A composition and its preparation method and application in preparing products with caries prevention effect

Abstract

The present invention relates to the field of biomedicine technology, and specifically discloses a composition and a preparation method thereof and an application thereof in preparing a product having a dental caries prevention effect. The composition is prepared using natural plants Inula japonica and Viola odorata as raw materials; the preparation method specifically comprises the following steps: taking the whole herb of Inula japonica and Viola odorata, extracting them with an ethanol aqueous solution, concentrating and drying the extract after the extraction to obtain an Inula japonica-Viola odorata alcohol extract; taking the Inula japonica-Viola odorata alcohol extract to obtain the composition. Studies have shown that the composition has an anti-Streptococcus mutans effect; therefore, using it as an active ingredient to prepare medicines, foods or daily chemicals having a dental caries prevention effect has important application value.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technology, and in particular to a composition and a preparation method thereof, and application of the composition in preparing a product having a caries prevention effect. Background Art

[0002] Dental caries is a bacterial infection disease that occurs in the hard tissues of teeth and has the highest incidence rate among oral diseases. Streptococcus mutans is the main pathogen of dental caries. It adheres to teeth layer by layer and forms a "biofilm" on the teeth. It consumes sugars to produce acid, which corrodes the enamel layer of teeth to cause cavities and then forms dental caries. Therefore, developing active ingredients that can resist Streptococcus mutans is a research direction for developing products to prevent dental caries.

[0003] Inula japonica, scientific name Inulajaponica Thunb., is a plant of the Asteraceae family and the genus Inula; its dried capitula can be used as a traditional Chinese medicine, which has the effects of lowering qi, eliminating phlegm, promoting diuresis, and stopping vomiting. Viola odorata, scientific name Viola odorata L., is a perennial herb of the genus Viola of the Violaceae family, which has the effects of dispelling wind and heat, relieving cough and expectoration, calming, and stopping diarrhea. However, there is no research report on the anti-streptococcal mutans effect of Inula japonica and Viola odorata in the prior art; therefore, using Inula japonica and Viola odorata as raw materials to develop active ingredients with anti-streptococcal mutans effect has important application value for the prevention of dental caries. Summary of the invention

[0004] In order to overcome at least one of the technical problems existing in the prior art, the present invention first provides a composition and a preparation method thereof and an application thereof in preparing a product having a caries prevention effect.

[0005] The invention firstly provides a preparation method of the composition, which is prepared by taking natural plants Inula japonica and Viola odorifera as raw materials.

[0006] The invention provides a method for preparing a new composition; studies have shown that the composition prepared with natural plants Inula japonica and Viola odorata as raw materials has an anti-streptococcal effect; therefore, using them as active ingredients to prepare products with a caries prevention effect has important application value.

[0007] Preferably, the method for preparing the composition comprises the following steps:

[0008] Take the whole herb of Inula japonica and the whole herb of Viola odorata, extract them with ethanol aqueous solution, and after the extraction, take the extract and concentrate and dry it to obtain the Inula japonica-Viola odorata ethanol extract;

[0009] The composition is obtained by taking the ethanol extract of Inula japonica-Viola odorata.

[0010] Preferably, the weight ratio of the whole herb of Inula japonica to the whole herb of Viola odorata is 1-3:1-3.

[0011] Most preferably, the weight ratio of the whole herb of Inula japonica to the whole herb of Viola odorata is 1:1.

[0012] Preferably, the ethanol aqueous solution refers to an ethanol aqueous solution with an ethanol volume fraction of 50% to 95%.

[0013] Preferably, the usage ratio of the total weight of the whole herb of Inula japonica and the whole herb of Viola odorata to the ethanol aqueous solution is 1 kg: 5-15 L.

[0014] Most preferably, the ratio of the total weight of the whole herb of Inula japonica and the whole herb of Viola odorata to the ethanol aqueous solution is 1 kg:12 L.

[0015] Preferably, the preparation method further comprises the following steps:

[0016] The alcohol extract of Inula japonica-Viola odorata is placed on a macroporous resin column, and impurities are first eluted with an ethanol aqueous solution having a volume fraction of 53-55%, and then eluted with an ethanol aqueous solution having a volume fraction of 66-68%, and the eluate eluted with the ethanol aqueous solution having a volume fraction of 66-68% is collected, and the eluate is concentrated and dried to obtain the Inula japonica-Viola odorata macroporous resin elution fraction;

[0017] The composition is obtained by taking the elution part of the Inula japonica-Viola ol macroporous resin.

[0018] Most preferably, the preparation method further comprises the following steps:

[0019] The alcohol extract of Inula japonica-Viola odorata is placed on a macroporous resin column, and impurities are first eluted with a 54% ethanol aqueous solution by volume; then eluted with a 67% ethanol aqueous solution by volume, and the eluate eluted with the 67% ethanol aqueous solution by volume is collected, concentrated and dried to obtain the Inula japonica-Viola odorata macroporous resin elution fraction;

[0020] The composition is obtained by taking the elution part of the Inula japonica-Viola ol macroporous resin.

[0021] The inventors found in further research that in the process of preparing the composition using natural plants Inula japonica and Viola odorata as raw materials, the preparation step is very critical; the study shows that on the basis of the Inula japonica-Viola odorata ethanol extract, the Inula japonica-Viola odorata macroporous resin elution fraction prepared by the above method of the present invention has a much higher anti-mutan Streptococcus than the Inula japonica-Viola odorata ethanol extract. Therefore, the composition using the Inula japonica-Viola odorata macroporous resin elution fraction as the composition has a much higher anti-mutan Streptococcus than the composition using the Inula japonica-Viola odorata ethanol extract as the composition.

[0022] In addition, the inventors need to emphasize that in the elution process of the macroporous resin column, the elution conditions of the macroporous resin column are very critical; on the basis of the Inula japonica-Viola odorata ethanol extract, only the Inula japonica-Viola odorata macroporous resin elution fraction prepared by the above-mentioned macroporous resin elution conditions of the present invention can have an anti-Streptococcus mutans effect significantly higher than that of the Inula japonica-Viola odorata ethanol extract; however, the Inula japonica-Viola odorata macroporous resin elution fraction prepared by other macroporous resin elution conditions cannot have an anti-Streptococcus mutans effect significantly higher than that of the Inula japonica-Viola odorata ethanol extract.

[0023] Preferably, the macroporous resin column is an AB-8 macroporous resin column.

[0024] Preferably, the preparation method further comprises the following steps:

[0025] The eluted portion of the Inula japonica-Viola odorata macroporous resin is further placed on a silica gel column, and impurities are first eluted with a mixed solvent consisting of cyclohexane and ethyl acetate in a volume ratio of 83-85:17-15; then eluted with a mixed solvent consisting of cyclohexane and ethyl acetate in a volume ratio of 76-78:24-22, and the eluate eluted with the mixed solvent consisting of cyclohexane and ethyl acetate in a volume ratio of 76-78:24-22 is collected, and the Inula japonica-Viola odorata silica gel eluted portion is obtained after concentrating and drying;

[0026] The composition is obtained by taking the silica gel elution part of Inula japonica-Viola odorata.

[0027] Most preferably, the preparation method further comprises the following steps:

[0028] The Inula-Viola macroporous resin elution part is further placed on a silica gel column, and first eluted with a mixed solvent consisting of cyclohexane and ethyl acetate in a volume ratio of 84:16 to remove impurities; then eluted with a mixed solvent consisting of cyclohexane and ethyl acetate in a volume ratio of 77:23, and the eluate eluted with the mixed solvent consisting of cyclohexane and ethyl acetate in a volume ratio of 77:23 is collected, and concentrated and dried to obtain the Inula-Viola silica gel elution part;

[0029] The composition is obtained by taking the silica gel elution part of Inula japonica-Viola odorata.

[0030] The inventors found in further research that, based on the elution part of the Inula-Viola macroporous resin, the silica gel elution part of the Inula-Viola prepared by the above-mentioned silica gel column elution conditions of the present invention has a much higher anti-mutan Streptococcus than the macroporous resin elution part of the Inula-Viola. Therefore, the anti-mutan Streptococcus effect of the composition using the silica gel elution part of the Inula-Viola is much higher than the composition using the macroporous resin elution part of the Inula-Viola; so that the composition has a more excellent anti-mutan Streptococcus effect.

[0031] Similarly, in the elution process of the silica gel column, the elution conditions of the silica gel column are very critical; on the basis of the elution site of the Inula japonica-Sweetia odorata macroporous resin, only the Inula japonica-Sweetia odorata silica gel elution site prepared by the above-mentioned silica gel column elution conditions of the present invention can further significantly increase the anti-Streptococcus mutans effect compared with the Inula japonica-Sweetia odorata macroporous resin elution site; however, the Inula japonica-Sweetia odorata silica gel elution site prepared by other silica gel column elution conditions cannot further significantly increase the anti-Streptococcus mutans effect compared with the Inula japonica-Sweetia odorata macroporous resin elution site.

[0032] Preferably, the composition is a composition having an anti-Streptococcus mutans effect.

[0033] The present invention also provides a composition prepared by the above preparation method.

[0034] The present invention also provides an application of the composition in preparing a product having an anti-mutan streptococcus effect.

[0035] The present invention also provides an application of the composition in preparing a product having the function of preventing dental caries.

[0036] Preferably, the product is medicine, food or daily chemical product.

[0037] More preferably, the food is chewing gum.

[0038] More preferably, the daily chemical products include toothpaste, tooth powder and mouthwash.

[0039] Beneficial effects: The present invention provides a new composition prepared with natural plants Inula japonica and Viola odorata as raw materials; studies have shown that the composition has an anti-Streptococcus mutans effect; therefore, using it as an active ingredient in the preparation of medicines, foods or daily chemicals with the effect of preventing dental caries has important application value. DETAILED DESCRIPTION

[0040] The present invention is further explained below in conjunction with specific examples, but the examples do not limit the present invention in any form.

[0041] Example 1 Preparation of the composition

[0042] The dried whole herb of Inula japonica and the dried whole herb of Viola odorata are mixed in a weight ratio of 1:1, and an ethanol aqueous solution with a volume fraction of 95% is added, and reflux extraction is performed at 78° C. for 1 hour; after the extraction is completed, the extract is separated, and the extract is concentrated and dried to obtain an Inula japonica-Viola odorata ethanol extract; wherein the total weight ratio of the dried whole herb of Inula japonica and the dried whole herb of Viola odorata to the ethanol aqueous solution is 1kg:12L;

[0043] The composition is obtained by taking the ethanol extract of Inula japonica-Viola odorata.

[0044] Example 2 Preparation of the composition

[0045] (1) Mixing dried whole herb of Inula japonica and dried whole herb of Viola odorata in a weight ratio of 1:1, adding 95% ethanol aqueous solution by volume, and performing reflux extraction at 78° C. for 1 h; separating the extract after the extraction, concentrating and drying the extract to obtain an Inula japonica-Viola odorata ethanol extract; wherein the total weight ratio of dried whole herb of Inula japonica and dried whole herb of Viola odorata to the ethanol aqueous solution is 1 kg:12 L;

[0046] (2) The alcohol extract of Inula japonica-Viola odorata is loaded onto a macroporous resin column (the macroporous resin column is filled with AB-8 type macroporous resin; the weight of the AB-8 type macroporous resin is 40 times the weight of the alcohol extract of Inula japonica-Viola odorata), and the impurities are first eluted with 5 times the column volume of 54% ethanol aqueous solution; then, the impurities are eluted with 5 times the column volume of 67% ethanol aqueous solution, and the eluate eluted with 67% ethanol aqueous solution is collected, and the eluate is concentrated and dried to obtain the elution fraction of the Inula japonica-Viola odorata macroporous resin;

[0047] The composition is obtained by taking the elution part of the Inula japonica-Viola ol macroporous resin.

[0048] Example 3 Preparation of the composition

[0049] (1) Mixing dried whole herb of Inula japonica and dried whole herb of Viola odorata in a weight ratio of 1:1, adding 95% ethanol aqueous solution by volume, and performing reflux extraction at 78° C. for 1 h; separating the extract after the extraction, concentrating and drying the extract to obtain an Inula japonica-Viola odorata ethanol extract; wherein the total weight ratio of dried whole herb of Inula japonica and dried whole herb of Viola odorata to the ethanol aqueous solution is 1 kg:12 L;

[0050] (2) The alcohol extract of Inula japonica-Viola odorata is loaded onto a macroporous resin column (the macroporous resin column is filled with AB-8 type macroporous resin; the weight of the AB-8 type macroporous resin is 40 times the weight of the alcohol extract of Inula japonica-Viola odorata), and the impurities are first eluted with 5 times the column volume of 54% ethanol aqueous solution; then, the impurities are eluted with 5 times the column volume of 67% ethanol aqueous solution, and the eluate eluted with 67% ethanol aqueous solution is collected, and the eluate is concentrated and dried to obtain the elution fraction of the Inula japonica-Viola odorata macroporous resin;

[0051] (3) The eluted portion of the Inula japonica-Sweet Viola macroporous resin is further subjected to a silica gel column (the silica gel column is filled with 200-300 mesh silica gel; the weight of the silica gel is 30 times the weight of the eluted portion of the Inula japonica-Sweet Viola macroporous resin), and impurities are first eluted with a mixed solvent consisting of cyclohexane and ethyl acetate in a volume ratio of 84:16; then, the mixture is eluted with a mixed solvent consisting of cyclohexane and ethyl acetate in a volume ratio of 77:23, and the eluate eluted with the mixed solvent consisting of cyclohexane and ethyl acetate in a volume ratio of 77:23 is collected, and the Inula japonica-Sweet Viola silica gel eluted portion is obtained after concentration and drying;

[0052] The composition is obtained by taking the silica gel elution part of Inula japonica-Viola odorata.

[0053] Comparative Example 1 Preparation of the composition

[0054] (1) Mixing dried whole herb of Inula japonica and dried whole herb of Viola odorata in a weight ratio of 1:1, adding 95% ethanol aqueous solution by volume, and performing reflux extraction at 78° C. for 1 h; separating the extract after the extraction, concentrating and drying the extract to obtain an Inula japonica-Viola odorata ethanol extract; wherein the total weight ratio of dried whole herb of Inula japonica and dried whole herb of Viola odorata to the ethanol aqueous solution is 1 kg:12 L;

[0055] (2) The alcohol extract of Inula japonica-Viola odorata is loaded onto a macroporous resin column (the macroporous resin column is filled with AB-8 type macroporous resin; the weight of AB-8 type macroporous resin is 40 times the weight of the alcohol extract of Inula japonica-Viola odorata), and impurities are first eluted with 5 times the column volume of 30% ethanol aqueous solution; then, eluted with 5 times the column volume of 50% ethanol aqueous solution, and the eluate eluted with 50% ethanol aqueous solution is collected, and the eluate is concentrated and dried to obtain the elution fraction of the Inula japonica-Viola odorata macroporous resin;

[0056] The composition is obtained by taking the elution part of the Inula japonica-Viola ol macroporous resin.

[0057] The difference between Comparative Example 1 and Example 2 is that the elution conditions of the macroporous resin column are different.

[0058] Comparative Example 2 Preparation of the composition

[0059] (1) Mixing dried whole herb of Inula japonica and dried whole herb of Viola odorata in a weight ratio of 1:1, adding 95% ethanol aqueous solution by volume, and performing reflux extraction at 78° C. for 1 h; separating the extract after the extraction, concentrating and drying the extract to obtain an Inula japonica-Viola odorata ethanol extract; wherein the total weight ratio of dried whole herb of Inula japonica and dried whole herb of Viola odorata to the ethanol aqueous solution is 1 kg:12 L;

[0060] (2) The alcohol extract of Inula japonica-Viola odorata is loaded onto a macroporous resin column (the macroporous resin column is filled with AB-8 type macroporous resin; the weight of AB-8 type macroporous resin is 40 times the weight of the alcohol extract of Inula japonica-Viola odorata), and impurities are first eluted with 5 times the column volume of 60% ethanol aqueous solution; then, eluted with 5 times the column volume of 80% ethanol aqueous solution, and the eluate eluted with 80% ethanol aqueous solution is collected, and after concentration and drying, the elution fraction of Inula japonica-Viola odorata macroporous resin is obtained;

[0061] The composition is obtained by taking the elution part of the Inula japonica-Viola ol macroporous resin.

[0062] The difference between Comparative Example 2 and Example 2 is that the elution conditions of the macroporous resin column are different.

[0063] Comparative Example 3 Preparation of Composition

[0064] (1) Mixing dried whole herb of Inula japonica and dried whole herb of Viola odorata in a weight ratio of 1:1, adding 95% ethanol aqueous solution by volume, and performing reflux extraction at 78° C. for 1 h; separating the extract after the extraction, concentrating and drying the extract to obtain an Inula japonica-Viola odorata ethanol extract; wherein the total weight ratio of dried whole herb of Inula japonica and dried whole herb of Viola odorata to the ethanol aqueous solution is 1 kg:12 L;

[0065] (2) The alcohol extract of Inula japonica-Viola odorata is loaded onto a macroporous resin column (the macroporous resin column is filled with AB-8 type macroporous resin; the weight of the AB-8 type macroporous resin is 40 times the weight of the alcohol extract of Inula japonica-Viola odorata), and the impurities are first eluted with 5 times the column volume of 54% ethanol aqueous solution; then, the impurities are eluted with 5 times the column volume of 67% ethanol aqueous solution, and the eluate eluted with 67% ethanol aqueous solution is collected, and the eluate is concentrated and dried to obtain the elution fraction of the Inula japonica-Viola odorata macroporous resin;

[0066] (3) The eluted portion of the Inula japonica-Sweet Viola macroporous resin is further subjected to a silica gel column (the silica gel column is filled with 200-300 mesh silica gel; the weight of the silica gel is 30 times the weight of the eluted portion of the Inula japonica-Sweet Viola macroporous resin), and impurities are first eluted with a mixed solvent consisting of cyclohexane and ethyl acetate in a volume ratio of 95:5; then, the mixture is eluted with a mixed solvent consisting of cyclohexane and ethyl acetate in a volume ratio of 85:15, and the eluate eluted with the mixed solvent consisting of cyclohexane and ethyl acetate in a volume ratio of 85:15 is collected, and the Inula japonica-Sweet Viola silica gel eluted portion is obtained after concentration and drying;

[0067] The composition is obtained by taking the silica gel elution part of Inula japonica-Viola odorata.

[0068] The difference between Comparative Example 3 and Example 3 is that the elution conditions of the silica gel column are different.

[0069] Comparative Example 4 Preparation of Composition

[0070] (1) Mixing dried whole herb of Inula japonica and dried whole herb of Viola odorata in a weight ratio of 1:1, adding 95% ethanol aqueous solution by volume, and performing reflux extraction at 78° C. for 1 h; separating the extract after the extraction, concentrating and drying the extract to obtain an Inula japonica-Viola odorata ethanol extract; wherein the total weight ratio of dried whole herb of Inula japonica and dried whole herb of Viola odorata to the ethanol aqueous solution is 1 kg:12 L;

[0071] (2) The alcohol extract of Inula japonica-Viola odorata is loaded onto a macroporous resin column (the macroporous resin column is filled with AB-8 type macroporous resin; the weight of the AB-8 type macroporous resin is 40 times the weight of the alcohol extract of Inula japonica-Viola odorata), and the impurities are first eluted with 5 times the column volume of 54% ethanol aqueous solution; then, the impurities are eluted with 5 times the column volume of 67% ethanol aqueous solution, and the eluate eluted with 67% ethanol aqueous solution is collected, and the eluate is concentrated and dried to obtain the elution fraction of the Inula japonica-Viola odorata macroporous resin;

[0072] (3) The eluted portion of the Inula japonica-Sweet Viola macroporous resin is further subjected to a silica gel column (the silica gel column is filled with 200-300 mesh silica gel; the weight of the silica gel is 30 times the weight of the eluted portion of the Inula japonica-Sweet Viola macroporous resin), and impurities are first eluted with a mixed solvent consisting of cyclohexane and ethyl acetate in a volume ratio of 75:25; then, the mixture is eluted with a mixed solvent consisting of cyclohexane and ethyl acetate in a volume ratio of 60:40, and the eluate eluted with the mixed solvent consisting of cyclohexane and ethyl acetate in a volume ratio of 60:40 is collected, and the Inula japonica-Sweet Viola silica gel eluted portion is obtained after concentration and drying;

[0073] The composition is obtained by taking the silica gel elution part of Inula japonica-Viola odorata.

[0074] The difference between Comparative Example 4 and Example 3 is that the elution conditions of the silica gel column are different.

[0075] Experimental Example 1

[0076] Add 100 μL of 1×10 6 cfu / ml of mutans Streptococcus mutans solution; then add the test sample solution diluted in multiple ratios to the test group; make the concentration of the test sample in the test well 256, 128, 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25, 0.125 μg / ml; at the same time, set up a blank control group without adding the test sample solution; after the addition of the drug, put the 96-well plate into a cell culture incubator and culture it at 37°C for 24 hours; after the culture, use an enzyme marker to detect the absorbance value in each small hole at 600nm, and calculate the inhibition rate of the cells under each test sample concentration condition; inhibition rate = (1-test group / blank control group) × 100%, and the obtained data are used to calculate the IC value of the test sample to mutans Streptococcus mutans using SPSS statistical software 50 The test results are shown in Table 1.

[0077] The samples to be tested are the compositions prepared in Examples 1 to 3 and Comparative Examples 1 to 4; the mutans streptococci used are mutans streptococci UA159.

[0078] Table 1. Experimental results of the composition of the present invention against Streptococcus mutans

[0079] <![CDATA[IC 50 Value]]> The composition prepared in Example 1 8.47μg / ml Example 2 Composition prepared 2.16μg / ml Example 3 Composition prepared 0.28μg / ml The composition prepared in Comparative Example 1 7.64μg / ml The composition prepared in Comparative Example 2 6.35μg / ml The composition prepared in Comparative Example 3 3.48μg / ml The composition prepared in Comparative Example 4 1.83μg / ml

[0080] It can be seen from the experimental results in Table 1 that the composition prepared in Example 1 has an IC value of 1.0 for anti-Streptococcus mutans. 50 The value is 8.47 μg / ml; this shows that the composition prepared by using natural plants Inula japonica and Viola odorata as raw materials has a good anti-Streptococcus mutans effect.

[0081] It can be seen from the experimental results in Table 1 that the composition prepared in Example 2 has an IC value of 1.0447 against Streptococcus mutans. 50 The value is much smaller than the composition prepared in Example 1; this shows that: in the process of preparing the composition using natural plants Inula japonica and Viola odorata as raw materials, the preparation step is very critical; the study shows that on the basis of the Inula japonica-Viola odorata ethanol extract, the Inula japonica-Viola odorata macroporous resin elution fraction prepared by the macroporous resin elution method of the present invention has a much higher anti-mutan Streptococcus than the Inula japonica-Viola odorata ethanol extract. It can be seen that the composition using the Inula japonica-Viola odorata macroporous resin elution fraction as the composition has a much higher anti-mutan Streptococcus than the composition using the Inula japonica-Viola odorata ethanol extract as the composition.

[0082] It can be seen from the experimental results in Table 1 that the compositions prepared in Comparative Examples 1 and 2 have an IC value of 0.04477 g / cm3 against Streptococcus mutans. 50 Compared with the composition prepared in Example 1, the value is reduced, but the reduction is not large; the reduction is much smaller than that of the composition prepared in Example 2. This shows that: in the elution process of the macroporous resin column, the elution conditions of the macroporous resin column are very critical; on the basis of the ethanol extract of Inula japonica-Viola odorata, only the elution part of the macroporous resin of Inula japonica-Viola odorata prepared by the above-mentioned macroporous resin elution conditions of the present invention can have a significantly higher anti-Streptococcus mutans effect than the ethanol extract of Inula japonica-Viola odorata; however, the elution part of the macroporous resin of Inula japonica-Viola odorata prepared by other macroporous resin elution conditions cannot have a significantly higher anti-Streptococcus mutans effect than the ethanol extract of Inula japonica-Viola odorata.

[0083] It can be seen from the experimental results in Table 1 that the composition prepared in Example 3 has an IC value of 1.0447 against Streptococcus mutans. 50 The value is further significantly smaller than that of the composition prepared in Example 2; it has a very low IC 50 Value. This indicates that: on the basis of the elution fraction of the Inula japonica-Sweet Viola macroporous resin, the elution fraction of the Inula japonica-Sweet Viola silica gel prepared by the above-mentioned silica gel column elution conditions of the present invention has a further significantly higher anti-mutan Streptococcus effect than the elution fraction of the Inula japonica-Sweet Viola macroporous resin. It can be seen that the anti-mutan Streptococcus effect of the composition using the elution fraction of the Inula japonica-Sweet Viola silica gel as the composition is further significantly higher than the anti-mutan Streptococcus effect of the composition using the elution fraction of the Inula japonica-Sweet Viola macroporous resin as the composition; so that the composition has a more excellent anti-mutan Streptococcus effect.

[0084] From the experimental results in Table 1, it can be seen that the composition prepared in Comparative Example 3 has an IC value of 1.0447 against Streptococcus mutans. 50 Compared with the composition prepared in Example 2, the IC value of the composition prepared in Comparative Example 4 against Streptococcus mutans was not reduced but increased. 50 Compared with the composition prepared in Example 2, the value is reduced, but the reduction is not large; the reduction is much smaller than that of the composition prepared in Example 3. This shows that: in the elution process of the silica gel column of the present invention, the elution conditions of the silica gel column are very critical; on the basis of the elution part of the Inula-Sweet Viola macroporous resin, only the Inula-Sweet Viola silica gel elution part prepared by the above-mentioned silica gel column elution conditions of the present invention can further significantly increase the anti-mutan Streptococcus effect than the Inula-Sweet Viola macroporous resin elution part; however, the Inula-Sweet Viola silica gel elution part prepared by other silica gel column elution conditions cannot further significantly increase the anti-mutan Streptococcus effect than the Inula-Sweet Viola macroporous resin elution part.

Claims

1. A method for preparing a composition, characterized in that: It is prepared with natural plants Inula japonica and Viola odorata as raw materials; The preparation method of the composition specifically comprises the following steps: Take the whole herb of Inula japonica and the whole herb of Viola odorata, extract them with ethanol aqueous solution, and after the extraction, take the extract and concentrate and dry it to obtain the Inula japonica-Viola odorata ethanol extract; The composition is obtained by taking the alcohol extract of Inula japonica-Viola odorata; The weight ratio of the whole herb of Inula japonica to the whole herb of Viola odorata is 1-3:1-3.

2. The method for preparing the composition according to claim 1, characterized in that: The weight ratio of the whole herb of Inula japonica to the whole herb of Viola odorata is 1:

1.

3. The method for preparing the composition according to claim 1, characterized in that: The ethanol aqueous solution refers to an ethanol aqueous solution with an ethanol volume fraction of 50% to 95%.

4. The method for preparing the composition according to claim 1, characterized in that: The usage ratio of the total weight of the whole herb of Inula japonica and the whole herb of Viola odorata to the ethanol aqueous solution is 1kg:5-15L.

5. The method for preparing the composition according to claim 1, characterized in that: The usage ratio of the total weight of the whole herb of Inula japonica and the whole herb of Viola odorata to the ethanol aqueous solution is 1kg:12L.

6. The method for preparing the composition according to claim 1, characterized in that: The preparation method further comprises the following steps: The alcohol extract of Inula japonica-Viola odorata is placed on a macroporous resin column, and impurities are first eluted with an ethanol aqueous solution having a volume fraction of 53-55%, and then eluted with an ethanol aqueous solution having a volume fraction of 66-68%, and the eluate eluted with the ethanol aqueous solution having a volume fraction of 66-68% is collected, and the eluate is concentrated and dried to obtain the Inula japonica-Viola odorata macroporous resin elution fraction; The composition is obtained by taking the elution part of the Inula japonica-Viola ol macroporous resin.

7. Use of the composition prepared by the preparation method according to any one of claims 1 to 6 in preparing a product having an anti-Streptococcus mutans effect.

8. Use of the composition prepared by the preparation method according to any one of claims 1 to 6 in preparing a product having the effect of preventing dental caries.