CN112138033A - Ginseng ointment with effect of reducing uric acid activity and preparation method thereof - Google Patents
Ginseng ointment with effect of reducing uric acid activity and preparation method thereof Download PDFInfo
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- CN112138033A CN112138033A CN202011179673.XA CN202011179673A CN112138033A CN 112138033 A CN112138033 A CN 112138033A CN 202011179673 A CN202011179673 A CN 202011179673A CN 112138033 A CN112138033 A CN 112138033A
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- ginseng
- enzyme
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- water
- preparation
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- 238000012360 testing method Methods 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 229940126680 traditional chinese medicines Drugs 0.000 description 1
- 150000003648 triterpenes Chemical class 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
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Abstract
The invention provides a ginseng cream with the function of reducing the activity of uric acid and a preparation method thereof, wherein the content of Compound K and 20(S) -F2 in the ginseng cream accounts for more than 1 percent of the proportion of total saponins of ginseng. The ginseng cream contains 20(S) -F2 and Compound K, and contains ginseng polysaccharide and other components, and has the effect of reducing uric acid. The product can be directly applied to food and health care products, is suitable for industrialization and has strong applicability.
Description
Technical Field
The invention belongs to the field of ginseng deep processing, and particularly relates to ginseng cream with the function of reducing uric acid activity and a preparation method thereof.
Background
Ginseng, a famous and precious traditional Chinese medicine, enjoys the reputations of "Baicaozhiwang" and "northeast Sanbao", has been used for treating diseases and daily nourishing by oriental countries such as China, Korea and Japan. The plant classification belongs to the genus of Panax in Araliaceae, and the same genus of American ginseng and pseudo-ginseng are common Chinese herbal medicines. The main active component of ginseng is ginsenoside, which can be divided into tetracyclic triterpene dammarane type saponin and oleanolic acid type saponin according to the difference of aglycon structure. Different amounts and different types of glycosyl groups are connected at different positions of different aglycones to form different types of ginsenosides, and more than 180 types of ginsenosides are found at present; the ginsenosides can be classified into polyglycosyl saponins containing 3-4 glycosyl groups and low glycosyl saponins containing 1-2 glycosyl groups according to the amount of glycosyl groups.
In ancient times, ginseng used by people is wild ginseng, but the wild ginseng is more and more rare due to factors such as resource limitation, demand and the like, so that artificially cultivated ginseng becomes the mainstream of the market. The main active ingredients of the planted ginseng and the wild ginseng are ginsenoside, and the difference is that the planted ginseng only contains polysaccharide-based ginsenoside, while the wild ginseng contains a small amount of ginsenoside with low glycosyl. The polysaccharide-based saponin has low human body absorption rate and poor activity, and the low-sugar-based saponin has high human body absorption rate and strong activity. The average absorption rate of the polysaccharide-based saponin in the digestive system of a human body is only about 5 percent, and the polysaccharide-based saponin can be absorbed only by being metabolized into low-sugar-based saponin, but the conversion rate is extremely low, and the low-sugar-based saponin can be directly absorbed by the human body, which is the reason that the pharmacological activity of the wild ginseng is obviously higher than that of the cultivated ginseng. If the polysaccharide-based saponin is converted into the low-glycosyl saponin before being taken by a human body, the value of the ginseng is improved by several times or even tens of times.
Although the chemical synthesis industry has been rapidly developed and the level of the chemical synthesis industry is higher and higher in recent years, due to the structural complexity of ginsenoside, most of the ginsenoside cannot be prepared by the chemical synthesis route at present, and the synthesis of ginsenoside can be completed only in a laboratory, so that the industrialization cannot be realized. However, the presence of glycosidic linkages makes it possible to prepare oligosaponins from polyglucosides, and many reports on the in vitro preparation of oligosaponins from polyglucosides include, for example, the acid-base method for Rg3, Rh1 and Rh2, but the acid-base method does not allow the preparation of monosaccharide-based Compound K (20-O-. beta. -D-glucopyranosyl-20(S) -protopanaxadiol) and disaccharide-based 20(S) -F2 (3-O-. beta. -D-glucopyranosyl-20(S) -protopanaxadiol). However, except for the very individual ginsenoside which is obtained from the medicine (Rg3) and the health product batch, other saponin products cannot be directly applied to the medicine, the food and the health product at present, and the change of the saponin products into the commodity has the disadvantages of huge investment, long period, low success rate and long way. Therefore, the change of the contained components through the innovation of the process on the basis of the existing dosage form and on the premise of legal use is a very realistic matter.
Gout is known by human for thousands of years, is often seen in the wealthy class in ancient times, and is a common disease with the rapid development of social economy and the change of life style of people. Gout is associated with blood Uric Acid (Serum Uric Acid) levels. Uric acid is an intermediate product in purine metabolic process, and if a human body cannot successfully produce uricase, uric acid cannot be converted into water-soluble final product allantoin. Generally, 30% of uric acid is from food, 70% of uric acid is from human metabolism, and is normally discharged from kidney and digestive system, and if a certain link is in trouble, hyperuricemia is caused, and when the blood uric acid exceeds a certain concentration, crystals are precipitated and deposited in joints and tissues, so that a series of symptoms are caused. At present, the acute gout is mainly treated by using medicaments, such as non-steroidal anti-inflammatory drugs, glucocorticoids, colchicine and the like. For frequent authors, uric acid lowering therapy is required, including drugs inhibiting uric acid production (e.g., febuxostat, allopurinol), and drugs promoting uric acid excretion (probenecid, benzbromarone), however, some people have adverse reactions and side effects when taking these drugs. Therefore, it is very important to take natural, effective and safe components to prevent and treat hyperuricemia.
The ginseng cream is a traditional ginseng taking preparation form, and researches on the ginseng extract are more, but the ginseng cream is almost directly extracted by using an organic solvent or water, or is compounded with other traditional Chinese medicines in a surrounding way, the conversion of the components is very little, and reports on the components of Compound K and 20(S) -F2 are not found. The structural formulas of Compound K and 20(S) -F2 are as follows:
for example, the invention patent application "an antitumor pharmaceutical composition and a preparation method thereof" (publication number: CN103977060B) discloses a method for preparing ginseng extract, astragalus extract and kurarinone into extract or injection, which removes macromolecules such as polysaccharide and reduces the possibility of allergy, but only extracts and purifies natural saponin, and does not relate to the conversion of saponin components. The invention patent application of ginseng tuckahoe paste and its production technology (publication number: CN102273575B) discloses a production technology, which mixes ginseng extract, tuckahoe water extract, white sugar, honey, sodium alginate, pectin and potassium sorbate, and ginseng is concentrated after being extracted by ethanol, and does not relate to the change of saponin component. The invention patent application, Ginseng radix colla Corii Asini paste (Notification No. 101647838B), discloses a method, grinding Ginseng radix, sieving, mixing with melted colla Corii Asini, the method only pulverizes Ginseng radix into powder, and does not involve component change.
Disclosure of Invention
So far, no report of ginseng paste containing ginsenoside Compound K and 20(S) -F2 is found, and the traditional ginseng paste mainly contains polyglycosyl saponin with 3-4 glycosyl groups. Structurally, the ginsenosides Compound K and 20(S) -F2 are diol ginsenosides containing mono-and di-glycosyl groups, and thus can be obtained by hydrolyzing diol ginsenosides containing 3-4 glycosyl groups to remove 1-2 glycosyl groups, but the conversion process cannot extract the ginsenosides so as to be directly commercialized, but directly react in the state of ginseng paste. Therefore, the application provides a ginseng cream containing Compound K and 20(S) -F2 and a preparation method thereof, and the ginseng cream can be processed to contain ginsenoside Compound K and 20(S) -F2. In order to realize this process, microorganisms capable of producing glucosidase, arabinopyranosidase and arabinofuranosidase are screened from the natural world, and as specified in GB2760 Aspergillus niger, Aspergillus oryzae and Rhizopus oryzae can be used for producing enzyme preparation food additives, so that Aspergillus niger, Aspergillus oryzae or Rhizopus oryzae can be screened for producing the above enzymes and can be directly applied to product production. The transformation mechanism is shown in FIG. 1 (solid arrows are the primary transformation pathways and open arrows are the secondary transformation pathways):
in order to achieve the purpose, the technical scheme of the invention is realized as follows:
a ginseng cream with uric acid reducing activity contains Compound K and 20(S) -F2 in an amount of more than 1% of total saponins of ginseng.
A preparation method of ginseng cream with uric acid reducing activity comprises the following steps:
s1, screening strains: screening out strains which can simultaneously produce glucosidase, arabinopyranosidase and arabinofuranosidase;
s2, preparing enzyme solution: culturing the screened strains to obtain an enzyme solution containing the glucosidase, arabinopyranosidase and arabinofuranosidase obtained in the step S1;
s3, extraction: extracting ginsenoside from raw material ginseng by adopting a mode of mixed extraction of alcohol and water or total water extraction;
s4, enzyme conversion: mixing the ginsenoside extracted in the step S3 with the enzyme prepared in the step S2, reacting for 10-48 hours to convert the natural diol saponins into 20(S) -F2 and Compound K, raising the temperature to 90-150 ℃ after the reaction is finished, keeping the temperature for 5-40 min, and lowering the temperature to 50-70 ℃ after the enzyme is inactivated;
s5, paste preparation: mixing the water soluble components extracted in step S3 with the ginseng extract containing Compound K and 20(S) -F2 after enzyme deactivation of S4, stirring uniformly, and concentrating to above 55Brix to obtain the product.
Further, the strain of step S1 is obtained by screening from nature, and the specific steps are as follows:
(1) inoculating bacteria from soil containing dead ginseng tissue in the planting field of Panax plants to a plate culture medium for culturing;
(2) selecting mould of a single bacterial colony;
(3) culturing in a ginseng culture medium of single colony bacteria,
(4) detecting the enzyme activity of Compound K and 20(S) -F2 generated by hydrolyzing the ginsenoside PPD mixture, and screening out the bacteria which can simultaneously generate glucosidase, arabinopyranosidase and arabinofuranosidase and have the best enzyme activity.
Further, the strain in step S1 is aspergillus niger, aspergillus oryzae, or rhizopus oryzae.
Further, the enzyme solution of step S2 is prepared by liquid fermentation culture, and the specific steps are as follows: preparing a liquid culture medium, inoculating bacteria, fermenting and culturing, shaking a bottle or introducing sterile air in the culture process, culturing at 28-35 ℃ for 7-14 days, centrifuging or filtering to remove thalli, wherein the supernatant is an enzyme solution which can be directly used for enzyme conversion reaction or used for reaction after enrichment and concentration.
Further, the enzyme solution of step S2 is prepared by solid fermentation culture, and the specific steps are as follows: inoculating the screened bacteria into a sterilized ginseng culture medium, culturing at 28-33 ℃ for 4-8 days, soaking in 2-4 times of buffer solution with the pH value of 4.5-6.5, and filtering, wherein the supernatant is an enzyme solution which can be directly used for enzyme conversion reaction or used for reaction after enrichment and concentration.
Further, when liquid fermentation culture is adopted, the liquid culture medium in the step S2 is an extract obtained by extracting a solid fermentation culture medium with 5-10 times of water at 95-100 ℃ for 1-3 hours, adjusting the content of soluble solids to 5-12 Brix with water, and sterilizing the extract for use.
Further, the plate medium in step S1 is a chai' S medium to which 5% of an aqueous extract of ginseng is added.
Further, when solid fermentation culture is adopted, the ginseng culture medium in the step S2 is prepared by uniformly stirring 1-30% of roots, stems, leaves or pollen of the ginseng plant or an extract thereof, 0-30% of wheat straw, 0-30% of wheat bran, 0-15% of sophora flower, 0-15% of soybean meal, 0-5% of rice hull and 40-60% of water, and sterilizing.
Further, the enzyme solution prepared in the step S2 is precipitated with one of ammonium sulfate and ethanol, and a precipitate fraction having a high enzyme content is collected and dissolved in a buffer solution having an amount of 5 to 15% of the original enzyme solution. The purpose of enrichment and concentration is achieved.
Further, the buffer solution is one of 0.001-0.1M acetic acid buffer solution, phosphoric acid buffer solution or citric acid buffer solution.
Further, the step of mixing and extracting the alcohol and the water in the step S3 is as follows:
(1) extracting ginsenoside in raw material ginseng by using 70-95% edible alcohol, extracting for 3-8 h at 40-70 ℃, filtering, repeating for 2-3 times, and combining extracting solutions;
(2) extracting with 40-50% edible alcohol at 40-70 deg.C for 4-8 h 1-2 times, filtering, mixing the filtrate with the above extractive solution, concentrating until the solid content is 50-70 Brix,
(3) adding water which is 1-5 times of the raw material parameter, continuously concentrating, repeating for 1-3 times to reduce the ethanol content, and obtaining a component rich in natural ginsenoside;
(4) and heating the filter residue extracted in the last step to 90-100 ℃ by using 5-10 times of water, extracting for 5-8 h, repeating for 2-3 times, combining the extracting solutions and concentrating.
Further, the step of extracting the whole water in the step S3 is:
(1) mixing 6-12 times of raw material parameters of water with the raw material ginseng, extracting for 1-8 hours at 70-100 ℃, repeating for 2-3 times to obtain a mixed solution containing natural ginsenoside, and rapidly extracting a component rich in natural ginsenoside under the condition of not activating or less activating a ginseng autocatalytic substance;
(2) extracting the mixed solution for 6-12 hours at 90-100 ℃ by using 6-10 times of water, repeating for 1-2 times, and extracting the polysaccharide-rich component with low saponin content.
The invention also provides application of the ginseng cream in preparing a medicine for reducing uric acid activity.
Compared with the prior art, the invention has the following advantages:
the ginseng cream containing 20(S) -F2 and Compound K is prepared for the first time by the preparation method of the ginseng cream, contains ginseng polysaccharide and other components, and has the effect of reducing uric acid. The product can be directly applied to food and health care products, is suitable for industrialization and has strong applicability.
Drawings
FIG. 1 is a diagram showing the mechanism of conversion of saponins of the present invention to Compound K and 20(S) -F2;
FIG. 2 is an HPLC chromatogram of natural ginsenoside;
FIG. 3 is HPLC detection spectrum of ginseng cream;
FIG. 4 is an HPLC detection profile of ginseng cream;
FIG. 5 is HPLC detection profile of ginseng cream;
Detailed Description
Unless defined otherwise, technical terms used in the following examples have the same meanings as commonly understood by one of ordinary skill in the art to which the present invention belongs. The test reagents used in the following examples, unless otherwise specified, are all conventional biochemical reagents; the experimental methods are conventional methods unless otherwise specified.
The invention will be described in detail with reference to the following examples.
Experimental example 1: HPLC method for detecting ginsenoside
The instrument comprises the following steps: waters e2996 liquid chromatograph: 2998 with photodiode array (PDA) detector;
a workstation: empower 3;
a chromatographic column: a C18 column (5 μm,
)；
mobile phase: acetonitrile (a), water (B);
column temperature: 35 ℃;
flow rate: 0.5-1 mL/min;
detection wavelength: 203 nm;
sample introduction amount: 10 mu L of the solution;
total saponin elution procedure: 0-20 min, 20% A; 20-31 min, 20% -32% A; 31-40 min, 32-43% A; 40-70 min, 43-100% A;
main saponin elution procedure: 0-35 min, 19% A; 35-55 min, 19-29% of A; 55-65 min, 29-40% A; 65-95 min, 40-100% A.
Detection of natural ginsenoside: taking 2g of ginseng sample, adding 50mL of methanol for reflux extraction for 6h, extracting for 3 times, combining extracting solutions, concentrating to remove the methanol, adding 30mL of water for dissolution, degreasing for 2 times by using 20mL of petroleum ether, extracting for 3 times by using n-butanol, combining n-butanol layers, concentrating under reduced pressure to remove a solvent, dissolving saponin by using 10mL of ethanol, mixing with 90mL of the mixture, putting the mixture on a pretreated macroporous adsorption resin column, washing with water to remove water-soluble components, eluting by using 95% of ethanol, evaporating to dryness and dissolving in methanol to detect components, and obtaining the result shown in figure 2, wherein the natural ginsenoside only contains Rg1, Re, Rf, Rb1, Rc, Rb2 and Rd and does not contain Compound K and 20(S) -F2.
Example 1
Screening bacteria: preparing a plate culture medium: extracting 200g of ginseng fragments with 1L of water at 95-100 ℃ for 3h each time, extracting for 3 times, combining water extracts, concentrating until the content of soluble solids is 5Brix, taking 1L of ginseng extract, adding 3g of sodium nitrate, 1g of dipotassium hydrogen phosphate, 0.5g of magnesium sulfate, 0.5g of potassium chloride, 0.01g of ferrous sulfate, 30g of cane sugar and 20g of agar, heating for dissolving, sterilizing, pouring into a flat plate, and preparing into a flat plate culture medium.
Collecting soil containing ginseng dead tissue (collection time of 7 months in 2018, collection place of northern wasteland farm in Sunwu county, Black river, Black dragon Jiang province) 5g from Ginseng radix planting field, adding into 45mL sterilized physiological saline, and repeatedly shaking for 10min to obtain 10-1The microbial dilution of (4); measuring 1mL10-1The microbial diluent is mixed with 9mL of sterilized normal saline to prepare 10-2Microbial dilutions made in this order 10-3、10-4、10-5、10-6And (4) diluting the solution. 0.1mL of each dilution was applied to the plate and incubated at 30 ℃. After 4 days of culture, 6 Aspergillus niger colonies were selected by microscopic observation and cultured in ginseng medium. The ginseng culture medium contains 10g of ginseng powder, 20g of wheat bran, 5g of rice hulls, 5g of soybean meal and 40g of water, and is uniformly stirred and sterilized at 120 ℃ for 20 min. Culturing at 28 deg.C for 7 days, soaking in 300mL 0.02M phosphate buffer solution (pH6.0), filtering for 2 hr, separating, adding 5% ginsenoside PPD (containing Rb1, Rb2, Rc, and Rd) into the supernatant, reacting at 40 deg.C for 24 hr, detecting enzyme activity by TLC, and selecting the bacteria with the highest conversion abilityKHAN008。
Culturing and extracting enzyme: preparing 10kg of ginseng culture medium, inoculating KHAN008 bacteria, culturing for 8 days, extracting 300L of enzyme solution, adding 9kg of ammonium sulfate into the enzyme solution, precipitating for 3h, centrifuging to remove the precipitate, adding 7.5kg of ammonium sulfate, dissolving the precipitate containing the target enzyme in 3L0.02M pH6.0 phosphate buffer solution, and keeping.
Extraction: taking 10kg of dry ginseng, beating into 10-20 meshes of fragments, putting into a stirring kettle, adding 100L of 70% edible ethanol solution, soaking for 2h, heating to 50 ℃, stirring and extracting for 6h, filtering and separating solid and liquid, continuously extracting filter residue for 2 times by using 80L of 70% edible ethanol, filtering, extracting the filter residue for 1 time at 60 ℃ by using 80L of 50% edible ethanol solution, combining the extracting solutions of four times, concentrating until the content of soluble solid matters is 15Brix, adding 20kg of water, and continuously concentrating until the content of the soluble solid matters is 20 Brix.
Extracting a polysaccharide-rich component: heating the residue with 60kg water to slightly boil for 4 hr, and repeating for 2 times to obtain polysaccharide-rich component
And (3) transformation: mixing the enzyme solution 1L with the saponin-rich component, reacting at 40 deg.C for 30h, and detecting to obtain the final product containing saponins Compound K and 20(S) -F2, which accounts for 1% of total saponins of Ginseng radix. Heating the reaction solution to 110 deg.C, maintaining for 30min, inactivating enzyme, sterilizing, cooling to 60 deg.C, adding the components rich in polysaccharide, stirring for 20min, and concentrating under reduced pressure to solid content of 65Brix to obtain high activity Ginseng radix extract rich in Compound K and 20(S) -F2.
Taking 2g of ginseng paste sample, adding 18g of water for dissolving, degreasing for 2 times by using 20mL of petroleum ether, extracting for 3 times by using n-butanol, 10mL of each time, combining n-butanol layers, concentrating under reduced pressure to remove a solvent, dissolving saponin by using 10mL of ethanol, mixing with 90mL of the solvent, putting the mixture on a pretreated macroporous adsorption resin column, washing by using water to remove water-soluble components, eluting by using 95% of ethanol, dissolving the mixture in methanol after evaporating to dryness, and detecting the components by using the method in the experimental example 1, wherein the content of Compound K and 20(S) -F2 is 57.8 percent as shown in figure 3.
Example 2
Preparing a ginseng culture medium: according to the extract of the stems and leaves of the American ginseng: wheat bran: rice hull: and (3) flos sophorae: the culture medium was prepared in a ratio of 25:20:2:3:45 with water (9.5 kg), and sterilized at 110 ℃ for 35min with stirring. The KHAN008 strain isolated in example 1 was inoculated, cultured at 30 ℃ for 8 days, soaked in 25L of phosphate buffer (pH5.50.005M) for 2 hours, centrifuged to remove solid impurities, and the enzyme solution was used.
Extraction: weighing 20kg of American ginseng, beating into 10-20 meshes of fragments, putting into a stirring kettle, adding 150L of water, soaking for 2h, heating to 80 ℃, stirring and extracting for 3h, filtering and separating solid and liquid, continuously extracting filter residue with 120L of water for 2 times, combining extracting solutions, and concentrating until the content of soluble solid is 20Brix, namely the saponin-rich component. Extracting the residue with 100L water at 95 deg.C for 2 times, each for 6 hr, repeating for 2 times, and mixing to obtain polysaccharide-rich fraction.
And (3) transformation: mixing the prepared 21.5L enzyme solution with the component rich in saponin, reacting at 45 deg.C for 36h, heating to 95 deg.C, maintaining for 30min to inactivate enzyme, cooling to 60 deg.C, adding the component rich in polysaccharide, stirring for 20min, and concentrating under reduced pressure to solid content of 70Brix to obtain the high-activity radix Panacis Quinquefolii extract rich in Compound K and 20(S) -F2. As a result of HPLC, the sum of the contents of Compound K and 20(S) -F2 in the paste was about 60.22%, as shown in FIG. 4.
Example 3
The strain CICC40338 is purchased from China industrial microorganism strain preservation management center and is used for producing enzyme by fermentation. The culture medium is prepared from notoginseng and ginseng flower: wheat straw: mixing wheat bran at a ratio of 2:25:25, adding 10.4kg of the mixture into 300 parts of water, heating to slightly boil for extraction for 1h, filtering to remove solid impurities, adding water to adjust the content of soluble solids to 10Brix, pouring about 35L of the mixture into a fermentation tank, heating to 125 ℃, sterilizing for 18min, cooling to 35 ℃, inoculating a fermentation strain CICC40338, fermenting and culturing at 28 ℃ for 12d, introducing sterile air, centrifuging to remove thalli, adding ethanol with the volume of 4 times of the volume of fermentation liquid to precipitate zymoprotein, centrifuging to collect enzyme precipitate, and dissolving in 3L of acetic acid buffer solution with the pH value of 5.00.01M to prepare enzyme solution.
Extraction: taking 10kg of dry ginseng, beating into 10-20 meshes of fragments, putting into a stirring kettle, adding 80L of 80% edible ethanol solution, soaking for 2h, heating to 55 ℃, stirring and extracting for 8h, filtering and separating solid and liquid, continuously extracting the filter residue for 2 times by using 60L of 80% edible ethanol, filtering, extracting the filter residue for 2 times at 65 ℃ by using 50L of 50% edible ethanol solution, combining the five times of extracting solutions, concentrating to the soluble solid content of 12Brix, adding 20kg of water, continuously concentrating to the soluble solid content of 15Brix, adding 20kg of water, and concentrating to the soluble solid content of 20 Brix. The saponin-rich component is obtained.
Heating the residue with 50kg water to slightly boil, extracting for 5 hr, repeating for 2 times, and removing residue to obtain polysaccharide-rich component.
And (3) transformation: mixing the obtained 3L enzyme solution with saponin-rich component, reacting at 42 deg.C for 24h, heating to 130 deg.C, maintaining for 10min to inactivate enzyme, cooling to 60 deg.C, adding polysaccharide-rich component, and concentrating under reduced pressure to 70Brix to obtain high activity Ginseng radix extract rich in Compound K and 20(S) -F2. As a result of HPLC, the sum of the contents of Compound K and 20(S) -F2 in the paste was about 12.44%, as shown in FIG. 5.
Experimental example 2
The influence of the ginseng cream on uric acid is examined by taking a hyperuricemia mouse as a model. Experimental materials: 32 Kunming mice weighing 18-22g, hermaphrodite, allopurinol, ginseng cream in example 1 herein, general ginseng cream (ginseng cream without Compound K and 20(S) -F2 made by conventional method).
The experimental method comprises the following steps: the 40 mice were randomly divided into 4 groups including a blank group, a model group, an allocatal group, a ginseng cream group, and a general ginseng cream group. The administration volume is 0.2ml, the allopurinol dosage is 0.5mg/10g, and the ginseng ointment is diluted by equal volume of water and then is irrigated. And (3) filling 0.5% of sodium carboxymethylcellulose (CMC) into the blank group, continuously filling for 5 days, then molding, injecting physiological saline into the blank group, and injecting 3mg/10g of oteracil potassium salt into the other groups. 1h after injection of each group of mice, taking eyeballs, taking blood, separating serum, and measuring the level of hematuria acid; the mouse liver was mixed with 9 times of physiological saline and homogenized, and then the xanthine oxidase was measured. Uric acid and xanthine oxidase were measured using an ELISA kit, and the results are shown in Table 1:
TABLE 1 Effect of Ginseng extract on uric acid
Note: p <0.01 compared to model group; , #, p <0.01, compared to blank
The experimental results show that the model group and the blank group have obvious difference, which indicates that the molding is successful. The ginseng ointment group can obviously reduce the uric acid level and xanthine oxidase of a model mouse, and the action effect is close to that of the medicine group and is far higher than that of the common ginseng ointment. The results show that the ginseng cream in the application has a remarkable effect of reducing blood uric acid.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and should not be taken as limiting the invention, so that any modifications, equivalents, improvements and the like, which are within the spirit and principle of the present invention, should be included in the scope of the present invention.
Claims (10)
1. The ginseng ointment with the function of reducing the activity of uric acid is characterized in that: the content of Compound K and 20(S) -F2 in the ginseng cream accounts for more than 1% of the proportion of the total saponins of ginseng.
2. The preparation method of ginseng cream as claimed in claim 1, which is characterized in that: the method comprises the following steps:
s1, screening strains: screening out strains which can simultaneously produce glucosidase, arabinopyranosidase and arabinofuranosidase;
s2, preparing enzyme solution: culturing the screened strains to obtain an enzyme solution containing the glucosidase, arabinopyranosidase and arabinofuranosidase obtained in the step S1;
s3, extraction: extracting ginsenoside from raw material ginseng by adopting a mode of mixed extraction of alcohol and water or total water extraction;
s4, enzyme conversion: mixing the ginsenoside extracted in the step S3 with the enzyme prepared in the step S2, reacting for 10-48 hours to convert the natural diol saponins into 20(S) -F2 and Compound K, raising the temperature to 90-150 ℃ after the reaction is finished, keeping the temperature for 5-40 min, and lowering the temperature to 50-70 ℃ after the enzyme is inactivated;
s5, paste preparation: mixing the water soluble components extracted in step S3 with the ginseng extract containing Compound K and 20(S) -F2 after enzyme deactivation of S4, stirring uniformly, and concentrating to above 55Brix to obtain the product.
3. The preparation method of ginseng cream as claimed in claim 1, which is characterized in that: the strain of step S1 is obtained by screening from nature, and the specific steps are as follows:
(1) inoculating bacteria from soil containing dead ginseng tissue in the planting field of Panax plants to a plate culture medium for culturing;
(2) selecting mould of a single bacterial colony;
(3) culturing in a ginseng culture medium of single colony bacteria,
(4) detecting the enzyme activity of Compound K and 20(S) -F2 generated by hydrolyzing the ginsenoside PPD mixture, and screening out the bacteria which can simultaneously generate glucosidase, arabinopyranosidase and arabinofuranosidase and have the best enzyme activity.
4. The preparation method of ginseng cream as claimed in claim 1, which is characterized in that: the strain of step S1 is Aspergillus niger, Aspergillus oryzae or Rhizopus oryzae.
5. The preparation method of ginseng cream as claimed in claim 1, which is characterized in that: the enzyme liquid of the step S2 is prepared by liquid fermentation culture, and the specific steps are as follows: and (3) inoculating bacteria to perform fermentation culture after preparing a liquid culture medium, shaking the bottle or introducing sterile air in the culture process, culturing for 7-14 days at 28-35 ℃, centrifuging or filtering to remove bacteria, wherein the supernatant is an enzyme solution.
6. The preparation method of ginseng cream as claimed in claim 1, which is characterized in that: the enzyme liquid of the step S2 is prepared by solid fermentation culture, and the specific steps are as follows: inoculating the screened bacteria into a sterilized ginseng culture medium, culturing at 28-33 ℃ for 4-8 days, soaking in 2-4 times of buffer solution with the pH value of 4.5-6.5, and filtering to obtain supernatant as enzyme solution.
7. The preparation method of ginseng cream as claimed in claim 1, which is characterized in that: when liquid fermentation culture is adopted, the liquid culture medium in the step S2 is an extract obtained by extracting a solid fermentation culture medium with 5-10 times of water at 95-100 ℃ for 1-3 h, adjusting the content of soluble solids to 5-12 Brix by using water, and sterilizing for use; preferably, the plate medium in step S1 is a chai' S medium to which 5% of an aqueous extract of ginseng is added; preferably, when solid fermentation culture is adopted, the ginseng culture medium in the step S2 is prepared by uniformly stirring 1-30% of roots, stems, leaves or pollen of the panax plants or extracts thereof, 0-30% of wheat straw, 0-30% of wheat bran, 0-15% of flos sophorae, 0-15% of soybean meal, 0-5% of rice hulls and 40-60% of water, and sterilizing.
8. The preparation method of ginseng cream as claimed in claim 1, which is characterized in that: and precipitating the enzyme solution prepared in the step S2 by using one of ammonium sulfate and ethanol, collecting a precipitate part with high enzyme content, and dissolving the precipitate part by using a buffer solution with 5-15% of the original enzyme solution. Preferably, the buffer solution is one of 0.001-0.1M acetic acid buffer solution, phosphate buffer solution or citric acid buffer solution.
9. The preparation method of ginseng cream as claimed in claim 1, which is characterized in that: the step of alcohol-water mixed extraction in the step S3 is as follows:
(1) extracting ginsenoside in raw material ginseng by using 70-95% edible alcohol, extracting for 3-8 h at 40-70 ℃, filtering, repeating for 2-3 times, and combining extracting solutions;
(2) extracting with 40-50% edible alcohol at 40-70 deg.C for 4-8 h 1-2 times, filtering, mixing the filtrate with the above extractive solution, concentrating until the solid content is 50-70 Brix,
(3) adding water which is 1-5 times of the raw material parameter, continuously concentrating, repeating for 1-3 times to reduce the ethanol content, and obtaining a component rich in natural ginsenoside;
(4) and heating the filter residue extracted in the last step to 90-100 ℃ by using 5-10 times of water, extracting for 5-8 h, repeating for 2-3 times, combining the extracting solutions and concentrating.
10. The preparation method of ginseng cream as claimed in claim 1, which is characterized in that: the step of total water extraction in the step S3 is as follows:
(1) mixing 6-12 times of raw material parameters of water with the raw material parameters, extracting for 1-8 h at 70-100 ℃, and repeating for 2-3 times to obtain a mixed solution containing natural ginsenoside;
(2) extracting the mixed solution for 6-12 hours at 90-100 ℃ by using 6-10 times of water, repeating for 1-2 times, and extracting the polysaccharide-rich component with low saponin content.
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| CN107308195A (en) * | 2017-06-30 | 2017-11-03 | 肖永坤 | A kind of method that high activity ginsenoside is prepared by solid dynamic fermentation technology |
| CN108367037A (en) * | 2014-12-15 | 2018-08-03 | 庆星大学校产学协力团 | Contain composition of the ginsenoside as active ingredient |
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| CN108367037A (en) * | 2014-12-15 | 2018-08-03 | 庆星大学校产学协力团 | Contain composition of the ginsenoside as active ingredient |
| CN107308195A (en) * | 2017-06-30 | 2017-11-03 | 肖永坤 | A kind of method that high activity ginsenoside is prepared by solid dynamic fermentation technology |
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