Small molecule ginseng paste with anti-aging effect and preparation method thereof

Abstract

The present invention belongs to the technical field of ginseng processing, and mainly relates to a small molecule ginseng paste and a preparation method thereof, and specifically relates to a small molecule ginseng paste with anti-aging efficacy and a preparation method thereof. The present invention develops a ginseng paste, and selects ultrafiltered small molecule ginseng active liquid and wolfberry extract to improve performance through synergistic fermentation of Bifidobacterium bifidum CICC-6171 and Lactobacillus crispatus CICC-24879 to achieve the efficacy of anti-oxidation, anti-aging and inhibiting gastric bleeding.

Description

Technical Field

[0001] The invention belongs to the technical field of ginseng processing, and mainly relates to a small molecule ginseng paste and a preparation method thereof, and specifically relates to a small molecule ginseng paste with anti-aging effect and a preparation method thereof. Background Art

[0002] Ginseng is the dried root and rhizome of the plant Panax ginseng CAMeyer in the Araliaceae family. It is a traditional Chinese medicinal resource with dual medicinal and nutritional values. It is known as the "king of herbs". It is sweet in nature and slightly bitter in taste. It enters the spleen, lung and heart meridians. It has the effects of calming the mind and improving intelligence, replenishing vital energy and tonifying the spleen and lungs. Long-term use can enhance its own immunity and delay aging. Its main chemical components are ginsenosides, and it also includes ginseng polysaccharides, amino acids and flavonoids. It is the material basis for ginseng to exert its multiple pharmacological effects. In recent years, as ginseng has been listed as a new resource food, its edible value has gradually attracted widespread attention, and ginseng series products have emerged, such as Jinxing ginseng capsules and ginseng beverages. According to the unique processing methods and medicinal value of ginseng, ginseng processed products have also been gradually applied, further enriching the development value of ginseng. At the same time, using ginseng deep-processing products to improve the body's defense and antiviral capabilities is more in line with the modern health concept of "food for nourishment and food for treatment". Therefore, the development of ginseng functional foods has important practical significance for promoting the in-depth development of ginseng-related products. Summary of the invention

[0003] In view of this, the present invention develops a ginseng paste, which selects ultrafiltered small molecule ginseng active liquid and wolfberry extract to improve the performance through synergistic fermentation by Bifidobacterium bifidum CICC-6171 and Lactobacillus crispatus CICC-24879 to achieve antioxidant and anti-aging effects.

[0004] In a first aspect, the present invention provides a method for preparing a small molecule ginseng paste, the specific steps of which are as follows:

[0005] S1. Preparation of small molecule ginseng active liquid: select 20g of high-quality ginseng, wash it and slice it, add 75% ethanol with a liquid-to-solid ratio of 1:10, distill and reflux to extract twice, and recover the ethanol under reduced pressure; then add 10% ethanol with a liquid-to-solid ratio of 1:10 and reflux to extract once, combine the extracts and concentrate them to 100mL; filter the concentrated extract through a polyethersulfone filter membrane, and then pour the filtrate into a BIE050D010 hollow fiber column for ultrafiltration, and retain small molecular substances below 10,000 units to prepare small molecule ginseng active liquid for use; in this step, the ultrafiltration process is used to prepare the small molecule ginseng active liquid, which retains the effective ingredients in ginseng to the maximum extent and improves the purity and quality of ginseng products.

[0006] S2. Preparation of processed pig bile powder: weigh 20-25g of pig bile powder, put it into a casserole, add refined honey and water, fry until it turns yellow and does not stick to the hands, take it out, let it cool, crush it, and set it aside; in this step, the hay is processed with refined honey. After the processing and modification, the pig bile powder has a sweet taste and a warm property, which enhances its effect of tonifying the middle and replenishing qi;

[0007] S3, preparation of wolfberry extract: take 15 g of washed wolfberry, add 100 mL of water, put into a water bath, extract at 90° C. for 3 h to obtain wolfberry extract, and set aside;

[0008] S4. Preparation of bacterial liquid: Bifidobacterium bifidum (preservation number: CICC-6171, purchased from China Industrial Microbiological Culture Collection Center) was inoculated into RCM agar medium and cultured in a 37°C incubator for 48 h to obtain activated Bifidobacterium bifidum. The activated Bifidobacterium bifidum was transferred to a shake flask containing 500-1000 mL RCM liquid medium and fermented for 6 h to obtain a concentration of 1.2-1.5×10 8 cfu / mL of Bifidobacterium bifidum bacterial liquid; Lactobacillus crispatus (preservation number: CICC-24879, purchased from China Industrial Microbiological Culture Collection Management Center) was inoculated into MRS agar medium and cultured in a 37°C incubator for 48 hours to obtain activated Lactobacillus crispatus, which was inoculated into a shake flask with 500-1000 mL MRS liquid medium and fermented for 6 hours to obtain a concentration of 2.2-2.4×10 8 cfu / mL of Lactobacillus crispatus bacterial liquid, and both bacterial liquids were placed in a bacterial liquid storage tank for later use;

[0009] S5. Preparation of ginseng fermentation product: Weigh 20-30 mL of the small molecule ginseng active liquid in step S1 and 15-20 mL of the wolfberry extract in step S3, place them in a fermentation tank containing 12 mL of Bifidobacterium bifidum CICC-6171 bacterial liquid and 12 mL of Lactobacillus crispatus CICC-24879 bacterial liquid prepared in step S4 and 12 mL of basic fermentation medium, and ferment them anaerobically at 37°C for 20 h. After the fermentation is completed, add 3% H 2 O 2 The solution is used to inactivate the Bifidobacterium bifidum and Lactobacillus crispatus in the obtained fermentation product to obtain a ginseng fermentation product for standby use; in this step, Bifidobacterium bifidum and Lactobacillus crispatus are used to synergistically ferment the small molecule ginseng active liquid and the wolfberry extract, effectively enhancing the anti-aging effect of the paste;

[0010] S6. Preparation of ginseng paste: weigh 15-25 mL of the ginseng fermentation product prepared in step S5 and 2 g of the processed pig bile powder prepared in step S2, put them in a ceramic casserole, add 150 mL of 25°C constant temperature soak for 30 min, boil and simmer on low heat for 30 min, filter with gauze, put into a clean porcelain, leave overnight (water is soaked and becomes a paste), and then cover to obtain ginseng paste; the main purpose of using 25°C water in this step is to prevent the drug ingredients from being destroyed.

[0011] Secondly, under fermentation conditions, the fermentation products of small molecule ginseng and wolfberry extracts have anti-aging and antioxidant effects.

[0012] Thirdly, processed pig bile powder has the effect of inhibiting gastric bleeding.

[0013] Preferably: the ginseng selected in step S1 of the present invention is produced in Changbai Mountain;

[0014] Preferably: the microporous filter membrane in step S1 of the present invention is a 0.2 μm polyethersulfone filter membrane;

[0015] Preferably: in step S2 of the present invention, the pig bile powder is fried slowly over low heat;

[0016] Preferably: in step S2 of the present invention, 5-6 g of refined honey and 15-20 mL of water are added;

[0017] Preferably: The RCM agar medium used in step S4 of the present invention comprises: beef extract powder 10.0 g, protein chen, 5.0 g, yeast extract powder 3.0 g, glucose 5.0 g, soluble starch 1.0 g, NaCl 5.0 g, NaAc 3.0 g, L-cysteine ​​hydrochloride 0.5 g, agar 15.0 g, distilled water 1000.0 mL, pH 6.8;

[0018] Preferably: The components of the MRS agar medium used in step S4 of the present invention are: 10 g peptone, 10 g beef extract powder, 5 g yeast extract powder, 20 g glucose, 1 g Tween, 5 g sodium acetate, 2 g triammonium citrate, 2 g dipotassium hydrogen phosphate, 0.2 g magnesium sulfate heptahydrate, 0.05 g sulfuric acid tetrahydrate, 15 g agar, 1000 mL distilled water, and the pH is adjusted to 6.2-6.5;

[0019] Preferably: the volumes of the RCM liquid culture medium and the MRS culture medium in step S4 of the present invention are both 800 mL;

[0020] Preferably: in step S5 of the present invention, 25 mL of small molecule ginseng active liquid and 18 mL of wolfberry extract are used;

[0021] Preferably, the basic fermentation medium in step S5 of the present invention comprises: 20 g / L glucose, 3.5 g / L peptone, 3 g / L yeast extract, MgSO 4 7H 2 0 1.5 g / L, KH 2 PO 4 3 g / L.

[0022] The present invention has the following beneficial effects:

[0023] 1. The present invention selects small molecule ginseng and wolfberry extracts to improve the performance through synergistic fermentation of Bifidobacterium bifidum CICC-6171 and Lactobacillus crispatus CICC-24879 to achieve anti-aging and antioxidant effects.

[0024] 2. In the present invention, refined honey is used to modify the pig bile powder, and the modified pig bile powder and ginseng fermentation product have a synergistic effect of inhibiting gastric bleeding.

[0025] 3. The present invention adopts ultrafiltration technology to prepare small molecule ginseng active liquid, which retains the effective ingredients in ginseng to the maximum extent and improves the purity and quality of ginseng products.

[0026] 4. In the present invention, the pig bile powder is processed with refined honey, and the pig bile powder after processing and modification has a sweet taste and a warm property, thereby enhancing its effect of tonifying the middle and replenishing qi;

[0027] 5. When preparing the ginseng paste in the present invention, constant temperature immersion in water at 25°C can prevent the destruction of the medicinal ingredients and at the same time slow down the phenomenon of easy corruption in summer.

[0028] 6. The ginseng paste prepared by the present invention uses natural plants as raw materials, does not contain chemical additives, pigments and other chemical synthetic substances, is healthier and safer, and is suitable for consumption by the elderly and children. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0030] Figure 1 It is a comparison chart of the effects of ginseng paste of Example 2 and Comparative Examples 2-7 on the antioxidant activity in nematodes.

[0031] Figure 2 It is a comparison chart of the effects of ginseng paste of Example 2 and Comparative Examples 2-7 on the life span of nematodes.

[0032] Figure 3It is a comparison chart of the effects of ginseng paste of Example 3 and Comparative Examples 8-9 on the comprehensive coagulation value of gastric bleeding in mice.

[0033] Figure 4 This is a schematic diagram of the ginseng paste product. DETAILED DESCRIPTION

[0034] The following describes the embodiments of the present invention by specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that the following embodiments and features in the embodiments can be combined with each other without conflict.

[0035] Ginseng was provided by Jilin Yuanling Ginseng and Deer Antler Co., Ltd.; pig bile powder was provided by Xi'an Heshun Biotechnology Co., Ltd.; wolfberry was provided by Zaokang Wolfberry Co., Ltd.; hollow fiber column (BIE050D010) was provided by Tianjin Jingbichun Biotechnology Co., Ltd. Example

[0036] This example demonstrates that the small molecule ginseng active liquid can retain the effective ingredients in ginseng to the maximum extent and improve the purity and quality of ginseng products.

[0037] This embodiment provides a small molecule ginseng cream with anti-aging effect and a preparation method thereof, and the specific steps are as follows:

[0038] S1. Preparation of small molecule ginseng active liquid: select 20g of high-quality ginseng, clean and slice it, add 75% ethanol with a liquid-to-solid ratio of 1:10, distill and reflux to extract twice, and recover the ethanol under reduced pressure; then add 10% ethanol with a liquid-to-solid ratio of 1:10 and reflux to extract once, combine the extracts and concentrate them to 100mL; filter the concentrated extract through a 0.2μm polyethersulfone filter membrane, and then pour the filtrate into a BIE050D010 hollow fiber column for ultrafiltration, and retain small molecular substances below 10,000 units to prepare small molecule ginseng active liquid for use; in this step, the ultrafiltration process is used to prepare the small molecule ginseng active liquid, which retains the effective ingredients in ginseng to the maximum extent and improves the purity and quality of ginseng products.

[0039] S2, preparation of processed pig bile powder: weigh 20g of pig bile powder, put it into a casserole, add 5g of refined honey and 15mL of water, fry until yellow and non-sticky, take out, cool, crush and set aside; in this step, the hay is processed with refined honey, and the pig bile powder after processing and modification has a sweet taste and a warm property, which enhances its effect of tonifying the middle and replenishing qi;

[0040] S3, preparation of wolfberry extract: take 15 g of washed wolfberry, add 100 mL of water, put into a water bath, extract at 90° C. for 3 h to obtain wolfberry extract, and set aside;

[0041] S4. Preparation of bacterial solution: Bifidobacterium bifidum (preservation number: CICC-6171, purchased from China Industrial Microbiological Culture Collection Center) was inoculated into RCM agar medium and cultured in a 37°C incubator for 48 h to obtain activated Bifidobacterium bifidum. The activated Bifidobacterium bifidum was transferred to a shake flask with 800 mL RCM liquid medium and fermented for 6 h to obtain a concentration of 1.2 × 10 8 cfu / mL of Bifidobacterium bifidum bacterial liquid; Lactobacillus crispatus (preservation number: CICC-24879, purchased from China Industrial Microbiological Culture Collection Management Center) was inoculated into MRS agar medium and cultured in a 37°C incubator for 48 hours to obtain activated Lactobacillus crispatus, which was inoculated into a shake flask with 800 mL MRS liquid medium and fermented for 6 hours to obtain a concentration of 2.2×10 8 cfu / mL of Lactobacillus crispatus bacterial liquid, and both bacterial liquids were placed in a bacterial liquid storage tank for later use;

[0042] S5. Preparation of ginseng fermentation product: Weigh 20 mL of the small molecule ginseng active liquid in step S1 and 15 mL of the wolfberry extract in step S3, place them in a fermentation tank containing 12 mL of Bifidobacterium bifidum CICC-6171 bacterial liquid and 12 mL of Lactobacillus crispatus CICC-24879 bacterial liquid prepared in step S4 and 12 mL of basic fermentation medium, and ferment them anaerobically at 37°C for 20 h. After the fermentation, add 3% H 2 O 2 The solution is used to inactivate the Bifidobacterium bifidum and Lactobacillus crispatus in the obtained fermentation product to obtain a ginseng fermentation product for standby use; in this step, Bifidobacterium bifidum and Lactobacillus crispatus are used to synergistically ferment the small molecule ginseng active liquid and the wolfberry extract, effectively enhancing the anti-aging effect of the paste;

[0043] S6. Preparation of ginseng paste: weigh 15 mL of the ginseng fermentation product prepared in step S5 and 2 g of the processed pig bile powder prepared in step S2, put them in a ceramic casserole, add 150 mL of 25°C constant temperature and soak for 30 min, boil and simmer for 30 min on low heat, filter with gauze, put into a clean porcelain, leave overnight (water is soaked and becomes a paste), and then cover to obtain ginseng paste; the main purpose of using 25°C water in this step is to prevent the drug ingredients from being destroyed.

[0044] Comparative Example 1: In step S1, the ultrafiltration process is not performed, and the ginseng active liquid filtered through the microporous filter membrane is directly used, and the remaining steps are the same as those in Example 1.

[0045] The active ingredients in ginseng mainly include ginsenosides, total phenols, polysaccharides and other substances. The specific determination method is as follows:

[0046] Determination of ginsenosides (vanillin-glacial acetic acid method): Weigh 40uL of sample (the small molecule ginseng active solution obtained in step S1 of Example 1 and the ginseng active solution obtained in step S1 of Comparative Example 1), evaporate the solvent in a constant temperature water bath at 80°C, add 20uL of 5% vanillin-glacial acetic acid solution and 80μL of perchloric acid, bathe in a 60°C water bath for 15 min, add 200μL of glacial acetic acid after an ice bath for 5 min to stop the reaction, measure the absorbance at a wavelength of 540 nm, and measure the absorbance of ginsenoside solutions with mass concentrations of 50, 100, 150, 200, 250, and 300μg / mL under the same conditions, draw a standard curve, and calculate the ginsenoside content. The experimental results are shown in Table 1 below;

[0047] Determination of total phenol (Folin-phenol colorimetric method): Weigh 50 μL of sample (the small molecule ginseng active liquid obtained in step S1 of Example 1 and the ginseng active liquid obtained in step S1 of Comparative Example 1), add 125 uL of 0.1 mol / mol Folin-phenol solution, and finally add 100 uL of 75 g / mL sodium carbonate solution. After reacting in the dark for 30 min, measure the absorbance at a wavelength of 765 nm. Under the same conditions, measure the absorbance when the mass concentration of gallic acid is 10, 20, 40, 60, 80, and 100 μg / mL, draw a standard curve, and calculate the total phenol content. The experimental results are shown in Table 1 below;

[0048] Determination of polysaccharides (phenol-sulfuric acid method): Weigh 50 μL of sample (the small molecule ginseng active liquid obtained in step S1 of Example 1 and the ginseng active liquid obtained in step S1 of Comparative Example 1), add 50 uL of 5% phenol solution, then slowly add 250 uL of concentrated sulfuric acid, keep constant temperature in a water bath at 40°C for 10 min, cool to room temperature, and measure the absorbance at a wavelength of 490 nm. Under the same conditions, measure the absorbance of D(+)-anhydrous glucose solutions with mass concentrations of 20, 40, 60, 80, 100, 150, and 200 μg / mL, draw a standard curve, and calculate the total polysaccharide content. The experimental results are shown in Table 1 below;

[0049] Table 1

[0050] Content (mg / g) Ginsenosides Total phenols Polysaccharides Ginseng active essence 141.3 16.2 344.2 Small molecule ginseng active liquid 214.5 22.7 450.7

[0051] The experimental results show that compared with ginseng active liquid, the total phenol content of small molecule ginseng active liquid increased from 16.2 mg / g to 22.7 mg / g, an increase of 40.1%. The content of polysaccharides increased from 334.2 mg / g to 450.7 mg / g, an increase of 34.9%. The content of ginsenosides increased from 141.3 mg / g to 214.5 mg / g, an increase of 51.80%. Compared with ginseng active liquid, small molecule ginseng active liquid has a higher content of active substances and more effective ingredients. Example

[0052] This example demonstrates that under fermentation conditions, the synergistic effect of small molecule ginseng active liquid and wolfberry extract has good antioxidant and anti-aging effects.

[0053] This embodiment provides a small molecule ginseng cream with anti-aging effect and a preparation method thereof, and the specific steps are as follows:

[0054] S1. Preparation of small molecule ginseng active liquid: select 20g of high-quality ginseng, clean and slice it, add 75% ethanol with a liquid-to-solid ratio of 1:10, distill and reflux to extract twice, and recover the ethanol under reduced pressure; then add 10% ethanol with a liquid-to-solid ratio of 1:10 and reflux to extract once, combine the extracts and concentrate them to 100mL; filter the concentrated extract through a 0.2μm polyethersulfone filter membrane, and then pour the filtrate into a BIE050D010 hollow fiber column for ultrafiltration, and retain small molecular substances below 10,000 units to prepare small molecule ginseng active liquid for use; in this step, the ultrafiltration process is used to prepare the small molecule ginseng active liquid, which retains the effective ingredients in ginseng to the maximum extent and improves the purity and quality of ginseng products.

[0055] S2, preparation of processed pig bile powder: weigh 22g of pig bile powder, put it into a casserole, add 5.5g of refined honey and 17mL of water, fry until yellow and non-sticky, take out, cool, crush and set aside; in this step, the hay is processed with refined honey, and the pig bile powder after processing and modification has a sweet taste and a warm property, which enhances its effect of tonifying the middle and replenishing qi;

[0056] S3, preparation of wolfberry extract: take 15g of washed wolfberry, add 100mL of water, put into a water bath, extract at 90°C for 3h to obtain wolfberry extract, and set aside; S4, preparation of bacterial liquid: Bifidobacterium bifidum (preservation number: CICC-6171, purchased from China Industrial Microbiological Culture Collection Management Center) was inoculated into RCM agar medium, cultured in a 37°C incubator for 48h to obtain activated Bifidobacterium bifidum, transferred to a shake flask with 800mL RCM liquid culture medium and fermented for 6h to obtain a concentration of 1.3×10 8cfu / mL of Bifidobacterium bifidum bacterial liquid; Lactobacillus crispatus (preservation number: CICC-24879, purchased from China Industrial Microbiological Culture Collection Management Center) was inoculated into MRS agar medium and cultured in a 37°C incubator for 48 hours to obtain activated Lactobacillus crispatus, which was inoculated into a shake flask with 800 mL MRS liquid medium and fermented for 6 hours to obtain a concentration of 2.3×10 8 cfu / mL of Lactobacillus crispatus bacterial liquid, and both bacterial liquids were placed in a bacterial liquid storage tank for later use;

[0057] S5. Preparation of ginseng fermentation product: Weigh 25 mL of the small molecule ginseng active liquid in step S1 and 17 mL of the wolfberry extract in step S3, place them in a fermentation tank containing 12 mL of Bifidobacterium bifidum CICC-6171 bacterial liquid and 12 mL of Lactobacillus crispatus CICC-24879 bacterial liquid prepared in step S4 and 12 mL of basic fermentation medium, and ferment anaerobically at 37°C for 20 h. After the fermentation, add 3% H 2 O 2 The solution is used to inactivate the Bifidobacterium bifidum and Lactobacillus crispatus in the obtained fermentation product to obtain a ginseng fermentation product for standby use; in this step, Bifidobacterium bifidum and Lactobacillus crispatus are used to synergistically ferment the small molecule ginseng active liquid and the wolfberry extract, effectively enhancing the anti-aging effect of the paste;

[0058] S6. Preparation of ginseng paste: weigh 20 mL of the ginseng fermentation product prepared in step S5 and 2 g of the processed pig bile powder prepared in step S2, put them in a ceramic casserole, add 150 mL of 25°C constant temperature soak for 30 min, boil and simmer on low heat for 30 min, filter with gauze, put into a clean porcelain, leave overnight (water is soaked and becomes a paste), cover and obtain ginseng paste; the main purpose of using 25°C water in this step is to prevent the drug ingredients from being destroyed.

[0059] Comparative Example 2: Except that the small molecule ginseng active liquid is not added in step S5, the remaining steps are the same as those in Example 2.

[0060] Comparative Example 3: Except that wolfberry extract is not added in step S5, the remaining steps are the same as those in Example 2.

[0061] Comparative Example 4: Except that Lactobacillus crispatus CICC-24879 is not added in step S5, the remaining steps are the same as those in Example 2.

[0062] Comparative Example 5: Except that Bifidobacterium bifidum CICC-6171 was not added in step S5, the remaining steps were the same as those in Example 2.

[0063] Comparative Example 6: Except that Lactobacillus crispatus CICC-24879 was replaced by Lactobacillus crispatus CICC-25191 (purchased from China Industrial Microbiological Culture Collection Center) in step S5, the remaining steps were the same as those in Example 2.

[0064] Comparative Example 7: Except that Bifidobacterium bifidum CICC-6171 was replaced by Bifidobacterium bifidum CICC-10395 (purchased from China Industrial Microbiological Culture Collection Center) in step S5, the remaining steps were the same as those in Example 2.

[0065] 80 N2 wild-type Caenorhabditis elegans lines provided by Beijing Krisbo Biotechnology Co., Ltd. were used as nematodes and randomly divided into 8 groups, 10 in each group, namely, a blank group, Example 2 group, Comparative Example 2 group, Comparative Example 3 group, Comparative Example 4 group, Comparative Example 5 group, Comparative Example 6 group, and Comparative Example 7 group. Except for the blank group, no ginseng paste was added, and the corresponding ginseng paste was added to the other groups.

[0066] The culture, subculture and synchronization of nematodes were carried out according to the methods in the literature (Wang Kai, Sun Jianchao, Zhao Jun, et al. Immunomodulatory activity and protective effect of Caenorhabditis elegans capsular polysaccharide in fermented Lactobacillus [J]. Chinese Journal of Biotechnology, 2016, 33(7): 127443-127456.).

[0067] Antioxidant activity experiment in nematodes: Preparation of nematode homogenate: All the adults grown to the L4 stage synchronously were transferred to the Nematode Growth Medium (NGM) plate containing ginseng paste (100ug / mL), basic food OP50 and a final concentration of 50 umol / L 5-fluorouracil. The plates were changed once a day and cultured for 4 days. The plates were rinsed with M9 to collect the nematodes. The washed worm bodies were transferred to a centrifuge tube, homogenized with a micro-homogenizer, and the volume was adjusted to 500 uL with 0.8% Chaps buffer. Centrifuged at 12000 r / min in a refrigerated centrifuge for 10 min, and the supernatant was taken as the sample to be tested. The T-AOC and T-SOD activities were determined according to the instructions of the kit. The results are as follows: Figure 1 shown.

[0068] Nematode lifespan experiment: transfer the synchronized L4 nematodes to NGM plates containing ginseng paste (100ug / mL), about 40 per plate, and transfer to new NGM culture dishes and count them every day. After all the nematodes die, use SPSS software to draw a survival curve and analyze the survival of the nematodes. The experimental results are as follows Figure 2 Shown

[0069] Figure 1It is a comparison chart of the effects of ginseng paste of Example 2 and Comparative Examples 2-7 on the antioxidant activity in nematodes. It can be clearly seen from the bar chart that the T-AOC and T-SOD activities of Example 2 group are significantly better than those of Comparative Examples 2-7 group, and have better antioxidant capacity.

[0070] Figure 2 This is a comparison chart of the effects of ginseng cream on the life span of nematodes in Example 2 and Comparative Examples 2-7: From the bar chart, it can be seen intuitively that the average life span of nematodes in Example 2 is the longest, proving that Example 2 can effectively prolong the life span of nematodes

[0071] In summary, this example demonstrates that the small molecule ginseng active liquid and wolfberry extract have good antioxidant and anti-aging effects under the specific fermentation conditions of Bifidobacterium bifidum CICC-6171 and Lactobacillus crispatus CICC-24879. Example

[0072] This example demonstrates that the modified pig bile powder and ginseng fermentation product have the effect of inhibiting gastric bleeding through synergistic effect.

[0073] This embodiment provides a small molecule ginseng cream with anti-aging effect and a preparation method thereof, and the specific steps are as follows:

[0074] S1. Preparation of small molecule ginseng active liquid: select 20g of high-quality ginseng, clean and slice it, add 75% ethanol with a liquid-to-solid ratio of 1:10, distill and reflux to extract twice, and recover the ethanol under reduced pressure; then add 10% ethanol with a liquid-to-solid ratio of 1:10 and reflux to extract once, combine the extracts and concentrate them to 100mL; filter the concentrated extract through a 0.2μm polyethersulfone filter membrane, and then pour the filtrate into a BIE050D010 hollow fiber column for ultrafiltration, and retain small molecular substances below 10,000 units to prepare small molecule ginseng active liquid for use; in this step, the ultrafiltration process is used to prepare the small molecule ginseng active liquid, which retains the effective ingredients in ginseng to the maximum extent and improves the purity and quality of ginseng products.

[0075] S2, preparation of processed pig bile powder: weigh 25g of pig bile powder, put it into a casserole, add 6g of refined honey and 20mL of water, stir-fry until yellow and non-sticky, take it out, cool it, crush it, and set it aside; in this step, the hay is processed with refined honey, and the pig bile powder after processing and modification has a sweet taste and a warm property, which enhances its effect of tonifying the middle and replenishing qi;

[0076] S3, preparation of wolfberry extract: take 15g of washed wolfberry, add 100mL of water, put into a water bath, extract at 90°C for 3h to obtain wolfberry extract, and set aside; S4, preparation of bacterial liquid: Bifidobacterium bifidum (preservation number: CICC-6171, purchased from China Industrial Microbiological Culture Collection Management Center) was inoculated into RCM agar medium, cultured in a 37°C incubator for 48h to obtain activated Bifidobacterium bifidum, transferred to a shake flask with 800mL RCM liquid medium and fermented for 6h to obtain a concentration of 1.5×10 8 cfu / mL of Bifidobacterium bifidum bacterial liquid; Lactobacillus crispatus (preservation number: CICC-24879, purchased from China Industrial Microbiological Culture Collection Management Center) was inoculated into MRS agar medium and cultured in a 37°C incubator for 48 h to obtain activated Lactobacillus crispatus, which was inoculated into a shake flask with 800 mL MRS liquid medium and fermented for 6 h to obtain a concentration of 2.4×10 8 cfu / mL of Lactobacillus crispatus bacterial liquid, and both bacterial liquids were placed in a bacterial liquid storage tank for later use;

[0077] S5. Preparation of ginseng fermentation product: 30 mL of the small molecule ginseng active liquid in step S1 and 20 mL of the wolfberry extract in step S3 were weighed and placed in a fermentation tank containing 12 mL of Bifidobacterium bifidum CICC-6171 bacterial liquid and 12 mL of Lactobacillus crispatus CICC-24879 bacterial liquid prepared in step S4 and 12 mL of basic fermentation medium, and fermented anaerobically at 37°C for 20 h. After the fermentation was completed, 3% H 2 O 2 The solution is used to inactivate the Bifidobacterium bifidum and Lactobacillus crispatus in the obtained fermentation product to obtain a ginseng fermentation product for standby use; in this step, Bifidobacterium bifidum and Lactobacillus crispatus are used to synergistically ferment the small molecule ginseng active liquid and the wolfberry extract, effectively enhancing the anti-aging effect of the paste;

[0078] S6. Preparation of ginseng paste: weigh 25 mL of the ginseng fermentation product prepared in step S5 and 2 g of the processed pig bile powder prepared in step S2, put them in a ceramic casserole, add 150 mL of 25°C constant temperature and soak for 30 min, boil and simmer for 30 min on low heat, filter with gauze, put into a clean porcelain, leave overnight (water is soaked and becomes a paste), cover and obtain ginseng paste; the main purpose of using 25°C water in this step is to prevent the drug ingredients from being destroyed.

[0079] Comparative Example 8: Except that the processed pig bile powder is not added in step S6, the remaining steps are the same as those in Example 3.

[0080] Comparative Example 9: Except that Lactobacillus crispatus CICC-24879 and Bifidobacterium bifidum CICC-6171 were not added for fermentation in step S5, the remaining steps were the same as those in Example 3.

[0081] Grouping: 60 SPF SD rats (purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd.), weighing 200±20g, were randomly divided into 6 groups, 10 rats in each group, namely, a normal group (deionized water), a model group, Example 3 group (ginseng paste with a concentration of 0.1g / mL), a comparative example group 8-9 group (ginseng paste with a concentration of 0.1g / mL), and a positive control group (Yunnan Baiyao with a concentration of 0.03 g / mL, Yunnan Baiyao was purchased from Yunnan Baiyao Group Wenshan Qihua Co., Ltd. with a specification of 10mL / bottle); except for the normal group, the mice in other groups were subjected to modeling treatment.

[0082] Modeling: Mice were subcutaneously injected with reserpine 0.35 mg / kg daily for 7 consecutive days, and indomethacin suspension 70 mg / kg was injected intraperitoneally on the 7th day. Rats showed symptoms of spleen deficiency and gastric bleeding, and the modeling was successful. The drug administration treatment was carried out from the 7th day. The Example 3 group and the comparative example 7-8 group (dosage was 1g / kg), the positive control group (dosage was 0.15g / kg), and the normal group were gavaged once in the morning and evening for 3 consecutive days, and the gavage volume was 10mL / kg. After 24 hours of the first gavage, 20% urethane (6 mL / kg) was injected intraperitoneally for anesthesia, and abdominal aortic blood was drawn. According to the ratio of 9 parts of venous blood and 1 part of anticoagulant, it was added to the sodium citrate anticoagulant tube, and it was inverted until it was fully mixed without clots and foam was not formed. After sampling, the specimen was centrifuged at 3000r / min for 15 min to stratify the plasma and take the upper plasma. The plasma thrombin time (TT), prothrombin time (PT), and activated partial thromboplastin time (APTT) were measured by automatic coagulation analyzer, and the comprehensive coagulation value of gastric bleeding was calculated.

[0083]

[0084]

[0085] Figure 3 It is a comparison chart of the effect of ginseng paste of Example 3 and Comparative Examples 8-9 on the comprehensive coagulation value of gastric bleeding in mice. It can be intuitively seen from the figure that compared with the blank group mice, the comprehensive coagulation value of gastric bleeding in the model group mice is significantly increased, and the coagulation time is increased; the comprehensive coagulation value of gastric bleeding in the model group mice is not much different from that in the Comparative Example 8 and Comparative Example 9 groups of mice, and the comprehensive coagulation value of gastric bleeding in the Example 3 group of mice is significantly reduced compared with the Example 3 group of mice, indicating that the Example 3 group has a good coagulation effect on mice after gastric bleeding, and the Comparative Example 8 and Comparative Example 9 groups of mice have no obvious coagulation effect after gastric bleeding. It is confirmed that the modified pig bile powder and ginseng fermentation product have the synergistic effect of inhibiting gastric bleeding and coagulating gastric bleeding.

[0086] The above-mentioned embodiments only express the specific implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the protection scope of the present invention. It should be pointed out that, for those skilled in the art, without departing from the concept of the present invention, any other changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principle of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A small molecule ginseng cream with anti-aging effect, characterized in that: The main components of the ginseng paste are ginseng fermentation product and processed pig bile powder; the ginseng fermentation product is obtained by fermenting a ginseng composition through a composite strain, the ginseng composition comprises 20-30 parts of ginseng active liquid and 15-20 parts of wolfberry extract by volume, and the composite strain comprises Bifidobacterium bifidum CICC-6171 and Lactobacillus crispatus CICC-24879; the processed pig bile powder comprises 20-25 parts of pig bile powder, 5-6 parts of refined honey, and 15-20 parts of water by weight; The preparation method of the small molecule ginseng paste with anti-aging effect is as follows: S1. Preparation of small molecule ginseng active liquid: select 20g of high-quality ginseng, clean and slice it, add 75% ethanol with a liquid-to-solid ratio of 1:10, distill and reflux extract twice, and recover ethanol under reduced pressure; then add 10% ethanol with a liquid-to-solid ratio of 1:10 and reflux extract once, combine the extracts and concentrate to 100mL; filter the concentrated extract through a polyethersulfone filter membrane, and then pour the obtained filtrate into a BIE050D010 hollow fiber column for ultrafiltration, and retain small molecular substances below 10,000 units to prepare small molecule ginseng active liquid for use; S2. Preparation of processed pig bile powder: weigh 20-25g of pig bile powder, put it into a casserole, add refined honey and water, fry until yellow and not sticky, take out, cool, crush and set aside; S3, preparation of wolfberry extract: take 15 g of washed wolfberry, add 100 mL of water, put into a water bath, extract at 90° C. for 3 h to obtain wolfberry extract, and set aside; S4. Preparation of bacterial liquid: Bifidobacterium bifidum CICC-6171 was inoculated into RCM agar medium and cultured in a 37°C incubator for 48 h to obtain activated Bifidobacterium bifidum, which was then transferred to a shake flask containing 500-1000 mL RCM liquid medium and fermented for 6 h to obtain a concentration of 1.2-1.5×10 8 cfu / mL of Bifidobacterium bifidum bacterial liquid; Lactobacillus crispatus CICC-24879 was inoculated into MRS agar medium and cultured in a 37°C incubator for 48 hours to obtain activated Lactobacillus crispatus, which was inoculated into a shake flask with 500-1000 mL MRS liquid medium and fermented for 6 hours to obtain a concentration of 2.2-2.4×10 8 cfu / mL of Lactobacillus crispatus bacterial liquid, and both bacterial liquids were placed in a bacterial liquid tank for later use; S5, preparation of ginseng fermented product: weigh 20-30mL of the small molecule ginseng active liquid in step S1 and 15-20mL of the wolfberry extract in step S3, place them in a fermentation tank containing 12mL of Bifidobacterium bifidum CICC-6171 bacterial liquid and 12mL of Lactobacillus crispatus CICC-24879 bacterial liquid prepared in step S4 and 12mL of basic fermentation medium, and ferment them anaerobically at a temperature of 37°C for 20h. After the fermentation is completed, the Bifidobacterium bifidum and Lactobacillus crispatus bacterial liquid in the obtained fermented product are inactivated with 3% H2O2 solution to obtain ginseng fermented product for standby use; S6. Preparation of ginseng paste: weigh 15-25 mL of the ginseng fermentation product prepared in step S5 and 2 g of the processed pig bile powder prepared in step S2, put them in a ceramic casserole, add 150 mL of the mixture and soak at a constant temperature of 25°C for 30 min, boil and simmer on low heat for 30 min, filter with gauze, put into a clean porcelain container, leave overnight, and cover to obtain ginseng paste.

2. The small molecule ginseng cream with anti-aging effect according to claim 1, characterized in that: The ginseng active liquid adopts ultrafiltration technology to intercept small molecular substances below 10,000 units to prepare small molecule ginseng active liquid.

3. The small molecule ginseng paste with anti-aging efficacy according to claim 2, wherein the small molecule ginseng paste has the efficacy of anti-aging, anti-oxidation and inhibiting gastric bleeding.

4. The method for preparing a small molecule ginseng paste with anti-aging effect according to claim 3, characterized in that: The ginseng selected in step S1 is produced in Changbai Mountain.

5. The method for preparing a small molecule ginseng cream with anti-aging effect according to claim 4, characterized in that: The microporous filter membrane in step S1 is a 0.2 μm polyethersulfone filter membrane.

6. The method for preparing a small molecule ginseng paste with anti-aging effect according to claim 5, characterized in that: In step S2, the pig bile powder should be fried slowly over a low heat.

7. The method for preparing a small molecule ginseng paste with anti-aging effect according to claim 6, characterized in that: In step S2, 5-6 g of refined honey and 15-20 mL of water are added.

8. The method for preparing a small molecule ginseng cream with anti-aging effect according to claim 7, characterized in that: In step S4, the volumes of the RCM liquid culture medium and the MRS culture medium are both 800 mL.

9. The method for preparing a small molecule ginseng paste with anti-aging effect according to claim 8, characterized in that: In the step S5, 25 mL of small molecule ginseng active liquid and 18 mL of wolfberry extract are used.

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