Traditional Chinese medicine compound extract for relieving physical fatigue and preparation method thereof

Abstract

The invention discloses a traditional Chinese medicine compound extract for relieving physical fatigue and a preparation method thereof. The traditional Chinese medicine compound extract is prepared from the following traditional Chinese medicine decoction pieces in parts by weight: 1-3 parts of ginseng traditional Chinese medicine decoction pieces, 3-10 parts of astragalus membranaceus traditional Chinese medicine decoction pieces, 4-13 parts of salvia miltiorrhiza traditional Chinese medicine decoction pieces and 2-7 parts of lucid ganoderma traditional Chinese medicine decoction pieces. Experiments show that the traditional Chinese medicine compound extract can prolong the load swimming time of mice, promote hepatic glycogen storage of the mice and reduce the area under a blood lactic acid curve of the mice after exercise, and has the effect of relieving physical fatigue.

Description

Technical Field

[0001] The present invention relates to the technical field of Chinese medicine extracts, and in particular to a Chinese medicine compound extract for relieving physical fatigue and a preparation method thereof. Background Art

[0002] Fatigue is defined as a feeling of extreme physical or mental exhaustion that can negatively impact work performance, family life, and social relationships. It's a complex phenomenon that may be present in many diseases. Fatigue is caused by the depletion of energy sources, including decreased blood sugar levels and liver glycogen depletion; the accumulation of fatigue end products; a disrupted internal environment; and impaired metabolic control of the nervous system, enzymes, and hormones during exercise. With economic development and an accelerated pace of life, the number of people experiencing fatigue is increasing. If chronic fatigue is not promptly alleviated, it can lead to aging, anxiety, insomnia, depression, physical dysfunction, cognitive impairment, energy imbalance, and even serious diseases related to biological regulation and the immune system. Epidemiological data indicate that approximately 20% of people worldwide experience exercise-induced fatigue each year. One way to alleviate exercise-induced fatigue is to replenish nutrients promptly. Consequently, the consumption of sports nutrition foods related to post-exercise recovery has seen rapid growth in recent years. Currently, a wide variety of sports nutrition foods, including beverages, tablets, energy bars, and capsules, have been developed both domestically and internationally. According to reports, the overall size of my country's sports nutrition market was US$329 million in 2018. In the past five years, the industry's retail sales have grown at a compound annual growth rate of 40%. In the next five years, this figure is expected to reach 24%, far higher than the 11% of the global sports nutrition industry. It is estimated that in 2023, the size of the Chinese market will reach US$859 million. In future development, the sports nutrition food industry still has a large room for development and broad market potential. Summary of the Invention

[0003] The present invention aims to provide a traditional Chinese medicine compound extract for relieving physical fatigue and a preparation method thereof. The traditional Chinese medicine compound extract has a precise formulation and good safety as confirmed by animal testing. Functional evaluation (animal function) confirms that it has a significant health-promoting function of relieving physical fatigue and has good market development prospects.

[0004] The technical solution adopted in the present invention is as follows: A method for preparing a traditional Chinese medicine compound extract for relieving physical fatigue comprises the following steps: 1) mixing Chinese medicinal slices of ginseng, astragalus, salvia miltiorrhiza and ganoderma lucidum according to a proportion to obtain a Chinese medicinal slice mixture; 2) extracting the mixture of Chinese herbal medicine slices obtained in step 1) with water as an extraction solvent, filtering to obtain a filtrate 1 and a filter residue 1; extracting the filter residue 1 with water as an extraction solvent, filtering to obtain a filtrate 2 and a filter residue 2; combining the filtrate 1 and the filtrate 2, concentrating, precipitating with alcohol, and standing to separate to obtain an alcohol supernatant and an alcohol precipitate; freeze-drying and pulverizing the alcohol precipitate to obtain component A; 3) recovering ethanol from the alcohol precipitation supernatant of step 2), adsorbing the supernatant to saturation using a macroporous adsorption resin column, eluting the column with ethanol after adsorption, and collecting the eluate until the effluent is clarified; concentrating the collected eluate, freeze-drying, and pulverizing the column to obtain component B; 4) extracting the filter residue 2 from step 2) with an ethanol solution as an extraction solvent, filtering to obtain a filtrate 3 and a filter residue 3; extracting the filter residue 3 with an ethanol solution as an extraction solvent, filtering to obtain a filtrate 4 and a filter residue 4; combining the filtrate 3 and the filtrate 4, concentrating, freeze-drying, and pulverizing to obtain component C; 5) Component A from step 2), component B from step 3) and component C from step 4) are mixed to obtain a Chinese herbal compound extract.

[0005] Furthermore, the above-mentioned Chinese herbal medicine slice mixture is composed of the following Chinese herbal medicine slices in parts by weight: 1 to 3 parts of ginseng Chinese herbal medicine slices, 3 to 10 parts of astragalus Chinese herbal medicine slices, 4 to 13 parts of salvia miltiorrhiza Chinese herbal medicine slices, and 2 to 7 parts of ganoderma lucidum Chinese herbal medicine slices.

[0006] Furthermore, the preparation steps of the above-mentioned component A are specifically as follows: extracting the mixture of Chinese herbal medicine pieces with water as the extraction solvent, the weight volume ratio of the mixture of Chinese herbal medicine pieces to the extraction solvent is 1:10-15, the extraction time is 1-2 hours, the extraction temperature is 95-100°C, filtering to obtain filtrate 1 and filter residue 1; extracting the filter residue 1 with water as the extraction solvent, the weight volume ratio of the filter residue 1 to the extraction solvent is 1:10-15, the extraction time is 1-2 hours, the extraction temperature is 95-100°C, filtering to obtain filtrate 2 and filter residue 2; combining filtrate 1 and filtrate 2, and concentrating to a density of 1.05-1.15 g / cm 3 , add 95% ethanol solution by volume to the obtained concentrated solution, with the volume ratio of the concentrated solution to the ethanol solution being 1:4, and carry out alcohol precipitation at 1-5°C for more than 12 hours. Let it stand and separate to obtain an alcohol precipitation supernatant and an alcohol precipitation precipitate; freeze-dry and pulverize the alcohol precipitation precipitate to obtain a crude polysaccharide component with a moisture content of less than 5%, which is recorded as component A.

[0007] Furthermore, the preparation steps of the above-mentioned component B are specifically as follows: the supernatant of the alcohol precipitation is subjected to ethanol recovery treatment until there is no alcohol taste, and then adsorbed to saturation using a macroporous adsorption resin chromatography column, the model of the macroporous adsorption resin is D101C, and the adsorption rate is 2-3 BV / h. After the adsorption is completed, it is eluted with an ethanol solution with a volume fraction of 70-75% at an elution rate of 1 BV / h, and the eluate is collected until the effluent is clear; the collected eluate is concentrated until there is no alcohol taste, and the obtained concentrated solution is freeze-dried and pulverized to obtain a total saponin component with a moisture content of less than 5%, which is recorded as component B.

[0008] Furthermore, the preparation steps of the above-mentioned component C are specifically as follows: extracting the filter residue 2 with an ethanol solution with a volume fraction of 60% to 80% as the extraction solvent, the weight-to-volume ratio of the filter residue 2 to the extraction solvent is 1:10-15, the extraction time is 1 to 2 hours, the extraction temperature is 60 to 70°C, filtering to obtain filtrate 3 and filter residue 3; extracting the filter residue 3 with an ethanol solution with a volume fraction of 60% to 80% as the extraction solvent, the weight-to-volume ratio of the filter residue 3 to the extraction solvent is 1:10-15, the extraction time is 1 to 2 hours, the extraction temperature is 60 to 70°C, filtering to obtain filtrate 4 and filter residue 4; combining the filtrate filtrate 3 and the filtrate 4, concentrating until there is no alcohol taste, freeze-drying and crushing the obtained concentrated liquid to obtain an alcohol extract of the traditional Chinese medicine mixture with a moisture content of less than 5%, which is recorded as component C.

[0009] A traditional Chinese medicine compound extract for relieving physical fatigue is prepared by the above-mentioned preparation method.

[0010] Application of the above-mentioned Chinese herbal compound extract in the preparation of a product for relieving physical fatigue.

[0011] Furthermore, the above products include medicines and health foods.

[0012] The significant advantages of the present invention are: The present invention discloses a traditional Chinese medicine compound extract for relieving physical fatigue and its preparation method. The compound extract is made from four traditional Chinese medicines: ginseng, astragalus, salvia miltiorrhiza, and ganoderma. The four herbs are mixed in a suitable ratio and first subjected to water extraction and alcohol precipitation to produce a crude polysaccharide component A. The supernatant of the alcohol precipitation is concentrated and purified with a macroporous resin to produce a total saponin component B. The residues of the four herbs are then subjected to alcohol extraction to produce an alcohol extract C containing characteristic components such as salvianolic acid B and astragaloside IV. The crude polysaccharide component A, total saponin component B, and alcohol extract C are then mixed to produce the compound extract. This compound extract is the first of its kind for relieving physical fatigue, featuring precise characteristic components and filling a gap in the industry.

[0013] The compound Chinese herbal extract of the present invention is extracted by mixing four Chinese herbal pieces. Compared with the extraction and purification of individual pieces, it is simpler, requires fewer steps, and is more economical. Safety toxicology evaluation and animal functional evaluation tests support the compound Chinese herbal extract, demonstrating that it is not only safe for consumption but also has significant health benefits in alleviating physical fatigue.

[0014] The preparation process of the traditional Chinese medicine compound extract of the present invention adopts water extraction and alcohol precipitation, supernatant concentration, purification, and alcohol extraction of medicinal residues to prepare three components A / B / C, and the extraction utilization rate of the effective components of the medicinal materials is high. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 : Effects of Chinese herbal compound extracts on body weight in mice subjected to weight-bearing swimming experiment.

[0016] Figure 2 : Effects of Chinese herbal compound extracts on body weight in experimental mice tested for serum urea.

[0017] Figure 3 : Effects of Chinese herbal compound extracts on body weight of mice in the liver glycogen content determination experiment.

[0018] Figure 4 : Effects of Chinese herbal compound extracts on body weight in mice subjected to blood lactate determination experiments.

[0019] Figure 5 : Effects of Chinese herbal compound extracts on weighted swimming time in mice in weighted swimming experiment.

[0020] Figure 6 : Effects of Chinese herbal compound extracts on serum urea content in mice tested for serum urea.

[0021] Figure 7 : Effects of Chinese herbal compound extracts on liver glycogen content in experimental mice.

[0022] Figure 8 : Effects of Chinese herbal compound extracts on blood lactate levels in mice tested for blood lactate. DETAILED DESCRIPTION

[0023] To make the contents of the present invention easier to understand, the technical solutions of the present invention are further described below in conjunction with specific implementation methods, but the present invention is not limited thereto.

[0024] Example 1: A method for preparing a traditional Chinese medicine compound extract for relieving physical fatigue comprises the following steps performed in sequence: 1) 1 part by weight of ginseng Chinese medicinal slices, 3 parts by weight of astragalus Chinese medicinal slices, 4 parts by weight of salvia miltiorrhiza Chinese medicinal slices and 2 parts by weight of ganoderma lucidum Chinese medicinal slices are mixed to obtain a Chinese medicinal slice mixture.

[0025] 2) extracting the Chinese herbal medicine slice mixture in step 1) with water as the extraction solvent, the weight volume ratio of the Chinese herbal medicine slice mixture to the extraction solvent is 1:15, the extraction time is 2 hours, the extraction temperature is 95-100° C., filtering to obtain a filtrate 1 and a filter residue 1; extracting the filter residue 1 with water as the extraction solvent, the weight volume ratio of the filter residue 1 to the extraction solvent is 1:12, the extraction time is 1.5 hours, the extraction temperature is 95-100° C., filtering to obtain a filtrate 2 and a filter residue 2; combining the filtrate 1 and the filtrate 2, and concentrating at 65° C. and -0.1 MPa to a density of 1.05 g / cm 3 , add 95% ethanol solution by volume to the obtained concentrated solution, with the volume ratio of the concentrated solution to the ethanol solution being 1:4, and subject to alcohol precipitation at 3°C ​​for 15 hours. Let it stand for separation to obtain an alcohol precipitation supernatant and an alcohol precipitation precipitate; freeze-dry and pulverize the alcohol precipitation precipitate to obtain a crude polysaccharide component with a moisture content of <5%, which is recorded as component A.

[0026] 3) The supernatant of the alcohol precipitation in step 2) was subjected to ethanol recovery treatment at 65° C. and −0.08 MPa until the alcohol taste was eliminated, and then adsorbed to saturation using a macroporous adsorption resin column (model D101C (Xi'an Lanxiao Technology New Materials Co., Ltd.)) at an adsorption rate of 2 BV / h. After adsorption, the supernatant was eluted with a 70% by volume ethanol solution at a rate of 1 BV / h, and the eluate was collected until the effluent was clear; the collected eluate was concentrated at 70° C. and −0.1 MPa until the alcohol taste was eliminated, and the resulting concentrated solution was freeze-dried and pulverized to obtain a total saponin fraction with a moisture content of less than 5%, which was designated as component B.

[0027] 4) extracting the filter residue 2 from step 2) with a 70% ethanol solution by volume as the extraction solvent, the weight-to-volume ratio of the filter residue 2 to the extraction solvent being 1:15, the extraction time being 2 h, the extraction temperature being 65° C., and filtering to obtain a filtrate 3 and a filter residue 3; extracting the filter residue 3 with a 70% ethanol solution by volume as the extraction solvent, the weight-to-volume ratio of the filter residue 3 to the extraction solvent being 1:12, the extraction time being 1.5 h, the extraction temperature being 65° C., and filtering to obtain a filtrate 4 and a filter residue 4; combining the filtrate 3 and the filtrate 4, and concentrating at 70° C. and −0.1 MPa until there is no alcohol taste, freeze-drying the obtained concentrate, and pulverizing to obtain an alcohol extract of the traditional Chinese medicine mixture having a moisture content of <5%, which is recorded as component C; 5) Component A from step 2), component B from step 3) and component C from step 4) are mixed to obtain a Chinese herbal compound extract. The extraction yield of the Chinese herbal compound extract is 17.5%.

[0028] Example 2: A method for preparing a traditional Chinese medicine compound extract for relieving physical fatigue comprises the following steps performed in sequence: 1) 2 parts by weight of ginseng Chinese medicinal slices, 6.5 parts by weight of astragalus Chinese medicinal slices, 7.5 parts by weight of salvia miltiorrhiza Chinese medicinal slices and 4.5 parts by weight of ganoderma lucidum Chinese medicinal slices are mixed to obtain a Chinese medicinal slice mixture.

[0029] 2) extracting the Chinese herbal medicine slice mixture in step 1) with water as the extraction solvent, wherein the weight-to-volume ratio of the Chinese herbal medicine slice mixture to the extraction solvent is 1:12, the extraction time is 1.5 hours, the extraction temperature is 95-100° C., filtering to obtain a filtrate 1 and a filter residue 1; extracting the filter residue 1 with water as the extraction solvent, wherein the weight-to-volume ratio of the filter residue 1 to the extraction solvent is 1:10, the extraction time is 1 hour, the extraction temperature is 95-100° C., filtering to obtain a filtrate 2 and a filter residue 2; combining the filtrate 1 and the filtrate 2, and concentrating at 70° C. and −0.1 MPa to a density of 1.1 g / cm 3 , add 95% ethanol solution by volume to the obtained concentrated solution, with the volume ratio of the concentrated solution to the ethanol solution being 1:4, and subject to alcohol precipitation at 4°C for 15 hours. Let it stand for separation to obtain an alcohol precipitation supernatant and an alcohol precipitation precipitate; freeze-dry and pulverize the alcohol precipitation precipitate to obtain a crude polysaccharide component with a moisture content of <5%, which is recorded as component A.

[0030] 3) The supernatant of the alcohol precipitation in step 2) was subjected to ethanol recovery treatment at 65° C. and −0.08 MPa until the alcohol taste was eliminated, and then adsorbed to saturation using a macroporous adsorption resin column, the macroporous adsorption resin being D101C (Xi'an Lanxiao Technology New Materials Co., Ltd.) at an adsorption rate of 3 BV / h. After adsorption, the supernatant was eluted with a 75% by volume ethanol solution at an elution rate of 1 BV / h, and the eluate was collected until the effluent was clear; the collected eluate was vacuum concentrated at 70° C. and −0.1 MPa until the alcohol taste was eliminated, and the resulting concentrate was freeze-dried and pulverized to obtain a total saponin fraction with a moisture content of less than 5%, which was designated as component B.

[0031] 4) extracting the filter residue 2 from step 2) with a 65% volume fraction ethanol solution as the extraction solvent, the weight volume ratio of the filter residue 2 to the extraction solvent being 1:12, the extraction time being 1.5 h, the extraction temperature being 70° C., filtering to obtain a filtrate 3 and a filter residue 3; extracting the filter residue 3 with a 70% volume fraction ethanol solution as the extraction solvent, the weight volume ratio of the filter residue 3 to the extraction solvent being 1:10, the extraction time being 1 h, the extraction temperature being 70° C., filtering to obtain a filtrate 4 and a filter residue 4; combining the filtrate 3 and the filtrate 4, and concentrating under vacuum at 75° C. and −0.1 MPa until there is no alcohol taste, freeze-drying the obtained concentrate, and pulverizing to obtain an alcohol extract of the traditional Chinese medicine mixture with a moisture content of <5%, recorded as component C; 5) Component A from step 2), component B from step 3) and component C from step 4) are mixed to obtain a Chinese herbal compound extract. The extraction yield of the Chinese herbal compound extract is 15%.

[0032] Example 3: Pharmaceutical research was carried out using the Chinese herbal compound extract prepared in Example 2: Trial grouping and dosage design: 160 male SPF Kunming mice weighing 18-22g were randomly divided into four experimental groups, each containing 40 mice. Each experimental group was further randomly divided into three dose groups and a negative (pure water) control group, each containing 10 mice. Experimental group 1 performed a weighted swimming test, experimental group 2 performed a serum urea test, experimental group 3 performed a blood lactate test, and experimental group 4 performed a liver glycogen test. Dosage design: Three dose groups were set up: 1.59g / kg.BW (high dose), 1.06g / kg.BW (medium dose), and 0.53g / kg.BW (low dose); Sample preparation: 3.20g, 2.14g, and 1.06g of the Chinese herbal compound extract were weighed and added to 40mL of pure water to make suspensions with mass concentrations of 8%, 5.4%, and 2.6% as high, medium, and low doses, respectively. The Chinese herbal compound extract was administered by gavage at a volume of 20mL / kg.BW, and the negative control group was given the same volume of pure water for continuous gavage for 30 days.

[0033] Experimental methods: Weighted swimming test: Thirty minutes after the last administration of the Chinese herbal compound extract, mice, each bearing a lead weight of 5% of their body weight at the base of their tails, were placed in a swimming tank at a temperature of 25±1°C and a water depth of at least 30 cm. The time from the start of swimming to death was recorded for each group of mice, which was used as the weighted swimming time. Death was determined when the mice sank to the bottom of the tank and ceased breathing through their mouths. If the swimming time of the dosed group was significantly longer than that of the negative control group, and the difference was statistically significant (P<0.05), the Chinese herbal compound extract was considered to have prolonged the weighted swimming time of the mice.

[0034] Serum urea test: Thirty minutes after the last administration of the Chinese herbal compound extract, the mice swam unweighted in water at 30±1°C for 90 minutes. After a 60-minute rest, 0.5 mL of blood was drawn from their eyeballs. The serum was separated and then measured for serum urea using an automated biochemical analyzer. If the serum urea content in the dosed group was significantly lower than that in the negative control group, and the difference was statistically significant (P<0.05), it was determined that the Chinese herbal compound extract had the effect of reducing urea production in fatigued mice.

[0035] Liver glycogen test: Thirty minutes after the last administration of the TCM compound extract, mice were sacrificed and their livers removed. Liver glycogen levels were measured according to the kit instructions, and the results from each dose group were compared with those from the negative control group using analysis of variance. Liver glycogen levels were measured using a kit provided by the Nanjing Jiancheng Bioengineering Research Institute. If the liver glycogen content in the dose group was significantly higher than that in the negative control group, and the difference was statistically significant (P < 0.05), the TCM compound extract was considered to have an effect on promoting liver glycogen storage in mice.

[0036] Blood lactate test: Blood was collected 30 minutes after the last administration of the Chinese herbal compound extract. Mice were allowed to swim unweighted in 30±1°C water for 10 minutes before blood was collected. After resting for 20 minutes, blood was collected three times (from the inner canthus) to measure blood lactate levels. The area under the blood lactate curve at these three time points was used as an indicator, and the results from each dose group were compared with those of the negative control group through analysis of variance. Blood lactate was measured using a kit provided by the Nanjing Jiancheng Bioengineering Research Institute. If the area under the blood lactate curve in the dose group was smaller than that in the negative control group, and the difference was significant (P < 0.05), it was determined that the Chinese herbal compound extract reduced the area under the blood lactate curve in mice after exercise.

[0037] Result judgment: If the result of the weighted swimming test is positive and any two of the three biochemical indicators of blood lactate, serum urea and liver glycogen are positive, it can be determined that the Chinese herbal compound extract has the effect of relieving physical fatigue.

[0038] Experimental results: Effects of Chinese herbal compound extracts on the body weight of experimental mice Figures 1 to 4 As shown in Tables 1 to 4.

[0039] Table 1 Effects of Chinese herbal compound extracts on body weight of mice in weight-bearing swimming experiment Table 2 Effects of Chinese herbal compound extracts on body weight of mice in serum urea determination experiment Table 3 Effects of Chinese herbal compound extracts on the body weight of mice in the liver glycogen content determination experiment Table 4 Effects of Chinese herbal compound extracts on body weight of mice in blood lactate determination experiment Depend on Figures 1 to 4As shown in Tables 1 to 4, after administration of the Chinese herbal compound extract, there was no significant difference in body weight and total weight gain of mice in each dose group compared with the negative control group (P>0.05).

[0040] Effects of Chinese herbal compound extracts on weight-bearing swimming time in mice Figure 5 and as shown in Table 5.

[0041] Table 5 Effects of Chinese herbal compound extracts on weight-bearing swimming time in mice Depend on Figure 5 As shown in Table 5, after administration of the Chinese herbal compound extract, the weighted swimming time of mice in each dose group was significantly prolonged compared with the negative control group, with significant differences between the medium and high dose groups (P < 0.001). This indicates that the Chinese herbal compound extract has the effect of prolonging the weighted swimming time of mice.

[0042] Effects of Chinese herbal compound extracts on serum urea during exercise in mice Figure 6 and as shown in Table 6.

[0043] Table 6 Effects of Chinese herbal compound extracts on serum urea during exercise in mice Depend on Figure 6 As shown in Table 6, after administration of the Chinese herbal compound extract, there was no significant difference in the serum urea content of mice in each dose group compared with the negative control group (P>0.05).

[0044] Effects of Chinese herbal compound extracts on liver glycogen content in mice Figure 7 and as shown in Table 7.

[0045] Table 7 Effects of Chinese herbal compound extracts on liver glycogen content in mice Depend on Figure 7 As shown in Table 7, after administration of the Chinese herbal compound extract, liver glycogen content increased in all dose groups compared with the negative control group, with significant differences between the medium and high dose groups (P < 0.001). This indicates that the Chinese herbal compound extract has the effect of promoting liver glycogen storage in mice.

[0046] Effects of Chinese herbal compound extracts on blood lactate in mice Figure 8 As shown in Tables 8 to 11.

[0047] Table 8 Effects of Chinese herbal compound extracts on blood lactate in mice before swimming Table 9 Effects of Chinese herbal compound extracts on blood lactate in mice after swimming for 10 minutes Table 10 Effects of Chinese herbal compound extracts on blood lactate in mice after 20 minutes of swimming rest Table 11 Effects of Chinese herbal compound extracts on the area under the curve of blood lactate in mice Depend on Figure 8 As shown in Tables 8 to 11, after administration of the Chinese herbal compound extract, the blood lactate content of each dose group decreased compared with the negative control group, and the area under the blood lactate curve was significantly reduced. As shown in Tables 8 to 9, there was no significant difference between the dose groups in the effect of the Chinese herbal compound extract on the blood lactate level of mice before swimming and 10 minutes after swimming (p>0.05). As shown in Tables 10 to 11, there was a significant difference in the blood lactate level and the area under the blood lactate curve of mice after swimming and resting for 20 minutes (P<0.01) between the negative control group and the high-dose group. It can be determined that the Chinese herbal compound extract has the effect of reducing the blood lactate level and the area under the blood lactate curve of mice after exercise.

[0048] summary: The Chinese herbal compound extract was administered to mice orally at doses of 1.59g / kg.BW, 1.06g / kg.BW, and 0.53g / kg.BW, once a day for 30 consecutive days. The test results are as follows: Effect on the body weight of mice: After administration of the Chinese herbal compound extract, there was no significant difference in the body weight and total weight gain of mice in each dose group compared with the negative control group (P>0.05).

[0049] Mouse weighted swimming test: After administration of the Chinese herbal compound extract, the weighted swimming time of mice in each dose group was significantly prolonged compared with the negative control group, and the difference between the medium and high dose groups was significant (P<0.001). The result was positive.

[0050] Determination of mouse serum urea: After administration of the Chinese herbal compound extract, there was no significant difference in the serum urea content of mice in each dose group compared with the negative control group (P>0.05). The result was negative.

[0051] Mouse liver glycogen determination: After administration of the Chinese herbal compound extract, the liver glycogen content of each dose group increased compared with the negative control group, and the liver glycogen content of the medium and high dose groups increased significantly (P<0.001). The result was positive.

[0052] Blood lactate determination in mice: After administration of the Chinese herbal compound extract, the blood lactate content in each dose group decreased compared with the negative control group, and the area under the blood lactate curve was significantly reduced, and the difference in the high dose was significant (P<0.01). The result was positive.

[0053] To sum up: the results of the weighted swimming test were positive, and the two indicators of blood lactate and liver glycogen were positive. According to the "Evaluation Method for Relieving Physical Fatigue Function" in the "Health Food Functional Testing and Evaluation Methods (2023 Edition)", it can be determined that the Chinese medicine compound extract has the effect of relieving physical fatigue.

[0054] Example 4: Component C of Example 2 was determined with reference to Volume 1 of the Pharmacopoeia of the People's Republic of China (2020 edition), astragaloside IV was determined with reference to the method under "Huangqi Shengmai Granules", and salvianolic acid B was determined with reference to the method under "Danshen". The results showed that component C contained astragaloside IV and salvianolic acid B.

[0055] Table 12 Astragaloside IV standard curve data and linear regression equation Table 13 Salvianolic acid B standard curve data and linear regression equation Table 14 Test data of three batches of astragaloside IV and salvianolic acid B in component C The above description is only a preferred embodiment of the present invention. All equivalent changes and modifications made according to the scope of the patent application of the present invention should fall within the scope of the present invention.

Claims

1. A method for preparing a Chinese herbal compound extract for relieving physical fatigue, characterized in that: The following steps are involved: 1) Mixing the Chinese herbal medicine slices of ginseng, astragalus, salvia miltiorrhiza and ganoderma lucidum according to a proportion to obtain a Chinese herbal medicine slice mixture; 2) extracting the mixture of Chinese herbal medicine slices from step 1) with water as an extraction solvent, filtering to obtain a filtrate 1 and a filter residue 1; extracting the filter residue 1 with water as an extraction solvent, filtering to obtain a filtrate 2 and a filter residue 2; combining the filtrate 1 and the filtrate 2, concentrating, precipitating with alcohol, and standing to separate to obtain an alcohol supernatant and an alcohol precipitate; freeze-drying and pulverizing the alcohol precipitate to obtain component A; 3) recovering ethanol from the alcohol precipitation supernatant of step 2), adsorbing the supernatant to saturation using a macroporous adsorption resin column, eluting the column with ethanol after adsorption, and collecting the eluate until the effluent is clear; concentrating the collected eluate, freeze-drying it, and pulverizing it to obtain component B; 4) extracting the filter residue 2 from step 2) with an ethanol solution as an extraction solvent, filtering to obtain a filtrate 3 and a filter residue 3; extracting the filter residue 3 with an ethanol solution as an extraction solvent, filtering to obtain a filtrate 4 and a filter residue 4; combining the filtrate 3 and the filtrate 4, concentrating, freeze-drying, and pulverizing to obtain component C; 5) Mixing component A from step 2), component B from step 3) and component C from step 4) to obtain a Chinese herbal compound extract.

2. The preparation method according to claim 1, wherein: The Chinese herbal medicine slice mixture consists of the following Chinese herbal medicine slices in parts by weight: 1-3 parts of ginseng Chinese herbal medicine slices, 3-10 parts of astragalus Chinese herbal medicine slices, 4-13 parts of salvia miltiorrhiza Chinese herbal medicine slices, and 2-7 parts of ganoderma lucidum Chinese herbal medicine slices.

3. The preparation method according to claim 1, wherein: The preparation steps of component A are specifically as follows: extracting the mixture of Chinese herbal medicine pieces with water as the extraction solvent, the weight volume ratio of the mixture of Chinese herbal medicine pieces to the extraction solvent is 1:10-15, the extraction time is 1-2 hours, the extraction temperature is 95-100°C, filtering to obtain filtrate 1 and filter residue 1; extracting the filter residue 1 with water as the extraction solvent, the weight volume ratio of the filter residue 1 to the extraction solvent is 1:10-15, the extraction time is 1-2 hours, the extraction temperature is 95-100°C, filtering to obtain filtrate 2 and filter residue 2; combining the filtrate 1 and the filtrate 2, and concentrating to a density of 1.05-1.15 g / cm 3 , add 95% ethanol solution by volume to the obtained concentrated solution, with the volume ratio of the concentrated solution to the ethanol solution being 1:4, and subject the product to alcohol precipitation at 1-5°C for more than 12 hours. Allow the product to stand for separation to obtain an alcohol precipitation supernatant and an alcohol precipitation precipitate. The alcohol precipitation precipitate is freeze-dried and pulverized to obtain a crude polysaccharide component with a moisture content of less than 5%, which is recorded as component A.

4. The preparation method according to claim 1, wherein: The preparation steps of the component B are specifically as follows: subjecting the alcohol precipitation supernatant to ethanol recovery treatment until there is no alcohol taste, then adsorbing to saturation using a macroporous adsorption resin chromatography column, wherein the model of the macroporous adsorption resin is D101C and the adsorption rate is 2-3 BV / h; after adsorption, eluting with an ethanol solution with a volume fraction of 70-75% at an elution rate of 1 BV / h, and collecting the eluate until the effluent is clear; concentrating the collected eluate until there is no alcohol taste, freeze-drying and pulverizing the obtained concentrated solution to obtain a total saponin component with a moisture content of less than 5%, which is recorded as component B.

5. The preparation method according to claim 1, wherein: The preparation steps of the component C are specifically as follows: extracting the filter residue 2 with an ethanol solution with a volume fraction of 60% to 80% as an extraction solvent, the weight-to-volume ratio of the filter residue 2 to the extraction solvent is 1:10-15, the extraction time is 1-2 hours, the extraction temperature is 60-70°C, filtering to obtain a filtrate 3 and a filter residue 3; extracting the filter residue 3 with an ethanol solution with a volume fraction of 60% to 80% as an extraction solvent, the weight-to-volume ratio of the filter residue 3 to the extraction solvent is 1:10-15, the extraction time is 1-2 hours, the extraction temperature is 60-70°C, filtering to obtain a filtrate 4 and a filter residue 4; combining the filtrate 3 and the filtrate 4, concentrating until there is no alcohol taste, freeze-drying the obtained concentrated solution, and pulverizing to prepare an alcohol extract of a traditional Chinese medicine mixture with a moisture content of less than 5%, which is recorded as component C.

6. A Chinese herbal compound extract for relieving physical fatigue, characterized in that: Prepared by the preparation method according to any one of claims 1 to 5.

7. Use of the Chinese herbal compound extract according to claim 6 in preparing a product for relieving physical fatigue.

8. The use according to claim 7, characterized in that: The products include medicines and health foods.

Images