An oral liquid preparation containing ginseng stem cells and preparation method thereof

Abstract

The present invention belongs to the field of preparation of oral liquid containing ginseng stem cells, and in particular to an oral liquid preparation containing ginseng stem cells and a preparation method thereof. The present invention adopts ultrasonic crushing after cultivation of growth-promoting bacteria, and cooperates with nutrient slurry and calcium chloride to promote the rapid formation of cell walls of pure ginseng stem cell protoplasts, and the cells have high growth vitality. The plant extract obtained by extraction improves the synthesis and accumulation of ginsenosides in pure ginseng stem cell groups; the nutrient absorption-promoting nanoemulsion obtained by high-pressure homogenization can improve the solubility of each component, and adhere to the surface of small intestinal epithelial cells, and play a long-term absorption-promoting role; the ginseng stem cell mixture is mixed with a flavor-correcting and stabilizing solvent, and the prepared oral liquid preparation containing ginseng stem cells contains rich ginsenosides, is easy to be absorbed by the small intestine, has strong stability, and will not precipitate and stratify after long-term storage.

Description

Technical Field

[0001] The present invention belongs to the field of preparation of oral liquid containing ginseng stem cells, and specifically refers to an oral liquid preparation containing ginseng stem cells and a preparation method thereof. Background Art

[0002] As we age, the body's various physiological functions gradually decline, and we are prone to problems such as decreased physical strength, memory loss, and decreased immunity. Ginseng stem cell oral liquid is an oral preparation with ginseng as the main ingredient. Ginseng stem cells refer to cells with regenerative ability extracted from ginseng. The growth factors and nutrients contained in ginseng stem cells help promote cell regeneration and repair, which is beneficial for wound healing and tissue repair. It also contains active ingredients that can regulate the immune system, which helps to improve the body's resistance to disease, reduce fatigue, enhance physical strength and endurance, and the antioxidant ingredients it contains help slow down cell aging, improve skin health, reduce wrinkles and pigmentation, and reduce the body's inflammatory response.

[0003] The preparation of ginseng stem cell oral liquid involves multiple steps, including the planting and harvesting of ginseng, cleaning and disinfection, the extraction of ginseng stem cells using biotechnology methods, and cultivation under suitable conditions. Finally, ginseng stem cell oral liquid containing active ingredients is prepared. Adding ginseng stem cells to the oral liquid ensures that all active ingredients of ginseng can be obtained, not just a single chemical component.

[0004] At present, the existing oral liquid preparation technology containing ginseng stem cells has the following problems: First, the ginsenoside content in the ginseng stem cells extracted in the existing technology is low, the operation is complicated, and it is easy to be contaminated; Second, ginsenosides will not be hydrolyzed in the stomach, but after entering the small intestine, they will be affected by digestive enzymes, decomposed and absorbed, but the complex glycoside structure of ginsenosides and their low solubility in the intestine make them poorly absorbed in the small intestine; Third, the existing ginseng stem cell oral liquid has low stability, and precipitation and stratification occur after long-term storage. Summary of the invention

[0005] In view of the above situation, in order to overcome the defects of the prior art, the present invention provides an oral liquid preparation containing ginseng stem cells and a preparation method thereof. In order to solve the problems of low ginsenoside content, complex operation, poor absorbability of ginsenosides and low stability of the existing oral liquid containing ginseng stem cells, the present invention uses induced ginseng stem cells to be evenly dispersed in a nutrient absorption promoting nanoemulsion and mixed with mogroside, sodium gluconate, disodium ethylenediaminetetraacetic acid and ultrapure water. The ginseng stem cells in the prepared oral liquid containing ginseng stem cells contain a large amount of ginsenosides, are easily absorbed by the small intestine and have strong stability.

[0006] In order to achieve the above object, the technical solution adopted by the present invention is as follows: the present invention proposes an oral liquid preparation containing ginseng stem cells, comprising the following components in parts by weight:

[0007] The invention comprises 70-90 parts of induced ginseng stem cells, 60-80 parts of nutrient absorption promoting nanoemulsion, 7-10 parts of mogroside, 5-8 parts of sodium gluconate, 3-5 parts of disodium edetate and 20-30 parts of ultrapure water.

[0008] Preferably, the method for preparing induced ginseng stem cells specifically comprises the following steps:

[0009] S1. Weigh 200-300 parts by weight of fresh ginseng rhizomes, soak them in 70% ethanol solution in a 28°C clean bench for 20-30 seconds, disinfect them in 0.1% mercuric chloride solution for 10-12 minutes, rinse them with ultrapure water for 3-5 times, cut the ginseng rhizomes into slices with a thickness of 0.1 mm, and cut the cambium tissue in the slices to obtain the ginseng rhizome cambium;

[0010] S2, weigh 3-5 parts of cellulase and 3-5 parts of pectinase, 100-130 parts of 0.6 mol / L sucrose solution and the ginseng rhizome cambium prepared in S1, put them into a stirring tank with a power of 1.2-1.5 kW, the stirring temperature is 25-28°C, the stirring time is 8-12 min, the stirring speed is 110-130 r / min, soak for 30-50 min after stirring, put them into a high-speed centrifuge with a power of 2.5-2.8 kW, the centrifugal temperature is 2°C, the centrifugal time is 10-15 min, the centrifugal speed is 12000 r / min, and after centrifugation, rinse with ultrapure water 2-3 times to obtain pure ginseng stem cell protoplasts;

[0011] S3, weighing 40-60 parts of giant kelp and 50-70 parts of red hair algae, putting them into a pulping machine with a power of 1.8-2.2 kW, refining temperature of 18-23 ° C, refining time of 15-20 min, refining speed of 130-150 r / min, mixing and refining to obtain nutrient slurry;

[0012] S4, weigh 10-12 parts by weight of growth-promoting bacteria and 170-210 parts of sterile LB liquid culture medium and put them into a constant temperature shaking incubator with a power of 1.2-1.6 kW, the constant temperature shaking culture temperature is 33-37°C, the constant temperature shaking culture time is 24-30 h, the constant temperature shaking culture speed is 230-260 r / min, after the constant temperature shaking culture, use an ultrasonic cell disruptor with a power of 1.2-1.5 kW, the ultrasonic disruption frequency is 20 kHz, the ultrasonic disruption temperature is 0°C, the ultrasonic disruption time is 10-13min, after disruption, filter with a 0.22 mm sterile filter membrane, and put them into a stirring tank with a power of 1.2-1.5 kW with the nutrient slurry prepared in S3, 15-20 parts of inositol, 8-12 parts of calcium chloride, and 130-160 parts of sterile MS liquid culture medium, the stirring temperature is 25-28°C, the stirring time is 15-20 min, the stirring speed is 110-130 r / min, and the mixture is evenly mixed to obtain a cell growth promoting nutrient solution;

[0013] S5, placing the purified ginseng stem cell protoplasts prepared in S2 and the cell growth promoting nutrient solution prepared in S4 into a constant temperature shaking incubator with a power of 1.2-1.6 kW, the constant temperature shaking culture temperature is 25-28°C, the constant temperature shaking culture time is 40-45 d, the constant temperature shaking culture speed is 50-80 r / min, after mixed culture, placing in a high-speed centrifuge with a power of 2.5-2.8 kW, the centrifugation temperature is 2°C, the centrifugation time is 10-15 min, the centrifugation speed is 12000 r / min, and after centrifugation, washing with ultrapure water 2-3 times to obtain a purified ginseng stem cell population;

[0014] S6, weighing 30-50 parts of Aralia dasyphylla, 20-40 parts of Fenugreek, and 80-100 parts of anhydrous ethanol, and putting them into an ultrasonic extractor with a power of 1.3-1.6 kW, the extraction pressure is 1.5 MPa, the extraction frequency is 25 kHz, the extraction temperature is 40-45°C, and the extraction time is 8-13 min. After extraction, put them into a vacuum dryer with a power of 1.6-2.2 kW, the vacuum drying pressure is 0.3 MPa, the vacuum drying temperature is 55-60°C, and the vacuum drying time is 1.5 h, and the anhydrous ethanol is removed to obtain a plant extract;

[0015] S7. Place the plant extract prepared in S6, 180-220 parts by weight of sterile MS liquid culture medium, and the pure ginseng stem cell population prepared in S5 into a constant temperature shaking incubator with a power of 1.2-1.6 kW, the constant temperature shaking culture temperature is 35-37°C, the constant temperature shaking culture time is 5-7 d, the constant temperature shaking culture speed is 60-90 r / min, after mixed culture, place in a high-speed centrifuge with a power of 2.5-2.8 kW, the centrifugation temperature is 2°C, the centrifugation time is 10-15 min, the centrifugation speed is 12000 r / min, and after centrifugation, rinse with ultrapure water 2-3 times to obtain induced ginseng stem cells.

[0016] Furthermore, the composition of the growth-promoting bacterial agent includes: Actinomycetes aurantifolia, Gluconacetobacter xylosus, and Pseudomonas betae, and the weight ratio of the mixture is 1-2.5:1-2:1.

[0017] Furthermore, the aurantiacus coralliform actinomycetes were purchased from the China General Microbiological Culture Collection Center with a collection number of CGMCC4.2187; the xylosyl gluconoacetobacter was purchased from the China General Microbiological Culture Collection Center with a collection number of CGMCC1.2706; and the beet Pseudomonas aeruginosa was purchased from the China General Microbiological Culture Collection Center with a collection number of CGMCC1.8810.

[0018] Preferably, the method for preparing the nanoemulsion for promoting nutrient absorption comprises the following steps:

[0019] L1, weighing 30-40 parts of narcissus glycosides, 15-20 parts of betaine and 90-110 parts of triacetin into a stirrer with a power of 1.2-1.5 kW, stirring at a temperature of 25-28° C., stirring for 15-20 min, stirring at a speed of 130-160 r / min, and mixing to obtain a transport-promoting solution;

[0020] L2, weigh 60-80 parts of lactoglobulin, 50-70 parts of potassium lauryl ether phosphate, and 30-50 parts of ultrapure water, put them into the transport-promoting solution prepared in L1, stir at a temperature of 25-28°C, for 13-18 min, at a speed of 130-160 r / min, and mix well to obtain a mixture that promotes nutrient absorption;

[0021] L3. Place the nutrient absorption promoting mixture prepared in L2 into an ultra-high pressure homogenizer with a power of 2.5-2.8 kW, a pressure of 150 MPa, and a high-speed air flow rate of 1200-1500 m / s for ultra-high pressure homogenization to form a nutrient absorption promoting nanoemulsion.

[0022] The present invention also proposes a method for preparing an oral liquid preparation containing ginseng stem cells, which specifically comprises the following steps:

[0023] Step 1: Place the induced ginseng stem cells in a vacuum freeze dryer with a power of 5.5-7.5 kW, a vacuum degree of 60-80 Pa, a freezing temperature of -20°C, and a freezing time of 2-3 h, then heat to 2-4°C, dry for 1.2-1.5 h, and then place them in an ultrasonic disperser with a nutrient absorption-promoting nanoemulsion with a power of 1.6-1.9 kW, with an ultrasonic dispersion frequency of 25 kHz, an ultrasonic dispersion temperature of 10-15°C, and an ultrasonic dispersion time of 8-10 min. After ultrasonic dispersion, a mixed solution containing ginseng stem cells is obtained;

[0024] Step 2: Put mogroside, sodium gluconate, disodium ethylenediaminetetraacetic acid and ultrapure water into a stirrer with a power of 1.2-1.5 kW, stir at a temperature of 25-28° C., stir for 10-15 min, stir at a speed of 130-160 r / min, and mix to obtain a flavor-correcting and stabilizing solvent;

[0025] Step 3: Place the mixed solution containing ginseng stem cells prepared in step 1 into the flavor correcting and stabilizing solvent prepared in step 2, stir at a temperature of 25-28°C, for 20-30 min, and at a speed of 130-160 r / min to obtain an oral liquid preparation containing ginseng stem cells.

[0026] The beneficial effects achieved by the present invention are as follows:

[0027] The present invention adopts a nutrient slurry containing giant algae and red hair algae, which are rich in cellulose, hemicellulose and pectin. The growth-promoting bacteria contained in the coralline actinomycetes, xylose gluconate acetobacter and beet pseudomonas can release a large amount of auxin, cytokinin, phenylacetic acid, succinic acid and cellulose synthase after being cultured and ultrasonically broken, and cooperate with the nutrient slurry and calcium chloride to promote the synthesis of expansin in the cell wall. Expansin can break the non-covalent bonds between cellulose and hemicellulose in the cell wall to promote them. The mutual displacement between them causes the cell wall to stretch, so that the pure ginseng stem cell protoplasts quickly form cell walls, and the cells have high growth vitality and can grow and divide efficiently; Aralia ternata contains dimethylallyl pyrophosphate and dammarenediol, and fenugreek contains isoprene pyrophosphate, 2,3-oxidized squalene, and lanosterol. The plant extracts obtained by extracting Aralia ternata and fenugreek provide a variety of precursor substances for the biosynthesis of ginsenosides, which promotes the synthesis and accumulation of ginsenosides in the pure ginseng stem cell population. The content of ginsenosides in ginseng stem cells was effectively increased after induction; narcissin, betaine and triacetin were mixed, narcissin could inhibit the ATPase activity of efflux transporter in the small intestine and reduce the pumping of ginsenosides from intestinal cells, betaine could promote the activity of ABC transporter in small intestinal epithelial cells, and the two synergistically promoted the absorption of ginsenosides by the small intestine, and the transport-promoting solution was mixed with lactoglobulin, potassium lauryl ether phosphate and ultrapure water to form a nutrient absorption-promoting mixture, which was pressurized by a high-pressure pump to form a micro-liquid The induced ginseng stem cells are made into freeze-dried powder, ultrasonically dispersed in the nutrient absorption promoting nanoemulsion, and mixed with a flavor correcting and stabilizing solvent. The ginseng stem cells in the prepared oral liquid preparation contain rich ginsenosides, which are easy for the small intestine in the body to absorb the effective ingredients. The oral liquid preparation has strong stability and will not precipitate or stratify after long-term storage. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] In order to more clearly illustrate the present invention or the technical solutions in the prior art, the following will be described in a clear and understandable manner in conjunction with the accompanying drawings. Obviously, the drawings described below are only for the present invention, and for ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0029] Figure 1 This is a graph showing the results of the content of PPD-type ginsenosides in the samples described in Experimental Example 1 of the present invention;

[0030] Figure 2 This is a graph showing the results of the immunoglobulin IgG content in mouse plasma described in Experimental Example 2 of the present invention;

[0031] Figure 3 This is a graph showing the results of the immunoglobulin IgM content in mouse plasma described in Experimental Example 2 of the present invention;

[0032] Figure 4 This is a graph showing the results of the immunoglobulin IgA content in mouse plasma as described in Experimental Example 2 of the present invention. DETAILED DESCRIPTION

[0033] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments; based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0034] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein may be applied to the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only and are not intended to limit the content of this application.

[0035] The experimental methods in the following examples are conventional methods unless otherwise specified; the experimental materials and test strains used in the following examples are purchased from commercial channels unless otherwise specified.

[0036] Example 1: This example provides an oral liquid preparation containing ginseng stem cells and a preparation method thereof. The oral liquid preparation containing ginseng stem cells comprises the following components in parts by weight:

[0037] 70 parts of induced ginseng stem cells, 60 parts of nutrient absorption promoting nanoemulsion, 7 parts of mogroside, 5 parts of sodium gluconate, 3 parts of disodium edetate, and 20 parts of ultrapure water.

[0038] The method for preparing induced ginseng stem cells specifically comprises the following steps:

[0039] S1. Weigh 200 parts by weight of fresh ginseng rhizomes, soak them in 70% ethanol solution for 20 seconds in a 28°C clean bench, disinfect them in 0.1% mercuric chloride solution for 10 minutes, rinse them with ultrapure water for 3 times, cut the ginseng rhizomes into slices with a thickness of 0.1 mm, and cut the cambium tissue in the slices to obtain the ginseng rhizome cambium;

[0040] S2, weigh 3 parts of cellulase and 3 parts of pectinase, 100 parts of 0.6 mol / L sucrose solution and the ginseng rhizome cambium prepared in S1, put them into a stirring tank with a power of 1.2 kW, the stirring temperature is 25°C, the stirring time is 8 min, the stirring speed is 110r / min, soak for 30 min after stirring, put them into a high-speed centrifuge with a power of 2.5 kW, the centrifugal temperature is 2°C, the centrifugal time is 10 min, the centrifugal speed is 12000 r / min, and after centrifugation, rinse with ultrapure water twice to obtain pure ginseng stem cell protoplasts;

[0041] S3, weighing 40 parts of giant kelp and 50 parts of red hair algae, putting them into a pulping machine with a power of 1.8 kW, the pulping temperature is 18°C, the pulping time is 15 min, the pulping speed is 130 r / min, and mixing and grinding to obtain a nutrient slurry;

[0042] S4, weighing 10 parts by weight of the growth-promoting bacterial agent and 170 parts of sterile LB liquid culture medium, and putting them into a constant temperature shaking incubator with a power of 1.2 kW, the constant temperature shaking culture temperature is 33°C, the constant temperature shaking culture time is 24 h, and the constant temperature shaking culture speed is 230 r / min. After the constant temperature shaking culture, use an ultrasonic cell disruptor with a power of 1.2 kW, the ultrasonic disruption frequency is 20 kHz, the ultrasonic disruption temperature is 0°C, and the ultrasonic disruption time is 10 min. After disruption, use a 0.22 mm sterile filter membrane to filter, and put them into a stirring tank with a power of 1.2 kW with the nutrient slurry prepared in S3, 15 parts of inositol, 8 parts of calcium chloride, and 130 parts of sterile MS liquid culture medium, the stirring temperature is 25°C, the stirring time is 15 min, and the stirring speed is 110 r / min, and mix them evenly to obtain a cell growth-promoting nutrient solution;

[0043] S5. Place the purified ginseng stem cell protoplasts prepared in S2 and the cell growth promoting nutrient solution prepared in S4 into a constant temperature shaking incubator with a power of 1.2 kW, the constant temperature shaking culture temperature is 25°C, the constant temperature shaking culture time is 40 days, the constant temperature shaking culture speed is 50 r / min, after mixed culture, place in a high-speed centrifuge with a power of 2.5 kW, the centrifugal temperature is 2°C, the centrifugal time is 10 min, the centrifugal speed is 12000 r / min, and after centrifugation, rinse with ultrapure water twice to obtain a purified ginseng stem cell population;

[0044] S6, weighing 30 parts of Aralia dasyphylla, 20 parts of Fenugreek, and 80 parts of anhydrous ethanol, and putting them into an ultrasonic extractor with a power of 1.3 kW, the extraction pressure is 1.5 MPa, the extraction frequency is 25 kHz, the extraction temperature is 40°C, and the extraction time is 8 min. After extraction, put them into a vacuum dryer with a power of 1.6 kW, the vacuum drying pressure is 0.3 MPa, the vacuum drying temperature is 55°C, and the vacuum drying time is 1.5 h, and the anhydrous ethanol is removed to obtain a plant extract;

[0045] S7. Place the plant extract prepared in S6, 180 parts by weight of sterile MS liquid culture medium, and the pure ginseng stem cell population prepared in S5 into a constant temperature shaking incubator with a power of 1.2 kW. The constant temperature shaking culture temperature is 35°C, the constant temperature shaking culture time is 5 days, and the constant temperature shaking culture speed is 60 r / min. After mixed culture, place it in a high-speed centrifuge with a power of 2.5 kW, the centrifugation temperature is 2°C, the centrifugation time is 10 min, and the centrifugation speed is 12000 r / min. After centrifugation, rinse it twice with ultrapure water to obtain induced ginseng stem cells.

[0046] The growth-promoting bacterial agent comprises: Actinomycetes aurantifolia, Gluconacetobacter xylosus and Pseudomonas betae, and the mixing ratio by weight is 1:1:1.

[0047] The aurantiacus coralliformis was purchased from the China General Microbiological Culture Collection Center with a collection number of CGMCC4.2187; the xylosyl gluconoacetobacter was purchased from the China General Microbiological Culture Collection Center with a collection number of CGMCC1.2706; and the beet Pseudomonas was purchased from the China General Microbiological Culture Collection Center with a collection number of CGMCC1.8810.

[0048] The preparation method of the nanoemulsion for promoting nutrient absorption specifically comprises the following steps:

[0049] L1. Weigh 30 parts of narcissus glycoside, 15 parts of betaine and 90 parts of triacetin into a 1.2 kW stirrer, stir at 25°C for 15 min, and stir at 130 r / min to obtain a transport-promoting solution.

[0050] L2, weigh 60 parts of lactoglobulin, 50 parts of potassium lauryl ether phosphate, and 30 parts of ultrapure water, put them into the transport-promoting solution prepared in L1, stir at 25°C, stir for 13 min, stir at 130 r / min, and mix well to obtain a mixture that promotes nutrient absorption;

[0051] L3. Place the nutrient absorption promoting mixture prepared in L2 into an ultra-high pressure homogenizer with a power of 2.5 kW, a pressure of 150 MPa, and a high-speed air flow rate of 1200 m / s for ultra-high pressure homogenization to form a nutrient absorption promoting nanoemulsion.

[0052] This embodiment also provides a method for preparing an oral liquid preparation containing ginseng stem cells, which specifically comprises the following steps:

[0053] Step 1: Place the induced ginseng stem cells in a vacuum freeze dryer with a power of 5.5 kW, a vacuum degree of 60 Pa, a freezing temperature of -20°C, and a freezing time of 2 h, then heat to 2°C, dry for 1.2 h, and then place them in an ultrasonic disperser with a nutrient absorption-promoting nanoemulsion with a power of 1.6 kW, with an ultrasonic dispersion frequency of 25 kHz, an ultrasonic dispersion temperature of 10°C, and an ultrasonic dispersion time of 8 min. After ultrasonic dispersion, a mixed solution containing ginseng stem cells is obtained;

[0054] Step 2: Put mogroside, sodium gluconate, disodium ethylenediaminetetraacetic acid and ultrapure water into a 1.2 kW stirrer at a stirring temperature of 25° C. for 10 min at a stirring speed of 130 r / min, and mix well to obtain a flavor-correcting and stabilizing solvent;

[0055] Step 3: Place the mixed solution containing ginseng stem cells prepared in step 1 into the flavor-correcting and stabilizing solvent prepared in step 2, stir at a temperature of 25° C., for 20 min, at a speed of 130 r / min to obtain an oral liquid preparation containing ginseng stem cells.

[0056] Example 2: This example provides an oral liquid preparation containing ginseng stem cells and a preparation method thereof. The oral liquid preparation containing ginseng stem cells comprises the following components in parts by weight:

[0057] 80 parts of induced ginseng stem cells, 70 parts of nutrient absorption promoting nanoemulsion, 8 parts of mogroside, 6 parts of sodium gluconate, 4 parts of disodium edetate, and 25 parts of ultrapure water.

[0058] The method for preparing induced ginseng stem cells specifically comprises the following steps:

[0059] S1. Weigh 250 parts by weight of fresh ginseng rhizomes, soak them in 70% ethanol solution in a 28°C clean bench for 25 seconds, disinfect them in 0.1% mercuric chloride solution for 11 minutes, rinse them with ultrapure water for 4 times, cut the ginseng rhizomes into slices with a thickness of 0.1 mm, and cut the cambium tissue in the slices to obtain the ginseng rhizome cambium;

[0060] S2, weigh 4 parts of cellulase and 4 parts of pectinase, 120 parts of 0.6 mol / L sucrose solution and the ginseng rhizome cambium prepared in S1, put them into a stirring tank with a power of 1.3 kW, the stirring temperature is 26°C, the stirring time is 10 min, the stirring speed is 120 r / min, soak for 40 min after stirring, put them into a high-speed centrifuge with a power of 2.6 kW, the centrifugal temperature is 2°C, the centrifugal time is 13 min, the centrifugal speed is 12000 r / min, and after centrifugation, rinse with ultrapure water 3 times to obtain pure ginseng stem cell protoplasts;

[0061] S3, weighing 50 parts of giant kelp and 60 parts of red hair algae, putting them into a pulping machine with a power of 1.9 kW, the pulping temperature is 20°C, the pulping time is 18 min, the pulping speed is 140 r / min, and mixing and grinding to obtain a nutrient slurry;

[0062] S4, weighing 11 parts by weight of the growth-promoting bacterial agent and 190 parts of sterile LB liquid culture medium, and putting them into a constant temperature shaking incubator with a power of 1.4 kW, the constant temperature shaking culture temperature is 35°C, the constant temperature shaking culture time is 28 h, the constant temperature shaking culture speed is 240 r / min, after the constant temperature shaking culture, use an ultrasonic cell disruptor with a power of 1.4 kW, the ultrasonic disruption frequency is 20 kHz, the ultrasonic disruption temperature is 0°C, the ultrasonic disruption time is 11 min, and after disruption, use a 0.22 mm sterile filter membrane to filter, and put them into a stirring tank with a power of 1.3 kW with the nutrient slurry prepared in S3, 18 parts of inositol, 10 parts of calcium chloride, and 140 parts of sterile MS liquid culture medium, the stirring temperature is 26°C, the stirring time is 18 min, the stirring speed is 120 r / min, and mix them evenly to obtain a cell growth-promoting nutrient solution;

[0063] S5. Place the purified ginseng stem cell protoplasts prepared in S2 and the cell growth promoting nutrient solution prepared in S4 into a constant temperature shaking incubator with a power of 1.4 kW, the constant temperature shaking culture temperature is 26°C, the constant temperature shaking culture time is 43 days, the constant temperature shaking culture speed is 60 r / min, after mixed culture, place in a high-speed centrifuge with a power of 2.6 kW, the centrifugation temperature is 2°C, the centrifugation time is 13 min, the centrifugation speed is 12000 r / min, and after centrifugation, rinse twice with ultrapure water to obtain a purified ginseng stem cell population;

[0064] S6, weighing 40 parts of Aralia dasyphylla, 30 parts of Fenugreek, and 90 parts of anhydrous ethanol, and putting them into an ultrasonic extractor with a power of 1.4 kW, the extraction pressure is 1.5 MPa, the extraction frequency is 25 kHz, the extraction temperature is 43°C, and the extraction time is 10 min. After extraction, put them into a vacuum dryer with a power of 1.9 kW, the vacuum drying pressure is 0.3 MPa, the vacuum drying temperature is 58°C, and the vacuum drying time is 1.5 h, and the anhydrous ethanol is removed to obtain a plant extract;

[0065] S7. Place the plant extract prepared in S6, 200 parts by weight of sterile MS liquid culture medium, and the pure ginseng stem cell population prepared in S5 into a constant temperature shaking incubator with a power of 1.4 kW. The constant temperature shaking culture temperature is 36°C, the constant temperature shaking culture time is 6 days, and the constant temperature shaking culture speed is 70 r / min. After mixed culture, place it in a high-speed centrifuge with a power of 2.6 kW, the centrifugation temperature is 2°C, the centrifugation time is 13 min, and the centrifugation speed is 12000 r / min. After centrifugation, rinse it twice with ultrapure water to obtain induced ginseng stem cells.

[0066] The growth-promoting bacterial agent comprises: Actinomycetes aurantifolia, Gluconacetobacter xylosus, and Pseudomonas betae, and the weight ratio of the mixture is 2:1.5:1.

[0067] The aurantiacus coralliformis was purchased from the China General Microbiological Culture Collection Center with a collection number of CGMCC4.2187; the xylosyl gluconoacetobacter was purchased from the China General Microbiological Culture Collection Center with a collection number of CGMCC1.2706; and the beet Pseudomonas was purchased from the China General Microbiological Culture Collection Center with a collection number of CGMCC1.8810.

[0068] The preparation method of the nanoemulsion for promoting nutrient absorption specifically comprises the following steps:

[0069] L1, weighing 35 parts of narcissus glycosides, 18 parts of betaine and 100 parts of triacetin, putting them into a stirrer with a power of 1.3 kW, stirring at a temperature of 26°C, stirring for 18 min, and stirring at a speed of 150 r / min, mixing to obtain a transport-promoting solution;

[0070] L2, weigh 70 parts of lactoglobulin, 60 parts of potassium lauryl ether phosphate and 40 parts of ultrapure water, put them into the transport-promoting solution prepared in L1, stir at 26°C for 15 min, stir at 140 r / min, and mix well to obtain a mixture that promotes nutrient absorption;

[0071] L3. Place the nutrient absorption promoting mixture prepared in L2 into an ultra-high pressure homogenizer with a power of 2.6 kW, a pressure of 150 MPa, and a high-speed air flow rate of 1300 m / s for ultra-high pressure homogenization to form a nutrient absorption promoting nanoemulsion.

[0072] This embodiment also provides a method for preparing an oral liquid preparation containing ginseng stem cells, which specifically comprises the following steps:

[0073] Step 1: Place the induced ginseng stem cells in a vacuum freeze dryer with a power of 6.5 kW, a vacuum degree of 70 Pa, a freezing temperature of -20°C, and a freezing time of 2.5 h, then heat to 3°C, dry for 1.3 h, and then place them in an ultrasonic disperser with a nutrient absorption-promoting nanoemulsion with a power of 1.7 kW, with an ultrasonic dispersion frequency of 25 kHz, an ultrasonic dispersion temperature of 13°C, and an ultrasonic dispersion time of 9 min. After ultrasonic dispersion, a mixed solution containing ginseng stem cells is obtained;

[0074] Step 2: Put mogroside, sodium gluconate, disodium ethylenediaminetetraacetic acid and ultrapure water into a 1.3 kW stirrer at a stirring temperature of 26° C. for 13 min at a stirring speed of 150 r / min, and mix well to obtain a flavor-correcting and stabilizing solvent;

[0075] Step 3: Place the mixed solution containing ginseng stem cells prepared in step 1 into the flavor-correcting and stabilizing solvent prepared in step 2, stir at a temperature of 26° C., for 25 min, at a speed of 140 r / min to obtain an oral liquid preparation containing ginseng stem cells.

[0076] Example 3: This example provides an oral liquid preparation containing ginseng stem cells and a preparation method thereof. The oral liquid preparation containing ginseng stem cells comprises the following components in parts by weight:

[0077] 90 parts of induced ginseng stem cells, 80 parts of nutrient absorption promoting nanoemulsion, 10 parts of mogroside, 8 parts of sodium gluconate, 5 parts of disodium edetate, and 30 parts of ultrapure water.

[0078] The method for preparing induced ginseng stem cells specifically comprises the following steps:

[0079] S1. Weigh 300 parts by weight of fresh ginseng rhizomes, soak them in 70% ethanol solution for 30 seconds in a 28°C clean bench, disinfect them in 0.1% mercuric chloride solution for 12 minutes, rinse them with ultrapure water for 5 times, cut the ginseng rhizomes into slices with a thickness of 0.1 mm, and cut the cambium tissue in the slices to obtain the ginseng rhizome cambium;

[0080] S2, weigh 5 parts of cellulase and 5 parts of pectinase, 130 parts of 0.6 mol / L sucrose solution and the ginseng rhizome cambium prepared in S1, put them into a stirring tank with a power of 1.5 kW, the stirring temperature is 28°C, the stirring time is 12 min, the stirring speed is 130 r / min, soak for 50 min after stirring, put them into a high-speed centrifuge with a power of 2.8 kW, the centrifugal temperature is 2°C, the centrifugal time is 15 min, the centrifugal speed is 12000 r / min, and after centrifugation, rinse with ultrapure water 3 times to obtain pure ginseng stem cell protoplasts;

[0081] S3, weighing 60 parts of giant kelp and 70 parts of red hair algae, putting them into a pulping machine with a power of 2.2 kW, the pulping temperature is 23°C, the pulping time is 20 min, the pulping speed is 150 r / min, and mixing and grinding to obtain a nutrient slurry;

[0082] S4, weigh 12 parts by weight of growth-promoting bacteria agent and 210 parts of sterile LB liquid culture medium, put them into a constant temperature shaking incubator with a power of 1.6 kW, the constant temperature shaking culture temperature is 37°C, the constant temperature shaking culture time is 30 h, the constant temperature shaking culture speed is 260 r / min, after the constant temperature shaking culture, use an ultrasonic cell disruptor with a power of 1.5 kW, the ultrasonic disruption frequency is 20 kHz, the ultrasonic disruption temperature is 0°C, the ultrasonic disruption time is 13 min, and after disruption, filter with a 0.22 mm sterile filter membrane, and put them into a stirring tank with a power of 1.5 kW with the nutrient slurry prepared in S3, 20 parts of inositol, 12 parts of calcium chloride, and 160 parts of sterile MS liquid culture medium, the stirring temperature is 28°C, the stirring time is 20 min, the stirring speed is 130 r / min, and mix them evenly to obtain a cell growth-promoting nutrient solution;

[0083] S5, placing the purified ginseng stem cell protoplasts prepared by S2 and the cell growth promoting nutrient solution prepared by S4 into a constant temperature shaking incubator with a power of 1.6 kW, the constant temperature shaking culture temperature is 28°C, the constant temperature shaking culture time is 45 days, the constant temperature shaking culture speed is 80 r / min, after mixed culture, placing in a high-speed centrifuge with a power of 2.8 kW, the centrifugation temperature is 2°C, the centrifugation time is 15 min, the centrifugation speed is 12000 r / min, and after centrifugation, washing with ultrapure water for 3 times to obtain a purified ginseng stem cell population;

[0084] S6, weighing 50 parts of Aralia dasyphylla, 40 parts of Fenugreek, and 100 parts of anhydrous ethanol, and putting them into an ultrasonic extractor with a power of 1.6 kW, the extraction pressure is 1.5 MPa, the extraction frequency is 25 kHz, the extraction temperature is 45°C, and the extraction time is 13 min. After extraction, put them into a vacuum dryer with a power of 2.2 kW, the vacuum drying pressure is 0.3 MPa, the vacuum drying temperature is 60°C, and the vacuum drying time is 1.5 h, and the anhydrous ethanol is removed to obtain a plant extract;

[0085] S7. Place the plant extract prepared in S6, 220 parts by weight of sterile MS liquid culture medium, and the pure ginseng stem cell population prepared in S5 into a constant temperature shaking incubator with a power of 1.6 kW. The constant temperature shaking culture temperature is 37°C, the constant temperature shaking culture time is 7 days, and the constant temperature shaking culture speed is 90 r / min. After mixed culture, place it in a high-speed centrifuge with a power of 2.8 kW, the centrifugation temperature is 2°C, the centrifugation time is 15 min, and the centrifugation speed is 12000 r / min. After centrifugation, rinse it with ultrapure water 3 times to obtain induced ginseng stem cells.

[0086] The growth-promoting bacterial agent comprises: Actinomycetes aurantifolia, Gluconacetobacter xylosus, and Pseudomonas betae, and the weight ratio of the mixture is 2.5:2:1.

[0087] The aurantiacus coralliformis was purchased from the China General Microbiological Culture Collection Center with a collection number of CGMCC4.2187; the xylosyl gluconoacetobacter was purchased from the China General Microbiological Culture Collection Center with a collection number of CGMCC1.2706; and the beet Pseudomonas was purchased from the China General Microbiological Culture Collection Center with a collection number of CGMCC1.8810.

[0088] The preparation method of the nanoemulsion for promoting nutrient absorption specifically comprises the following steps:

[0089] L1, weighing 40 parts of narcissus glycosides, 20 parts of betaine and 110 parts of triacetin, putting them into a 1.5 kW stirrer, stirring at 28°C, stirring for 20 min, stirring at 160 r / min, and mixing to obtain a transport-promoting solution;

[0090] L2, weigh 80 parts of lactoglobulin, 70 parts of potassium lauryl ether phosphate, and 50 parts of ultrapure water, put them into the transport-promoting solution prepared in L1, stir at 28°C for 18 min, stir at 160 r / min, and mix well to obtain a nutrient absorption-promoting mixture;

[0091] L3. Place the nutrient absorption promoting mixture prepared in L2 into an ultra-high pressure homogenizer with a power of 2.8 kW, a pressure of 150 MPa, and a high-speed air flow rate of 1500 m / s for ultra-high pressure homogenization to form a nutrient absorption promoting nanoemulsion.

[0092] This embodiment also provides a method for preparing an oral liquid preparation containing ginseng stem cells, which specifically comprises the following steps:

[0093] Step 1: Place the induced ginseng stem cells in a vacuum freeze dryer with a power of 7.5 kW, a vacuum degree of 80 Pa, a freezing temperature of -20°C, and a freezing time of 3 h, then heat to 4°C, dry for 1.5 h, and then place them in an ultrasonic disperser with a nutrient absorption-promoting nanoemulsion with a power of 1.9 kW, with an ultrasonic dispersion frequency of 25 kHz, an ultrasonic dispersion temperature of 15°C, and an ultrasonic dispersion time of 10 min. After ultrasonic dispersion, a mixed solution containing ginseng stem cells is obtained;

[0094] Step 2: Put mogroside, sodium gluconate, disodium ethylenediaminetetraacetic acid and ultrapure water into a 1.5 kW stirrer at a stirring temperature of 28° C. for 15 min at a stirring speed of 160 r / min, and mix well to obtain a flavor-correcting and stabilizing solvent;

[0095] Step 3: Place the mixed solution containing ginseng stem cells prepared in step 1 into the flavor-correcting and stabilizing solvent prepared in step 2, stir at a temperature of 28° C., for 30 min, at a speed of 160 r / min to obtain an oral liquid preparation containing ginseng stem cells.

[0096] Example 4: This example provides an oral liquid preparation containing ginseng stem cells and a preparation method thereof. The oral liquid preparation containing ginseng stem cells comprises the following components in parts by weight:

[0097] 75 parts of induced ginseng stem cells, 65 parts of nutrient absorption promoting nanoemulsion, 7 parts of mogroside, 5 parts of sodium gluconate, 3 parts of disodium edetate, and 23 parts of ultrapure water.

[0098] The method for preparing induced ginseng stem cells specifically comprises the following steps:

[0099] S1. Weigh 230 parts by weight of fresh ginseng rhizomes, soak them in 70% ethanol solution for 23 seconds in a 28°C clean bench, disinfect them in 0.1% mercuric chloride solution for 10 minutes, rinse them with ultrapure water for 3 times, cut the ginseng rhizomes into slices with a thickness of 0.1 mm, and cut the cambium tissue in the slices to obtain the ginseng rhizome cambium;

[0100] S2, weigh 4 parts of cellulase and 3 parts of pectinase, 115 parts of 0.6 mol / L sucrose solution and the ginseng rhizome cambium prepared in S1, put them into a stirring tank with a power of 1.2 kW, the stirring temperature is 26°C, the stirring time is 9 min, the stirring speed is 110 r / min, soak for 35 min after stirring, put them into a high-speed centrifuge with a power of 2.5 kW, the centrifugal temperature is 2°C, the centrifugal time is 12 min, the centrifugal speed is 12000 r / min, and after centrifugation, rinse with ultrapure water twice to obtain pure ginseng stem cell protoplasts;

[0101] S3, weighing 45 parts of giant kelp and 55 parts of red hair algae, putting them into a pulping machine with a power of 1.8 kW, the pulping temperature is 19°C, the pulping time is 17 min, the pulping speed is 130 r / min, and mixing and grinding to obtain a nutrient slurry;

[0102] S4, weighing 10 parts by weight of the growth-promoting bacterial agent and 180 parts of sterile LB liquid culture medium, and putting them into a constant temperature shaking incubator with a power of 1.3 kW, the constant temperature shaking culture temperature is 34°C, the constant temperature shaking culture time is 26 h, and the constant temperature shaking culture speed is 230 r / min. After the constant temperature shaking culture, use an ultrasonic cell disruptor with a power of 1.3 kW, the ultrasonic disruption frequency is 20 kHz, the ultrasonic disruption temperature is 0°C, and the ultrasonic disruption time is 11 min. After disruption, use a 0.22 mm sterile filter membrane to filter, and put it into a stirring tank with a power of 1.3 kW with the nutrient slurry prepared in S3, 17 parts of inositol, 9 parts of calcium chloride, and 135 parts of sterile MS liquid culture medium, the stirring temperature is 26°C, the stirring time is 17 min, and the stirring speed is 120 r / min, and mix them evenly to obtain a cell growth-promoting nutrient solution;

[0103] S5. Place the purified ginseng stem cell protoplasts prepared in S2 and the cell growth promoting nutrient solution prepared in S4 into a constant temperature shaking incubator with a power of 1.3 kW, the constant temperature shaking culture temperature is 25°C, the constant temperature shaking culture time is 42 days, the constant temperature shaking culture speed is 50 r / min, after mixed culture, place in a high-speed centrifuge with a power of 2.5 kW, the centrifugal temperature is 2°C, the centrifugal time is 12 min, the centrifugal speed is 12000 r / min, and after centrifugation, rinse twice with ultrapure water to obtain a purified ginseng stem cell population;

[0104] S6, weighing 35 parts of Aralia dasyphylla, 25 parts of Fenugreek, and 85 parts of anhydrous ethanol, and putting them into an ultrasonic extractor with a power of 1.3 kW, the extraction pressure is 1.5 MPa, the extraction frequency is 25 kHz, the extraction temperature is 42°C, and the extraction time is 9 minutes. After extraction, put them into a vacuum dryer with a power of 1.8 kW, the vacuum drying pressure is 0.3 MPa, the vacuum drying temperature is 56°C, and the vacuum drying time is 1.5 h, and the anhydrous ethanol is removed to obtain a plant extract;

[0105] S7. Place the plant extract prepared in S6, 190 parts by weight of sterile MS liquid culture medium, and the pure ginseng stem cell population prepared in S5 into a constant temperature shaking incubator with a power of 1.3 kW. The constant temperature shaking culture temperature is 36°C, the constant temperature shaking culture time is 6 d, and the constant temperature shaking culture speed is 60 r / min. After mixed culture, place it in a high-speed centrifuge with a power of 2.5 kW, the centrifugation temperature is 2°C, the centrifugation time is 12 min, and the centrifugation speed is 12000 r / min. After centrifugation, rinse it twice with ultrapure water to obtain induced ginseng stem cells.

[0106] The growth-promoting bacterial agent comprises: Actinomycetes aurantifolia, Gluconacetobacter xylosus, and Pseudomonas betae, and the mixing ratio by weight is 1.5:1.3:1.

[0107] The aurantiacus coralliformis was purchased from the China General Microbiological Culture Collection Center with a collection number of CGMCC4.2187; the xylosyl gluconoacetobacter was purchased from the China General Microbiological Culture Collection Center with a collection number of CGMCC1.2706; and the beet Pseudomonas was purchased from the China General Microbiological Culture Collection Center with a collection number of CGMCC1.8810.

[0108] The preparation method of the nanoemulsion for promoting nutrient absorption specifically comprises the following steps:

[0109] L1, weighing 33 parts of narcissus glycosides, 17 parts of betaine and 95 parts of triacetin, putting them into a 1.2 kW stirrer, stirring at 26°C, stirring for 17 min, stirring at 140 r / min, and mixing to obtain a transport-promoting solution;

[0110] L2, weighing 65 parts of lactoglobulin, 55 parts of potassium lauryl ether phosphate and 35 parts of ultrapure water, and adding them into the transport-promoting solution prepared in L1, stirring at 26°C, stirring for 14 min, stirring at 130 r / min, and mixing to obtain a nutrient absorption-promoting mixture;

[0111] L3. Place the nutrient absorption promoting mixture prepared in L2 into an ultra-high pressure homogenizer with a power of 2.6 kW, a pressure of 150 MPa, and a high-speed air flow rate of 1200 m / s for ultra-high pressure homogenization to form a nutrient absorption promoting nanoemulsion.

[0112] This embodiment also provides a method for preparing an oral liquid preparation containing ginseng stem cells, which specifically comprises the following steps:

[0113] Step 1: Place the induced ginseng stem cells in a vacuum freeze dryer with a power of 5.8 kW, a vacuum degree of 60 Pa, a freezing temperature of -20°C, and a freezing time of 2.3 h, then heat to 3°C, dry for 1.3 h, and then place them in an ultrasonic disperser with a nutrient absorption-promoting nanoemulsion with a power of 1.6 kW, an ultrasonic dispersion frequency of 25 kHz, an ultrasonic dispersion temperature of 12°C, and an ultrasonic dispersion time of 8 min. After ultrasonic dispersion, a mixed solution containing ginseng stem cells is obtained;

[0114] Step 2: Put mogroside, sodium gluconate, disodium ethylenediaminetetraacetic acid and ultrapure water into a 1.2 kW stirrer at a stirring temperature of 25° C. for 12 min at a stirring speed of 140 r / min, and mix well to obtain a flavor-correcting and stabilizing solvent;

[0115] Step 3: Place the mixed solution containing ginseng stem cells prepared in step 1 into the flavor-correcting and stabilizing solvent prepared in step 2, stir at a temperature of 26° C., for 23 min, at a speed of 130 r / min to obtain an oral liquid preparation containing ginseng stem cells.

[0116] Example 5: This example provides an oral liquid preparation containing ginseng stem cells and a preparation method thereof. The oral liquid preparation containing ginseng stem cells comprises the following components in parts by weight:

[0117] 85 parts of induced ginseng stem cells, 75 parts of nutrient absorption promoting nanoemulsion, 9 parts of mogroside, 7 parts of sodium gluconate, 5 parts of disodium edetate, and 28 parts of ultrapure water.

[0118] The method for preparing induced ginseng stem cells specifically comprises the following steps:

[0119] S1. Weigh 280 parts by weight of fresh ginseng rhizomes, soak them in 70% ethanol solution in a 28°C clean bench for 28 seconds, disinfect them in 0.1% mercuric chloride solution for 12 minutes, rinse them with ultrapure water for 5 times, cut the ginseng rhizomes into slices with a thickness of 0.1 mm, and cut the cambium tissue in the slices to obtain the ginseng rhizome cambium;

[0120] S2, weigh 5 parts of cellulase and 4 parts of pectinase, 125 parts of 0.6 mol / L sucrose solution and the ginseng rhizome cambium prepared in S1, put them into a stirring tank with a power of 1.4 kW, the stirring temperature is 27°C, the stirring time is 11 min, the stirring speed is 120 r / min, soak for 45 min after stirring, put them into a high-speed centrifuge with a power of 2.7 kW, the centrifugal temperature is 2°C, the centrifugal time is 14 min, the centrifugal speed is 12000 r / min, and after centrifugation, rinse with ultrapure water 3 times to obtain pure ginseng stem cell protoplasts;

[0121] S3, weighing 55 parts of giant kelp and 65 parts of red hair algae, putting them into a pulping machine with a power of 2.1 kW, the pulping temperature is 21°C, the pulping time is 19 min, the pulping speed is 140 r / min, and mixing and grinding to obtain a nutrient slurry;

[0122] S4, weigh 12 parts by weight of growth-promoting bacteria agent and 200 parts of sterile LB liquid culture medium, put them into a constant temperature shaking incubator with a power of 1.5 kW, the constant temperature shaking culture temperature is 36 ° C, the constant temperature shaking culture time is 29 h, the constant temperature shaking culture speed is 250 r / min, after the constant temperature shaking culture, use an ultrasonic cell disruptor with a power of 1.5 kW, the ultrasonic disruption frequency is 20 kHz, the ultrasonic disruption temperature is 0 ° C, the ultrasonic disruption time is 12 min, and after disruption, use a 0.22 mm sterile filter membrane to filter, and put them into a stirring tank with a power of 1.4 kW with the nutrient slurry prepared in S3, 19 parts of inositol, 11 parts of calcium chloride, and 150 parts of sterile MS liquid culture medium, the stirring temperature is 27 ° C, the stirring time is 19 min, the stirring speed is 120 r / min, and mix them evenly to obtain a cell growth-promoting nutrient solution;

[0123] S5. Place the purified ginseng stem cell protoplasts prepared in S2 and the cell growth promoting nutrient solution prepared in S4 into a constant temperature shaking incubator with a power of 1.5 kW, the constant temperature shaking culture temperature is 27°C, the constant temperature shaking culture time is 44 d, the constant temperature shaking culture speed is 70 r / min, after mixed culture, place in a high-speed centrifuge with a power of 2.7 kW, the centrifugal temperature is 2°C, the centrifugal time is 14 min, the centrifugal speed is 12000 r / min, and after centrifugation, rinse with ultrapure water 3 times to obtain a purified ginseng stem cell population;

[0124] S6, weighing 45 parts of Aralia dasyphylla, 35 parts of Fenugreek, and 95 parts of anhydrous ethanol, and putting them into an ultrasonic extractor with a power of 1.5 kW, the extraction pressure is 1.5 MPa, the extraction frequency is 25 kHz, the extraction temperature is 44°C, and the extraction time is 12 min. After extraction, put them into a vacuum dryer with a power of 2.1 kW, the vacuum drying pressure is 0.3 MPa, the vacuum drying temperature is 59°C, and the vacuum drying time is 1.5 h, and the anhydrous ethanol is removed to obtain a plant extract;

[0125] S7. Place the plant extract prepared in S6, 210 parts by weight of sterile MS liquid culture medium, and the pure ginseng stem cell population prepared in S5 into a constant temperature shaking incubator with a power of 1.5 kW. The constant temperature shaking culture temperature is 36°C, the constant temperature shaking culture time is 6 days, and the constant temperature shaking culture speed is 80 r / min. After mixed culture, place it in a high-speed centrifuge with a power of 2.7 kW, the centrifugation temperature is 2°C, the centrifugation time is 14 min, and the centrifugation speed is 12000 r / min. After centrifugation, rinse it with ultrapure water 3 times to obtain induced ginseng stem cells.

[0126] The growth-promoting bacterial agent comprises: Actinomycetes aurantifolia, Gluconacetobacter xylosus, and Pseudomonas betae, and the weight ratio of the mixture is 2.3:1.8:1.

[0127] The aurantiacus coralliformis was purchased from the China General Microbiological Culture Collection Center with a collection number of CGMCC4.2187; the xylosyl gluconoacetobacter was purchased from the China General Microbiological Culture Collection Center with a collection number of CGMCC1.2706; and the beet Pseudomonas was purchased from the China General Microbiological Culture Collection Center with a collection number of CGMCC1.8810.

[0128] The preparation method of the nanoemulsion for promoting nutrient absorption specifically comprises the following steps:

[0129] L1, weighing 38 parts of narcissus glycosides, 19 parts of betaine and 105 parts of triacetin, putting them into a 1.4 kW stirrer, stirring at 27°C, stirring for 19 min, stirring at 160 r / min, and mixing to obtain a transport-promoting solution;

[0130] L2, weigh 75 parts of lactoglobulin, 65 parts of potassium lauryl ether phosphate and 45 parts of ultrapure water, put them into the transport-promoting solution prepared in L1, stir at 27°C for 17 min, stir at 150 r / min, and mix well to obtain a mixture that promotes nutrient absorption;

[0131] L3. Place the nutrient absorption promoting mixture prepared in L2 into an ultra-high pressure homogenizer with a power of 2.7 kW, a pressure of 150 MPa, and a high-speed air flow rate of 1400 m / s for ultra-high pressure homogenization to form a nutrient absorption promoting nanoemulsion.

[0132] This embodiment also provides a method for preparing an oral liquid preparation containing ginseng stem cells, which specifically comprises the following steps:

[0133] Step 1: Place the induced ginseng stem cells in a vacuum freeze dryer with a power of 7.2 kW, a vacuum degree of 80 Pa, a freezing temperature of -20°C, and a freezing time of 2.8 h, then heat to 4°C, dry for 1.4 h, and then place them in an ultrasonic disperser with a nutrient absorption-promoting nanoemulsion with a power of 1.8 kW, with an ultrasonic dispersion frequency of 25 kHz, an ultrasonic dispersion temperature of 14°C, and an ultrasonic dispersion time of 10 min. After ultrasonic dispersion, a mixed solution containing ginseng stem cells is obtained;

[0134] Step 2: Put mogroside, sodium gluconate, disodium ethylenediaminetetraacetic acid and ultrapure water into a 1.4 kW stirrer at a stirring temperature of 27° C. for 14 min at a stirring speed of 160 r / min, and mix well to obtain a flavor-correcting and stabilizing solvent;

[0135] Step 3: Place the mixed solution containing ginseng stem cells prepared in step 1 into the flavor correcting and stabilizing solvent prepared in step 2, stir at a temperature of 27° C., for 28 min, at a speed of 150 r / min to obtain an oral liquid preparation containing ginseng stem cells.

[0136] Experimental Example 1: Test for determining the content of PPD-type ginsenosides in ginseng stem cells.

[0137] The test steps for determining the content of PPD-type ginsenosides in the oral liquid preparation containing ginseng stem cells prepared in Examples 1-5 of the present invention are as follows:

[0138] (1) Take 1 mL of the oral liquid preparation containing ginseng stem cells prepared in Examples 1-5 as the experimental group sample, and take 1 mL of ordinary ginseng stem cell oral liquid as the CK control group sample, and take 6 parallel samples for each group;

[0139] (2) The oral liquid preparation containing ginseng stem cells prepared in Example 1-5 and the ordinary ginseng stem cell oral liquid were extracted with 80% methanol at a volume ratio of 1:40 at 60°C. Ultrasonic assisted extraction was used during the extraction process. The operation was fully performed 3 times, each time for 10 min. The extract was evaporated to dry methanol at 60°C, extracted with water-saturated n-butanol, and the n-butanol layer was collected. The n-butanol was then evaporated in a 60°C water bath, and then dissolved with an appropriate amount of methanol to obtain a ginsenoside extract for subsequent HPLC analysis and detection;

[0140] (3) The ginsenoside content in the oral liquid preparation containing ginseng stem cells prepared in Examples 1-5 and the ordinary ginseng stem cell oral liquid sample extract was determined by high performance liquid chromatography (HPLC). The mobile phase for determination was a mixture of acetonitrile and 1% formic acid. The flow rate of the mobile phase was set to 1.0 mL / min, the column temperature was controlled at 35°C, the injection volume was 3 mL, and the detection wavelength was set to 202 nm. In order to accurately determine the content of PPD-type ginsenosides in the oral liquid samples, ginsenosides Rb1, Rd, Rg3, Rh2 and F2 were used as standards to determine the content of each saponin monomer in the sample, and then the sum of the content of each saponin monomer represented the total content of PPD-type ginsenosides in ginseng cells. The average value was taken and recorded. The obtained data were statistically analyzed using IBM SPSS Statistics 22.0 software, and graphics were drawn using Origin 2022.

[0141] Experimental Example 2: Mouse absorption test.

[0142] The experimental steps for determining the absorption of the oral liquid preparation containing ginseng stem cells prepared in Examples 1-5 of the present invention on mice are as follows:

[0143] (1) 42 Kunming mice weighing 25 g were selected and randomly divided into 6 groups, with 7 mice in each group. Experimental groups 1-5 were gavaged with 3.5 mL of the oral liquid preparation containing ginseng stem cells prepared in Examples 1-5 every day for 10 consecutive days. The mice that consumed 3.5 mL of ordinary ginseng stem cell oral liquid every day were the CK control group, which consumed it for 10 consecutive days.

[0144] (2) Starting from the first feeding of the oral liquid preparation containing ginseng stem cells and ordinary ginseng stem cell oral liquid prepared in Examples 1-5, 0.6 mL of sterile venous blood was collected from each Kunming mouse every 1 day, and heparin was used to coagulate to prepare plasma. The immunoglobulin IgG assay kit (ELISA, catalog number: M1245L96), immunoglobulin IgM assay kit (ELISA, catalog number: M1246L96), and immunoglobulin IgA assay kit (ELISA, catalog number: M1244L96) were used. The contents of immunoglobulins IgG, IgM and IgA in the plasma of each group of mice were measured using an automatic blood biochemical analyzer. The measurements were continued until the 10th day of feeding, and the average values ​​were recorded. The obtained data were statistically analyzed using IBM SPSS Statistics 22.0 software, and the graphics were drawn using Origin 2022.

[0145] Experimental Example 3: Stability test of ginseng stem cell oral liquid preparation.

[0146] The experimental steps for determining the stability of the oral liquid preparation containing ginseng stem cells prepared in Examples 1-5 of the present invention are as follows:

[0147] (1) Take 10 mL of the oral liquid preparation containing ginseng stem cells prepared in Examples 1-5 as the experimental group sample, and take 10 mL of ordinary ginseng stem cell oral liquid as the CK control group sample, and take 6 parallel samples for each group;

[0148] (2) Store in a light-proof refrigerator at 4°C. After 30, 60, and 90 days of storage, score the samples in the experimental and control groups and record the results.

[0149] Table 1. Oral liquid sample scoring standard table

[0150]

[0151] Table 2. Scoring table of oral solution samples on the 30th day of storage

[0152]

[0153] Table 3. Scoring table of oral solution samples on the 60th day of storage

[0154]

[0155] Table 4. Scoring table of oral solution samples on the 90th day of storage

[0156]

[0157] Result analysis: As shown in Tables 2, 3 and 4, the oral liquid preparations containing ginseng stem cells prepared in Examples 1-5 were stored in a 4°C refrigerator away from light. At the 30th, 60th and 90th days of storage in the dark, the appearance, smell and taste scores were all 5 points, while the ordinary ginseng stem cell oral liquid in the CK control group was stored in a 4°C refrigerator away from light. At the 90th day of storage in the dark, the appearance score was 2.85 points, the smell score was 2.73 points, and the taste score was 2.69 points. The oral liquid preparations containing ginseng stem cells prepared in Examples 1-5 had higher appearance, smell and taste scores than ordinary ginseng stem cell oral liquid, indicating that the addition of the nutrient absorption promoting nanoemulsion and the flavor correcting and stabilizing solvent makes the appearance, smell and taste of the oral liquid preparation containing ginseng stem cells described in the present invention stable after long-term storage.

[0158] Result analysis:

[0159] Figure 1 This is a graph showing the results of the content of PPD-type ginsenosides in the samples described in Experimental Example 1 of the present invention. As shown in the figure, the oral liquid preparations containing ginseng stem cells described in Examples 1-5 were measured by HPLC, and the PPD-type ginsenosides contained were 5.92 mg / mL, 6.08 mg / mL, 6.21 mg / mL, 5.98 mg / mL, and 6.07 mg / mL, while the ordinary ginseng stem cell oral liquid in the control group was measured by HPLC, and the PPD-type ginsenosides contained were 0.91 mg / mL, indicating that in the oral liquid preparation containing ginseng stem cells described in the present invention, the induced ginseng stem cells synthesized and accumulated abundant ginsenosides.

[0160] Figure 2This is a graph showing the results of the immunoglobulin IgG content in mouse plasma as described in Experimental Example 2 of the present invention. As shown in the figure, the mice in groups 1 to 5 of Examples were fed with the oral liquid preparation containing ginseng stem cells described in groups 1 to 5 of Examples for 10 consecutive days by gavage. The immunoglobulin IgG content in the plasma of the mice in group 3 of Example was 9.17 mg / mL, 10.76 mg / mL, 11.73 mg / mL, 12.74 mg / mL, 13.78 mg / mL, 14.02 mg / mL, 14.97 mg / mL, 15.55 mg / mL, 16.93 mg / mL, and 17.49 mg / mL, while the mice in the control group were fed with ordinary ginseng stem cell oral liquid for 10 consecutive days by gavage, and the immunoglobulin IgG content in the mouse plasma was 9.12 mg / mL, 9.17 mg / mL, 9.23 mg / mL, 9.27 mg / mL, 9.34 mg / mL, 9.41 mg / mL, 9.57 mg / mL, 9.7 mg / mL, 9.78 mg / mL, 9.84 mg / mL; Figure 3 This is a graph showing the results of the immunoglobulin IgM content in mouse plasma as described in Experimental Example 2 of the present invention. As shown in the figure, the mice in groups 1 to 5 of Examples were fed with the oral liquid preparation containing ginseng stem cells described in groups 1 to 5 of Examples for 10 consecutive days by gavage. The immunoglobulin IgM content in the plasma of mice in group 3 of Example was 0.63 mg / mL, 0.96 mg / mL, 1.12 mg / mL, 1.37 mg / mL, 1.61 mg / mL, 1.77 mg / mL, 1.84 mg / mL, 1.91 mg / mL, 2.1 mg / mL, and 2.24 mg / mL, while the mice in the control group were fed with ordinary ginseng stem cell oral liquid for 10 consecutive days by gavage, and the immunoglobulin IgM content in the mouse plasma was 0.61 mg / mL, 0.64 mg / mL, 0.66 mg / mL, 0.68 mg / mL, 0.68 mg / mL, 0.71 mg / mL, 0.73 mg / mL, 0.75 mg / mL, 0.77 mg / mL, 0.81 mg / mL; Figure 4This is a graph showing the results of the immunoglobulin IgA content in mouse plasma as described in Experimental Example 2 of the present invention. As shown in the figure, the mice in groups 1 to 5 of Examples were fed with the oral liquid preparation containing ginseng stem cells described in groups 1 to 5 of Examples for 10 consecutive days by gavage. The immunoglobulin IgA content in the plasma of mice in group 3 of Example was 1.24 mg / mL, 1.61 mg / mL, 1.91 mg / mL, 2.26 mg / mL, 2.47 mg / mL, 2.63 mg / mL, 2.91 mg / mL, 3.19 mg / mL, 3.37 mg / mL, and 3.58 mg / mL, while the mice in the control group were fed with ordinary ginseng stem cell oral liquid for 10 consecutive days by gavage. The immunoglobulin IgA content in the mouse plasma was 1.21 mg / mL, 1.28 mg / mL, 1.34 mg / mL, 1.37 mg / mL, 1.44 mg / mL, 1.53 mg / mL, 1.57 mg / mL, 1.61 mg / mL, 1.67 mg / mL, and 1.71 mg / mL, indicating that in the oral liquid preparation containing ginseng stem cells described in the present invention, the nutrient absorption promoting nanoemulsion improves the solubility of each component, can adhere to the surface of small intestinal epithelial cells, and promotes the absorption of ginsenosides in the small intestine for a long time with high efficiency.

[0161] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

[0162] The present invention and its implementation methods are described above, which is not restrictive. The drawings are only one of the implementation methods of the present invention, and the actual application is not limited thereto. In short, if ordinary technicians in the field are inspired by it and design methods and embodiments similar to the technical solution without creativity without departing from the purpose of the invention, they should all fall within the protection scope of the present invention.

Claims

1. An oral liquid preparation containing ginseng stem cells, characterized in that: The oral liquid preparation containing ginseng stem cells is composed of the following ingredients: 70-90 parts of induced ginseng stem cells, 60-80 parts of nutrient absorption promoting nanoemulsion, 7-10 parts of mogroside, 5-8 parts of sodium gluconate, 3-5 parts of disodium edetate, and 20-30 parts of ultrapure water; The method for preparing induced ginseng stem cells specifically comprises the following steps: S1, weighing 200-300 parts by weight of fresh ginseng rhizomes, disinfecting them in a clean bench, rinsing them with ultrapure water for 3-5 times, cutting the ginseng rhizomes into slices with a thickness of 0.1 mm, and cutting the cambium tissue in the slices to obtain the ginseng rhizome cambium; S2, weighing 3-5 parts of cellulase and 3-5 parts of pectinase, 100-130 parts of 0.6 mol / L sucrose solution and the ginseng rhizome cambium prepared in S1, stirring and soaking, centrifuging and washing with ultrapure water 2-3 times to obtain pure ginseng stem cell protoplasts; S3, weighing 40-60 parts by weight of giant kelp and 50-70 parts by weight of red hair algae, mixing and grinding to obtain a nutrient slurry; S4, weighing 10-12 parts by weight of a growth-promoting bacterial agent and 170-210 parts of a sterile LB liquid culture medium, culturing at a constant temperature with shaking, performing ultrasonic disruption, and filtering using a 0.22 mm sterile filter membrane, and mixing with the nutrient slurry prepared in S3, 15-20 parts of inositol, 8-12 parts of calcium chloride, and 130-160 parts of a sterile MS liquid culture medium to obtain a cell growth-promoting nutrient solution; S5, mixing the purified ginseng stem cell protoplasts prepared in S2 and the cell growth promoting nutrient solution prepared in S4, culturing at a constant temperature and shaking, centrifuging and then washing with ultrapure water 2-3 times to obtain purified ginseng stem cell populations; S6, weighing 30-50 parts by weight of Aralia dasyphylla, 20-40 parts by weight of Fenugreek, and 80-100 parts by weight of anhydrous ethanol for mixed extraction, and then drying under reduced pressure to remove the anhydrous ethanol to obtain a plant extract; S7, mixing the plant extract prepared in S6 with 180-220 parts by weight of a sterile MS liquid culture medium and the purified ginseng stem cell population prepared in S5, centrifuging at high speed and washing with ultrapure water 2-3 times to obtain induced ginseng stem cells; The preparation method of the nanoemulsion for promoting nutrient absorption specifically comprises the following steps: L1, weighing 30-40 parts by weight of narcissus glycoside, 15-20 parts by weight of betaine and 90-110 parts by weight of triacetin to obtain a transport-promoting solution; L2, weigh 60-80 parts by weight of lactoglobulin, 50-70 parts by weight of potassium lauryl ether phosphate, and 30-50 parts by weight of ultrapure water and the transport-promoting solution prepared in L1, and mix them to obtain a nutrient absorption-promoting mixture; L3, subjecting the nutrient absorption promoting mixture prepared in L2 to ultra-high pressure homogenization to form a nutrient absorption promoting nanoemulsion; The method for preparing the oral liquid preparation containing ginseng stem cells is characterized by comprising the following steps: Step 1: freeze-dry the induced ginseng stem cells, and then disperse them in a nutrient absorption-promoting nanoemulsion with ultrasonic waves to obtain a mixed solution containing ginseng stem cells; Step 2, mixing mogroside, sodium gluconate, disodium ethylenediaminetetraacetic acid and ultrapure water to obtain a flavor-correcting and stabilizing solvent; Step 3: Stir the mixed solution containing ginseng stem cells prepared in step 1 and the flavor correcting and stabilizing solvent prepared in step 2 to obtain an oral liquid preparation containing ginseng stem cells.

2. The oral liquid preparation containing ginseng stem cells according to claim 1, characterized in that: The composition of the growth-promoting bacterial agent in step S4 includes: Actinomycetes aurantifolia, Gluconacetobacter xylosus, and Pseudomonas betae, and the weight ratio is 1-2.5:1-2:1.

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