CN112791127B - Peony petal extract and preparation method and application thereof - Google Patents
Peony petal extract and preparation method and application thereof Download PDFInfo
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- CN112791127B CN112791127B CN202110187953.3A CN202110187953A CN112791127B CN 112791127 B CN112791127 B CN 112791127B CN 202110187953 A CN202110187953 A CN 202110187953A CN 112791127 B CN112791127 B CN 112791127B
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- extract
- peony petal
- peony
- solution
- ethyl acetate
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- A61K36/185—Magnoliopsida (dicotyledons)
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A61K36/18—Magnoliophyta (angiosperms)
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- A61K36/65—Paeoniaceae (Peony family), e.g. Chinese peony
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- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
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- A—HUMAN NECESSITIES
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- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
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Abstract
The invention discloses a peony petal extract and a preparation method and application thereof. The preparation method of the peony petal extract provided by the invention comprises the following steps: (1) drying and crushing peony petals, adding an organic acid-organic solvent-water mixed solvent for ultrasonic extraction, and collecting and combining extracting solutions to obtain a crude extracting solution; (2) concentrating the crude extract obtained in the step (1) under reduced pressure until no organic solvent smell exists, adding an aqueous alkali solution, adjusting the pH value of the solution to 11.0-13.0, filtering to remove insoluble substances, extracting the filtered solution with ethyl acetate, and removing the ethyl acetate extract to obtain an aqueous alkali extract; (3) acidifying the alkaline water extract obtained in the step (2) with acid water, extracting with ethyl acetate to obtain ethyl acetate extract, removing the solvent and drying to obtain the target extract.
Description
Technical Field
The invention relates to the technical field of biology, in particular to a peony petal extract and a preparation method and application thereof.
Background
The peony petals can be soaked in water for drinking, and have certain pharmacological effects. The theory of traditional Chinese medicine considers that the proper amount of peony petal tea can condition the health of human body, and the peony petal tea is a scented tea beverage with good efficacy. CN103494206B discloses a preparation process of a soft capsule containing peony petal isoflavone and peony seed essential oil, wherein the peony petal isoflavone is extracted by a biological method in the provided preparation process. CN108452035A discloses a peony petal polyphenol extraction method, which comprises the steps of raw material pretreatment, polyphenol extraction and crude liquid centrifugation, wherein peony petals are dried, milled and sieved, fine peony petal powder is uniformly mixed with an ethanol solution with the volume fraction of 50-90% according to the liquid-material ratio of 30: 1-50: 1, the mixed solution of the fine peony petal powder and the ethanol solution is placed in an ultrasonic cleaning machine for ultrasonic extraction, and the crude peony petal extraction liquid is obtained after ultrasonic extraction; filtering the petal extract, and centrifuging to obtain polyphenol crude extract. CN110063496A discloses a peony petal capsule and a preparation method thereof, the method is to wash, pre-cool, dry and crush peony petals in full bloom stage to prepare peony petals as capsule filling materials, starch is extracted from crops, plant gel is used as gel to prepare capsule shells. CN110050995A discloses peony petal electuary and a preparation method thereof.
The peony petal products or preparations disclosed above are all based on the traditional knowledge and efficacy of peony petals, the preparation method is also simpler, and the medicinal value analysis of peony petal extracts is lacked.
Disclosure of Invention
In view of the above, the main object of the present invention is to provide a peony petal extract, a preparation method and applications thereof.
In order to achieve the above objects, according to one aspect of the present invention, there is provided a method for preparing a peony petal extract.
The preparation method of the peony petal extract provided by the invention comprises the following steps:
(1) drying and crushing peony petals, adding an organic acid-organic solvent-water mixed solvent for ultrasonic extraction, and collecting and combining extracting solutions to obtain a crude extracting solution;
(2) concentrating the crude extract obtained in the step (1) under reduced pressure until no organic solvent smell exists, diluting with water until the relative density is 1.10-1.20, adding an alkaline water solution, adjusting the pH value of the solution to 11.0-13.0, filtering to remove insoluble substances, extracting the filtered solution with ethyl acetate, and removing the ethyl acetate extract to obtain an alkaline water extract; the alkaline aqueous extract refers to the solution which is left after removing the acetic acid extraction solution;
(3) acidifying the alkaline water extract obtained in the step (2) with acid water, extracting with ethyl acetate to obtain ethyl acetate extract, removing the solvent and drying to obtain the target extract.
The organic acid-organic solvent-water mixed solvent added in the step (1) is directly mixed with the organic acid, the organic solvent and the water; wherein the organic acid can make the extraction solvent acid, and the polyphenol compounds in the peony petals can be kept in a free state under the environment, so that the solubility in the organic solvent is increased. Peony petals are different from compounds contained in other parts of peony, for example, peony seeds are rich in fatty acid, peony roots contain simple aromatic compounds paeonol and terpenoid compounds paeoniflorin, and peony petals are rich in polyphenol compounds mainly containing flavone.
Adjusting the pH value with alkaline water in the step (2), and extracting with ethyl acetate, wherein the main purpose is to enrich the acidic polyphenol compounds in the peony petals in an alkaline water layer, and transfer neutral or alkaline organic compounds to an ethyl acetate layer; then, in step (3), the reaction mixture is further acidified with an acid solution and extracted with ethyl acetate, so that the acidic polyphenol compounds are transferred to the ethyl acetate layer in a free state.
The purpose of adding the aqueous alkali solution in the step (2) is to dissolve the target component in the aqueous alkali and to dissolve impurities in ethyl acetate.
The purpose of the acidification in the step (3) is to allow the target component to exist in a free state and to be more easily transferred to the ethyl acetate layer.
The organic acid-organic solvent-water mixed solvent in the step (1), wherein the organic acid is selected from any one of malic acid, acetic acid and formic acid, and the volume percentage concentration of the organic acid is 3-5%; wherein the organic solvent is ethanol, and the volume percentage concentration of the organic solvent is 67-80%;
the material-liquid ratio of the peony petals to the organic acid-organic solvent-water mixed solvent is 1 g: 12 ml-1 g: 20 ml.
The ultrasonic extraction conditions in the step (1) are as follows: ultrasonic extraction is carried out for 1 to 3 times at the temperature of between 20 and 30 ℃, and each time lasts for 10 to 30 min.
The alkaline water in the step (2) is sodium hydroxide or potassium hydroxide aqueous solution; the volume percentage concentration of the alkaline water solution is 1-3%;
the vacuum concentration in the step (2) is carried out under the conditions that the temperature is 30-60 ℃ and the pressure is 0.01-0.1 MPa until no alcohol smell exists.
In the step (3), the acid water is 0.1-1M hydrochloric acid water solution, and is acidified to pH of 1.0-3.0.
The peony petal extract prepared by the preparation method of the peony petal extract also belongs to the protection scope of the invention.
According to another aspect of the present invention, there is provided a fatty acid synthase inhibitor.
The active component of the fatty acid synthase inhibitor provided by the invention is the peony petal extract prepared by the method.
The application of the peony petal extract prepared by the method in preparing products or medicines for inducing cancer cell apoptosis also belongs to the protection scope of the invention.
The cancer cell is breast cancer cell MDA-MB-231 or MCF-7.
The application of the peony petal extract in preparing medicines, cosmetics or foods with weight-losing or anticancer effects also belongs to the protection scope of the invention.
The preparation method of the peony pistil extract provided by the invention has the following characteristics or advantages:
(1) the method for preparing the peony petal extract uses an organic solvent containing organic acid for ultrasonic extraction, adjusts the pH value and then uses ethyl acetate for extraction, and is the most key for preparing the peony petal extract. Wherein, the ultrasonic extraction with the solvent containing organic acid is to avoid the dissociation of acid components in the peony petals and complete the extraction of the effective components with the maximum efficiency under the condition of relatively low temperature, and fat-soluble impurities can be effectively removed by using ethyl acetate to extract after the pH value is adjusted by using alkaline water and acid water.
(2) Unlike conventional extraction methods, the present method takes full account of the effect of temperature on the chemical components of the extract, and therefore all operations are performed at temperatures not higher than 60 ℃.
(3) The invention has another advantage that the phenolic acid components in the peony petals are fully enriched by combining repeated acid-base treatment and extraction operation of an organic solvent and an aqueous solution.
Drawings
For purposes of illustration and not limitation, the present invention will now be described in accordance with its preferred embodiments, particularly with reference to the accompanying drawings, in which:
FIG. 1 is a graph showing the results of comparing the inhibitory ability of the extract of the present invention against three currently known inhibitors for the activity of fatty acid synthase; wherein 1 is the extract a of the invention of example 1, 2 is the extract B of the invention of example 2, 3 is the extract C of the invention of example 3, 4 is cerulenin, 5 is EGCG; the unit of ordinate is microgram/ml.
FIG. 2 shows the effect of the extract of the present invention on the activity of breast cancer cells MDA-MB-231; wherein 1 is DMSO-treated group, 2 is inventive extract A, 3 is inventive extract B, and 4 is inventive extract C.
FIG. 3 shows the effect of the extract of the present invention on the activity of MCF-7 in breast cancer cells; wherein 1 is DMSO-treated group, 2 is inventive extract A, 3 is inventive extract B, and 4 is inventive extract C.
Detailed Description
The present invention will be further described with reference to the following specific examples.
Example 1 preparation of peony petal extract A
Preparation method of peony petal extract A
The preparation method of the peony petal extract A comprises the following steps:
(1) drying and crushing peony petals, and mixing the materials according to a material-liquid ratio of 1 g: adding malic acid-ethanol-water solution at volume ratio of 3:67:30 into 15ml, performing ultrasonic extraction at 20 deg.C for 30min for 2 times, and collecting and mixing extractive solutions to obtain crude extractive solution;
(2) concentrating the crude extract obtained in step (1) under reduced pressure at 50 deg.C and 0.05MPa until no alcohol smell exists, diluting with water to relative density of 1.10, adding 1% by volume sodium hydroxide aqueous solution, adjusting pH to 13.0, filtering to remove insoluble substances, extracting the filtered solution with ethyl acetate for 3 times, and removing ethyl acetate extract to obtain alkaline water extract;
(3) acidifying the alkaline water extract obtained in the step (2) with a hydrochloric acid aqueous solution with the concentration of 0.1M, adjusting the pH of the aqueous solution to 1.0 by using a pH meter, extracting for 3 times by using ethyl acetate, combining the extraction liquids for 3 times to obtain an ethyl acetate extraction liquid, removing the solvent and drying to obtain the target peony petal extract A.
Secondly, the inhibition effect of the peony petal extract A on the activity of Fatty Acid Synthase (FAS)
A proper amount of the peony petal extract A is dissolved in DMSO to be used as a test sample solution to prepare a mother solution of 1mg/mL, and the mother solution is diluted into samples with different concentrations by the DMSO to carry out the test by using a standard determination method of the activity of fatty acid synthase. The method for measuring the activity of the fatty acid synthase is a conventional method for measuring the activity of the enzyme in the tumor cells in vitro. After the breast Cancer cells MDA-MB-231 (purchased from the cell bank of the China academy of sciences type culture Collection) or MCF-7 (purchased from the cell bank of the China academy of sciences type culture Collection) were disrupted, the determination was carried out by a standard assay method using acetyl coenzyme A and malonyl coenzyme A, NADPH as substrates (Tian W.X.et al, 1985.J.biol.chem.,260: 11375-11387; Li P., et al, 2014.Mol Cancer 13: 138). The specific method is that breast cancer cells MDA-MB-231 or MCF-7 are inoculated on a cell culture dish (the diameter is 10cm) at the density of 70-80%, and after the cells adhere to the wall overnight, culture media containing different drug concentrations are added. After 24h drug treatment, the adherent cells were washed twice with pre-cooled PBS and transferred to centrifuge tubes with a cell scraper on ice. Cells were sonicated until clear. Collecting the crushed liquid, centrifuging at 13000rpm for 20min at 4 ℃, taking the supernatant, and measuring the enzyme activity. FAS was monitored continuously at 37 ℃ for 120 seconds at 340nm in the UV range using the method reported previously. Wherein 500. mu.L of the assay system contains 25mM KH2PO4-K2HPO4Buffer for assay (pH 7), 0.25mM EDTA, 0.25mM DTT, 30. mu.M acetyl-CoA, 100. mu.M malonyl-CoA and 350. mu.M NADPH the total volume was made up with the supernatantFill to 500. mu.L. And (3) absorbing by taking malonyl coenzyme A which is not added as a background, and finally determining the total protein concentration by using a BCA method, wherein each sample is repeatedly determined for more than three times. The viability of FAS was calculated as nmol NADPH consumption per mg total protein per min. All experiments were repeated three times.
The degree of inhibition of Fatty Acid Synthase (FAS) activity is expressed in terms of relative activity, and is calculated as: the fatty acid synthase activity value was determined as a control by adding only the corresponding extraction solvent to the substrate, and this value was represented by A0 and was set to 100%. The test group added with the fatty acid synthase inhibitor extract liquid has the fatty acid synthase activity value of 50% of A0, and the concentration of the inhibitor is IC50 value.
The half inhibitory concentrations of the two currently known inhibitors were compared against the fatty acid synthase inhibitory activity, which was determined by the conventional method for determining the activity of the enzyme in tumor cells in vitro, as described above. Both controls were not pre-treated before activity measurements were made, but were dissolved in DMSO at concentrations that varied depending on the activity.
Control 1: cerulenin. Cerulenin is a widely used inhibitor of natural Fatty Acid Synthase (FAS). Cerulenin is produced by the fungus cephalosporium caeruleueus. Cerulenin for use in the present invention is available from Sigma-Aldrich, Cat. 219557.
Control 2: EGCG. EGCG (epigallocatechin gallate), is the main component of green tea polyphenols, and is a catechin monomer separated from tea leaves. EGCG for use in the present invention is available from Sigma-Aldrich under catalog number 93894.
The results of comparison of the peony petal extract A of this example 1 with the above-mentioned two currently known inhibitors at half inhibitory concentrations for the fatty acid synthase inhibitory activity are shown in FIG. 1. Wherein 1 is the extract A of the invention of example 1, 4 is cerulenin, 5 is EGCG; the unit of ordinate is microgram/ml. The half inhibitory concentration (IC50) of the peony petal extract A of example 1 against fatty acid synthase was 5.30. mu.g/mL, while the half inhibitory concentrations (IC50) of control 1(Cerulenin) and control 2(EGCG) against fatty acid synthase were 24.2. mu.g/mL and 25.8. mu.g/mL, respectively, indicating that the peony petal extract A of example 1 strongly inhibits the activity of fatty acid synthase and that the required effective dose is low.
Thirdly, the influence of the peony petal extract A on the activities of human breast cancer MDA-MB-231 cells and MCF-7 cells with high fatty acid synthase expression
The cell viability was determined by the CCK-8 method. Inoculating the breast cancer cells MDA-MB-231 or MCF-7 into a 96-well plate, making the cell density to be 70% -80% after the cells are attached to the wall overnight, removing a culture medium containing serum, adding a serum-free culture medium containing different drug concentrations, and setting 6 multiple wells for each concentration of drug. After the cells are treated with the drug for 24 hours, the culture medium is aspirated, 100 mu L of fresh serum-free culture medium and 10 mu L of CCK-8 solution are added into each well, incubation is carried out for 1-2 hours at 37 ℃, and then the light absorption value under 450nm is detected by using an enzyme-linked immunosorbent assay. Data are the average of 6 replicates, and all experiments were repeated three more times. Relative viability of the cells of the peony petal extract A treatment group is calculated by taking the viability of the cells of the DMSO treatment group as 100%.
The results of the effect of peony petal extract A on the activity of MDA-MB-231 cells of human breast cancer with high fatty acid synthase expression are shown in FIG. 2, in which the ordinate of FIG. 2 is the relative activity of MDA-MB-231 cells, and in the abscissa: the DMSO-treated group was 1, and the extract A of the present invention was 2 (sample concentrations were 10. mu.g/ml). The results show that the relative viability of the cells in the DMSO-treated group was 100% in MDA-MB-231 cells, and that the relative viability of the cells after the treatment with the extract A of the invention was 45.0%. Compared with the DMSO-treated group, the peony petal extract A of the embodiment can significantly reduce the MDA-MB-231 cell activity (p <0.05) at the concentration of 10 micrograms/ml.
The effect of the peony petal extract A on MCF-7 cell viability is shown in FIG. 3, in which the vertical axis of FIG. 3 is the relative MCF-7 cell viability, and the horizontal axis is: the DMSO-treated group was 1, and the extract A of the present invention was 2 (sample concentrations were 10. mu.g/ml). The results showed that the relative viability of the cells in the DMSO-treated group was 100% and that the relative viability of the cells in the extract A of the present invention was 39.3% in MCF-7 cells. Compared with the DMSO-treated group, the peony petal extract A of the embodiment can significantly reduce MCF-7 cell activity (p <0.05) at a concentration of 10 micrograms/ml.
Example 2 preparation of peony petal extract B
Preparation method of peony petal extract B
The preparation method of the peony petal extract B comprises the following steps:
(1) drying and crushing peony petals, and mixing the materials according to a material-liquid ratio of 1 g: adding 20ml of acetic acid-ethanol-water solution at a volume ratio of 5:80:15, performing ultrasonic extraction at 25 deg.C for 3 times (30 min, 20min, 10min each time), collecting the extractive solutions, and mixing to obtain crude extractive solution;
(2) concentrating the crude extract obtained in the step (1) under reduced pressure at 30 ℃ and 0.1MPa until no alcohol smell exists, diluting with water to a relative density of 1.20, adding a potassium hydroxide aqueous solution with a volume percentage concentration of 3%, adjusting the pH value of the solution to 11.0, filtering to remove insoluble substances, extracting the filtered solution with ethyl acetate for 3 times, and removing the ethyl acetate extract to obtain an alkaline water extract;
(3) acidifying the alkaline water extract obtained in the step (2) with 0.5M hydrochloric acid aqueous solution, adjusting the pH of the aqueous solution to 2.0 by using a pH meter, extracting for 3 times by using ethyl acetate, combining the extraction solutions for 3 times to obtain ethyl acetate extraction solution, removing the solvent and drying to obtain the target extract B.
Secondly, the inhibition effect of the peony petal extract B on the activity of Fatty Acid Synthase (FAS)
An appropriate amount of the peony petal extract B was taken as a test sample solution in DMSO, and the test was performed by the standard measurement method of the fatty acid synthase activity described above.
The results are shown in FIG. 1, which is a comparison of the half inhibitory concentrations of the peony petal extract and the above-mentioned two currently known inhibitors for the fatty acid synthase inhibitory activity. In figure 1, 2 is the peony petal extract B, 4 is cerulenin and 5 is EGCG of the invention of example 2; the unit of ordinate is microgram/ml. The half inhibitory concentration (IC50) of the peony petal extract B of example 2 against fatty acid synthase was 5.63. mu.g/mL, while the half inhibitory concentrations (IC50) of control 1(Cerulenin) and control 2(EGCG) against fatty acid synthase were 24.2. mu.g/mL and 25.8. mu.g/mL, respectively, indicating that the peony petal extract B of example 2 strongly inhibits the activity of fatty acid synthase and that the required effective dose is low.
Influence of peony petal extract B on breast cancer MDA-MB-231 cell and MCF-7 cell viability
The cell viability was measured by the CCK-8 method, which was the same as in example 1.
The results of the effect of the peony petal extract B on the activity of MDA-MB-231 cells of human breast cancer with high fatty acid synthase expression are shown in FIG. 2, wherein the ordinate of FIG. 2 is the relative activity of MDA-MB-231 cells, and the abscissa is: 1 is DMSO-treated group, 3 is inventive extract B (sample concentration is 10. mu.g/ml). The results showed that the relative viability of the cells in the DMSO-treated group was 100% in MDA-MB-231 cells, and that the relative viability of the cells after the treatment with extract B of the invention was 56.3%. Compared with the DMSO-treated group, the peony petal extract B of the embodiment can significantly reduce the MDA-MB-231 cell activity (p <0.05) at the concentration of 10 micrograms/ml.
The effect of the peony petal extract B on MCF-7 cell viability is shown in FIG. 3, in which the vertical axis of FIG. 3 is the relative MCF-7 cell viability, and the horizontal axis is: 1 is DMSO-treated group, 3 is inventive extract B (sample concentration is 10. mu.g/ml). The results showed that the relative viability of the cells in the DMSO-treated group was 100% and that the relative viability of the cells in the extract B of the present invention was 43.2% in MCF-7 cells. Compared with the DMSO-treated group, the peony petal extract B of the embodiment can significantly reduce MCF-7 cell activity (p <0.05) at a concentration of 10 micrograms/ml.
Example 3 preparation of peony petal extract C
Preparation method of peony petal extract C
The preparation method of the peony petal extract C comprises the following steps:
(1) drying and crushing peony petals, and mixing the materials according to a material-liquid ratio of 1 g: adding 12ml formic acid-ethanol-water solution with volume ratio of 4:76:20, performing ultrasonic extraction at 30 deg.C for 3 times, each time for 15min, collecting and mixing extractive solutions to obtain crude extractive solution;
(2) concentrating the crude extract obtained in the step (1) under reduced pressure at 60 ℃ and 0.01MPa until no alcohol smell exists, diluting with water to a relative density of 1.10, adding a 2% by volume potassium hydroxide aqueous solution, adjusting the pH value of the solution to 12.0, filtering to remove insoluble substances, extracting the filtered solution with ethyl acetate for 3 times, and removing the ethyl acetate extract to obtain an alkaline water extract;
(3) acidifying the alkaline water extract obtained in the step (2) with 1M hydrochloric acid aqueous solution, adjusting the pH of the aqueous solution to 3.0 by using a pH meter, extracting for 3 times by using ethyl acetate, combining the 3 times of extract liquor to obtain ethyl acetate extract liquor, removing the solvent and drying to obtain the target peony petal extract C.
Secondly, the inhibition effect of peony petal extract C on Fatty Acid Synthase (FAS) activity
An appropriate amount of the peony petal extract C was dissolved in DMSO as a test sample solution, and the test was performed by the standard measurement method of the fatty acid synthase activity described above.
The results are shown in FIG. 1, which is a comparison of the half inhibitory concentrations of the peony petal extract and the above-mentioned two currently known inhibitors for the fatty acid synthase inhibitory activity. In FIG. 1, 3 is the peony petal extract C, 4 is cerulenin and 5 is EGCG of the invention in example 3; the unit of ordinate is microgram/ml. The half inhibitory concentration (IC50) of the peony petal extract C of example 3 against fatty acid synthase was 5.72. mu.g/mL, while the half inhibitory concentrations (IC50) of control 1(Cerulenin) and control 2(EGCG) against fatty acid synthase were 24.2. mu.g/mL and 25.8. mu.g/mL, respectively, indicating that the peony petal extract C of example 3 strongly inhibits the activity of fatty acid synthase and that the required effective dose is low.
Influence of peony petal extract C on breast cancer MDA-MB-231 cell and MCF-7 cell viability
The cell viability was measured by the CCK-8 method, which was the same as in example 1.
The results of the effect of peony petal extract C on the activity of MDA-MB-231 cells of human breast cancer with high fatty acid synthase expression are shown in FIG. 2, in which the ordinate of FIG. 2 is the relative activity of MDA-MB-231 cells, and in the abscissa: the DMSO-treated group was 1, and the extract C of the present invention was 4 (sample concentrations were 10. mu.g/ml). The results showed that the relative viability of the cells in the DMSO-treated group was 100% in MDA-MB-231 cells, and 47.5% after the treatment with extract C of the invention. Compared with the DMSO-treated group, the peony petal extract C of the embodiment can significantly reduce the MDA-MB-231 cell activity (p <0.05) at the concentration of 10 micrograms/ml.
The effect of peony petal extract C on MCF-7 cell viability is shown in FIG. 3, in which the vertical axis of FIG. 3 is MCF-7 cell relative viability, and the horizontal axis is: the DMSO-treated group was 1, and the extract C of the present invention was 4 (sample concentrations were 10. mu.g/ml). The results showed that the relative viability of the cells in the DMSO-treated group was 100% and that the relative viability of the cells in the extract C of the present invention was 36.8% in MCF-7 cells. Compared with the DMSO-treated group, the peony petal extract C of the embodiment can significantly reduce MCF-7 cell activity (p <0.05) at a concentration of 10 micrograms/ml.
The application of the peony petal extract in the embodiments 1-3 in preparing the medicine with the weight-losing or anticancer effect.
The application of the peony petal extract in the embodiments 1-3 in preparing a health food with weight-losing or anticancer effects.
The application of the peony petal extract in the embodiments 1-3 in preparing functional cosmetics with weight-losing or anticancer effects.
The above-described embodiments should not be construed as limiting the scope of the invention. Those skilled in the art will appreciate that various modifications, combinations, sub-combinations, and substitutions can occur, depending on design requirements and other factors. Any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (7)
1. The preparation method of the peony petal extract comprises the following steps:
(1) drying and crushing peony petals, adding an organic acid-organic solvent-water mixed solvent for ultrasonic extraction, and collecting and combining extracting solutions to obtain a crude extracting solution;
(2) concentrating the crude extract obtained in the step (1) under reduced pressure until no organic solvent smell exists, diluting with water until the relative density is 1.10-1.20, adding an alkaline water solution, adjusting the pH value of the solution to 11.0-13.0, filtering to remove insoluble substances, extracting the filtered solution with ethyl acetate, and removing the ethyl acetate extract to obtain an alkaline water extract;
(3) acidifying the alkaline water extract obtained in the step (2) with acid water until the pH is 1.0-3.0, extracting with ethyl acetate to obtain ethyl acetate extract, removing the solvent and drying to obtain a target extract;
the organic acid-organic solvent-water mixed solvent in the step (1) is selected from any one of malic acid, acetic acid and formic acid, and the volume percentage concentration of the organic acid is 3% -5%; wherein the organic solvent is ethanol, and the volume percentage concentration of the organic solvent is 67-80%;
the material-liquid ratio of the peony petals to the organic acid-organic solvent-water mixed solvent is 1 g: 12 ml-1 g: 20 ml.
2. The method for preparing a peony petal extract according to claim 1, wherein the method comprises the steps of:
the ultrasonic extraction conditions in the step (1) are as follows: ultrasonic extraction is carried out for 1-3 times at the temperature of 20-30 ℃, and each time lasts for 10-30 min.
3. The method for preparing a peony petal extract according to claim 1, wherein the method comprises the steps of:
the alkaline water in the step (2) is sodium hydroxide or potassium hydroxide aqueous solution; the volume percentage concentration of the alkaline water solution is 1-3%;
the vacuum concentration in the step (2) is carried out under the conditions that the temperature is 30-60 ℃ and the pressure is 0.01-0.1 MPa until no alcohol smell exists.
4. The method for preparing a peony petal extract according to claim 1, wherein the method comprises the steps of:
the acid water in the step (3) is hydrochloric acid water solution with the concentration of 0.1M-1M.
5. A peony petal extract produced by the method of any one of claims 1 to 4.
6. Use of a peony petal extract produced by the method of any one of claims 1-4 in the preparation of a product for inducing apoptosis in cancer cells; the cancer cell is breast cancer cell MDA-MB-231 or MCF-7.
7. Use of the peony petal extract prepared by the method of any one of claims 1 to 4 in the preparation of a medicament or cosmetic having an anti-breast cancer effect.
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