Preparation process of peony seed extract with high paeoniflorin and resveratrol content

Abstract

The invention discloses a preparation process of peony seed extract with high paeoniflorin and resveratrol content, and belongs to the technical field of paeoniflorin and resveratrol extraction. The technical problem to be solved is to improve the content of resveratrol and paeoniflorin in the peony seed extract. According to the technical scheme, peony seeds are smashed, soaked, subjected to alcohol extraction, filtered, concentrated and dried; a solvent for alcohol extraction is an ethanol solution with the volume fraction of 70-80%, and the solid-liquid ratio of alcohol extraction is 1g: (15-20) g.

Description

Technical Field

[0001] This invention provides a preparation process for a peony seed extract high in paeoniflorin and resveratrol, and designs the field of paeoniflorin and resveratrol extraction technology. Background Technology

[0002] For understanding the technical content of this invention: peony( Paeonia suffruticosaAndr Peony (Paeonia lactiflora) is a perennial deciduous shrub belonging to the Ranunculaceae family and the Paeonia genus. It contains various active ingredients, mainly including flavonoids, polyphenols, proteins, and volatile oils. Golden camellia (Paeonia suffruticosa) is a traditional medicinal and edible plant; related studies have shown that peony has antibacterial, antioxidant, antithrombotic, and antitumor effects.

[0003] Relevant patent documents retrieved: This document, published in China (CN 106265977 A) on January 4, 2017, discloses peony seed extract, its preparation method, and applications, including a topical skin preparation. The preparation method includes: Method 1, mixing pulverized peony seeds with a 60-80% ethanol aqueous solution, extracting at 10-30°C for 0.5-2 hours, separating the solid and liquid phases to obtain a filtrate, and removing the solvent to obtain a crude peony seed extract; Method 2, loading the crude peony seed extract from Method 1 onto a macroporous resin column, and sequentially eluting with deionized water, a 10-25% ethanol aqueous solution, a 30-45% ethanol aqueous solution, a 50-65% ethanol aqueous solution, a 70-85% ethanol aqueous solution, and a 95% or higher ethanol aqueous solution, collecting the eluent and removing the solvent to obtain the respective eluted fractions. However, this method cannot increase the content of resveratrol and paeoniflorin.

[0004] Relevant non-patent literature retrieved: The journal or book title is "Food and Nutritional Science," and the document title is "Analysis of Components and Activity of Ethanol Extract from Peony Processing Residue, a New Resource Food," volume number 2021, 10(2), 133-147, published on May 26, 2021. This document discloses the following method: 40.0 g of peony seed shell powder was mixed with a certain volume of ethanol solution in a round-bottom flask, refluxed for extraction, and centrifuged at 5000 r / min for 8 min after a certain time. The supernatant was then removed and the solvent ethanol was evaporated under negative pressure using a rotary evaporator. The residue was identified as peony seed shell extract, labeled as PEO, and stored at -20℃ for later use. This document also tested the contents of resveratrol and paeoniflorin, and conducted experiments on the effects of ethanol concentration, material-to-liquid ratio, extraction temperature, and extraction time on the PEO yield.

[0005] Experiments showed that the PEO yield increased continuously with increasing ethanol concentration. 95% ethanol was selected as the optimal extraction concentration. Within a certain range, the PEO yield increased first with increasing solid-liquid ratio, reaching its highest value at a ratio of 1:10. The PEO yield showed a positive correlation with temperature, meaning higher temperatures resulted in higher yields, peaking at 90℃ and stabilizing at 100℃. Between 2 and 4 hours, the PEO yield showed a positive correlation with time, reaching its maximum at 4 hours; further increases in extraction time did not lead to further yield increases. However, while this extraction process increased the PEO yield, it neglected to improve the content of resveratrol and paeoniflorin. The resveratrol content in PEO was only 0.014%, and the paeoniflorin content was only 1.40%, which were relatively low.

[0006] The prior art represented by the aforementioned literature has at least the following unresolved technical problems or defects: Existing technologies cannot increase the content of resveratrol and paeoniflorin in peony seed extract. Relevant evidence is that the resveratrol content in the study "Analysis of Components and Activity of Ethanol Extract from Peony Processing Residues (New Resource Food)" is 0.014% and the paeoniflorin content is 1.40%, which are relatively low. Summary of the Invention

[0007] The purpose of this invention is to provide: A preparation process for a peony seed extract high in paeoniflorin and resveratrol, and related technologies, to solve technical problems such as increasing the content of resveratrol and paeoniflorin in peony seed extract, or combinations thereof.

[0008] Terminology Explanation: Unless otherwise defined, all technical terms in this document have the same meanings as commonly understood by one of ordinary skill in the art to which the subject matter of the claims pertains. Unless otherwise stated, all patents, patent inventions, and publications cited in this document are incorporated herein by reference in their entirety. If multiple definitions exist for terms in this document, the definitions in this chapter shall prevail.

[0009] It should be understood that the above brief description and the following detailed description are exemplary and for illustrative purposes only, and do not limit the subject matter of the invention in any way. In this invention, the singular is used in conjunction with the plural unless otherwise specifically stated. It should also be noted that, unless otherwise stated, the use of “or” or “or” means “and / or”. Furthermore, the use of the term “comprising” and other forms such as “including,” “containing,” and “contains” are not limiting.

[0010] The definitions of standard chemical terms can be found in the reference "Pharmacopoeia of the People's Republic of China (2020 Edition): China Medical Science and Technology Press: May 2020: 1st Edition."

[0011] Unless otherwise specified, conventional methods within the scope of the art, such as DPPH radical inhibition assays and HPLC detection, shall be used.

[0012] Unless specifically defined herein, the use of all commercially available products described herein follows standard techniques. For example, they may be implemented using the manufacturer's instructions for use with the kit, or in accordance with methods known in the art or the description of this invention. Generally, references can be made to the numerous general and more specific references cited and discussed in this specification.

[0013] The term "extraction" as used in this article refers to the process of separating a target substance from a mixture and transferring it to another solvent by utilizing the difference in solubility of the substance in different solvents.

[0014] The term "paeoniflorin" as used in this article refers to: 4'-O-demethylpaeoniflorin, with the molecular formula C0. 23 H 28 O 11 Its molecular weight is 456.46.

[0015] The term "resveratrol" as used in this article refers to a natural polyphenol compound with the chemical name 3,5,4'-trihydroxystilbene and the molecular formula C2. 14 H 12 O3 has a molecular weight of 228.24. It is mainly found in plants such as grapes, Japanese knotweed, and mulberries.

[0016] The term "peony seed" as used in this article refers to the seed of the peony (Paeonia suffruticosa Andr.), which belongs to the genus Paeonia in the family Paeoniaceae. Peony seeds are usually oval or ovate in shape, with a wrinkled surface, and are dark brown or blackish-brown in color.

[0017] As used herein, the term "normal temperature" refers to: In this invention, "normal temperature" refers to a typical ambient temperature, ranging from 20°C to 30°C. In some embodiments, "normal temperature" refers to a temperature ranging from 20°C to 30°C; in other embodiments, "normal temperature" refers to a temperature ranging from 25°C to 30°C; and in still other embodiments, "normal temperature" refers to 10°C, 15°C, 20°C, 25°C, 30°C, etc.

[0018] In a first aspect, the present invention provides a process for preparing a peony seed extract high in paeoniflorin and resveratrol, comprising the following steps: pulverizing, soaking, extracting with alcohol, filtering, concentrating and drying the peony seeds; wherein the solvent for the alcohol extraction is an ethanol solution with a volume fraction of 70-80%, and the material-to-liquid ratio for the alcohol extraction is 1g:(15-20)g.

[0019] The preferred technical feature of the ethanol solution is an aqueous solution containing 70-80% by volume of ethanol.

[0020] The preferred embodiment of the technical feature is an ethanol solution containing an aqueous solution of 75-80% by volume.

[0021] The preferred embodiment of the technical feature is an ethanol solution containing an aqueous solution with a volume fraction of 75% ethanol.

[0022] Among them, the technical feature of soaking is: the soaking time is 20-30 hours and the temperature is 20-30℃.

[0023] The soaking time for the technical features is further preferably 20h, 21h, 22h, 23h, 24h, 25h, 26h, 27h, 28h, 29h, 30h, or any combination thereof.

[0024] The soaking time for the technical features is further preferably 24 hours.

[0025] Among them, the technical feature extraction: the temperature for alcohol extraction is 80-100℃.

[0026] The preferred temperatures for alcohol extraction are 80℃, 85℃, 90℃, and 100℃.

[0027] The preferred temperature for alcohol extraction is 80℃.

[0028] Among them, the technical feature is alcohol extraction: the alcohol extraction time is 200-250 min.

[0029] The preferred extraction time for alcohol extraction is 210-250 min.

[0030] The extraction time for the technical features is further optimized to be 210 min, 220 min, 230 min, 240 min, and 250 min.

[0031] The preferred extraction time for alcohol extraction is 210 min.

[0032] Among the technical features, the alcohol extraction process involves a mass ratio of peony seeds to solvent of 1:(10-20). Preferably, the mass ratio of peony seeds to solvent is 1:(15-20).

[0033] The preferred mass ratio of peony seeds to solvent is 1:15, 1:16; 1:17, 1:18, 1:19, 1:20.

[0034] The preferred technical feature is a mass ratio of peony seeds to solvent of 1:20.

[0035] Secondly, the present invention provides a process for preparing a peony seed extract with high levels of paeoniflorin and resveratrol, comprising the following steps: pulverizing peony seeds, soaking, water extraction, alcohol precipitation, filtering, and drying. The ratio of material to liquid in the water extraction is 1g:(15-20)g.

[0036] Among them, the technical feature of soaking is: the soaking time is 20-30 hours and the temperature is 20-30℃.

[0037] The soaking time for the technical features is further preferably 20h, 21h, 22h, 23h, 24h, 25h, 26h, 27h, 28h, 29h, 30h, or any combination thereof.

[0038] The soaking time for the technical features is further preferably 24 hours.

[0039] Among them, the technical feature is water extraction: the water extraction temperature is 80-100℃.

[0040] The preferred temperatures for water extraction are 80℃, 85℃, 90℃, and 100℃.

[0041] The preferred temperature for water extraction is 80℃.

[0042] Among them, the technical feature extraction: the water extraction time is 200-250 min.

[0043] The preferred water extraction time is 210-250 min.

[0044] The water extraction time for the technical characteristics is further optimized to be 210 min, 220 min, 230 min, 240 min, and 250 min.

[0045] The preferred water extraction time is 210 min.

[0046] Among the technical features, water extraction: In water extraction, the mass ratio of peony seeds to solvent is 1:(10-20). Preferably, the mass ratio of peony seeds to solvent is 1:(15-20).

[0047] The preferred mass ratio of peony seeds to solvent is 1:15, 1:16; 1:17, 1:18, 1:19, 1:20.

[0048] The preferred technical feature is a mass ratio of peony seeds to solvent of 1:20.

[0049] Among them, the technical feature of concentration is that the filtrate is concentrated to 1 / 10 to 1 / 8 of its original volume.

[0050] Among them, the technical feature is alcohol precipitation: ethanol is added and the mixture is allowed to stand at low temperature.

[0051] Among them, the amount of ethanol is 2-6 times the mass of peony seeds.

[0052] Among them, the technical characteristic is low temperature: 0-4℃.

[0053] Among them, the technical feature is that the settling time is 24-48 hours.

[0054] Thirdly, the present invention provides: a peony seed extract, comprising at least one of the peony seed extracts prepared by the above-described preparation process.

[0055] The mass ratio of the peony seed extract prepared by the first preparation process to the peony seed extract prepared by the second preparation process is (1-2):(1-2).

[0056] Fourthly, the present invention provides the application of the above-mentioned preparation process in improving the quality of peony seed extract.

[0057] Among them, the technical feature that improves the quality of peony seed extract is to increase the content of paeoniflorin and resveratrol in peony seed extract.

[0058] The beneficial effects of this invention are as follows: The present invention has at least the following beneficial effects: 1. Compared with the prior art, the present invention has better technical effects in increasing the content of resveratrol and paeoniflorin in peony seed extract.

[0059] According to experimental tests, the present invention increases the resveratrol content and paeoniflorin content from 0.014% and 1.40% in the prior art to over 0.123% and 3.7%.

[0060] Furthermore, based on the present invention: 1. Based on the comparison of Example 1 and Comparative Examples 1-5, the present invention employs a combination of specific solvent conditions for alcohol precipitation, specific ethanol volume concentration, and specific material-to-liquid ratio to achieve a resveratrol content of 0.208% and a paeoniflorin content of over 3.7%. The combined technical effect is superior to the sum of the effects of each individual technical method. Detailed Implementation

[0061] The following non-limiting embodiments are intended to enable those skilled in the art to gain a more comprehensive understanding of the present invention, but do not limit the invention in any way. The following content is merely an exemplary description of the scope of protection claimed by the present invention, and those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, and such changes should also fall within the scope of protection claimed by the present invention.

[0062] The present invention will be further described below by way of specific embodiments. Unless otherwise specified, all instruments, devices, equipment, reagents, products, etc., used in the embodiments of the present invention are obtained through conventional commercial means.

[0063] Unless otherwise specified, the ethanol described in this invention refers to an aqueous solution of ethanol with a corresponding volume percentage.

[0064] Example 1 (1) Take peony seeds and crush them, then collect the crushed powder.

[0065] (2) Take 20g of peony seed powder, add 75% ethanol at a material-to-liquid ratio of 1:20, soak at room temperature for 24h, extract by hot reflux at 80°C for 210min, extract by hot reflux three times, and combine the extracts.

[0066] (3) Use qualitative filter paper to filter out the medicinal residue.

[0067] (4) The filtrate is concentrated and dried to obtain a solid powder.

[0068] Example 2 (1) Take peony seeds and crush them, then collect the crushed powder.

[0069] (2) Take 20g of peony seed powder, add pure water at a material-to-liquid ratio of 1:20, soak at room temperature for 24h, extract by hot reflux at 100°C for 210min, extract by hot reflux three times, and combine the extracts.

[0070] (3) Use qualitative filter paper to filter out the medicinal residue.

[0071] (4) After the filtrate is concentrated to 1 / 10 of its original volume, add 4 times the weight of anhydrous ethanol of peony seeds, let it stand at 4°C for 48 hours, take the supernatant, concentrate and dry to obtain solid powder.

[0072] Example 3 (1) Take peony seeds and crush them, then collect the crushed powder.

[0073] (2) Take 20g of peony seed powder, add 75% ethanol at a material-to-liquid ratio of 1:20, soak at room temperature for 6 hours, reflux extract at 80°C for 210 minutes, reflux extract three times, and combine the extracts.

[0074] (3) Use qualitative filter paper to filter out the medicinal residue.

[0075] (4) The filtrate is concentrated and dried to obtain a solid powder.

[0076] Example 4 (1) Take peony seeds and crush them, then collect the crushed powder.

[0077] (2) Take 20g of peony seed powder, add 75% ethanol at a material-to-liquid ratio of 1:20, soak at room temperature for 12h, extract by hot reflux at 80°C for 210min, extract by hot reflux three times, and combine the extracts.

[0078] (3) Use qualitative filter paper to filter out the medicinal residue.

[0079] (4) The filtrate is concentrated and dried to obtain a solid powder.

[0080] Comparative Example 1 (1) Take peony seeds and crush them, then collect the crushed powder.

[0081] (2) Take 20g of peony seed powder, add pure water at a material-to-liquid ratio of 1:20, soak at room temperature for 24h, extract by hot reflux at 100°C for 210min, extract by hot reflux three times, and combine the extracts.

[0082] (3) Use qualitative filter paper to filter out the medicinal residue.

[0083] (4) The filtrate is concentrated and dried to obtain a solid powder.

[0084] Comparative Example 2 (1) Take peony seeds and crush them, then collect the crushed powder.

[0085] (2) Take 20g of peony seed powder, add 95% ethanol at a material-to-liquid ratio of 1:20, soak at room temperature for 24h, extract by hot reflux at 80°C for 210min, extract by hot reflux three times, and combine the extracts.

[0086] (3) Use qualitative filter paper to filter out the medicinal residue.

[0087] (4) The filtrate is concentrated and dried to obtain a solid powder.

[0088] Comparative Example 3 (1) Take peony seeds and crush them, then collect the crushed powder.

[0089] (2) Take 20g of peony seed powder, add pure water at a ratio of 1:10, soak at room temperature for 24h, extract by hot reflux at 100°C for 210min, extract by hot reflux three times, and combine the extracts.

[0090] (3) Use qualitative filter paper to filter out the medicinal residue.

[0091] (4) After the filtrate is concentrated to 1 / 10 of its original volume, add 4 times the weight of anhydrous ethanol of peony seeds, let it stand at 4°C for 48 hours, take the supernatant, concentrate and dry to obtain solid powder.

[0092] Comparative Example 4 (1) Take peony seeds and crush them, then collect the crushed powder.

[0093] (2) Take 20g of peony seed powder, add pure water at a material-to-liquid ratio of 1:20, soak at room temperature for 24h, extract by hot reflux at 100°C for 210min, extract by hot reflux three times, and combine the extracts.

[0094] (3) Use qualitative filter paper to filter out the medicinal residue.

[0095] (4) After the filtrate is concentrated to 1 / 10 of its original volume, 7 times the weight of anhydrous ethanol of peony seeds is added. The mixture is allowed to stand at 4°C for 48 hours. The supernatant is then collected, concentrated, and dried to obtain a solid powder.

[0096] Comparative Example 5 (1) Take peony seeds and crush them, then collect the crushed powder.

[0097] (2) Take 20g of peony seed powder, add pure water at a material-to-liquid ratio of 1:20, soak at room temperature for 24h, extract by hot reflux at 100°C for 210min, extract by hot reflux three times, and combine the extracts.

[0098] (3) Use qualitative filter paper to filter out the medicinal residue.

[0099] (4) Add the filtrate directly to anhydrous ethanol at a ratio of 4 times the mass of the peony seed powder, let it stand at a low temperature of 4°C for 48 hours, take the supernatant, concentrate and dry to obtain solid powder.

[0100] Comparative Example 6 (1) Take peony seeds and crush them, then collect the crushed powder.

[0101] (2) Take 20g of peony seed powder, add pure water at a material-to-liquid ratio of 1:20, soak at room temperature for 24h, extract by hot reflux at 100°C for 210min, extract by hot reflux three times, and combine the extracts.

[0102] (3) Use qualitative filter paper to filter out the medicinal residue.

[0103] (4) After the filtrate is concentrated to 1 / 10 of its original volume, add 4 times the weight of anhydrous ethanol of peony seeds, let it stand at 4°C for 12 hours, take the supernatant, concentrate and dry to obtain solid powder.

[0104] Comparative Example 7 (1) Take peony seeds and crush them, then collect the crushed powder.

[0105] (2) Take 20g of peony seed powder, add pure water at a material-to-liquid ratio of 1:20, soak at room temperature for 24h, extract by hot reflux at 100°C for 210min, extract by hot reflux three times, and combine the extracts.

[0106] (3) Use qualitative filter paper to filter out the medicinal residue.

[0107] (4) After the filtrate is concentrated to 1 / 10 of its original volume, add 4 times the weight of anhydrous ethanol of peony seeds, let it stand at 4°C for 24 hours, take the supernatant, concentrate and dry to obtain solid powder.

[0108] Comparative Example 8 (1) Take peony seeds and crush them, then collect the crushed powder.

[0109] (2) Take 20g of peony seed powder, add pure water at a material-to-liquid ratio of 1:20, soak at room temperature for 6 hours, extract by hot reflux at 100°C for 210 minutes, extract by hot reflux three times, and combine the extracts.

[0110] (3) Use qualitative filter paper to filter out the medicinal residue.

[0111] (4) After the filtrate is concentrated to 1 / 10 of its original volume, add 4 times the weight of anhydrous ethanol of peony seeds, let it stand at 4°C for 48 hours, take the supernatant, concentrate and dry to obtain solid powder.

[0112] Comparative Example 9 (1) Take peony seeds and crush them, then collect the crushed powder.

[0113] (2) Take 20g of peony seed powder, add pure water at a material-to-liquid ratio of 1:20, soak at room temperature for 12h, extract by hot reflux at 100°C for 210min, extract by hot reflux three times, and combine the extracts.

[0114] (3) Use qualitative filter paper to filter out the medicinal residue.

[0115] (4) After the filtrate is concentrated to 1 / 10 of its original volume, add 4 times the weight of anhydrous ethanol of peony seeds, let it stand at 4°C for 48 hours, take the supernatant, concentrate and dry to obtain solid powder.

[0116] Comparative Example 10 (1) Take peony seeds and crush them, then collect the crushed powder.

[0117] (2) Take 20g of peony seed powder, add pure water at a ratio of 1:20, soak at room temperature for 36h, extract by hot reflux at 100°C for 210min, extract by hot reflux three times, and combine the extracts.

[0118] (3) Use qualitative filter paper to filter out the medicinal residue.

[0119] (4) After the filtrate is concentrated to 1 / 10 of its original volume, add 4 times the weight of anhydrous ethanol of peony seeds, let it stand at 4°C for 48 hours, take the supernatant, concentrate and dry to obtain solid powder.

[0120] Example 1: In vitro antioxidant effect experiment (DPPH free radical inhibition experiment) (1) Preparation of DPPH solution: Weigh 20 mg of DPPH, dissolve it in anhydrous ethanol, and dilute to a final volume of 250 mL in a volumetric flask. The DPPH concentration is prepared to be 2 × 10⁻⁶ mg / mL. -4 mol / L; store protected from light at 0-4℃, prepare and use immediately, effective within 4 hours. (Positive control: Vitamin C: 1 mg / mL) (2) Preparation of the test solution: Comparative Examples 1, 3, 4, 5, and Example 2 used pure water as the solvent, while the remaining examples and comparative examples used ethanol as the solvent to prepare 1 mg / mL test solutions. (The type of solvent does not affect the detection results; however, for better dissolution, water was used for water extraction experiments, and ethanol was used for alcohol extraction experiments, and the same applies below.) (3) Experimental steps: 1) Take 1 mL of the test solution and 1 mL of 2×10⁻⁶ solution. -4 Mix the mol / L DPPH solution thoroughly (tube A). 2) Take 1 mL of solvent and 1 mL of 2×10 -4 Mix the mol / L DPPH solution thoroughly (tube B). 3) Mix 1 mL of solvent with 1 mL of the test solution (tube C). 4) After reacting in the dark for 30 minutes, measure the absorbance values ​​of tubes A, B, and C at 517 nm.

[0121] The reagent ratios for tubes A, B, and C are shown in Table 1.

[0122] Table 1

[0123] (4) DPPH free radical inhibition rate calculation formula: DPPH inhibition rate (%) = (B+CA) / B×100%, where A, B and C represent the absorbance values ​​of tubes A, B and C respectively.

[0124] (5) The results of the DPPH free radical inhibition experiment are shown in Table 2.

[0125] Table 2

[0126] The combination example is a mixture of 50 wt% Example 1 and 50 wt% Example 2.

[0127] Test Example 2: Detection of Paeoniflorin Content 1) Sample pretreatment methods for HPLC detection: 1. Comparative Examples 1, 3, 4, 5 and Example 2 were prepared into 1 mg / ml solutions using pure water as the solvent and filtered through a 0.22 μm organic filter membrane before testing.

[0128] 2. In the remaining examples and comparative examples, ethanol was used as the solvent to prepare a 1 mg / ml solution, which was then filtered through a 0.22 μm organic filter membrane before testing.

[0129] 2) Determination of standard curve: Chromatographic column: The packing material is octadecylsilane-bonded silica gel with a particle size of 5 μm; the column length is 250 mm and the inner diameter is 4.6 mm.

[0130] Mobile phase: Mobile phase A is pure water, mobile phase B is acetonitrile; gradient elution; injection volume 10 μl; flow rate 1 ml / min; column temperature 30℃; detection wavelength UV 230 nm; plot the standard curve. The gradient elution program is shown in Table 3.

[0131] Table 3

[0132] 3) Data processing The paeoniflorin content was calculated using a standard curve, as shown in Table 4.

[0133] Table 4

[0134] Test Example 3: Resveratrol Content Detection 1) Sample pretreatment methods for HPLC detection: 1. Comparative Examples 1, 3, 4, 5 and Example 2 were prepared into 1 mg / ml solutions using pure water as the solvent and filtered through a 0.22 μm organic filter membrane before testing.

[0135] 2. In the remaining examples and comparative examples, ethanol was used as the solvent to prepare a 1 mg / ml solution, which was then filtered through a 0.22 μm organic filter membrane before testing.

[0136] 2) Determination of standard curve: Chromatographic column: The packing material is octadecylsilane-bonded silica gel with a particle size of 5 μm; the column length is 250 mm and the inner diameter is 4.6 mm.

[0137] Mobile phase: Mobile phase A is pure water, mobile phase B is acetonitrile; gradient elution; injection volume 10 μl; flow rate 1 ml / min; column temperature 30℃; detection wavelength UV 230 nm; plot the standard curve. The gradient elution program is shown in Table 5.

[0138] Table 5

[0139] 3) Data processing: The resveratrol content was calculated using a standard curve, as shown in Table 6.

[0140] Table 6

[0141] Verification of technical effectiveness and / or analysis of technical problem solving The above experimental results demonstrate that, using the preparation process described in this invention, with peony seeds as raw material, the extraction process with the highest content of paeoniflorin and resveratrol was successfully selected as 75% ethanol extraction. This process yields ideal results, has a high content of active ingredients, and exhibits excellent antioxidant properties.

[0142] The peony seed extract obtained by mixing the alcohol extract and the water extract of the present invention was used to obtain a combination example. Under the same test solution concentration, the DPPH inhibition rate of the combination example was significantly improved.

[0143] Comparative Example 1 did not undergo alcohol precipitation after water extraction, and its resveratrol content was 0.18. Unlike Example 2, it did not enrich the target component by adding anhydrous ethanol and allowing it to stand at low temperature, which ultimately resulted in a low resveratrol content in the extract.

[0144] Comparative Example 2 changed the ethanol concentration of the alcohol extract, and its DPPH inhibition rate, paeoniflorin and resveratrol content were lower than those of the embodiments of the present invention.

[0145] Comparative Example 3 changed the material-to-liquid ratio, and its DPPH inhibition rate, paeoniflorin and resveratrol content were lower than those in the embodiments of the present invention.

[0146] Comparative Examples 4-5 changed the conditions of alcohol precipitation, and their DPPH inhibition rate, paeoniflorin and resveratrol content were lower than those in the embodiments of the present invention.

[0147] Comparative Examples 6-7, which varied the alcohol precipitation time, had lower DPPH inhibition rates, paeoniflorin content, and resveratrol content than those in the embodiments of the present invention.

[0148] Comparative Examples 8-10, which varied the soaking time, had lower DPPH inhibition rates, paeoniflorin content, and resveratrol content than the embodiments of the present invention.

[0149] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A preparation process for a peony seed extract high in paeoniflorin and resveratrol, characterized in that, Includes the following steps: Peony seeds are crushed, soaked, extracted with alcohol, filtered, concentrated and dried; the solvent for alcohol extraction is an ethanol solution with a volume fraction of 70-80%, and the material-to-liquid ratio for alcohol extraction is 1g:(15-20)g.

2. The preparation process according to claim 1, characterized in that, The soaking time is 20-30 hours, and the soaking temperature is 20-30°C; and / or The alcohol extraction is performed at a temperature of 80-100℃ for 200-250 minutes.

3. A preparation process for a peony seed extract high in paeoniflorin and resveratrol, characterized in that, Includes the following steps: The peony seeds were crushed, soaked, extracted with water, precipitated with alcohol, filtered, and dried. The ratio of material to liquid in the water extraction is 1g:(15-20)g.

4. The preparation process according to claim 3, characterized in that, The soaking time is 20-30 hours, and the soaking temperature is 20-30°C; and / or The water extraction temperature is 80-100℃, and the time is 200-250 minutes.

5. The preparation process according to claim 3, characterized in that, The alcohol precipitation is achieved by adding ethanol and allowing it to stand at a low temperature.

6. The preparation process according to claim 5, characterized in that, The amount of ethanol is 2-6 times the mass of the peony seeds.

7. The preparation process according to claim 5, characterized in that, The low temperature is 0-4°C; and / or The settling time is 24-48 hours.

8. A peony seed extract, characterized in that, It includes at least one of the peony seed extracts prepared by the preparation process according to any one of claims 1-2 and the peony seed extracts prepared by the preparation process according to any one of claims 3-7.

9. The application of the preparation process according to any one of claims 1-2 or the preparation process according to any one of claims 3-7 in improving the quality of peony seed extract.

10. The application according to claim 9, characterized in that, The improvement of the quality of peony seed extract refers to increasing the content of paeoniflorin and resveratrol in the peony seed extract.

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