4

[0107] The antioxidant activities in vitro of the camel placenta hydrolysates prepared in Examples 1 - 7 were evaluated:

[0108] The camel placenta hydrolysates prepared in Examples 1 - 7 were formulated into sample solutions, and the concentrations of the camel placenta hydrolysates in the sample solutions were 0.2, 0.4, 0.6, 0.8, and 1.0 mg / mL, respectively.

[0109] 0.1 mL of sample solutions with different concentrations was mixed with 1 mL of 6 mmol / L FeSO4 solution and 1 mL of 6 mmol / L H2O2 solution, reacted for 10 min, then 1 mL of 6 mmol / L salicylic acid solution was added, and heated and reacted at 37 °C for 30 min, and the absorbance at 510 nm was measured.

[0110] Distilled water was used to replace the sample solution as a blank control, and the hydroxyl radical scavenging rate was calculated according to the following formula.

[0111] Hydroxyl radical scavenging rate = (A0 - Ai) / A0 × 100%;

[0112] In the formula, A0: the absorbance at 510 nm of the blank control group; Ai: the absorbance at 510 nm of the sample group.

[0113] The results are shown in Table 3. It can be seen that the prepared camel placenta hydrolysates have significant scavenging activity against hydroxyl radicals and show a dose - dependence.

[0114] Table 3 Hydroxyl radical scavenging rates of camel placenta hydrolysates at different concentrations (%)

[0115] Concentration (mg / mL) 0.2 0.4 0.6 0.8 1 Clearance rate (%) <![CDATA[31.89±1.34 e > <![CDATA[50.57±3.75 d > <![CDATA[65.98±2.77 c > <![CDATA[75.24±1.97 b > <![CDATA[81.87±1.09 a >

[0116] Note: Different lowercase letters represent significant differences at the P < 0.01 level.

[0117] Example 5

[0118] The antioxidant activities in vitro of the camel placenta hydrolysates prepared in Examples 1 - 7 were evaluated:

[0119] The camel placenta hydrolysates (CPE) prepared in Examples 1 - 7 were formulated into sample solutions, and the concentrations of the camel placenta hydrolysates in the sample solutions were 1, 2, 3, 4, and 5 mg / mL, respectively.

[0120] Take 1 mL of sample solutions with different concentrations, and successively add 2.5 mL of 0.2 mol / L sodium phosphate buffer (pH 6.6) and 2.5 mL of 1% potassium ferricyanide solution by mass concentration. Mix well, react at 50 °C for 20 min, cool in an ice-water bath, add 2.5 mL of 10% trichloroacetic acid solution by mass concentration, and centrifuge (3000 r / min, 10 min). Take 1 mL of the supernatant, add 0.2 mL of 0.1% ferric chloride solution by mass concentration, mix well, add 1 mL of deionized water and shake well, keep in a constant temperature water bath at 50 °C for 10 min, zero with distilled water and perform colorimetric analysis at 700 nm.

[0121] Use distilled water to replace the sample solution as a blank control. The larger the absorbance value, the stronger the antioxidant ability.

[0122] The results are shown in Table 4. It can be seen that the prepared camel placenta hydrolysate has a significant scavenging activity on ferric ions and shows a dose-dependence.

[0123] Table 4 Fe 3+ Reducing power (%)

[0124] Concentration (mg / mL) 1 2 3 4 5 Absorbance value <![CDATA[0.1607±0.0047 e > <![CDATA[0.2177±0.0078 d > <![CDATA[0.272±0.0175 c > <![CDATA[0.3134±0.01 b > <![CDATA[0.3601±0.0115 a >

[0125] Note: Different lowercase letters represent significant differences at the P<0.01 level.

[0126] Example 6

[0127] Evaluation of the effect of the camel placenta hydrolysate prepared in Examples 1-7 on the food intake of Drosophila:

[0128] Blank group: Add 0.0% camel placenta hydrolysate to the culture medium to feed Drosophila. At the same time, add acid blue dye to the culture medium as a food tracer to quantify the amount of food ingested by Drosophila.

[0129] 0.25% hydrolysate peptide group (abbreviated as 0.25%): Add camel placenta hydrolysate with a mass concentration of 0.25% to the culture medium to feed Drosophila. At the same time, add acid blue dye to the culture medium as a food tracer to quantify the amount of food ingested by Drosophila.

[0130] 1% hydrolysate peptide group (abbreviated as 1%): Add camel placenta hydrolysate with a mass concentration of 1% to the culture medium to feed Drosophila. At the same time, add acid blue dye to the culture medium as a food tracer to quantify the amount of food ingested by Drosophila.

[0131] During the experiment, the fruit flies in each group were allowed to feed freely. There were 100 female fruit flies and 100 male fruit flies in each group. Through the feeding experiment, it was determined whether the extension of the lifespan of fruit flies by camel placental hydrolysate was related to a decrease in food intake. The results are shown in Table 5. For female fruit flies, compared with the control group, the food intake of 0.25% hydrolysate did not change significantly, while the food intake of female fruit flies with 1% hydrolysate increased significantly (P<0.05). For male fruit flies, the food intakes of both 0.25% hydrolysate and 1% hydrolysate increased significantly (P<0.01).

[0132] Table 5 Effects of camel placental hydrolysate on the food intake of fruit flies

[0133]

[0134] Note: Compared with the blank control group, * indicates a significant difference (P<0.05); compared with the blank control group, ## indicates a highly significant difference (P<0.01).

[0135] Example 7

[0136] The camel placental hydrolysate prepared in Examples 1-7 was evaluated for its effect on the crawling ability of fruit flies: The climbing ability of fruit flies can reflect the aging condition of fruit flies from the side.

[0137] The grouping was the same as in Example 6. 20 fruit flies were collected in each group and placed at the bottom of a small bottle. Then, a vertical distance of 6 cm was marked on the small bottle. The fruit fly tube was gently vibrated to make the fruit flies fall to the bottom of the tube while timing for 15 s, and the number of fruit flies that climbed to 6 cm was recorded. The crawling ability of fruit flies was expressed as the proportion of the number of fruit flies that climbed to 6 cm to the total number. The test was carried out at 0, 10, 20, 30, and 40 d. The results are as Figure 2 shown in Table 6. The camel placental hydrolysate can improve the decrease in the crawling ability of fruit flies caused by aging. At 30 d, 0.25% and 1% camel placental hydrolysate significantly improved the crawling ability of female fruit flies (P<0.05), and at 40 d, 0.25% and 1% camel placental hydrolysate more significantly improved the crawling ability of female fruit flies (P<0.01); at 30 d and 40 d, 0.25% camel placental hydrolysate significantly improved the crawling ability of male fruit flies (P<0.05), and at 40 d, the crawling ability of male fruit flies with 1% hydrolysate was more significantly improved compared with the control group (P<0.01). In summary, the camel placental hydrolysate has a restorative effect on the decline in motor ability caused by fruit fly aging. Figure 2 In the table, compared with the control group, * indicates a significant difference (P<0.05); ** indicates a highly significant difference (P<0.01).

[0138] Table 6 Effects of camel placental hydrolysate on the crawling ability of female fruit flies

[0139]

[0140]

[0141] Example 8

[0142] Evaluation of the effect of camel placenta hydrolysate prepared in Examples 1-7 on the lifespan of Drosophila melanogaster:

[0143] After female and male Drosophila melanogaster were cultured in the basal medium for about 10 days, the next generation of Drosophila melanogaster that laid eggs and reproduced was collected as experimental materials.

[0144] In the 0.25% dose group (0.25% group), Drosophila melanogaster was fed with camel placenta hydrolysate with a mass concentration of 0.25% added to the basal medium;

[0145] In the 1% dose group (1% group), Drosophila melanogaster was fed with camel placenta hydrolysate with a mass concentration of 1% added to the basal medium;

[0146] The blank control group was fed with Drosophila melanogaster using the basal medium;

[0147] The adult Drosophila melanogaster that emerged within 24 hours was anesthetized and then sorted by gender, and randomly divided into three groups. Each group had 10 tubes of male and female, with 20 Drosophila melanogaster in each tube. The three groups were the blank control group, the 0.25% dose group, and the 1% dose group. Drosophila melanogaster was raised in a climate incubator at a constant temperature of (25 ± 1) °C and a constant humidity of (60 ± 5)%, with 12 hours of light and 12 hours of darkness alternating per day. The same medium was replaced every 3 days and the number of dead Drosophila melanogaster was recorded, as shown in Table 7, and the survival curve is shown in Figure 3 . When all Drosophila melanogaster died, the survival experiment ended.

[0148] In the first 20 days of the experiment, the lifespan curves of each group were basically the same. However, after the 20th day of the experiment, compared with the control group, the lifespan of Drosophila melanogaster treated with camel placenta hydrolysate increased, and the lifespan curve shifted to the right, indicating that the effect of camel placenta hydrolysate on extending the lifespan of Drosophila melanogaster mainly occurred in the middle and late stages of Drosophila melanogaster survival.

[0149] As can be seen from Table 7, both the camel placenta hydrolyzate dosage groups (0.25% and 1%) can extend the maximum lifespan, median death time, and average lifespan of female and male Drosophila melanogaster. Among them, in female Drosophila melanogaster, the average lifespan of the 0.25% dosage group of female Drosophila melanogaster was significantly extended by 9.10% compared with the control group, and the average lifespan of the 1% dosage group of female Drosophila melanogaster was extended by 14.26% compared with the control group; 0.25% camel placenta hydrolyzate extended the median death time and maximum lifespan of female Drosophila melanogaster by 8.07% and 6.59% respectively compared with the control group; 1% camel placenta hydrolyzate extended the median death time and maximum lifespan of female Drosophila melanogaster by 11.98% and 9.97% respectively compared with the control group. In male Drosophila melanogaster, 0.25% and 1% camel placenta hydrolyzate extended the average lifespan of male Drosophila melanogaster by 8.74% and 12.53% respectively compared with the control group, the median death time by 5.13% and 9.97% respectively compared with the control group, and the maximum lifespan by 5.15% and 8.91% respectively compared with the control group. Camel placenta hydrolyzate can extend the lifespan of Drosophila melanogaster and delay the aging process of Drosophila melanogaster.

[0150] Table 7 Effects of Camel Placenta Hydrolyzate on the Lifespan of Drosophila melanogaster

[0151]

[0152]

[0153] Maximum lifespan: The average lifespan of the last 10% of the surviving Drosophila melanogaster, which is the arithmetic mean of the lifespans of the last 20 dead Drosophila melanogaster in each group.

[0154] Example 9

[0155] The 0.25% dosage group (abbreviated as 0.25%) fed Drosophila melanogaster with a basic medium supplemented with 0.25% camel placenta hydrolyzate by mass concentration;

[0156] The 1% dosage group (abbreviated as 1%) fed Drosophila melanogaster with a basic medium supplemented with 1% camel placenta hydrolyzate by mass concentration;

[0157] The blank control group (abbreviated as blank) fed Drosophila melanogaster with a basic medium;

[0158] Evaluation of the effect of camel placenta hydrolysate prepared in Examples 1-7 on the activity of T-SOD (total superoxide dismutase) in Drosophila melanogaster: Adult Drosophila melanogaster that emerged within 24 hours were anesthetized and sorted by gender, and then transferred to the blank control group, 0.25% dose group, and 1% dose group, respectively. There were 10 tubes for each gender in each group, with 20 flies in each tube. At 25 days and 45 days of culture, 5 tubes of flies were taken out, starved for 2 hours, anesthetized, weighed, and then sacrificed by quick freezing in liquid nitrogen. PBS was added according to the mass-to-volume ratio of 1:9 (w:v), and homogenized in an ice bath. Then, it was centrifuged at 4°C and 12,000 r / min for 10 minutes to obtain 10% tissue homogenate. The total SOD activity was measured according to the instructions of the total SOD activity detection kit. The results are shown in Figure 4 . Figure 4 The effect of camel placenta hydrolysate on the activity of T-SOD in Drosophila melanogaster (left: 25 days; right: 45 days), Figure 4 and in Table 8, compared with the blank control group, * indicates a significant difference (P < 0.05), **, ## indicate a highly significant difference (P < 0.01).

[0159] Table 8 Effect of camel placenta hydrolysate on the activity of T-SOD (total superoxide dismutase) in Drosophila melanogaster (unit: U / mg prot)

[0160]

[0161] It can be seen that at 25 days of age, compared with the blank control group, the T-SOD activity in female Drosophila melanogaster in the 0.25% dose group was significantly increased, with a significant difference (P < 0.05), and the T-SOD in female Drosophila melanogaster in the 1% dose group was significantly increased, with a highly significant difference (P < 0.01). For male Drosophila melanogaster, at 25 days of age, compared with the blank control group, the T-SOD activity in male Drosophila melanogaster in the 0.25% dose group increased slightly, but the difference was not significant (P > 0.05), and the T-SOD in male Drosophila melanogaster in the 1% dose group was significantly increased, with a highly significant difference (P < 0.01); at 45 days of age, compared with the blank control group, the T-SOD activity in female and male Drosophila melanogaster in each dose group was significantly increased, with a highly significant difference (P < 0.01).

[0162] Example 10

[0163] Evaluation of the effect of camel placenta hydrolysate prepared in Examples 1-7 on the activity of Cu-Zn SOD (copper-zinc superoxide dismutase) in Drosophila melanogaster:

[0164] The method of raising and treating Drosophila melanogaster was the same as in Example 9. The results are shown in Figure 5 , Figure 5 The effect of camel placenta hydrolysate on the activity of Cu-Zn SOD in Drosophila melanogaster (left: 25 days; right: 45 days), Figure 5Compared with the blank control group, *, # indicate significant differences (P < 0.05), **, ## indicate extremely significant differences (P < 0.01).

[0165] From Figure 5 As can be seen from Table 9, at 25 days old, compared with the blank control group, the activities of Cu-Zn SOD in female and male Drosophila melanogaster in the 0.25% dose group were significantly increased (P < 0.05), and those in the 1% dose group were extremely significantly increased (P < 0.01). At 45 days old, compared with the blank control group, the activity of Cu-Zn SOD in female Drosophila melanogaster in the 0.25% dose group was significantly increased (P < 0.05), and the activity of Cu-Zn SOD in male Drosophila melanogaster increased slightly, but the difference was not significant (P > 0.05); the activity of Cu-Zn SOD in female Drosophila melanogaster in the 1% dose group was significantly increased (P < 0.01), and the activity of Cu-Zn SOD in male Drosophila melanogaster was significantly increased (P < 0.05).

[0166] Table 9 Effects of camel placenta hydrolysate on the activity of Cu-Zn SOD in Drosophila melanogaster (unit: U / mgprot)

[0167]

[0168] Example 11

[0169] Evaluation of the effects of the camel placenta hydrolysate prepared in Examples 1-7 on the activity of CAT (catalase) in Drosophila melanogaster:

[0170] The method of raising and treating Drosophila melanogaster was the same as that in Example 9.

[0171] The experimental results are shown in Figure 6 and Table 10, Figure 6 which is the effect of camel placenta hydrolysate on the activity of CAT in Drosophila melanogaster (left: 25d; right: 45d); Figure 6 In, compared with the blank control group, *, # indicate significant differences (P < 0.05), **, ## indicate extremely significant differences (P < 0.01);

[0172] Table 10 Effects of camel placenta hydrolysate on the activity of CAT in Drosophila melanogaster (unit: U / mg prot)

[0173]

[0174] It can be seen that feeding camel placental hydrolysate for 25 days and 45 days can increase the CAT activity in female and male Drosophila melanogaster. At 25 days, compared with the control group, the CAT activity of female Drosophila melanogaster with 0.25% hydrolysate increased slightly but not significantly, while the CAT activity of female Drosophila melanogaster with 1% hydrolysate increased significantly (P < 0.05). At 45 days, the CAT activity of female Drosophila melanogaster with 0.25% hydrolysate increased significantly (P < 0.05), and the CAT activity of female Drosophila melanogaster with 1% hydrolysate increased extremely significantly (P < 0.01).

[0175] For male Drosophila melanogaster, at 25 days, compared with the control group, the CAT activity of 0.25% hydrolysate increased significantly (P < 0.05), and the CAT activity of male Drosophila melanogaster with 1% hydrolysate increased extremely significantly (P < 0.01). At 45 days, compared with the control group, the CAT enzyme activities of male Drosophila melanogaster with 0.25% and 1% hydrolysates both increased significantly (P < 0.05). In summary, the increase in CAT activity in Drosophila melanogaster with hydrolysate indicates that adding camel placental hydrolysate to the diet can effectively increase the CAT activity in Drosophila melanogaster.

[0176] Example 12

[0177] Evaluation of the effect of camel placental hydrolysate prepared in Examples 1 - 7 on the MDA (malondialdehyde) level in Drosophila melanogaster:

[0178] The method of raising and treating Drosophila melanogaster was the same as in Example 9.

[0179] MDA is the degradation product of lipid peroxide, and the content of MDA in Drosophila melanogaster reflects the level of oxidative stress in Drosophila melanogaster.

[0180] The results are shown in Figure 7 and Table 11.

[0181] Table 11 Results of the effect of camel placental hydrolysate on the MDA level in Drosophila melanogaster (unit: U / mg prot)

[0182]

[0183]

[0184] Figure 7 shows the effect of camel placental hydrolysate on the MDA level in Drosophila melanogaster (left: 25 days; right: 45 days), Figure 7 in which, compared with the blank control group, *, # indicate significant differences (P < 0.05), **, ## indicate extremely significant differences (P < 0.01).

[0185] Figure 7It reflects the effect of camel placental hydrolysate on the MDA content in female and male Drosophila melanogaster. The results show that the MDA content in Drosophila melanogaster increases with the aging of Drosophila melanogaster. Compared with the control group, at 25 days and 45 days, the MDA content in female Drosophila melanogaster with 0.25% hydrolysate decreased significantly (P<0.05), while the MDA content in female Drosophila melanogaster with 1% hydrolysate decreased extremely significantly (P<0.01).

[0186] For male Drosophila melanogaster, at 25 days, compared with the control group, the MDA level of male Drosophila melanogaster with 0.25% hydrolysate decreased slightly but not significantly, while at 45 days, the MDA level of male Drosophila melanogaster with 0.25% hydrolysate decreased significantly compared with the control group (P<0.05); the MDA levels of male Drosophila melanogaster with 1% hydrolysate decreased significantly at both 25 days and 45 days (P<0.01). In summary, the decrease in the MDA content in Drosophila melanogaster with hydrolysate indicates that camel placental hydrolysate can reduce the degree of lipid oxidation in Drosophila melanogaster.

[0187] Example 13

[0188] Acute experiment of Drosophila melanogaster with camel placental hydrolysate prepared in Examples 1-7

[0189] Hydroxyl radical is one of the most reactive free radicals in the body. H2O2 can generate hydroxyl radicals under certain conditions, leading to the functional decline of the body and causing certain oxidative damage to the body. In this invention, a hydrogen peroxide oxidation damage model was established to study the protective effect of camel placental hydrolysate on Drosophila melanogaster in the oxidation damage group induced by hydroxyl radicals.

[0190] After anesthetizing the adult Drosophila melanogaster that emerged within 24 hours, distinguish between males and females and randomly divide them into a blank control group, a 0.25% group, and a 1% group. There are 10 tubes of female and male Drosophila melanogaster in each group, with 20 in each tube. After culturing Drosophila melanogaster for 20 days, subject them to starvation treatment for 2 hours, and then anesthetize and introduce them into the Drosophila melanogaster culture tubes with filter paper strips soaked in 30% hydrogen peroxide solution by volume. The filter paper is fully wetted. Supply an appropriate amount of glucose hydrogen peroxide solution with a syringe every 4 hours, and record the number of dead Drosophila melanogaster every 2 hours until all Drosophila melanogaster die. Draw a lifespan curve and calculate the average lifespan and maximum lifespan of Drosophila melanogaster. The results are shown in Figure 8 。

[0191] Figure 8 It is the effect of camel placental hydrolysate on the lifespan of Drosophila melanogaster with acute hydrogen peroxide injury. As Figure 8As shown, compared with the control group, in the case of acute oxidative damage, the lifespan of female and male Drosophila fed with camel placenta hydrolysate was significantly prolonged. Among them, the average survival time and the maximum survival time of female Drosophila in the 0.25% group were prolonged by 13.61% (P<0.05) and 10.74% respectively, and those of male Drosophila were prolonged by 7.22% and 7.25% respectively; the average survival time and the maximum survival time of female Drosophila in the 1% group were prolonged by 21.43% (P<0.01) and 16.71% (P<0.05) respectively, and those of male Drosophila were prolonged by 22.10% (P<0.01) and 13.33% respectively. This indicates that camel placenta hydrolysate has a protective effect on Drosophila with acute oxidative damage and can extend the survival time of Drosophila in an oxidative stress environment.

[0192] Comparative Example 1

[0193] Treatment 1: Homogenize 10 g of camel placenta at high speed. The parameters of high-speed homogenization are 12,000 rpm for 10 min. Then add alkaline protease with a mass of 0.52% of the mass of camel placenta, and the enzyme activity of the added alkaline protease is 1,000 U / g. Hydrolyze at 40 °C for 34 h, and the initial pH of hydrolysis is 10. After the enzymatic hydrolysis is completed, inactivate it by water bath at 95 °C for 10 min. After cooling to room temperature, centrifuge at 6,000 r / min for 10 min, and take the supernatant for measuring the degree of hydrolysis and antioxidant level of the camel placenta enzymatic hydrolysate.

[0194] Treatment 2 is the same as Treatment 1. The only difference is that papain is added. The mass of papain added is 0.1252% of the mass of camel placenta, and the enzyme activity of the added papain is 1,000 U / g; hydrolyze at 50 °C for 34 h, and the initial pH of hydrolysis is 7.

[0195] Treatment 3 is the same as Treatment 1. The only difference is that neutral protease is added. The mass of neutral protease added is 12% of the mass of camel placenta, and the enzyme activity of the added neutral protease is 1,000 U / g; hydrolyze at 40 °C for 34 h, and the initial pH of hydrolysis is 7.

[0196] Treatment 4 is the same as Treatment 1. The only difference is that bromelain is added. The mass of bromelain added is 0.332% of the mass of camel placenta, and the enzyme activity of the added bromelain is 1,000 U / g; hydrolyze at 50 °C for 4 h, and the initial pH of hydrolysis is 7.

[0197] Treatment 5 is the same as Treatment 1. The only difference is that flavor protease is added. The mass of flavor protease added is 52% of the mass of camel placenta, and the enzyme activity of the added flavor protease is 1,000 U / g; hydrolyze at 50 °C for 34 h, and the initial pH of hydrolysis is 7.

[0198] The results of Treatments 1 to 5 are shown inFigures 9 - 10 and Tables 12 - 13, it can be seen from Figure 9 and Table 12 that the hydrolysis effect of alkaline protease is the best, and the degree of hydrolysis is 29.67%. It can be seen from Figure 10 and Table 13 that the ABTS radical scavenging rate of camel placenta hydrolysate CPE after hydrolysis by alkaline protease is 87.54%, which is significantly higher than that of the other 4 enzymes (P < 0.01). When measuring the ABTS radical scavenging rate, the concentration of camel placenta hydrolysate CPE is 1 mg / mL.

[0199] The method for measuring the degree of hydrolysis is as follows:

[0200] Refer to the o - phthalaldehyde method (OPA) to measure the degree of hydrolysis. Use 0.9516 mmol / L serine as the standard solution and deionized water as the blank control. During the specific operation, transfer 3 mL of the chromogenic reagent into a cuvette, quickly add 400 μL of the test solution and mix well. Strictly control the chromogenic reaction time at 120 seconds under room temperature conditions, and then measure the absorbance value at a wavelength of 340 nm.

[0201] The calculation formula is as follows:

[0202]

[0203] In the formula, x: protein content in the sample;

[0204] h0: degree of hydrolysis of the sample before the hydrolysis reaction;

[0205] h1: degree of hydrolysis of the sample after the hydrolysis reaction;

[0206] h tot : constant of the degree of hydrolysis of food - derived protein.

[0207] The preparation method of the chromogenic reagent is as follows:

[0208] Solution A1. Dissolve 7.620 g of borax (CAS No. 1303 - 96 - 4) and 200 mg of sodium dodecyl sulfate (SDS) in 150 mL of deionized water. Only add other reagents after complete dissolution.

[0209] Solution A2. Dissolve 160 mg of o - phthalaldehyde (OPA, purity 97%) in 4 mL of absolute ethanol. After complete dissolution, transfer this OPA solution to the above - mentioned Solution A1, and rinse and transfer with deionized water.

[0210] Solution A3. Add 176 mg of 1,4 - dithiothreitol (DTT, purity 99%) to Solution A1, rinse with deionized water and transfer.

[0211] A4. Make up the volume of Solution A1 to 200 mL with deionized water. This reagent should be prepared and used immediately.

[0212] Table 12 Degree of hydrolysis (%) of camel placenta with different enzymes

[0213] Alkaline protease Neutral protease Papain Bromelain Flavourzyme Parallel 1 30.79217 27.54342 22.35488 23.34843 26.8022 Parallel 2 30.80794 25.05166 21.12477 23.12764 26.8022 Parallel 3 30.49253 24.72048 21.9133 24.20004 26.56564 Mean ± Standard deviation 30.70±0.15 25.77±1.26 21.80±0.51 23.56±0.46 26.72±0.11

[0214] Table 13 ABTS radical scavenging rate (%) of camel placenta hydrolysate CPE obtained differently

[0215]

[0216]

[0217] In summary, the present invention prepared camel placenta hydrolysate, and evaluated the co-antioxidant and anti-aging activities of camel placenta hydrolysate by different methods, including detailed verification through in vitro and in vivo experiments. It can be seen that camel placenta hydrolysate has good co-antioxidant and anti-aging effects.

[0218] Although the above embodiments have described the present invention in detail, they are only a part of the embodiments of the present invention, rather than all embodiments. People can also obtain other embodiments based on these embodiments without creative efforts, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A preparation method of camel placenta hydrolyzate, characterized in that, Comprising: Hydrolyzing camel placenta with alkaline protease to obtain a camel placenta extract; the mass ratio of the alkaline protease to the camel placenta is (0.025 - 0.1):

100.

2. The preparation method according to claim 1, characterized in that, The temperature of the hydrolysis is 40 °C, and the hydrolysis time is 2 - 4 h.

3. The preparation method according to claim 1, characterized in that, After the hydrolysis, it further includes inactivating the enzyme and centrifuging, and the obtained supernatant is the camel placenta hydrolyzate; the temperature of the inactivation treatment is 85 - 100 °C, and the time is 15 min. Before the camel placenta is hydrolyzed, it further includes homogenization; the rotation speed of the homogenization is 3000 - 8000 rpm, and the time is 5 - 15 min.

4. A preparation method of camel placenta hydrolyzate, characterized in that, Comprising: Concentrating and drying the camel placenta hydrolyzate in sequence to obtain a camel placenta hydrolyzate; the camel placenta hydrolyzate is obtained by the preparation method according to any one of claims 1 - 3.

5. The preparation method according to claim 4, characterized in that, The way of concentration includes vacuum concentration.

6. The preparation method according to claim 4, characterized in that, The way of drying includes freeze - drying; the temperature of the freeze - drying is - 55 - - 45 °C, and the vacuum degree is 10 - 30 Pa.

7. The camel placenta hydrolyzate prepared by the preparation method according to any one of claims 4 to 6, characterized in that, The protein content of the camel placenta hydrolyzate is 8% to 12%. 。 8. Use of the camel placenta hydrolyzate prepared by the preparation method according to any one of claims 1 - 3 or the camel placenta hydrolyzate prepared by the preparation method according to any one of claims 4 - 6 in the preparation of antioxidant products and / or the preparation of anti - aging products.

9. Use of the preparation method of the camel placenta hydrolyzate according to any one of claims 1 - 3 or the preparation method of the camel placenta hydrolyzate according to any one of claims 4 - 6 in any one or more than two of the following 1) - 3); 1) Improving the antioxidant property of the camel placenta hydrolyzate; 2) Improving the anti - aging performance of the camel placenta hydrolyzate; 3) Improving the hydrolysis degree of the camel placenta hydrolyzate.

10. A product for assisting antioxidation and / or anti-aging, characterized in that, Comprising the camel placenta hydrolyzate according to claim 7.

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