CN113855731B - Oral liquid with antifatigue and immunoregulation functions and preparation method thereof - Google Patents

Oral liquid with antifatigue and immunoregulation functions and preparation method thereof Download PDF

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CN113855731B
CN113855731B CN202111135821.2A CN202111135821A CN113855731B CN 113855731 B CN113855731 B CN 113855731B CN 202111135821 A CN202111135821 A CN 202111135821A CN 113855731 B CN113855731 B CN 113855731B
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冯荣权
胡增仁
邵良碧
缪国燕
余红伟
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Zhejiang Kangde Pharmaceutical Co ltd
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Abstract

The invention discloses an oral liquid with anti-fatigue and immunoregulation functions and a preparation method thereof. The oral liquid with antifatigue and immunoregulation effects comprises placenta Caprae Seu Ovis, radix Acanthopanacis Senticosi and fructus Jujubae as raw materials. The preparation method of the oral liquid with the functions of resisting fatigue and regulating immunity comprises the following steps: mixing Chinese date and acanthopanax, soaking by ethanol, and heating and extracting to obtain a mixed extracting solution; hydrolyzing sheep placenta to obtain sheep placenta hydrolysate; mixing the mixed extractive solution and placenta Caprae Seu Ovis hydrolysate, adding water, fine filtering, and packaging to obtain oral liquid. The oral liquid has the advantages of stable quality, high content of functional components, greatly improved anti-fatigue and immunoregulation effects, simple and convenient preparation method, and easy dissolution and absorption of effective components.

Description

Oral liquid with antifatigue and immunoregulation functions and preparation method thereof
Technical Field
The invention relates to the technical field of traditional Chinese medicines, in particular to an oral liquid with anti-fatigue and immunoregulation functions and a preparation method thereof.
Background
With the enhancement of the competitive consciousness of the modern society, the pace of social life is accelerated, and the number of patients who complain about mental stress and chronic fatigue due to the overstrain of long-term work and the over-high mental stress is increased.
However, there are currently more studies on products for enhancing immunity, and there are fewer reports on products for relieving physical fatigue. Although some products can achieve the effect of relieving physical fatigue, the raw materials of the products are complex, the preparation process is tedious, and the effect is not obvious.
Disclosure of Invention
Based on the above, the invention aims to provide the oral liquid with high content of effective components and anti-fatigue and immunoregulation functions and the preparation method thereof.
The above purpose of the invention is realized by the following technical scheme:
according to one aspect of the invention, the oral liquid with the functions of resisting fatigue and regulating immunity provided by the invention comprises the following raw materials of effective components in each 1000ml of the oral liquid in parts by weight:
160-250 parts of sheep placenta;
10-150 parts by weight of acanthopanax;
6 to 100 parts of Chinese date.
In the invention, the raw materials of the effective components in the oral liquid can also comprise one or more of sucrose, isomalt, sodium benzoate and citric acid. Wherein, the weight portion of sucrose is 150 to 230 portions or the weight portion of isomalt is 320 to 450 portions. 0.5 to 2.0 portions of sodium benzoate. 1.5 to 4.0 portions of citric acid.
Optionally, in each 1000ml of the oral liquid, the raw materials of the effective components include, by weight: 200g of sheep placenta, 100g of acanthopanax, 60g of Chinese date, 180g of cane sugar, 1.0g of sodium benzoate and 2.0g of citric acid.
According to another aspect of the present invention, there is provided a method for preparing an oral liquid having anti-fatigue and immunoregulatory effects, comprising:
s1, weighing each component in the raw materials according to a ratio;
step S2, mixing the Chinese dates and the acanthopanax, adding ethanol for soaking, extracting, filtering, recovering ethanol, performing cold precipitation, and extracting supernatant to obtain a mixed extracting solution;
s3, hydrolyzing the sheep placenta at 45-55 ℃, filtering, performing cold sedimentation, and extracting supernatant to obtain sheep placenta hydrolysate;
and S4, mixing the mixed extracting solution and the sheep placenta hydrolysate, adding water, adjusting the pH of the solution to 4.0-6.0, for example, adjusting the pH of the solution to 4.0, 4.2, 4.5, 5.0, 5.2, 5.5, 5.8, 6.0 and the like, finely filtering, and encapsulating.
Optionally, in step S2, the concentration of ethanol is 55 to 70%. The soaking time is 10 to 30 hours. The extraction time is 3-5 hours. The cold precipitation time is more than 20 hours.
Optionally, in step S3, the step of hydrolyzing the sheep placenta comprises: cleaning sheep placenta, mincing, adding 2-4 times of water, and uniformly stirring; heating to 45-55 deg.C, such as 45 deg.C, 47 deg.C, 50 deg.C, 52 deg.C, 55 deg.C, adding trypsin, hydrolyzing, and hydrolyzing for 2-4 hr.
Optionally, in step S3, the filtrate is cryoprecipitated for 20 to 30 hours.
Optionally, step S4, before the step of adding water, comprises: adding water into sucrose, and heating to dissolve to obtain sucrose solution; adding a mixed solution of the mixed extracting solution and the sheep placenta hydrolysate into the sucrose solution; adding sodium benzoate and citric acid, stirring, and mixing.
The sheep placenta in the oral liquid is sweet and salty in taste and warm in nature, and has the effects of tonifying qi, nourishing blood and benefiting essence. The sheep placenta contains various bioactive substances such as hormone, cytokine, etc., thus having the functions of resisting oxidation, resisting aging, regulating immunity, etc.
The acanthopanax root oral liquid has warm, pungent and slightly bitter taste, has the functions of invigorating qi and spleen, soothing nerves and tonifying kidney, and is mainly used for treating diseases such as spleen yang deficiency, body weakness and hypodynamia, inappetence, soreness of waist and knees, insomnia and dreaminess and the like. Stearic acid, b-sitosterol, daucosterol, sesamin, clove and some triterpenoid saponin compounds can be separated from acanthopanax senticosus, and the acanthopanax senticosus has the effects of resisting tumor, aging and fatigue, regulating immunity and improving various diseases of cardiovascular systems.
The Chinese dates in the oral liquid are sweet in nature and taste and warm in nature and enter spleen and stomach meridians. It is indicated for middle-jiao and Qi tonifying, blood nourishing and tranquilization. Can be used for treating spleen deficiency, anorexia, asthenia, loose stool, and hysteria of women. Besides abundant nutrient components such as jujube carotene, riboflavin, ascorbic acid, vitamin E and various mineral substances, the jujube contains various medicinal components such as jujube saponin, spina date seed saponin B and adenosine cyclophosphate, participates in the regulation of various physiological and biochemical processes in vivo, has the effect of increasing total serum protein and albumin, and has the auxiliary treatment effect on chronic diseases such as tumors, coronary heart diseases, myocardial infarction, hepatitis, dystrophic edema, hypertension and the like.
One or more of sucrose or isomalt, sodium benzoate and citric acid can also be added into the oral liquid. Wherein, the sucrose is a sweetener, which can improve the taste of the product; sodium benzoate is used as preservative to prevent microbial contamination; citric acid is used as pH regulator, and can control pH of the oral liquid within a proper range, so as to improve taste and stability of the product.
The invention has the beneficial effects that: the oral liquid has high content of functional components, stable product quality and obviously improved anti-fatigue and immunoregulation functions.
The preparation method improves the process on the basis of the improved formula of the oral liquid, so that the content of the functional components is increased under the combined action of the sheep placenta, the acanthopanax and the Chinese date, the stability of the quality of the oral liquid product is improved through the process control, the content of the functional components of the product is stable after three-month accelerated tests (at the temperature of 37-40 ℃ and the relative humidity of 75 percent), for example, the reduction rate of protein is only about 3.5 percent, and the reduction rate of total saponin is only about 6 percent.
The oral liquid has the advantages of few raw materials, simple and quick preparation method, and easy dissolution and absorption of effective components.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments of the present invention, and it should be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. The following description of at least one exemplary embodiment is merely illustrative in nature and is in no way intended to limit the invention, its application, or uses. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Examples 1 to 5
The formulations and processes of examples 1-5 are detailed in Table 1. The specific preparation steps of each example are as follows:
1) The raw materials were weighed according to the formula in table 1.
2) Mixing fructus Jujubae and radix Acanthopanacis Senticosi, soaking in 60% ethanol overnight, boiling and extracting for 4 hr, filtering, recovering ethanol from filtrate, cold precipitating for 24 hr, and collecting supernatant to obtain mixed extractive solution.
3) Cleaning sheep placenta, mincing, adding 3 times of water, uniformly stirring, adjusting the pH value to 7.5, heating at the temperature shown in table 1, adding a proper amount of trypsin, hydrolyzing, maintaining the pH value of the solution to be 7.4-7.6, hydrolyzing for 4 hours, filtering, performing cold sedimentation on the filtrate for 24 hours, and extracting supernatant to obtain sheep placenta hydrolysate;
4) Adding water into sucrose, heating to dissolve, and adding mixed extractive solution, placenta Caprae Seu Ovis hydrolysate, sodium benzoate and citric acid; stirring, adding water to a sufficient amount (example 5 adding water directly to a sufficient amount), adjusting the pH of the solution as shown in Table 1, fine filtering, and filling into 10mL bottles with 10mL of each bottle. The above operations were all carried out in a clean plant with a cleanliness of 10 ten thousand levels.
TABLE 1 formulations and Processes of the examples
Figure BDA0003281987700000041
Figure BDA0003281987700000051
As can be seen from Table 1: in the oral liquid prepared in the embodiments 1-5, the protein and the total saponin both meet the standard that the protein is more than or equal to 2.4g/100mL, and the total saponin is more than or equal to 4.2mg/100mL, and the oral liquid prepared by the invention has higher protein and total saponin contents.
Comparative examples 1 to 3
The formulations and processes of comparative examples 1-3 are shown in table 2. The method comprises the following specific steps: extracting radix Acanthopanacis Senticosi or fructus Jujubae with ethanol to obtain extractive solution; hydrolyzing sheep placenta to obtain sheep placenta hydrolysate; mixing the extractive solutions with placenta caprae seu ovis extractive solution, adding adjuvants and/or water, fine filtering, and packaging. The adjuvants are sucrose, sodium benzoate and citric acid. The steps are similar to the invention, and the differences are that the raw material composition, the enzymolysis sheep placenta temperature and the pH value are shown in table 2. The products obtained in comparative examples 1 to 3, three of which were each subjected to functional component identification according to the detection method of the present invention example, are shown in table 2.
TABLE 2 respective proportional formulations and Processes
Figure BDA0003281987700000052
As can be seen from table 2: in comparative examples 1-3, the protein content is not more than 2.4g/100mL, and the total saponin content is not more than 4.2mg/100mL. By comparing tables 1 and 2, it can be seen that the contents of the functional components of comparative examples 1 to 3 are significantly lower than those of examples 1 to 4 of the present invention.
The invention also carries out the detection of physicochemical index and microbial index of the oral liquid, toxicology test, stability test, immunoregulation test and anti-fatigue test.
Physical and chemical index and microorganism index detection
The physicochemical indexes of the oral liquid in the embodiments 1 to 4 are detected according to GB/T5009, the microbial indexes are detected according to GB 4789, and the results show that the physicochemical indexes and the microbial indexes of the oral liquid conform to the national standard.
Toxicology test
The results of examples 1 to 4 (5 times of concentrated solution) of the present invention were as follows, when examined in accordance with GB-15193:
acute toxicity: the dosage of the oral liquid is 2.15g/kg, 4.64g/kg, 10.0g/kg and 21.5g/kg, and each dosage is respectively used for testing female and male rats and mice. The results show that: during the test period, no obvious symptoms and no death of the mice and rats in each group are observed. The oral liquid of the invention is orally administered to rats and mice by LD 50 The male and female are both more than 21.5g/kg body weight, which is nontoxic.
Micronucleus test: the oral liquid of the invention is used for testing female and male mice respectively with the dosage of 5g/kg, 10g/kg and 20g/kg, the dosage of 0g/kg as a negative control group and the dosage of 0.04g/kg as a positive control group. The results show that: and (3) carrying out micronucleus test on the mice, wherein the micronucleus permillage rate of each dose of male and female is not obviously different compared with that of a negative control group, and the positive control group is obviously higher than that of the negative control group. The oral liquid of the invention has negative detection result on mouse marrow addicted to multiple staining erythrocyte micronucleus test.
Mouse teratospermia test: the oral liquid of the invention is adopted with the dosage of 5g/kg, 10g/kg and 20g/kg, the dosage of 0g/kg as a negative control group and the MMC dosage of 0.002g/kg as a positive control group, and the test is respectively carried out, and the result shows that: compared with the negative control group, the rate of the mouse with the teratospermia of each dose group has no significant difference, and the positive control group is obviously higher than the negative control group. The oral liquid of the invention has negative result on mouse teratospermia test.
Ames test detection: the dosage of the oral liquid is five dosage groups of 160 mug/dish, 800 mug/dish, 4000 mug/dish, 20000 g/dish and 100000 mug/dish, a negative control and a positive control are added, and the strain is TA 97a 、TA 98 、TA 100 、TA 102 . The number of colonies which became back was counted by incubating in an incubator at 37 ℃ for 48 hours using the standard plate incorporation method. The results show that: the bacteriostatic phenomenon appears at 100000 mu g/dish, the retrogradation colonies of the rest tested substances with different concentrations are similar to the negative control group, and the number of the positive control group is more than 2 times higher than that of the negative control retrogradation colonies. The oral liquid Ames test result is negative.
Rat 30 day feeding trial: the oral liquid of the invention is fed to rats with 1.7, 5.0 and 7.0g/kg body weight groups in 30 days, and is about 25, 75 and 100 times of the recommended amount of human body. The test result shows that the animals have no toxic symptoms, abnormal pathological changes are not seen in gross anatomy and histological observation, and the conventional and biochemical indexes of blood have no damaging effect.
Stability test
The stability tests of the examples 1 to 4 of the invention are respectively carried out, and the results show that the products of the examples 1 to 4 have stable quality.
The results of the four measurements will be described below by taking example 2 as an example. Wherein, the four tests are respectively 0 month, 1 month, 2 months and 3 months, as shown in tables 3-6.
Table 3 test results of 0 month test on 3 oral liquids of example 2
Figure BDA0003281987700000071
TABLE 4 test results of 1 month test on 3 oral liquids of example 2
Figure BDA0003281987700000081
TABLE 5 test results of 2-month test on 3 oral liquids of example 2
Figure BDA0003281987700000082
TABLE 6 3 oral liquid of example 2, test results in 3 months
Figure BDA0003281987700000091
As can be seen from tables 3-6 above: the oral liquid is stored for three months under the conditions of 37-40 ℃ and 75% of relative humidity, the detection is carried out once a month for four times, the detection result conforms to the standard of Q/ZKD0022002, the product quality is stable, and the oral liquid can be stored for 24 months. Specifically, after being stored for three months, the pH value, the relative density and the contents of arsenic, lead and mercury of the three oral liquids are basically unchanged; the average content of protein is only reduced by 3.95 percent, and the average content of total saponin is only reduced by 6.23 percent; coliform, mould and yeast are not increased, and salmonella, shigella, staphylococcus aureus and hemolytic streptococcus are not detected.
Immunomodulatory assays
The immunoregulation effect detection is carried out in the embodiments 1-4 of the invention, and the result shows that the oral liquid in the embodiments 1-4 has the immunoregulation effect.
The following will explain in detail by taking example 2 as an example. The method is carried out according to the immunoregulation test method in the evaluation program and test method of health food functionality. Namely, the influence of the oral liquid on the immune function of the mouse is evaluated by delayed allergy, serum hemolysin determination, carbon clearance test and the ratio of viscera to body. The change in body weight of the animals before and after the test is shown in Table 7. The effect of the oral liquid of the present invention on lymph organs is shown in table 8. The results of Dinitrofluorobenzene (DNFB) induced DTH (ear swelling method) in mice, serum hemolysin assay, and carbon clearance test in mice are shown in tables 9-11, respectively.
TABLE 7 animal body weight changes before and after the test
Figure BDA0003281987700000101
From the above table it can be seen that: after 30d of mouse experiments, there was no statistical difference in initial, intermediate and final body weights (P > 0.05) for each group.
TABLE 8 Effect of oral liquid on lymph organs
Figure BDA0003281987700000102
From the above table, it can be seen that: oral liquid, chest/body ratio and spleen/body ratio of each dose group have no significant difference (p is more than 0.05) through statistical test.
TABLE 9 Effect of oral solutions on DNFB Induction of mouse DTH
Figure BDA0003281987700000111
From the above table it can be seen that: the THD of the oral liquid in the high, medium and low dose groups is obviously higher than that of the control group (p is less than 0.01).
TABLE 10 Effect of oral liquid on the formation of serum hemolysin in mice
Figure BDA0003281987700000112
From the above table it can be seen that: the serum hemolysin value of the oral liquid high-dose group is obviously higher than that of a control group (p is less than 0.05).
TABLE 11 Effect of oral liquid on mouse carbon clearance
Figure BDA0003281987700000113
From the above table, it can be seen that: the phagocytosis index of the oral liquid in the high, medium and low dosage groups is higher than that of the control group (p is less than 0.01).
The result of the above immunoassay is as follows: the oral liquid is continuously fed for 30 days at 1.7ml/kg, 3.3ml/kg and 10ml/kg, and the result shows that the oral liquid has the capacity of promoting the formation of the hemolysin of the mice and enhancing the DTH and carbon clearance of the mice, so the oral liquid has the immunoregulation function.
Anti-fatigue detection
The anti-fatigue tests of the oral liquid of the examples 1 to 4 of the invention are carried out, and the results show that the oral liquid of the examples 1 to 4 has the anti-fatigue effect.
The following will explain in detail by taking example 2 as an example. The test method is carried out according to the immunoregulation test method in the evaluation program and test method of health food functionality. The method comprises the following steps: weight swimming test, blood lactic acid assay, serum urea nitrogen assay, liver glycogen assay. The breeding conditions are temperature: 20-22 ℃ and the relative humidity is 50-70%. Wherein, the animal body weight has no obvious difference before and after the test. The effects of the oral liquid of the present invention on the swimming time of mice, the blood lactic acid of mice, the serum urea nitrogen and the liver glycogen are shown in tables 12, 13 and 14, respectively.
TABLE 12 Effect of oral liquid on weight bearing swimming time of mice
Figure BDA0003281987700000121
As can be seen from the above table: the heavy swimming time of the mice in the high-dose group is obviously longer than that of the control group.
TABLE 13 Effect of oral liquid on blood lactic acid in mice
Figure BDA0003281987700000122
As can be seen from the above table: the blood lactic acid of mice in the high, medium and low dose groups and the high and medium dose groups at 0min after swimming is obviously lower than that of the control group (p is less than 0.05 or less than 0.01).
TABLE 14 Effect of oral liquid on serum urea nitrogen and hepatic glycogen of mice
Figure BDA0003281987700000123
As can be seen from the above table: the liver glycogen content of the high and medium-low dose groups is higher than that of the control group, and the liver glycogen content of the mice of the high dose group is obviously higher than that of the control group.
The anti-fatigue detection result is as follows: the anti-fatigue effect of the oral liquid is detected by adopting a load swimming test and biochemical index detection, and the high, medium and low doses are continuously infused into the stomach of a mouse for 30 days, and the result shows that: the heavy-load swimming time of the high-dose group mouse is obviously longer than that of the control group; the blood lactic acid of the mice in the high, medium and low dose group at 0min and the high, medium and high dose group at 30min after swimming is obviously lower than that of the control group; the liver glycogen content of mice in a high-dose group is obviously higher than that of a control group, and the oral liquid has an anti-fatigue effect.
The description of the present invention has been presented for purposes of illustration and description, and is not intended to be exhaustive or limited to the invention in the form disclosed. Many modifications and variations will be apparent to practitioners skilled in this art. The embodiment was chosen and described in order to best explain the principles of the invention and the practical application, and to enable others of ordinary skill in the art to understand the invention for various embodiments with various modifications as are suited to the particular use contemplated.

Claims (3)

1. The oral liquid with the functions of resisting fatigue and regulating immunity is characterized in that the oral liquid comprises the following raw materials of effective components and auxiliary materials in each 1000ml by weight:
160-250 parts of sheep placenta;
10-150 parts by weight of acanthopanax;
6-100 parts of Chinese date;
150-230 parts of sucrose;
0.5 to 2.0 weight portions of sodium benzoate;
1.5 to 4.0 portions of citric acid
The preparation method of the oral liquid comprises the following steps: weighing the components according to the proportion;
mixing the Chinese dates and the acanthopanax, adding 55-70% ethanol for soaking for 10-30 hours, extracting for 3-5 hours, filtering, recovering ethanol, performing cold precipitation for more than 20 hours, and extracting supernatant to obtain mixed extract;
cleaning and mincing sheep placenta, adding 2-4 times of water, stirring uniformly, adding trypsin at 45-55 ℃, hydrolyzing the sheep placenta for 2-4 hours, filtering, performing cold precipitation for 20-30 hours, and extracting supernatant to obtain sheep placenta hydrolysate;
adding water into sucrose, and heating to dissolve to obtain sucrose solution; adding the mixed extracting solution and the sheep placenta hydrolysate, adding sodium benzoate and citric acid, mixing, adding water, adjusting the pH of the solution to 4.0-5.2, finely filtering, and encapsulating to obtain the oral liquid;
the functional components of the oral liquid are that protein is more than or equal to 2.4g/100ml, and total saponin is more than or equal to 4.2mg/100ml.
2. The oral liquid with the effects of resisting fatigue and regulating immunity according to claim 1, wherein the oral liquid comprises the following raw materials of active ingredients and auxiliary materials in each 1000 ml: 200g of sheep placenta, 100g of acanthopanax, 60g of Chinese date, 180g of cane sugar, 1.0g of sodium benzoate and 2.0g of citric acid.
3. A method for preparing the oral liquid with anti-fatigue and immunoregulatory effects according to claim 1, comprising:
s1, weighing the components according to the proportion;
step S2, mixing the Chinese dates and the acanthopanax, adding 55-70% ethanol for soaking for 10-30 hours, extracting for 3-5 hours, filtering, recovering ethanol, performing cold precipitation for more than 20 hours, and extracting supernatant to obtain a mixed extracting solution;
step S3, cleaning and mincing the sheep placenta, adding 2-4 times of water and uniformly stirring; adding trypsin to hydrolyze sheep placenta for 2-4 hours at 45-55 ℃, filtering, performing cold precipitation for 20-30 hours, and extracting supernatant to obtain sheep placenta hydrolysate;
and S4, adding water into cane sugar, heating and dissolving to prepare a cane sugar solution, adding the mixed extracting solution and the sheep placenta hydrolysate, adding sodium benzoate and citric acid, mixing, adding water, adjusting the pH value of the solution to 4.0-5.2, finely filtering, and encapsulating.
CN202111135821.2A 2021-09-27 2021-09-27 Oral liquid with antifatigue and immunoregulation functions and preparation method thereof Active CN113855731B (en)

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1951434A (en) * 2006-09-29 2007-04-25 李志民 Soft capsule with broomrape and Chinese date having fatigue-resisting radiation-resisting function and preparation method thereof
CN109222110A (en) * 2018-07-17 2019-01-18 闵红岗 A kind of antifatigue soluble granule and preparation method of the polypeptide dry powder containing Goat Placenta

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CA2448044A1 (en) * 2001-06-02 2002-12-12 Kolon Ind. Inc. Korean acanthopanax senticosus extract, protein extract, crude protein-polysaccharide which were extracted from korean acanthopanax senticosus, and immunoregulating compositions comprising the same and use thereof
CN107549657A (en) * 2016-06-30 2018-01-09 贺兴荣 Goat Placenta capsule and preparation method thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1951434A (en) * 2006-09-29 2007-04-25 李志民 Soft capsule with broomrape and Chinese date having fatigue-resisting radiation-resisting function and preparation method thereof
CN109222110A (en) * 2018-07-17 2019-01-18 闵红岗 A kind of antifatigue soluble granule and preparation method of the polypeptide dry powder containing Goat Placenta

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