A composition containing Cistanche deserticola extract and its preparation method and application

Abstract

The present invention relates to the field of biotechnology, and in particular to a composition containing a Cistanche deserticola extract, a preparation method thereof and an application thereof. The composition comprises components: a Cistanche deserticola extract, a ginseng extract, a cashew protein peptide and a rose extract; the Cistanche deserticola extract is obtained by ultrasonic reflux, vacuum decompression concentration and freeze drying; the rose extract is obtained by ultrasonic extraction and concentration and drying. The composition of the Cistanche deserticola extract, the ginseng extract, the rose extract and the cashew protein peptide provided by the present invention has the function of enhancing immunity.

Description

Technical Field

[0001] The invention relates to the field of biotechnology, and in particular to a composition containing a Cistanche deserticola extract, and a preparation method and application thereof. Background Art

[0002] Immunity refers to the body's own defense mechanism, which is the body's ability to identify and eliminate any foreign invading substances (viruses, bacteria, etc.), deal with aging, damaged, dead, and degenerated cells of the body, and identify and deal with mutated cells and virus-infected cells in the body.

[0003] Immunity is the physiological response of the human body to identify and exclude "foreign" things. The immune system performs this function in the human body, which can be divided into innate immunity (also known as natural immunity) and adaptive immunity (also known as acquired immunity), among which adaptive immunity can be divided into humoral immunity and cellular immunity.

[0004] Immunity can be divided into nonspecific immunity and specific immunity. Nonspecific immunity is innate, including the first and second lines of defense of the human body, which can resist foreign invading pathogens, as well as phagocytes in the body. Specific immunity includes cellular immunity and humoral immunity, which refers to the antibodies produced by the human body after pathogens enter the human body, and plays an important role in resisting pathogen invasion and eliminating pathogens.

[0005] The strength of immunity can reflect the health status of the human body to a certain extent. People with strong immunity are usually better able to resist diseases, while people with weak immunity are more likely to get sick.

[0006] Cistanche deserticola, as a precious Chinese medicinal material, has a long history and wide application. In traditional Chinese medicine theory, Cistanche deserticola is believed to have multiple effects such as tonifying kidney yang, benefiting essence and blood, moistening intestines and relieving constipation. Modern research shows that Cistanche deserticola not only has important application value in the field of traditional medicine, but also shows significant effects in improving human immunity. However, when Cistanche deserticola extract is used alone, its efficacy is not ideal, and the dosage needs to be increased when the efficacy is enhanced, resulting in an impact on the human body when the dosage is too large. Therefore, it is necessary to match other components to promote the efficacy of Cistanche deserticola and improve its efficacy under the reasonable dosage that ensures that it meets the requirements. Summary of the invention

[0007] In view of the deficiencies of the prior art, the object of the present invention is to provide a composition with the function of enhancing immunity, which is composed of Cistanche deserticola extract, rose extract, ginseng extract and cashew protein peptide. Through scientific proportioning, these four ingredients play a mutually promoting role in enhancing immunity.

[0008] To achieve the above object, the present invention discloses the following technical solutions:

[0009] In a first aspect, the present invention provides a composition containing a Cistanche deserticola extract, comprising the following components by weight:

[0010]

[0011] Preferably, the preparation method of the Cistanche deserticola extract comprises the following steps:

[0012] Step (1), adding water to Cistanche deserticola for ultrasonic reflux extraction, and filtering to obtain an extract;

[0013] Step (2), concentrating the extract obtained in step (1) under vacuum to obtain a Cistanche deserticola concentrate;

[0014] Step (3), vacuum freeze-drying the Cistanche deserticola concentrate obtained in step (2) to obtain a freeze-dried extract;

[0015] Step (4), grinding the freeze-dried extract from step (3) into powder and sieving to obtain a Cistanche deserticola extract.

[0016] Further preferably, the amount of water added in step (1) is 30 times the mass of Cistanche deserticola, the number of reflux extractions in ultrasonic reflux extraction is 3 times, each time for 1.1 h, the ultrasonic power is 3 kW, and the reflux extraction temperature is 90°C.

[0017] Further preferably, the concentration temperature of the vacuum concentration in step (2) is 65° C., the vacuum degree is -0.095 MPa, and the relative density of the obtained Cistanche deserticola concentrate is 1.13 g / mL.

[0018] Further preferably, in the vacuum freeze-drying of step (3), the initial pre-freezing temperature is -25°C, the end point pre-freezing temperature is -70°C, the pre-freezing rate is 0.95°C / min, and the drying time is 20h.

[0019] Preferably, the preparation method of the rose extract comprises the following steps:

[0020] Step S1, grinding and sieving dried roses to obtain coarse rose powder;

[0021] Step S2, adding water to the rose coarse powder obtained in step S1, mixing, and soaking to obtain a mixture;

[0022] Step S3, raising the temperature of the mixture obtained in step S2 to 75-85°C, and ultrasonically extracting for ≥45 minutes to obtain an extract;

[0023] Step S4, filtering and concentrating the extract obtained in step S3 to obtain a concentrated solution;

[0024] Step S5, adding resistant dextrin to the concentrated solution of step S4, and drying to obtain rose extract.

[0025] Further preferably, in step S1, after grinding, the product is sieved through a sieve of ≥80 mesh;

[0026] The mass ratio of the rose coarse powder to water in step S2 is 1:20, and the soaking time is ≥30 min;

[0027] The ultrasonic frequency of step S3 is ≥30kHz;

[0028] The extract in step S4 is concentrated to 20% of the original volume to obtain a concentrated solution;

[0029] The amount of resistant dextrin added in step S5 is 25wt% of the concentrated solution, and the drying is spray drying.

[0030] Further preferably, the inlet air temperature of the spray drying is 150° C., the outlet air temperature is 75° C., and the rotation speed is 16000 rpm.

[0031] In a second aspect, the present invention provides use of the composition containing the Cistanche deserticola extract described in the first aspect in preparing a health food that helps to enhance immunity.

[0032] Preferably, the added amount of the composition containing the Cistanche deserticola extract is ≤10wt%.

[0033] In the present invention:

[0034] Cistanche extract enhances immunity. The polysaccharide components in Cistanche extract can increase cell proliferation response, thereby enhancing immune function, prolonging life and reducing the risk of death. It also has a strong laxative effect, can improve intestinal peristalsis and help the body detoxify. According to traditional Chinese medicine theory, Cistanche has the effect of replenishing kidney yang and benefiting essence and blood, which helps to improve symptoms such as male impotence. As a basic ingredient, Cistanche extract can better play its role in enhancing immunity and improving blood circulation when working together with other substances.

[0035] Rose extract is rich in various nutrients, including vitamin C, flavonoids, polyphenols, etc. These ingredients have the potential to enhance immunity. Among them, vitamin C in rose extract is a powerful antioxidant, which can enhance the function of white blood cells and help the body resist the invasion of bacteria and viruses, thereby improving immunity; flavonoids have also been shown to have anti-inflammatory, antioxidant and immunomodulatory effects, which can help reduce the body's inflammatory response while promoting the activity and proliferation of immune cells; polyphenols also have certain antioxidant and anti-inflammatory effects, which are equally important for improving immunity; in addition, rose extract also has certain anti-depressant and stress-relieving effects, which also help maintain the body's immune balance and health.

[0036] Ginsenosides and other ingredients in ginseng extract can enhance the body's resistance and improve the activity of immune cells. Ginseng extract also has the function of regulating endocrine, which helps to maintain the body's balance. When ginseng extract and Cistanche deserticola extract work together, they can further enhance the effect of immune regulation and improve overall immunity.

[0037] Cashew protein peptide has multiple biological activities such as anti-oxidation, anti-inflammatory, and cell repair promotion. It can be used as a food nutritional additive to improve the nutritional structure of food. The addition of cashew protein peptide can provide the composition with necessary amino acids and nutrients. At the same time, its antioxidant and anti-inflammatory effects can further promote the role of Cistanche extract in immune regulation and cell repair.

[0038] Beneficial effects of the present invention:

[0039] The composition provided by the invention, which consists of Cistanche deserticola extract, ginseng extract, rose extract and cashew protein peptide, has the function of enhancing immunity. DETAILED DESCRIPTION

[0040] In order to make the purpose, technical scheme and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with specific embodiments. However, the following embodiments are only preferred embodiments of the present invention, not all. For the sake of clarity, all features of the actual embodiments are not described.

[0041] Based on the embodiments in the implementation manner, other embodiments obtained by those skilled in the art without making any creative work shall all fall within the protection scope of the present invention.

[0042] In the present invention:

[0043] Cistanche tubulosa is Cistanche tubulosa (Schenk) R.Wight, a plant of the Orobanchaceae family, purchased from Bozhou Tengfengtang Pharmaceutical Co., Ltd.

[0044] Roses were Rosa rugosa Thunb., Rosaceae, purchased from Jinan Cuihe Rose Products Co., Ltd.;

[0045] Cashew protein peptide was purchased from Fufeng Snot Biotechnology Co., Ltd.;

[0046] Ginseng extract was purchased from Xi'an Haoyu Kangze Biotechnology Co., Ltd.;

[0047] The remaining raw materials are commercially available.

[0048] Preparation of Cistanche deserticola extract:

[0049] Step (1), placing the whole plant of Cistanche deserticola in an ultrasonic reflux extraction tank, performing reflux extraction for 3 times, each time for 1.1 h, adding water in an amount of 30 times the weight of the medicinal material, maintaining the ultrasonic power at 3 kW, maintaining the extraction temperature at 90° C., and filtering to obtain an extract;

[0050] Step (2), concentrating the extract under vacuum at a temperature of 65° C. and a vacuum degree of -0.095 MPa to obtain a Cistanche deserticola concentrate having a relative density of 1.13 g / mL;

[0051] Step (3), putting the Cistanche deserticola concentrate into a vacuum freeze drying device for drying, wherein the parameters are set as follows: pre-freezing initial temperature -25°C, pre-freezing rate 0.95°C / min, pre-freezing end temperature -70°C, drying chamber pressure 0.035MPa, heating plate temperature 30°C, drying time 20h, to obtain a freeze-dried extract;

[0052] Step (4), grinding the freeze-dried extract into powder by a universal grinder, and passing the powder through a 120-mesh sieve to obtain a Cistanche deserticola extract.

[0053] Preparation of rose flower extract:

[0054] Step 1, grind the rose stem (flower part) into powder, and pass it through an 80-mesh sieve to obtain rose coarse powder;

[0055] Step 2, mixing the obtained rose flower powder and pure water in a mass ratio of 1:20, and soaking at room temperature for 30 minutes to obtain a mixture;

[0056] Step 3, then raising the temperature of the mixture to 75-85° C., and performing ultrasonic assisted extraction at a frequency of 30 kHz for 45 minutes to obtain an extract;

[0057] Step 4, cooling the extract to room temperature, filtering to remove impurities, and concentrating to 20% of the original volume to obtain a concentrated solution;

[0058] Step 5, adding 25% by mass of resistant dextrin to the concentrated solution, and spray drying to obtain rose extract, wherein the inlet air temperature of the spray drying is 150° C., the outlet air temperature is 75° C., and the rotation speed is 16000 rpm.

[0059] Preparation of composition containing Cistanche deserticola extract

[0060] The raw materials were mixed according to the proportions in Table 1 to obtain a composition containing the Cistanche deserticola extract.

[0061] Table 1 Raw material ratio of composition 1-8

[0062] Composition 1 Composition 2 Composition 3 Composition 4 Composition 5 Composition 6 Composition 7 Composition 8 Cistanche deserticola extract 20 25 30 25 25 25 25 25 Rose Flower Extract 6 5 3 - 5 5 5 5 Ginseng Extract 1 0.50 0.26 0.50 - 0.50 0.50 0.50 Cashew Protein Peptide 7 9 10 9 9 - - - Soy protein peptide - - - - - - 9 - Wheat protein peptide - - - - - - - 9

[0063] Note: “-” in the table means not added.

[0064] Preparation of Examples

[0065] The above compositions were mixed with maltodextrin respectively to prepare an embodiment with a content of 10 wt %, as shown in Table 2.

[0066] Table 2 Mass percentage of raw materials in Examples 1-8

[0067] Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 Example 7 Example 8 Composition No. 1 2 3 4 5 6 7 8 Addition amount of composition / % 10 10 10 10 10 10 10 10 Maltodextrin addition amount / % 90 90 90 90 90 90 90 90

[0068] Evaluation of animal functional test for enhancing immunity

[0069] 1 Materials

[0070] The test sample is Example 1-8, and the recommended daily dose of the test sample is 10 g. According to an adult weighing 60 kg, the recommended dose of the sample is 0.167 g / kg.BW;

[0071] 1. In Examples 1-3, three dose groups and a negative control group (distilled water) were set up respectively. The low, medium and high doses were 0.833, 1.667 and 5.000 g / kg of the test sample, which were equivalent to 5, 10 and 30 times the recommended dose for humans;

[0072] 2. In each of the experiments of Examples 4-8, a medium dose group was set up, which was a 10-fold group, and a comparative experiment was conducted with Example 2. The amount of the test sample was the same as that of the medium dose group in Example 2, which was 1.667 g / kg;

[0073] Before the test, the test samples were prepared into various doses using distilled water as solvent and mixed well for testing;

[0074] For the three dosage groups, 8.33, 16.67, and 50.00 g of product powder were measured respectively, and distilled water was added to 100 mL to prepare samples with concentrations of 0.0833, 0.1667, and 0.5000 g / mL. The samples were administered by gavage, and the gavage volume was calculated as 0.1 mL / 10 g body weight.

[0075] The experiment used clean-grade ICR mice, female, weighing 20±2g, the test temperature was 20-24℃, the relative humidity was 40-70%, and the animals were adapted to the animal room environment for 4 days before the experiment. Different doses of the test samples were orally administered for 30-35 consecutive days, the negative control group was given the same volume of distilled water, and the rest were treated with the same dose group, with 10 mice in each group.

[0076] 2 Methods

[0077] (1) Antibody-producing cell detection:

[0078] Each group was gavaged with the sample once a day for 35 consecutive days. On the 30th day of the experiment, each mouse was intraperitoneally injected with 0.2 mL of 2% (v / v) packed sheep red blood cell suspension for immunization. Five days later, the spleen of each mouse was aseptically removed and placed in a small dish containing an appropriate amount of sterile Harks solution. It was gently torn with tweezers to make a single cell suspension, filtered through a 200-mesh screen, washed, counted, and finally adjusted to a cell concentration of 5×10 with RPMI1640 complete culture medium. 6 After the surface culture medium (1g agarose plus double distilled water to 100mL) was heated and dissolved, it was placed in a 45℃ water bath for heat preservation, mixed with an equal amount of pH 7.2-7.4 and 2-fold concentration of Harks solution, and dispensed into small test tubes, 0.5mL per tube, and then 50μL 10% SRBC (v / v, prepared with SA solution) and 20μL spleen cell suspension (5×10 6 / mL), mix quickly, pour on the glass slide brushed with a thin layer of agarose, make parallel slices, wait for the agar to solidify, put the glass slide horizontally on the slide rack, put it in a carbon dioxide incubator and incubate it for 1 to 1.5 hours, then add the complement diluted with physiological saline (1:8) into the groove of the slide rack, continue to incubate for 1 to 1.5 hours, and then count the number of hemolytic plaques.

[0079] (2) Mouse peritoneal macrophage phagocytosis of chicken red blood cells:

[0080] Each group was gavaged with samples once a day for 31 consecutive days. 30 minutes before the animals were killed, 1 mL of 20% (v / v) chicken red blood cell suspension was injected into the peritoneal cavity of each mouse, and 2 mL of normal saline was injected after the animals were killed. The peritoneal fluid droplets were taken and incubated in a 38°C incubator for 30 minutes, fixed, stained, and examined under a microscope. 100 macrophages were counted and the phagocytic rate and phagocytic index were calculated.

[0081] (3) Sheep red blood cells (SRBC) induced DTH in mice: Each group was gavaged with samples once a day for 35 consecutive days. On the 30th day of the experiment, each mouse was intraperitoneally injected with 0.2 mL of 2% (v / v) packed volume sheep red blood cell suspension for immunization. Four days after immunization, the thickness of the plantar part was measured, and then 20% (v / v) SRBC was subcutaneously injected at the measurement site, 20 μL per mouse. The thickness of the plantar part of the left and right feet was measured 24 hours after injection. The same site was measured three times and the average value was taken.

[0082] 3 Test results

[0083] (1) Effect of the product on antibody-producing cells: The results are shown in Table 1.

[0084] Table 1 Effects on mouse antibody-producing cells

[0085]

[0086] Note: # indicates significant difference compared with the negative control group (P < 0.05), ## indicates extremely significant difference compared with the negative control group (P < 0.01); b indicates significant difference compared with Example 2 (P < 0.05).

[0087] (2) Effect of the product on the phagocytosis of chicken red blood cells by mouse peritoneal macrophages: The results are shown in Table 2.

[0088] Table 2 Effects on phagocytosis of chicken red blood cells by mouse peritoneal macrophages

[0089]

[0090]

[0091] Note: # indicates significant difference compared with the negative control group (P < 0.05), ## indicates extremely significant difference compared with the negative control group (P < 0.01); b indicates significant difference compared with Example 2 (P < 0.05).

[0092] (3) Effect of the product on sheep erythrocyte-induced DTH in mice: The results are shown in Table 3.

[0093] Table 3 Effects of sheep red blood cells on induction of DTH in mice

[0094]

[0095] Note: # indicates significant difference compared with the negative control group (P < 0.05), ## indicates extremely significant difference compared with the negative control group (P < 0.01); b indicates significant difference compared with Example 2 (P < 0.05).

[0096] 4 Conclusion Analysis

[0097] (1) Compared with the negative control group, the number of hemolytic plaques in the low, medium and high dose groups of the test sample increased, and the differences were significant; compared with the negative control group, the percentage of phagocytosis of chicken red blood cells by peritoneal macrophages in the low, medium and high dose groups of the test sample increased, and the differences were significant; compared with the negative control group, the ConA-induced proliferation capacity of mouse spleen lymphocytes and the difference in thickness measurement values ​​of the plantar part of the left hind foot of the mouse at 24h and 0h in the low, medium and high dose groups of the test sample increased, and the differences were significant. The above results prove that the composition composed of Cistanche deserticola extract, ginseng extract, rose extract and cashew protein peptide provided by the present invention has the function of enhancing immunity.

[0098] (2) Compared with Example 2, the number of hemolytic plaques in the mice in the dose groups of Examples 4-8 was reduced, and the differences were significant; compared with Example 2, the percentage of phagocytosis of chicken red blood cells by peritoneal macrophages in the mice in the dose groups of Examples 4-8 was reduced, and the differences were significant; compared with Example 2, the proliferation capacity of mouse spleen lymphocytes induced by ConA and the difference in thickness measurement values ​​of the plantar part of the left hind foot of mice at 24h and 0h in the dose groups of Examples 4-8 were reduced, and the differences were significant. The above results show that the four components of the Cistanche extract, ginseng extract, rose extract and cashew protein peptide provided by the present invention have a certain mutual promotion effect when used in combination, and the reduction of any one component can lead to a decrease in the efficacy of the overall formula.

[0099] Although the specific embodiments of the present invention are described above, it should be understood by those skilled in the art that this is only for illustration and the protection scope of the present invention is defined by the appended claims. Those skilled in the art may make various changes or modifications to these embodiments without departing from the principles and essence of the present invention, but these changes and modifications all fall within the protection scope of the present invention.

Claims

1. A composition containing a Cistanche deserticola extract, characterized in that: By mass, it includes the following components: The preparation method of the Cistanche deserticola extract comprises the following steps: Step (1), adding water to Cistanche deserticola for ultrasonic reflux extraction, and filtering to obtain an extract; Step (2), concentrating the extract obtained in step (1) under vacuum to obtain a Cistanche deserticola concentrate; Step (3), vacuum freeze-drying the Cistanche deserticola concentrate obtained in step (2) to obtain a freeze-dried extract; Step (4), grinding the freeze-dried extract from step (3) into powder and sieving to obtain a Cistanche deserticola extract; The preparation method of the rose extract comprises the following steps: Step S1, grinding and sieving dried roses to obtain coarse rose powder; Step S2, adding water to the rose coarse powder obtained in step S1, mixing, and soaking to obtain a mixture; Step S3, raising the temperature of the mixture obtained in step S2 to 75-85°C, and ultrasonically extracting for ≥45 minutes to obtain an extract; Step S4, filtering and concentrating the extract obtained in step S3 to obtain a concentrated solution; Step S5, adding resistant dextrin to the concentrated solution of step S4, and drying to obtain rose extract.

2. The composition containing the Cistanche deserticola extract according to claim 1, characterized in that: The amount of water added in step (1) is 30 times the mass of Cistanche deserticola, the ultrasonic reflux extraction is performed 3 times, each time for 1.1 hours, the ultrasonic power is 3 kW, and the reflux extraction temperature is 90°C.

3. The composition containing the Cistanche deserticola extract according to claim 1, characterized in that: The vacuum concentration in step (2) is carried out at a concentration temperature of 65° C. and a vacuum degree of −0.095 MPa. The relative density of the obtained Cistanche deserticola concentrate is 1.13 g / mL.

4. The composition containing the Cistanche deserticola extract according to claim 1, characterized in that: In the vacuum freeze drying of step (3), the initial pre-freezing temperature is -25°C, the end point pre-freezing temperature is -70°C, the pre-freezing rate is 0.95°C / min, and the drying time is 20h.

5. The composition containing the Cistanche deserticola extract according to claim 1, characterized in that: After grinding in step S1, the mixture is sieved through a sieve with a mesh size of ≥80; The mass ratio of the rose coarse powder to water in step S2 is 1:20, and the soaking time is ≥30 min; The ultrasonic frequency of step S3 is ≥30kHz; The extract in step S4 is concentrated to 20% of the original volume to obtain a concentrated solution; The amount of resistant dextrin added in step S5 is 25wt% of the concentrated solution, and the drying is spray drying.

6. The composition containing the Cistanche deserticola extract according to claim 5, characterized in that: The spray drying has an inlet air temperature of 150°C, an outlet air temperature of 75°C, and a rotation speed of 16000 rpm.

7. Use of the composition containing the Cistanche deserticola extract according to any one of claims 1 to 6 in the preparation of health food that helps to enhance immunity.

8. The use according to claim 7, characterized in that: The addition amount of the composition containing the Cistanche deserticola extract is ≤10wt%.

Images