A peach blossom extract, its preparation method and application

Abstract

The present invention belongs to the technical field of deep processing of peach blossoms, and particularly relates to a peach blossom extract, a preparation method thereof and an application. Based on the traditional water extraction and alcohol precipitation method, the preparation method uses a specific protein mixed enzyme to hydrolyze the precipitation product after alcohol precipitation, and then uses an ultrafiltration membrane with a specific pore size to perform fractional ultrafiltration on the enzymolysis product, so as to enrich more active and effective components. The anti-inflammatory / soothing effect and firming effect of the obtained peach blossom extract are significantly improved, and it has broad application prospects in the fields of medicine, food or cosmetics preparation.

Description

Technical Field

[0001] The present invention belongs to the technical field of deep processing of peach blossoms. More specifically, it relates to a peach blossom extract, a preparation method thereof, and an application thereof. Background Art

[0002] Peach blossom is the flower of the peach tree of the Rosaceae family. When picked and dried during the spring flowering period, it can be used as a traditional Chinese medicine. It has the effects of inducing defecation and promoting diuresis to alleviate edema, and is mainly used for treating edema, ascites, and constipation.

[0003] Chinese Patent Application CN108014191A discloses a peach blossom extract obtained by ultrasonic extraction of peach blossoms with water or a hydrophilic solvent and its aqueous solution, followed by concentration under reduced pressure and evaporation to dryness. This peach blossom extract has a significant inhibitory effect on the activity of hyaluronidase, and it is stated that because it can inhibit the activity of hyaluronidase, it can relieve some inflammations associated with the degradation of hyaluronic acid by preventing the degradation of hyaluronic acid. However, this invention does not have relevant experimental data directly indicating that the extracted peach blossom extract can alleviate inflammatory symptoms, and there is no clear data available to reference and judge whether its effect can meet people's expectations. Moreover, its extract is obtained by a simple solvent extraction method, and different extraction methods will result in different active ingredients. Peach blossom extracts obtained by different extraction methods may produce better effects or even different effects. Therefore, developing different extraction processes for peach blossom extracts and studying the performance effects of peach blossom extracts obtained by specific processes have important research significance and application value. Summary of the Invention

[0004] The technical problem to be solved by the present invention is to overcome the deficiencies of the above-mentioned prior art in that the performance research on a single peach blossom extract is not deep enough, and there is no direct data indicating that it can alleviate inflammatory symptoms and has other performance effects, and to provide a preparation method of a peach blossom extract.

[0005] The object of the present invention is to provide a peach blossom extract prepared by the above-mentioned preparation method.

[0006] Another object of the present invention is to provide the application of the peach blossom extract in the preparation of drugs, foods, or cosmetics.

[0007] The above objects of the present invention are achieved by the following technical solutions:

[0008] The present invention protects a preparation method of a peach blossom extract, which comprises the following steps:

[0009] S1. Add water to peach blossoms, carry out heating and reflux extraction, then concentrate, add ethanol and mix well, fully stand, and then filter to obtain a precipitate;

[0010] S2. After the precipitate obtained in step S1 is fully dissolved in water, papain and neutral protease are added for sufficient enzymatic hydrolysis. After the enzymatic hydrolysis is completed, the enzymes are inactivated. The enzymatic hydrolysate is concentrated to obtain an enzymatic hydrolysis product.

[0011] S3. The enzymatic hydrolysis product obtained in step S2 is treated with a non-polar or weakly polar resin, eluted with water to obtain an eluate. The eluate is concentrated and first ultrafiltered with a 9.5 - 10.5 kDa ultrafiltration membrane for the first time. The filtrate obtained is then ultrafiltered with a 6.5 - 7.5 kDa ultrafiltration membrane for the second time. The retentate is concentrated to obtain a peach blossom extract.

[0012] The inventors found in previous research that the selection of enzymes in the enzymatic hydrolysis step of the present invention is crucial. The anti-inflammatory / soothing effects of peach blossom extracts prepared by enzymatic hydrolysis with different enzymes are different. The inventors creatively found in a large number of experiments that the peach blossom extract prepared by using a mixed enzyme composed of papain and neutral protease in the enzymatic hydrolysis step, compared with the peach blossom extract prepared by using single papain, single neutral protease, single bromelain or other mixed enzymes, synergistically improves the enzymatic hydrolysis efficiency, better removes proteins and increases the yield of polysaccharides, and jointly improves the anti-inflammatory / soothing effect of the prepared peach blossom extract.

[0013] The inventors further found in research that after elution with macroporous resin, the peach blossom extract prepared under the ultrafiltration parameter conditions of the present invention can enrich more active ingredients, and its anti-inflammatory / soothing effect and firming effect are further improved, making the prepared peach blossom extract have excellent anti-inflammatory / soothing and firming effects.

[0014] Preferably, in step S2, the weight ratio of the papain to the neutral protease is 1 - 3:1 - 3.

[0015] More preferably, the weight ratio of the papain to the neutral protease is 1 - 2:1 - 2.

[0016] Most preferably, the weight ratio of the papain to the neutral protease is 1:1.

[0017] Preferably, in step S2, the ratio of the total weight of the papain and the neutral protease to the weight of water is 3 - 6:100.

[0018] More preferably, in step S2, the ratio of the total weight of the papain and the neutral protease to the weight of water is 5:100.

[0019] Preferably, in step S3, the first ultrafiltration is carried out with a 9.5 - 10 kDa ultrafiltration membrane.

[0020] More preferably, in step S3, a 10 kDa ultrafiltration membrane is used for the first ultrafiltration.

[0021] Preferably, in step S3, a 6.5 - 7 kDa ultrafiltration membrane is used for the second ultrafiltration.

[0022] More preferably, in step S3, a 7 kDa ultrafiltration membrane is used for the second ultrafiltration.

[0023] Furthermore, in step S3, the treatment with non - polar or weakly polar resin is carried out by treating with a resin column filled with the non - polar or weakly polar resin.

[0024] Preferably, in step S3, the non - polar or weakly polar resin is AB - 8 or D101 macroporous resin.

[0025] Preferably, in step S3, the weight of the non - polar or weakly polar resin is 5 - 15 times the weight of the enzymolysis product.

[0026] More preferably, in step S3, the weight of the non - polar or weakly polar resin is 10 times the weight of the enzymolysis product.

[0027] Preferably, in step S3, the elution with water is carried out using 5 - 10 times the column volume of water.

[0028] More preferably, in step S3, the elution with water is carried out using 8 times the column volume of water.

[0029] Preferably, in steps S1 - S3, the temperature of concentration is 50 - 70 °C.

[0030] Preferably, in steps S1 - S3, the concentration is concentrated to a solid mass fraction of 25% - 40%.

[0031] Furthermore, in step S1, the addition of ethanol is such that the final volume concentration of ethanol is 75% - 85%.

[0032] Preferably, in step S1, the addition of ethanol is such that the final volume concentration of ethanol is 80%.

[0033] Preferably, in step S1, the weight ratio of peach blossom to water is 1:15 - 25.

[0034] More preferably, in step S1, the weight ratio of peach blossom to water is 1:20.

[0035] Preferably, in step S2, the weight ratio of the precipitate to water is 1:5 - 8.

[0036] More preferably, in step S2, the weight ratio of the precipitate to water is 1:6.

[0037] The present invention also protects the peach blossom extract prepared by the above preparation method.

[0038] Furthermore, the peach blossom extract also has the effect of promoting the expression of Eln1 gene.

[0039] Even further, the Eln1 gene is the Eln1 gene of zebrafish.

[0040] The present invention also protects the application of the peach blossom extract in the preparation of medicines, foods or cosmetics.

[0041] Furthermore, the application of the peach blossom extract in the preparation of cosmetics with a firming effect.

[0042] Furthermore, the application of the peach blossom extract in the preparation of cosmetics with an anti-inflammatory / soothing effect.

[0043] The present invention has the following beneficial effects: The present invention provides a new preparation method of peach blossom extract. On the basis of the traditional water extraction and alcohol precipitation method, a specific protein mixed enzyme is used to hydrolyze the precipitation product after alcohol precipitation, and then a specific pore size ultrafiltration membrane is used to perform fractional ultrafiltration on the enzymolysis product to enrich more active ingredients. The anti-inflammatory / soothing effect and firming effect of the obtained peach blossom extract are significantly improved, and it has a broad application prospect in the field of medicine, food or cosmetics preparation. Specific Embodiments

[0044] The following specific embodiments are used to further illustrate the present invention, but the embodiments do not limit the present invention in any form. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the technical field.

[0045] Unless otherwise specified, the reagents and materials used in the following examples are all commercially available.

[0046] Example 1 Preparation of Peach Blossom Extract

[0047] S1. Take peach blossoms, and then heat with 20 times the weight of water to 100 °C for reflux extraction for 1.5 h to obtain a peach blossom water extract. Concentrate the peach blossom water extract to a solid content mass fraction of 30% to obtain a concentrated water extract;

[0048] S2. Add ethanol with a volume concentration of 95% to the concentrated water extract so that the final volume concentration of ethanol is 80%. Let it stand for 12 h and then filter to obtain a precipitate;

[0049] S3. Take the precipitate, mix it with 6 times its weight of water, then add the enzyme for enzymatic hydrolysis. After the enzymatic hydrolysis is completed, heat it to 100 °C and maintain for 10 min to inactivate the enzyme. Take the enzymatic hydrolysate, concentrate and dry it to obtain the enzymatic hydrolysis product; among them, the weight ratio of the enzyme to water is 5:100; the enzyme is a mixed enzyme composed of papain (200,000 U / g) and neutral protease (200,000 U / g) with a weight ratio of 1:1.

[0050] S4. Load the enzymatic hydrolysis product onto an AB-8 macroporous resin column (the weight of the macroporous resin in the macroporous resin column is 10 times the weight of the enzymatic hydrolysis product), then elute with 8 times the column volume of water to obtain the macroporous resin eluate. Concentrate the macroporous resin eluate to a solid content mass fraction of 30% to obtain the elution concentrate.

[0051] S5. First ultrafilter the elution concentrate obtained in step S4 with a 10 kDa ultrafiltration membrane to obtain the retentate A and the filtrate A; then ultrafilter the filtrate A with a 7 kDa ultrafiltration membrane to obtain the retentate B and the filtrate B; take the retentate B, concentrate and dry it to obtain the peach blossom extract.

[0052] Preparation of peach blossom extract in Example 2

[0053] The main difference from Example 1 is that in step S5, during the ultrafiltration process, a 10.5 kDa ultrafiltration membrane and a 7.5 kDa ultrafiltration membrane are used successively.

[0054] Other steps and parameters refer to Example 1.

[0055] Preparation of peach blossom extract in Example 3

[0056] The main difference from Example 1 is that in step S5, during the ultrafiltration process, a 9.5 kDa ultrafiltration membrane and a 6.5 kDa ultrafiltration membrane are used successively.

[0057] Other steps and parameters refer to Example 1.

[0058] Preparation of peach blossom extract in Comparative Example 1

[0059] S1. Take peach blossoms, then heat with 20 times its weight of water to 100 °C for reflux extraction for 1.5 h to obtain the peach blossom water extract. Concentrate the peach blossom water extract to a solid content mass fraction of 30% to obtain the water extract concentrate.

[0060] S2. Add ethanol with a volume concentration of 95% to the water extract concentrate so that the final volume concentration of ethanol is 80%. Let it stand for 12 h and then filter to obtain the precipitate.

[0061] S3. Take the precipitate, mix it with 6 times its weight of water, then add the enzyme for enzymatic hydrolysis. After the enzymatic hydrolysis is completed, heat it to 100 °C and maintain for 10 min to inactivate the enzyme. Take the enzymatic hydrolysate, concentrate and dry it to obtain the enzymatic hydrolysis product; among them, the weight ratio of the enzyme to water is 5:100; the enzyme is papain (200,000 U / g);

[0062] S4. Load the enzymatic hydrolysis product onto an AB-8 macroporous resin column (the weight of the macroporous resin in the macroporous resin column is 10 times the weight of the enzymatic hydrolysis product), then elute with 8 column volumes of water to obtain the macroporous resin eluate; concentrate and dry the macroporous resin eluate to obtain the peach blossom extract.

[0063] The difference from Example 1 is that in step S3, papain is used for enzymatic hydrolysis; ultrafiltration is not carried out.

[0064] Preparation of peach blossom extract in Comparative Example 2

[0065] S1. Take peach blossoms, then heat with 20 times its weight of water to 100 °C for reflux extraction for 1.5 h to obtain the peach blossom water extract. Concentrate the peach blossom water extract to a solid content mass fraction of 30% to obtain the concentrated water extract;

[0066] S2. Add ethanol with a volume concentration of 95% to the concentrated water extract so that the final volume concentration of ethanol is 80%, let it stand for 12 h and then filter to obtain the precipitate;

[0067] S3. Take the precipitate, mix it with 6 times its weight of water, then add the enzyme for enzymatic hydrolysis. After the enzymatic hydrolysis is completed, heat it to 100 °C and maintain for 10 min to inactivate the enzyme. Take the enzymatic hydrolysate, concentrate and dry it to obtain the enzymatic hydrolysis product; among them, the weight ratio of the enzyme to water is 5:100; the enzyme is bromelain (200,000 U / g);

[0068] S4. Load the enzymatic hydrolysis product onto an AB-8 macroporous resin column (the weight of the macroporous resin in the macroporous resin column is 10 times the weight of the enzymatic hydrolysis product), then elute with 8 column volumes of water to obtain the macroporous resin eluate; concentrate and dry the macroporous resin eluate to obtain the peach blossom extract.

[0069] The difference from Example 1 is that in step S3, bromelain is used for enzymatic hydrolysis; ultrafiltration is not carried out.

[0070] Preparation of peach blossom extract in Comparative Example 3

[0071] S1. Take peach blossoms, then heat with 20 times its weight of water to 100 °C for reflux extraction for 1.5 h to obtain the peach blossom water extract. Concentrate the peach blossom water extract to a solid content mass fraction of 30% to obtain the concentrated water extract;

[0072] S2. Add ethanol with a volume concentration of 95% to the water extract concentrate so that the final volume concentration of ethanol is 80%. Let it stand for 12 h and then filter to obtain a precipitate.

[0073] S3. Take the precipitate, mix it with 6 times its weight of water, and then add an enzyme for enzymatic hydrolysis. After the enzymatic hydrolysis is completed, heat it to 100 °C and maintain for 10 min to inactivate the enzyme. Take the enzymatic hydrolysate, concentrate and dry it to obtain an enzymatic hydrolysis product. Among them, the weight ratio of the enzyme to water is 5:100; the enzyme is neutral protease (200,000 U / g).

[0074] S4. Load the enzymatic hydrolysis product onto an AB-8 macroporous resin column (the weight of the macroporous resin in the macroporous resin column is 10 times the weight of the enzymatic hydrolysis product), and then elute it with 8 column volumes of water to obtain a macroporous resin eluate. Concentrate and dry the macroporous resin eluate to obtain the peach blossom extract.

[0075] The difference from Example 1 is that in step S3, enzymatic hydrolysis is carried out using neutral protease; ultrafiltration is not carried out.

[0076] Preparation of peach blossom extract in Comparative Example 4

[0077] S1. Take peach blossoms, and then heat them with 20 times their weight of water to 100 °C for reflux extraction for 1.5 h to obtain a peach blossom water extract. Concentrate the peach blossom water extract to a solid content mass fraction of 30% to obtain a water extract concentrate.

[0078] S2. Add ethanol with a volume concentration of 95% to the water extract concentrate so that the final volume concentration of ethanol is 80%. Let it stand for 12 h and then filter to obtain a precipitate.

[0079] S3. Take the precipitate, mix it with 6 times its weight of water, and then add an enzyme for enzymatic hydrolysis. After the enzymatic hydrolysis is completed, heat it to 100 °C and maintain for 10 min to inactivate the enzyme. Take the enzymatic hydrolysate, concentrate and dry it to obtain an enzymatic hydrolysis product. Among them, the weight ratio of the enzyme to water is 5:100; the enzyme is a mixed enzyme composed of papain (200,000 U / g) and neutral protease (200,000 U / g) with a weight ratio of 1:1.

[0080] S4. Load the enzymatic hydrolysis product onto an AB-8 macroporous resin column (the weight of the macroporous resin in the macroporous resin column is 10 times the weight of the enzymatic hydrolysis product), and then elute it with 8 column volumes of water to obtain a macroporous resin eluate. Concentrate and dry the macroporous resin eluate to obtain the peach blossom extract.

[0081] The difference from Example 1 is that ultrafiltration is not carried out.

[0082] Preparation of peach blossom extract in Comparative Example 5

[0083] S1. Take peach blossoms, and then heat with 20 times the weight of water to 100 °C for reflux extraction for 1.5 h to obtain the aqueous extract of peach blossoms. Concentrate the aqueous extract of peach blossoms to a solid content mass fraction of 30% to obtain the concentrated aqueous extract;

[0084] S2. Add ethanol with a volume concentration of 95% to the concentrated aqueous extract so that the final volume concentration of ethanol is 80%. Let it stand for 12 h and then filter to obtain a precipitate;

[0085] S3. Take the precipitate, mix it with 6 times the weight of water, and then add an enzyme for enzymatic hydrolysis. After the enzymatic hydrolysis is completed, heat to 100 °C and maintain for 10 min to inactivate the enzyme. Take the enzymatic hydrolysate, concentrate and dry it to obtain the enzymatic hydrolysis product; among them, the weight ratio of the enzyme to water is 5:100; the enzyme is a mixed enzyme composed of papain (200,000 U / g) and bromelain (200,000 U / g) with a weight ratio of 1:1;

[0086] S4. Load the enzymatic hydrolysis product onto an AB-8 type macroporous resin column (the weight of the macroporous resin in the macroporous resin column is 10 times the weight of the enzymatic hydrolysis product), and then elute with 8 times the column volume of water to obtain the macroporous resin eluate; concentrate and dry the macroporous resin eluate to obtain the peach blossom extract.

[0087] The difference from Example 1 is that in step S3, the mixed enzyme used is papain and bromelain; ultrafiltration is not carried out.

[0088] Preparation of peach blossom extract in Comparative Example 6

[0089] S1. Take peach blossoms, and then heat with 20 times the weight of water to 100 °C for reflux extraction for 1.5 h to obtain the aqueous extract of peach blossoms. Concentrate the aqueous extract of peach blossoms to a solid content mass fraction of 30% to obtain the concentrated aqueous extract;

[0090] S2. Add ethanol with a volume concentration of 95% to the concentrated aqueous extract so that the final volume concentration of ethanol is 80%. Let it stand for 12 h and then filter to obtain a precipitate;

[0091] S3. Take the precipitate, mix it with 6 times the weight of water, and then add an enzyme for enzymatic hydrolysis. After the enzymatic hydrolysis is completed, heat to 100 °C and maintain for 10 min to inactivate the enzyme. Take the enzymatic hydrolysate, concentrate and dry it to obtain the enzymatic hydrolysis product; among them, the weight ratio of the enzyme to water is 5:100; the enzyme is a mixed enzyme composed of bromelain (200,000 U / g) and neutral protease with a weight ratio of 1:1;

[0092] S4. Load the enzymatic hydrolysis product onto an AB-8 type macroporous resin column (the weight of the macroporous resin in the macroporous resin column is 10 times the weight of the enzymatic hydrolysis product), and then elute with 8 times the column volume of water to obtain the macroporous resin eluate; concentrate and dry the macroporous resin eluate to obtain the peach blossom extract.

[0093] The difference from Example 1 is that in step S3, the mixed enzymes used are neutral protease and bromelain; ultrafiltration is not carried out.

[0094] Preparation of peach blossom extract for Comparative Example 7

[0095] Take peach blossoms, and then heat with 20 times the weight of water to 100 °C for reflux extraction for 1.5 h to obtain a water extract of peach blossoms. After concentrating and drying the water extract of peach blossoms, the peach blossom water extract is obtained, and thus the peach blossom extract is obtained.

[0096] Preparation of peach blossom extract for Comparative Example 8

[0097] S1. Take peach blossoms, and then heat with 20 times the weight of water to 100 °C for reflux extraction for 1.5 h to obtain a water extract of peach blossoms. Concentrate the water extract of peach blossoms to a solid content mass fraction of 30% to obtain a concentrated water extract;

[0098] S2. Add ethanol with a volume concentration of 95% to the concentrated water extract so that the final volume concentration of ethanol is 80%. Let it stand for 12 h and then filter to obtain a precipitate;

[0099] S3. Take the precipitate, mix it with 6 times the weight of water, and then add enzymes for enzymatic hydrolysis. After the enzymatic hydrolysis is completed, heat to 100 °C and maintain for 10 min to inactivate the enzymes. Take the enzymatic hydrolysate, concentrate and dry it to obtain an enzymatic hydrolysis product; among them, the weight ratio of the enzyme to water is 5:100; the enzyme used is a mixed enzyme composed of papain (200,000 U / g) and neutral protease (200,000 U / g) with a weight ratio of 1:1;

[0100] S4. Load the enzymatic hydrolysis product onto an AB-8 type macroporous resin column (the weight of the macroporous resin in the macroporous resin column is 10 times the weight of the enzymatic hydrolysis product), and then elute with 8 column volumes of water to obtain a macroporous resin eluate. Concentrate the macroporous resin eluate to a solid content mass fraction of 30% to obtain a concentrated eluate;

[0101] S5. First ultrafilter the concentrated eluate obtained in step S4 with a 20 kDa ultrafiltration membrane to obtain a retentate A and a filtrate A; then ultrafilter the filtrate A with a 15 kDa ultrafiltration membrane to obtain a retentate B and a filtrate B; take the retentate B, concentrate and dry it to obtain the peach blossom extract.

[0102] The difference from Example 1 is that in step S4, the ultrafiltration treatment is to first use a 20 kDa ultrafiltration membrane and then use a 15 kDa ultrafiltration membrane.

[0103] Preparation of peach blossom extract for Comparative Example 9

[0104] S1. Take peach blossoms, and then heat with 20 times the weight of water to 100 °C for reflux extraction for 1.5 h to obtain a water extract of peach blossoms. Concentrate the water extract of peach blossoms to a solid content mass fraction of 30% to obtain a concentrated water extract;

[0105] S2. Add ethanol with a volume concentration of 95% to the water extract concentrate so that the final volume concentration of ethanol is 80%. Let it stand for 12 h and then filter to obtain a precipitate.

[0106] S3. Take the precipitate, mix it with 6 times its weight of water, then add an enzyme for enzymatic hydrolysis. After the enzymatic hydrolysis is completed, heat it

[0107] to 100 °C and maintain for 10 min to inactivate the enzyme. Take the enzymatic hydrolysate, concentrate and dry it to obtain an enzymatic hydrolysis product. Among them, the weight ratio of the enzyme to water is 5:100; the enzyme is a mixed enzyme composed of papain (200,000 U / g) and neutral protease (200,000 U / g) with a weight ratio of 1:1.

[0108] S4. Load the enzymatic hydrolysis product onto an AB-8 macroporous resin column (the weight of the macroporous resin in the macroporous resin column is 10 times the weight of the enzymatic hydrolysis product), then elute with 8 times the column volume of water to obtain a macroporous resin eluate. Concentrate the macroporous resin eluate to a solid content mass fraction of 30% to obtain an elution concentrate.

[0109] S5. First ultrafilter the elution concentrate obtained in step S4 with a 15 kDa ultrafiltration membrane to obtain a retentate A and a filtrate A; then ultrafilter the filtrate A with a 10 kDa ultrafiltration membrane to obtain a retentate B and a filtrate B; Take the retentate B, concentrate and dry it to obtain the peach blossom extract.

[0110] The difference from Example 1 is that in step S4, the ultrafiltration treatment is to first use a 15 kDa ultrafiltration membrane and then use a 10 kDa ultrafiltration membrane.

[0111] Preparation of Peach Blossom Extract in Comparative Example 10

[0112] S1. Take peach blossoms, then heat with 20 times its weight of water to 100 °C for reflux extraction for 1.5 h to obtain a peach blossom water extract. Concentrate the peach blossom water extract to a solid content mass fraction of 30% to obtain a water extract concentrate.

[0113] S2. Add ethanol with a volume concentration of 95% to the water extract concentrate so that the final volume concentration of ethanol is 80%. Let it stand for 12 h and then filter to obtain a precipitate.

[0114] S3. Take the precipitate, mix it with 6 times its weight of water, then add an enzyme for enzymatic hydrolysis. After the enzymatic hydrolysis is completed, heat it

[0115] to 100 °C and maintain for 10 min to inactivate the enzyme. Take the enzymatic hydrolysate, concentrate and dry it to obtain an enzymatic hydrolysis product. Among them, the weight ratio of the enzyme to water is 5:100; the enzyme is a mixed enzyme composed of papain (200,000 U / g) and neutral protease (200,000 U / g) with a weight ratio of 1:1.

[0116] S4. Load the enzymatic hydrolysate onto an AB-8 macroporous resin column (the weight of the macroporous resin in the macroporous resin column is 10 times the weight of the enzymatic hydrolysate), then elute with 8 column volumes of water to obtain the macroporous resin eluate. Concentrate the macroporous resin eluate to a solid mass fraction of 30% to obtain the elution concentrate;

[0117] S5. Ultrafilter the elution concentrate obtained in step S4 first with an 8 kDa ultrafiltration membrane to obtain the retentate A and the filtrate A; then ultrafilter the filtrate A with a 5 kDa ultrafiltration membrane to obtain the retentate B and the filtrate B; Take the retentate B, concentrate and dry it to obtain the peach blossom extract.

[0118] The difference from Example 1 is that in step S4, the ultrafiltration treatment is to first use an 8 kDa ultrafiltration membrane and then a 5 kDa ultrafiltration membrane.

[0119] Experimental Example 1 Anti-inflammatory and Soothing Effect Experiment of Peach Blossom Extract

[0120] 1. Experimental Method

[0121] Select Tg(corola:EGFP) transgenic zebrafish at 3 dpf (days post fertilization) with normal development, pretreat with 1.00 mg / mL of the test sample for 1 h. The standard dilution water is used as the blank control group. Under a stereomicroscope, use a scalpel to excise 50% of the caudal fin of the 3 dpf zebrafish and place it in a 6-well cell culture plate, 14 fish per well. Add the test sample to each well of the experimental group to make its concentration 1.00 mg / mL. Use the zebrafish system culture water as the blank control group and continue to incubate in an incubator for 6 h. After incubating for 6 h, anesthetize the 3 dpf zebrafish with 0.02% tricaine, observe the aggregation of macrophages and neutrophils at the wound of the zebrafish caudal fin under a fluorescence microscope, take pictures and record, count the number of cells, and calculate the average number of inflammatory cells aggregated at the wound of each group of zebrafish.

[0122] Among them, the test samples are the peach blossom extracts prepared in Comparative Examples 1-10 and Example 1 respectively, and the test results are shown in Table 1.

[0123] 2. Experimental Results

[0124] Table 1 Anti-inflammatory and Soothing Test Results of the Peach Blossom Extract of the Present Invention

[0125] Group Average number of inflammatory cells aggregated at the wound site (cells) Example 1 6.8 Blank control group 16.7 Comparative example 1 14.3 Comparative example 2 15.0 Comparative example 3 14.8 Comparative example 4 10.7 Comparative example 5 14.6 Comparative example 6 14.9 Comparative example 7 16.0 Comparative example 8 16.7 Comparative example 9 15.3 Comparative example 10 11.5

[0126] As can be seen from Table 1, by comparing the data of the blank control and Comparative Examples 1-7, only the peach blossom extract prepared with a mixed enzyme composed of papain and neutral protease in the enzymatic hydrolysis step shows a significant improvement in its anti-inflammatory / soothing effect; while the peach blossom extract obtained only by water extraction or the peach blossom extract prepared by enzymatic hydrolysis with a mixed enzyme composed of other enzymes or a single enzyme shows a weak improvement in its anti-inflammatory / soothing effect and cannot produce a synergistic anti-inflammatory / soothing effect.

[0127] From the data of Comparative Examples 8-10, compared with the peach blossom extract prepared in Example 1, the improvement effect of the anti-inflammatory / soothing effect is weak. This shows that in the ultrafiltration step of the present invention, the selection of ultrafiltration conditions is very crucial. It is necessary to first ultrafilter with a 10 kDa ultrafiltration membrane and then ultrafilter with a 7 kDa ultrafiltration membrane. The peach blossom extract prepared from the retentate after ultrafiltration with a 7 kDa ultrafiltration membrane shows a significant improvement in its anti-inflammatory / soothing effect. The effects achieved by the peach blossom extracts prepared in Example 2 and Example 3 are basically the same as those in Example 1.

[0128] Experimental Example 2 Experiment on the firming effect of peach blossom extract

[0129] 1. Experimental method

[0130] Wild-type zebrafish larvae at 4 dpf (four days after fertilization) were collected. The larvae were placed in a 6-well plate, with 20 larvae in each well. 5 mL of sample solutions with different concentrations were added for treatment. The 20 larvae in the last well were treated with standard dilution water as the blank control group. After 72 h of treatment, all the larvae were collected and washed twice with sterilized double-distilled water. Subsequently, the larvae were transferred to sterilized 1.5 mL EP tubes marked respectively, and zebrafish RNA was extracted. Then the RNA was reverse transcribed into cDNA, and the change of Eln1 gene at the mRNA level was detected on a fluorescence quantitative PCR instrument.

[0131] The samples tested were the peach blossom extract prepared in Example 1 and the peach blossom extracts prepared in Comparative Examples 4, 7-9, with a mass concentration of 0.5 mg / mL; among which the positive control drug was 0.5 mg / mL polysaccharide of Dendrobium officinale. The test results are shown in Table 2.

[0132] 2. Experimental results

[0133] Table 2 Effects on the relative expression level of Eln1 gene in zebrafish

[0134] Relative expression level of Eln1 gene Example 1 2.99±0.28 Blank control group 1.00±0.15 Positive control group 1.91±0.10 Comparative example 4 1.78±0.11 Comparative example 7 1.65±0.17 Comparative example 8 1.44±0.13 Comparative example 9 1.31±0.15

[0135] As can be seen from Table 2, for the peach blossom extract prepared in Comparative Example 4, compared with the blank control group, the relative expression level of the Eln1 gene in zebrafish was increased. This indicates that the peach blossom extract obtained by enzymatic hydrolysis with papain and neutral protease has the effect of promoting the expression of the Eln1 gene, showing a certain firming effect. Based on Comparative Example 4, ultrafiltration under specific parameters was further carried out, and the obtained peach blossom extract (i.e., Example 1) further significantly increased the relative expression level of the Eln1 gene in zebrafish. Moreover, its relative expression level of the Eln1 gene in zebrafish was higher than that of the positive control drug, polysaccharide of Dendrobium officinale. This shows that by jointly carrying out enzymatic hydrolysis with papain and neutral protease and further enriching through specific ultrafiltration conditions, more firming active ingredients can be obtained, which can significantly increase the relative expression level of the Eln1 gene in zebrafish, and its relative expression level is also better than that of the positive control drug, polysaccharide of Dendrobium officinale.

[0136] From the data of Comparative Examples 7 to 9, it can be seen that compared with the peach blossom extract prepared in Example 1, it cannot significantly increase the relative expression level of the Eln1 gene in zebrafish. This shows that in the ultrafiltration step of the present invention, the selection of ultrafiltration conditions is very crucial. It is necessary to first carry out ultrafiltration with a 9.5 - 10.5 kDa ultrafiltration membrane, and then carry out ultrafiltration with a 6.5 - 7.5 kDa ultrafiltration membrane. The peach blossom extract prepared from the retentate after ultrafiltration with a 6.5 - 7.5 kDa ultrafiltration membrane can significantly improve its firming effect. The effects achieved by the peach blossom extracts prepared in Example 2 and Example 3 are basically the same as those in Example 1.

[0137] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited by the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications made without departing from the spirit and principle of the present invention shall be equivalent replacement methods and are all included in the protection scope of the present invention.

Claims

1. A peach blossom extract, characterized in that, The preparation method of the peach blossom extract comprises the following steps: S1. Add water to peach blossoms, carry out heating and reflux extraction, then concentrate, add ethanol and mix evenly, fully stand and then filter to obtain a precipitate; S2. Take the precipitate obtained in step S1, add water to fully dissolve it, add papain and neutral protease and carry out full enzymatic hydrolysis. After the enzymatic hydrolysis is completed, inactivate the enzymes. Take the enzymatic hydrolysate, concentrate it to obtain an enzymatic hydrolysis product; S3. Treat the enzymatic hydrolysis product obtained in step S2 with non-polar or weakly polar resin, elute with water to obtain an eluate. Concentrate the eluate, first carry out the first ultrafiltration with a 10 kDa ultrafiltration membrane, and then carry out the second ultrafiltration on the obtained filtrate with a 7 kDa ultrafiltration membrane. Take the retentate, concentrate it to obtain the peach blossom extract; In step S2, the weight ratio of the papain to the neutral protease is 1-3:1-3; In step S3, the non-polar or weakly polar resin is AB-8 or D101 macroporous resin.

2. The peach blossom extract according to claim 1, wherein, The weight ratio of the papain to the neutral protease is 1-2:1-2.

3. The peach blossom extract according to claim 1, characterized in that, In steps S1-S3, the temperature of the concentration is 50-70 °C.

4. The peach blossom extract according to claim 1, wherein In steps S1-S3, the concentration is to concentrate to a solid mass fraction of 25%-40%.

5. Use of the peach blossom extract according to any one of claims 1-4 in the preparation of a cosmetic having a firming effect and an anti-inflammatory and soothing effect.