Composition with effects of improving anemia and resisting fatigue and preparation method thereof

Abstract

The invention discloses a composition with the effects of improving anemia and resisting fatigue and a preparation method of the composition. The composition for improving anemia also has an anti-fatigue effect, and contains a ginseng fermentation product; wherein the ginseng fermentation product is obtained by fermenting a ginseng fermentation substrate by using a microbial strain and contains ginsenoside Rh4, and the ginseng fermentation product meets the following conditions: the content of the ginsenoside Rh4 is more than or equal to 7wt%; the total amount of ginsenoside in the ginseng fermentation product is 100% by weight. The composition provided by the invention has definite and prominent efficacy of improving anemia and / or resisting fatigue, and has the advantages of no side effect, good taste and high safety.

Description

Technical Field

[0001] The invention belongs to the field of biomedicine manufacturing or food biotechnology, and specifically relates to a composition rich in rare ginsenosides, which has the effects of improving anemia and resisting fatigue, and a preparation method thereof. Background Art

[0002] Anemia is a common blood disease, the main feature of which is a decrease in the number of red blood cells in the peripheral blood, resulting in insufficient oxygen supply to tissues and organs. The global prevalence of anemia is as high as 24.8%, affecting the health of 1.62 billion people. The causes of anemia can be attributed to reduced or insufficient red blood cell production, excessive destruction of red blood cells and blood loss, and may be affected by factors such as dietary habits, long-term and large-scale exposure to chemicals, viral infections, chronic diseases or malignant tumors. In anemia, the number of red blood cells in the blood decreases or the hemoglobin content decreases, resulting in a decrease in oxygen transport capacity. Various organs and tissues in the body do not get enough oxygen supply, especially the brain, heart and muscles, which have a high demand for oxygen. Brain hypoxia can cause symptoms such as dizziness, fatigue, and inattention, making people feel tired; in order to compensate for the lack of oxygen, the heart will beat faster, which will also increase fatigue in the long run; lack of sufficient oxygen in muscles will reduce exercise ability and make them easily fatigued.

[0003] Iron deficiency anemia is a common disease, mainly caused by an imbalance in iron supply and demand and reduced hemoglobin synthesis, accounting for 60% to 70% of anemic diseases. Iron supplementation alone is prone to adverse reactions. Traditional Chinese medicine often uses Chinese medicines such as ginseng, codonopsis, astragalus, acanthopanax, jujube, placenta, longan meat, etc. to replenish qi and blood, and the results are effective and have no side effects.

[0004] Ginseng has the effects of replenishing vital energy, strengthening the spleen and lungs, and restoring pulse and strengthening the body. The active ingredients it contains, such as saponins, polysaccharides, amino acids, volatile oils, polyacetylenes, and a variety of other trace elements, are the important material basis for its efficacy. The properties and pharmacological activities of different ginseng components are also different. The main active ingredient of ginseng is ginsenoside. Studies have found that the secondary metabolite derivatives of ginsenosides have stronger biological activity after transformation. This secondary metabolite derivative is named "rare ginsenoside" because of its rare content in natural ginseng. In contrast, ginsenosides extracted directly from Araliaceae plants are called "prototype ginsenosides."

[0005] By using probiotics to ferment ginseng with ginseng as a substrate, ginseng fermentation products such as fermented ginseng powder can be obtained. The fermentation process can convert prototype ginsenosides into rare ginsenosides, thereby improving their biological activity and utilization. The ginseng fermentation product has a better taste and can reduce the original bitterness of ginseng.

[0006] According to the literature research conducted by the inventors, ginsenoside Rh4 is the only rare ginsenoside that has a clear effect on improving anemia. However, the content of ginsenoside Rh4 in existing ginseng fermentation products is very low, and it is difficult to say that it has a clear and outstanding effect on improving anemia. Summary of the invention

[0007] In view of the above technical problems existing in the prior art, the object of the present invention is to provide a ginseng fermentation product rich in rare ginsenosides with enhanced anemia-improving effect, and a composition using the ginseng fermentation product as a raw material, wherein the composition has a clear and outstanding anemia-improving effect. Furthermore, the composition may also have a clear and outstanding anti-fatigue effect.

[0008] After literature research, the inventors found that among the rare ginsenosides, ginsenoside Rh4 is the one that can improve anemia. Based on this, the inventors believe that if the content of ginsenoside Rh4 in ginseng fermentation products can be increased, ginseng fermentation products with enhanced anemia-improving effects can be prepared. However, the technical difficulty is that even among the rare ginsenosides, ginsenoside Rh4 is difficult to obtain through microbial fermentation conversion, and its content in the composition of various rare ginsenosides produced by microbial fermentation is very low. If the total amount of ginsenosides in ginseng fermentation products is 100% by weight, the content of ginsenoside Rh4 generally does not exceed 1% by weight.

[0009] Furthermore, the inventors have found through in-depth research that fermenting ginseng matrix with a specific combination of Lactobacillus plantarum and Lactobacillus casei can produce a ginseng fermentation product with a high content of ginsenoside Rh4. Furthermore, the ginseng fermentation product can be used as a raw material to prepare a composition for improving anemia. Furthermore, because ginsenoside Rh4 also has anti-fatigue effects, the inventors have also found that by combining the above-mentioned ginseng fermentation product with refreshing and anti-fatigue ingredients such as honeysuckle powder, jasmine tea powder, B vitamins, taurine, etc., a composition with anemia improvement effect and anti-fatigue effect can be obtained.

[0010] The technical solution of the present invention includes:

[0011] 1. A ginseng fermentation product rich in ginsenoside Rh4, obtained by fermenting a ginseng fermentation substrate using a microbial strain, comprising ginsenoside Rh4, and satisfying:

[0012] The content of ginsenoside Rh4 is ≥7 wt%, preferably ≥8 wt%;

[0013] The total amount of ginsenosides in the ginseng fermentation product is taken as 100% by weight.

[0014] The ginseng fermentation product may be liquid or solid or a solid-liquid mixture, for example, fermented ginseng liquid or fermented ginseng powder.

[0015] 2. A bacterial composition, which consists of Lactobacillus plantarum and Lactobacillus casei; wherein:

[0016] The plant lactobacillus is plant lactobacillus GBFG 61, which is deposited in the General Microbiology Center of China National Microbiological Culture Collection Administration, with a deposit number of GGMCC NO: 31583 and a deposit date of August 14, 2024.

[0017] 3. The bacterial composition according to item 2, which can be used to ferment ginseng fermentation substrate to produce rare ginsenosides.

[0018] Here, the ginseng fermentation matrix refers to a fermentation matrix containing ginseng fines. The ginseng fines are obtained by reducing ginseng, preferably by physical methods such as extrusion, cutting, grinding, etc. The ginseng fines can be, for example, ginseng powder. The ginseng fermentation matrix can also contain other components commonly contained in the fermentation matrix, such as nitrogen source, carbon source, inorganic salt, pH regulator, etc.

[0019] 4. The bacterial composition according to item 2, wherein the dry weight ratio of Lactobacillus plantarum to Lactobacillus casei is 1:3-5.

[0020] 5. The bacterial composition according to item 2, wherein the Lactobacillus casei is Lactobacillus casei LC-12.

[0021] 6. A method for preparing a ginseng fermentation product rich in ginsenoside Rh4, comprising inoculating a bacterial composition into a ginseng fermentation substrate, and fermenting at 36-39°C, preferably 37°C, for 10-50 hours, preferably 15-40 hours, more preferably 20-30 hours, to obtain a ginseng fermentation product containing rare ginsenosides;

[0022] Wherein, the bacterial composition is the bacterial composition described in any one of items 2 to 5.

[0023] 7. The preparation method according to item 6, further comprising sterilizing the ginseng fermentation product.

[0024] 8. The preparation method according to item 6, wherein the ginseng fermentation product is the ginseng fermentation product according to item 1.

[0025] 9. The preparation method according to item 6, wherein the ginseng fermentation product is fermented ginseng liquid or fermented ginseng powder.

[0026] 10. A method for preparing a composition for improving anemia, comprising:

[0027] A ginseng fermentation product is prepared by the preparation method of any one of items 6 to 9; and

[0028] The ginseng fermentation product is mixed with other components of the anemia improving composition to obtain the anemia improving composition.

[0029] 11. A composition for improving anemia, comprising the ginseng fermentation product described in item 1.

[0030] 12. The composition for improving anemia according to item 13, further comprising a refreshing or anti-fatigue ingredient, and further having an anti-fatigue effect.

[0031] 13. A composition for improving anemia according to item 12, wherein the refreshing or anti-fatigue ingredients include one or more or all selected from honeysuckle, jasmine tea powder, B group vitamins (such as vitamin B1, vitamin B2, vitamin B6) and taurine.

[0032] 14. The composition for improving anemia according to item 12, comprising 15 to 300 parts by weight of the ginseng fermentation product, 30 to 54 parts by weight of honeysuckle, 30 to 54 parts by weight of jasmine tea powder, and vitamin B1

[0033] 0.2-0.25 weight part, vitamin B2 0.1-0.2 weight part, vitamin B6 0.1-0.2 weight part and taurine 0.05-0.07 weight part.

[0034] 15. The composition for improving anemia according to item 12, which is in liquid form.

[0035] One or more of sugars, sugar alcohols, non-sugar and non-sugar alcohol sweeteners, pectin, soluble dietary fiber, salts, other carbohydrates, carbon dioxide or other food additives may also be added to the anemia improving composition. Among them, the sugars include one or more of glucose, sucrose or fructose; the non-sugar and sugar alcohol sweeteners include one or more of stevia, aspartame, sucralose, monk fruit, glycyrrhizin, betaine or saccharin sodium; the sugar alcohols include one or more of xylitol, erythritol, maltitol, isomalt or sorbitol; the salts include one or more of sodium salt, potassium salt, phosphate; the other carbohydrates include oligosaccharides, starch and / or soluble and insoluble dietary fiber; the other food additives include one or more of flavors, pigments, malic acid, citric acid, sodium citrate, sorbic acid, potassium sorbate, benzoic acid, sodium benzoate, edible gum base, sucrose esters, soybean lecithin, sodium alginate, and the flavors and pigments are artificially synthesized or natural; the caffeine comes from cocoa, coffee, guarana, tea or chocolate.

[0036] 16. The method for preparing the anemia-improving composition according to item 11, comprising:

[0037] A ginseng fermentation product is prepared by the preparation method of any one of items 6 to 9; and

[0038] The ginseng fermentation product is mixed with other components of the anemia improving composition to obtain the anemia improving composition.

[0039] Advantageous Effects of the Invention

[0040] Ginseng fermentation products have the effects of invigorating qi and blood and resisting fatigue; honeysuckle contains a certain amount of iron, which can assist in iron supplementation, thereby preventing or treating iron deficiency anemia; jasmine tea has the effects of clearing away heat, relieving depression and calming the mind, refreshing the brain, and anti-oxidation; vitamin B1, B2, B6 and taurine can all promote iron absorption, increase the hemoglobin content in tissues, and also have the effect of resisting fatigue. Therefore, the anemia-improving composition of the present invention has clear and prominent anemia-improving and / or anti-fatigue effects, has no side effects, tastes good, and is safe to eat, and can be widely used in foods and medicines for improving anemia. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1 This is a high performance liquid chromatogram for determining the content of ginsenoside Rh4 in the fermented ginseng powder prepared in Example 1.

[0042] Figure 2 This is a high performance liquid chromatogram of the determination of the content of ginsenoside Rh4 in the fermented ginseng powder prepared in Comparative Example 3.

[0043] Figure 3 This is a graph showing the effect of the anemia-improving and anti-fatigue composition 1 on the weight-bearing swimming time of mice. DETAILED DESCRIPTION

[0044] The present invention is described in detail below in conjunction with specific embodiments. In the absence of conflict, the scientific terms in this specification have the meanings commonly understood by those skilled in the art. If there is a conflict, the definitions in this specification shall prevail.

[0045] Example

[0046] The plant lactobacillus (Lactobacillus plantarum) GBFG 61 is a lactobacillus strain that produces β-glucosidase isolated and cultivated from the soil of the ginseng planting base in Fusong County, Baishan City, Jilin Province. The lactobacillus strain was named plant lactobacillus GBFG 61, which is deposited in the General Microbiological Center of the China Microbiological Culture Collection Administration Committee, and the deposit number is CGMCC No.31583. The deposit date is August 14, 2024, and the deposit address is: Institute of Microbiology, Chinese Academy of Sciences, No. 3, No. 1, Beichen West Road, Chaoyang District, Beijing; the deposit postal code of the deposit is 100101.

[0047] The Lactobacillus casei LC-12 was purchased from Shandong Zhongke Jiayi Bioengineering Co., Ltd. The Lactobacillus casei LC-G11 was purchased from Runying Bioengineering (Shanghai) Co., Ltd. The Lactobacillus plantarum Lp90 was purchased from Weikang Probiotics (Suzhou) Co., Ltd. Ginseng was purchased from the Chinese herbal medicine market, and other raw materials and reagents were purchased from commercial channels.

[0048] Example 1 Preparation of fermented ginseng powder rich in rare ginsenoside Rh4

[0049] The steps include:

[0050] (1) Grinding of ginseng

[0051] Ginseng was crushed and ginseng powder with mesh size of 60-80 was selected as fermentation raw material.

[0052] (2) Preparation of culture medium

[0053] 40 wt% of ginseng powder, 1 wt% of glucose, 0.3 wt% of soy protein powder, 0.04 wt% of magnesium sulfate, 0.015 wt% of manganese sulfate and water were uniformly mixed to prepare a fermentation medium containing ginseng, which was sterilized at 121° C. for 15 min.

[0054] (3) Vaccination

[0055] After the temperature of the sterilized culture medium drops below 37° C., Lactobacillus plantarum GBFG 61 powder and Lactobacillus casei LC-12 powder are inoculated into the fermentation medium at a ratio of 1:3 and an inoculation amount of 3% by weight.

[0056] (4) Fermentation: The fermented liquid after inoculation is placed in an incubator at a fermentation temperature of 37° C. for 22 hours.

[0057] (5) Post-treatment: The fermented ginseng liquid was sterilized at 115°C for 20 min. The sterilized fermented liquid was centrifuged at 4500 rpm for 30 min to obtain a precipitate, which was freeze-dried to obtain fermented ginseng powder.

[0058] Example 2

[0059] The ratio of Lactobacillus plantarum GBFG 61 to Lactobacillus casei LC-12 in Example 1 was changed to 1:4, and the other conditions remained unchanged.

[0060] Example 3

[0061] The ratio of Lactobacillus plantarum GBFG 61 to Lactobacillus casei LC-12 in Example 1 was changed to 1:5, and the other conditions remained unchanged.

[0062] Comparative Example 1

[0063] In Example 1, only Lactobacillus plantarum GBFG 61 was inoculated, and the other conditions remained unchanged.

[0064] Comparative Example 2

[0065] Only Lactobacillus casei LC-12 was inoculated in Example 1, and the other conditions remained unchanged.

[0066] Comparative Example 3

[0067] The Lactobacillus plantarum GBFG 61 in Example 1 was replaced with Lactobacillus plantarum Lp90, and the other conditions remained unchanged.

[0068] Comparative Example 4

[0069] The Lactobacillus casei LC-12 in Example 1 was replaced by Lactobacillus casei LC-G11, and the other conditions remained unchanged.

[0070] Example 5 Determination of the content of rare ginsenoside Rh4

[0071] The fermented ginseng powders prepared in Examples 1-3 and Comparative Examples 1-4 were respectively dissolved in methanol and prepared into 80 mg / mL. 1 mL of the solution was respectively taken and filtered with a 0.45 μm microporous filter membrane to obtain the fermented ginseng powder clear solution. The total ginsenoside content and the content of various rare ginsenosides therein were determined, and the ginsenoside Rh4 content (wt %, with the total ginsenosides as 100 wt %) was calculated. The results are shown in Table 1.

[0072] The detection method of total ginsenosides was carried out according to the method for determining the content of total ginsenosides in the Pharmacopoeia of the People's Republic of China (2020 edition). 48 H 82 O 18 )count.

[0073] The detection method of rare ginsenosides is carried out according to the method for determining the content of rare ginsenosides in T / CNHFA 001-2021.

[0074] As an example, the HPLC chromatograms for determining the content of ginsenoside Rh4 in the fermented ginseng powder prepared in Example 1 and Comparative Example 3 are as follows: Figure 1 and Figure 2 shown.

[0075] Table 1. Content of rare ginsenoside Rh4 in fermented ginseng powder

[0076] Serial number Ginsenoside Rh4 content (weight %) Example 1 8.7 Example 2 7.9 Example 3 8.3 Comparative Example 1 <1 Comparative Example 2 <1 Comparative Example 3 1.6 Comparative Example 4 2.1

[0077] As can be seen from Table 1, when the plant lactobacillus GBFG 61 strain of the present invention and the lactobacillus casei LC-12 strain fermentation strain combination are selected, the content of rare ginsenoside Rh4 in the obtained fermented ginseng powder is significantly improved. This prompts that the obtained fermented ginseng powder has a strengthened effect of improving anemia. It should also be noted that, in addition to the plant lactobacillus Lp90 and the lactobacillus casei LC-G11 of the above-mentioned example comparison, the inventor has tried a variety of plant lactobacillus, lactobacillus casei and combinations thereof that are separated by themselves or purchased commercially, and all cannot achieve the technical effect that the content of rare ginsenoside Rh4 is more than 7% by weight.

[0078] Example 6 Effects of different fermented ginseng powders on iron deficiency anemia rats

[0079] Forty healthy weaned rats were selected and adapted to the experimental environment for 3 days, and then randomly divided into a normal control group of 10 rats and a low-iron experimental group of 30 rats. The normal group was fed with normal feed and deionized water, and the low-iron experimental group was fed with low-iron feed and deionized water. At the same time, blood was collected by tail-cutting method, once every 5 days, 0.4 ml each time. Starting from the third week, some rats were selected every week to collect tail blood for hemoglobin (Hb) measurement. Rats with an Hb mass concentration of about 60 g / L were selected as experimental animals. According to the Hb level and body weight of anemic rats, they were randomly divided into a low iron model group and two experimental groups. Each group continued to be fed with a low iron feed, and the low iron control group was given the corresponding solvent. The two experimental groups were the fermented ginseng powder group (50 mg / kg BW) in Example 1 and the fermented ginseng powder group (50 mg / kg BW) in Comparative Example 3. After 30 days of administration of the test sample, the abdominal aorta blood of the rats was taken, and the hemoglobin, red blood cell (RBC), and hematocrit (HCT) were detected using a fully automatic blood analyzer. The test results are shown in Table 2.

[0080] Table 2. Effects of each dose group on hemoglobin and red blood cell count in rats

[0081]

[0082] Note: Compared with the normal group, ***P<0.001; compared with the model group, ### P<0.001.

[0083] As shown in Table 2, as for the Example 1 group, after treatment, the Hb, RBC, and HCT levels of the anemic rats were significantly restored, with significant differences compared with the model group, and were close to the levels of the normal group, indicating that it has a significant effect on improving anemia; on the contrary, as for the Comparative Example 3 group, although the Hb, RBC, and HCT levels of the anemic rats were restored, the difference was not significant compared with the model group, but was significant compared with the normal group, indicating that its effect on improving anemia is limited.

[0084] Example 7 Preparation of Composition for Improving Anemia and Fighting Fatigue

[0085] The following components were weighed by weight: 30 parts of fermented ginseng powder, 40 parts of honeysuckle, 36 parts of jasmine tea powder, 0.225 parts of vitamin B1, 0.15 parts of vitamin B2, 0.15 parts of vitamin B6, and 0.065 parts of taurine. After the above raw materials were evenly mixed, 1500 parts of erythritol, 60 parts of citric acid and a certain amount of water were added, stirred evenly, and aseptically filled to obtain anemia-improving and anti-fatigue composition 1.

[0086] Example 8 Verification of the efficacy of the anemia-improving and anti-fatigue-improving composition in improving anemia

[0087] The anemia-improving anti-fatigue composition 1 was verified for its anemia-improving efficacy. The experimental operation was performed with reference to Example 6. The dosage of the anemia-improving anti-fatigue composition 1 was 80 mg / (kg BW). Erythritol, citric acid and water were not included in the effective ingredients of the anemia-improving anti-fatigue composition 1. The results of the determination of hemoglobin, red blood cell count and hematocrit are shown in Table 3.

[0088] Table 3. Effects of the administration of the composition on hemoglobin, red blood cell count, and hematocrit in rats

[0089]

[0090] Note: Compared with the normal group, ***P<0.001; compared with the model group, ### P<0.001.

[0091] As shown in Table 3, after treatment with the anemia-improving and anti-fatigue composition of the present invention, the levels of Hb, RBC and HCT of anemic rats were significantly restored, with significant differences compared with the model group, and no significant differences compared with the normal group, indicating that it has a good effect in improving anemia.

[0092] Example 9: Verification of the anti-fatigue efficacy of the anemia-improving anti-fatigue composition

[0093] The anti-fatigue effect of the anemia-improving and anti-fatigue composition 1 was verified.

[0094] Twenty SPF-grade Kunming male mice weighing 20-25g were adaptively raised for 7 days, and after daily swimming training, they were randomly divided into 2 groups, named as the anemia-improving and anti-fatigue composition 1 group and the control group, with 10 mice in each group. In addition to normal daily feeding, each group of mice was gavaged with 80mg / (kg BW) of the anemia-improving and anti-fatigue composition 1 and an equal amount of normal saline, and raised for 30 days. 30 minutes after the last administration, each group of mice was loaded with 5% lead blocks for swimming, and the swimming time of each group of mice was recorded. The weight-bearing swimming time test results are shown in Figure 3 shown.

[0095] Depend on Figure 3 It can be seen that the anemia-improving and anti-fatigue composition of the present invention has a good anti-fatigue effect (compared with the model group, ##P<0.01).

[0096] The above embodiments are preferred implementations of the present invention, but the implementations of the present invention are not limited to the above embodiments. For ordinary technicians in this technical field, other different forms of changes or modifications can be made without departing from the principles of the present invention. The derived changes are still within the scope of the creative protection of the present invention.

[0097] Industrial Applicability

[0098] The present invention provides a ginseng fermentation product containing rare ginsenosides and having enhanced anemia-improving effect, and a preparation method and application thereof.

Claims

1. A composition for improving anemia, comprising a ginseng fermentation product; wherein: The ginseng fermentation product is obtained by fermenting a ginseng fermentation substrate using a microbial strain, and contains ginsenoside Rh4, which satisfies: The content of ginsenoside Rh4 is ≥7 wt%, preferably ≥8 wt%; The total amount of ginsenosides in the ginseng fermentation product is taken as 100% by weight.

2. The composition for improving anemia according to claim 1, which also has an anti-fatigue effect, and further comprises a refreshing or anti-fatigue component.

3. The composition for improving anemia according to claim 2, wherein: The refreshing or anti-fatigue ingredients include one or more or all of honeysuckle, jasmine tea powder, B group vitamins and taurine.

4. The anemia-improving composition according to claim 1 or 2, comprising 15 to 300 parts by weight of the ginseng fermentation product, 30 to 54 parts by weight of honeysuckle, 30 to 54 parts by weight of jasmine tea powder, 0.2 to 0.25 parts by weight of vitamin B1, 0.1 to 0.2 parts by weight of vitamin B2, 0.1 to 0.2 parts by weight of vitamin B6 and 0.05 to 0.07 parts by weight of taurine. The composition for improving anemia according to claim 1 or 2, which is in liquid or solid form.

6. The method for preparing the anemia-improving composition according to claim 1, comprising: Inoculating the bacterial composition into a ginseng fermentation substrate, and fermenting at 36-39° C., preferably 37° C., for 10-50 hours, preferably 15-40 hours, more preferably 20-30 hours, to obtain a ginseng fermentation product containing rare ginsenoside Rh4; and The ginseng fermentation product is mixed with other components of the anemia improving composition to obtain the anemia improving composition.

7. The preparation method according to claim 6, wherein: The bacterial composition consists of Lactobacillus plantarum and Lactobacillus casei; The plant lactobacillus is plant lactobacillus (Lactobacillus plantarum) GBFG 61, which is deposited in the General Microbiological Center of China Microbiological Culture Collection Administration, with a deposit number of GGMCCNO: 31583 and a deposit date of August 14, 2024; The Lactobacillus casei is Lactobacillus casei LC-12.

8. The preparation method according to claim 7, wherein: The dry weight ratio of the Lactobacillus plantarum to the Lactobacillus casei is 1:3-5.

9. The preparation method according to claim 6, wherein: The ginseng fermentation product is fermented ginseng liquid or fermented ginseng powder.

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