CN114053339A

CN114053339A - Peony oil and preparation method and application thereof

Info

Publication number: CN114053339A
Application number: CN202111276443.XA
Authority: CN
Inventors: 翟春涛; 金敏蓉; 张大勇
Original assignee: Zhejiang Yige Enterprise Management Group Co ltd; Shanghai Zhina Biotechnology Co ltd
Current assignee: Zhejiang Yige Enterprise Management Group Co ltd; Shanghai Zhina Biotechnology Co ltd
Priority date: 2021-10-29
Filing date: 2021-10-29
Publication date: 2022-02-18

Abstract

The invention discloses peony oil and a preparation method and application thereof. The preparation method of the peony oil comprises the following steps: grinding peony petals and peony seeds, standing and squeezing; wherein the standing temperature is 40-70 ℃, and the standing time is 12-24 h. The preparation method has simple steps, is suitable for large-scale production requirements, and is safe. The prepared peony oil simultaneously reserves the components of peony petals and peony seeds, has good effect of repairing skin barrier, and can rapidly induce keratinocyte proliferation. The peony oil is applied to a skin external preparation, and the effect of repairing skin barriers is obviously improved.

Description

Peony oil and preparation method and application thereof

Technical Field

The invention relates to the technical field of traditional Chinese medicines, and particularly relates to peony oil and a preparation method and application thereof.

Background

Peony is a flower of Paeonia suffruticosa Andr, an important ornamental plant, and belongs to one of the ten famous flowers in China. Peony was used as a national flower in China, and has been called "the natural fragrance of China" because it is big and fragrant. Modern researches show that the peony extract has the skin care effects of resisting oxidation, whitening and the like. Most of peony extracts commonly used in cosmetics are peony hydrolat and peony water extract. The peony water extract only contains water-soluble components; the peony hydrolat is obtained by distilling peony with water, and only contains a small amount of peony volatile components.

The peony can be divided into oil peony and ornamental peony according to different purposes, and the peony seed oil mainly comes from the oil peony. The peony seed oil has extremely high nutritive value and no acute toxicity, hereditary toxicity and subacute toxicity, so the peony seed oil is known as 'liquid gold'. Peony seed oil contains mainly about 96% of triglyceride and 4% of diglyceride, and a part of free fatty acids, the main fatty acids consisting of palmitic acid, stearic acid, oleic acid, linoleic acid and linolenic acid. Meanwhile, the peony seed oil also contains unsaponifiable matters, such as sterol compounds, fat-soluble vitamin E, squalene and the like. Modern researches show that the peony seed oil has the main effects of resisting oxidation, preventing sunburn, preserving moisture, treating skin scald, repairing skin barrier and the like on skin.

The extraction method of peony essential oil in the prior art comprises distillation (Chinese patent document CN101130712B), petroleum ether extraction (Chinese patent document CN101130714B), and supercritical CO₂Extraction method (Chinese patent application CN101294119A, Chinese patent application CN1012391913A) and method for extracting peony oil with water (Chinese patent document CN 102766535B). However, these extraction methods are complicated in steps and harsh in conditionsThe extraction efficiency is not high, and meanwhile, organic solvent residue possibly exists, so that the method is not suitable for the requirements of mass production and cosmetic safety.

Therefore, the development of a preparation method of peony oil which has simple extraction steps, is suitable for large-scale production requirements, is safe and has good effects of skin barrier repair and keratinocyte proliferation induction is urgently needed.

Disclosure of Invention

The invention aims to solve the technical problems that the method for extracting the peony oil in the prior art is complicated in steps and harsh in conditions, and meanwhile, the method possibly has organic solvent residues, is not suitable for large-scale production and is safe for cosmetics, and provides the peony oil and the preparation method and application thereof.

The peony flower oil simultaneously retains the components of peony petals and peony seed oil, has good effect of repairing skin barrier, and can rapidly induce keratinocyte proliferation. The method for preparing the peony oil has simple steps, is suitable for large-scale production and is safe. The peony oil is applied to a skin external preparation, and the effect of repairing skin barriers is obviously improved.

The present invention provides the following technical solutions to solve the above technical problems.

The invention provides a preparation method of peony oil, which comprises the following steps:

grinding peony petals and peony seeds, standing and squeezing;

wherein the standing temperature is 40-70 ℃, and the standing time is 12-24 h.

In the present invention, the temperature of the standing is preferably 50 ℃ to 60 ℃, for example, 55 ℃.

In the present invention, the time for the standing is preferably 14h to 18h, for example 18 h.

In the invention, the weight portion of the peony petals can be 1-99 parts, and preferably 5-20 parts.

In the invention, the weight portion of the peony seeds can be 1-99 parts, and preferably 80-95 parts.

In the invention, the dosage ratio of the peony petals to the peony seeds in parts by weight can be (1-4): (16-19), preferably (1-2): (8-9), more preferably 1: 9.

in the invention, the peony petals are preferably cleaned and dried peony petals. Wherein the washing conditions may be conventional in the art, such as washing with water at normal temperature. The drying conditions may be conventional in the art, e.g. by wiping.

In the invention, the peony seeds are preferably cleaned and dried peony seeds. Wherein the washing conditions may be conventional in the art, such as washing with water at normal temperature. The drying conditions may be conventional in the art, for example air drying.

In the grinding process, the peony petals and the peony seeds can be respectively ground, or a mixture of the peony petals and the peony seeds can be directly ground.

In the present invention, the milling conditions may be conventional in the art, for example, milling with a colloid mill.

The colloid mill may be a conventional commercially available product, such as JM-F65, available from MacToron machines, Inc. of Winzhou.

In the present invention, the pressing conditions may be conventional in the art, for example, pressing with a static pressure type cold press. The static pressure cold Press may be a conventional commercial product, such as Carver Laboratory Press available from freds.

In the present invention, the pressing pressure is preferably 45 to 65MPa, for example, 55 MPa.

In the present invention, after the pressing, a step of oil-water separation of the pressed material is generally further included.

The water in the oil-water mixture to be subjected to oil-water separation is derived from the water in the peony petals.

The conditions for the oil-water separation treatment may be conventional in the art, for example, using a separatory funnel for oil-water separation.

In the invention, no solvent is preferably added in the preparation process of the peony oil. The solvent is generally water and/or an oily organic solvent.

In a preferred embodiment of the present invention, the preparation method of peony oil comprises the following steps:

step 1, washing 5-20 parts of peony petals and 80-95 parts of peony seeds with normal-temperature water, wiping the peony petals dry, and airing the peony seeds;

step 2, grinding and crushing the peony petals wiped in the step 1 and the aired peony seeds and uniformly mixing;

step 3, standing the mixture obtained in the step 2 at the temperature of 40-70 ℃ for 12-24 h;

and 4, squeezing the mixture obtained in the step 3, separating oil from water, and taking an oil phase part to obtain the peony oil.

The invention also provides the peony flower oil prepared by the preparation method.

The invention also provides application of the peony oil as an antioxidant active ingredient or an active ingredient for repairing skin barrier in preparing a skin external preparation.

In the present invention, the skin external preparation may be an external preparation such as essence, mask, cream, etc., which are conventional in the art.

In the skin external preparation, the dosage of the peony oil is 0.5-10 wt%, preferably 1-10 wt%, and the percentage is the weight percentage of the peony oil in the total amount of the raw materials of the skin external preparation.

In the present invention, the peony oil is preferably used as a skin barrier repair agent or a keratinocyte proliferation-inducing agent in the preparation of a skin external preparation.

The effect of peony oil as a skin barrier repair preparation can be studied by using a skin injury model of experimental animals and the influence on the proliferation and differentiation of HaCaT cells. For example, the peony oil prepared by the invention is used for a mouse back skin injury model and HaCaT cell proliferation and differentiation to research the effect on the skin barrier.

On the basis of the common knowledge in the field, the above preferred conditions can be combined randomly to obtain the preferred embodiments of the invention.

The reagents and starting materials used in the present invention are commercially available.

The positive progress effects of the invention are as follows:

(1) the peony flower oil prepared by the invention simultaneously reserves the components of peony petals and peony seeds, can quickly induce keratinocyte proliferation, and has obviously improved skin barrier repairing effect compared with peony flower water and peony seed oil prepared by the traditional squeezing method.

(2) The preparation method provided by the invention has simple steps, can be free from additional solvent, can meet the requirement of large-scale production and is safe.

(3) The peony selected by the invention is widely planted in China, the resource is easy to obtain, the price is low, and the preparation cost is low.

Drawings

FIG. 1 is a graph of experimental data of OD values of HaCaT cells at 370nm after inducing the proliferation of the HaCaT cells for 24h by peony flower oil prepared in blank groups, examples 1-3 and comparative examples 1-8.

Detailed Description

The invention is further illustrated by the following examples, which are not intended to limit the scope of the invention. The experimental methods without specifying specific conditions in the following examples were selected according to the conventional methods and conditions, or according to the commercial instructions.

In each of the following examples and comparative examples, peony petals and peony seeds were produced from lotus in Shandong.

The specific conditions of the treatment steps of the peony petals and the peony seeds in the following examples and comparative examples are as follows:

(1) grinding: grinding with colloid mill (model: JM-F65, Meltron mechanical Co., Ltd., Wenzhou);

(2) temperature maintenance of standing: an electric heating constant temperature water bath (model: HWS, Shanghai-constant technology Co., Ltd.) is adopted to keep the temperature of standing;

(3) squeezing: squeezing with static pressure type cold Press (model: Carver Laboratory Press, FREDS. CARVER. NC) at 55 MPa;

(4) oil-water separation: and (4) performing oil-water separation by using a separating funnel.

Example 1

Step 1, washing 5 parts of peony petals and 95 parts of peony seeds with normal-temperature water, wiping the peony petals dry, and airing the peony seeds;

step 3, standing the mixture obtained in the step 2 at 40 ℃ for 24 h;

Example 2

Step 1, washing 20 parts of peony petals and 80 parts of peony seeds with normal-temperature water, wiping off the peony petals, and airing the peony seeds;

step 3, standing the mixture obtained in the step 2 at 70 ℃ for 12 h;

Example 3

Step 1, washing 10 parts of peony petals and 90 parts of peony seeds with normal-temperature water, wiping the peony petals dry, and airing the peony seeds;

step 3, standing the mixture obtained in the step 2 at 55 ℃ for 18 h;

Comparative example 1

step 3, standing the mixture obtained in the step 2 at 30 ℃ for 6 h;

Comparative example 2

step 3, standing the mixture obtained in the step 2 at 80 ℃ for 36 h;

Comparative example 3

and 3, squeezing the mixture obtained in the step 2, separating oil from water, and taking an oil phase part to obtain the peony oil.

Comparative example 4

Step 1, washing 10 parts of peony petals with normal-temperature water, and wiping the peony petals dry;

step 2, grinding and crushing the peony petals wiped in the step 1;

step 3, adding 90 parts of peony seed oil into the peony petal grinding material obtained in the step 2, and standing the obtained mixture for 18h at 55 ℃;

Comparative example 5

Step 1, washing 100 parts of peony petals with normal-temperature water, and wiping the peony petals dry;

step 2, grinding and crushing the peony petals wiped in the step 1;

and 3, squeezing the peony petal grinding material obtained in the step 2, carrying out oil-water separation, and taking an oil phase part to obtain the peony oil.

Comparative example 6

Step 1, washing 100 parts of peony seeds with normal-temperature water, and airing the peony seeds;

step 2, grinding and crushing the peony seeds dried in the step 1;

and 3, squeezing the peony seed ground substance obtained in the step 2, performing oil-water separation, and taking an oil phase part to obtain the peony seed oil.

Comparative example 7

step 2, grinding and crushing the peony petals wiped in the step 1;

step 3, standing the peony petal grinding material obtained in the step 2 at 55 ℃ for 18 h;

Comparative example 8

step 2, grinding and crushing the peony seeds dried in the step 1;

step 3, standing the peony seed ground substance obtained in the step 2 at 50 ℃ for 18 h;

and 4, squeezing the mixture obtained in the step 3, separating oil from water, and taking an oil phase part to obtain the peony seed oil.

The process comparison ratios of the above examples and comparative examples are shown in table 1 below:

TABLE 1 comparison of the Processes

Effect example 1 skin barrier experiment

1. Experimental materials: healthy ICR mice (male, 18-22g) were purchased from shanghai SLAC laboratory animals ltd (shanghai, china), HaCaT cells were purchased from shanghai bridge, chainhou biotechnology ltd, Fetal Bovine Serum (FBS) (Gibco, new york life technologies, usa), DMEM (Gibco, new york life technologies, usa), BrdU kit (Roche, Mannheim, germany), and the like.

2. An experimental instrument: percutaneous water loss measuring instrument (Delfin Technologies, Finland), cell culture box (HEPA Class 100 CO)₂Incubator, Thermo Fisher scientific), clean bench, pipettor, row gun, and gun head, etc. The clean bench, the pipettor, the row gun, the gun head and the like are conventional experimental instruments in the field.

3. Experimental methods

3.1 animal experiments

Mice were housed at 22+1 ℃ for 12 hours in a light-dark cycle. Mice were randomly divided into 8 groups (n ═ 3, i.e. 3 mice per group): blank group (no treatment), example 1 group (tape stripping + example 1), example 2 group (tape stripping + example 2), example 3 group (tape stripping + example 3), comparative example 1 group (tape stripping + comparative example 1), comparative example 2 group (tape stripping + comparative example 2), comparative example 3 group (tape stripping + comparative example 3), comparative example 4 group (tape stripping + comparative example 4), comparative example 5 group (tape stripping + comparative example 5), comparative example 6 group (tape stripping + comparative example 6), comparative example 7 group (tape stripping + comparative example 7), comparative example 8 group (tape stripping + comparative example 8). Repeatedly peeling back skin with adhesive tape to destroy epidermis protective layer, and measuring percutaneous water loss (TEWL) to 35mg/cm per hour²The molding is successful. The test sample (peony oil prepared in example or comparative example) was applied to the skin surface of the skin lesion area on the back of the mouse at a rate of 100. mu.L/day for three days. The TEWL values were measured at 12, 24, 36, 48, 60 and 72 hours after the test sample (the peony oil prepared in example or comparative example) was used, and the recovery rate of the epidermal barrier was calculated according to the following formula:

epidermal barrier recovery (%) — (TEWL immediately after barrier disruption-TEWL at specific time)/(TEWL-baseline TEWL immediately after barrier disruption) × 100%.

3.2HaCaT cell culture and proliferation experiments

HaCaT cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) (Gibco, New York Life technologies, USA) and 1% penicillin/streptomycin in 5% CO₂At 37 ℃ in a cell culture chamber. HaCaT cells were plated at 8X 10³cell/well cell density was seeded in 96-well plates and incubated for 24 hours, and then the test sample (peony oil prepared in example or comparative example) was added (addition amount: 10. mu.L) to the cell culture medium to continue incubation for 24, 48, 72 hours, and cell proliferation was measured using BrdU kit (Roche, Mannheim, Germany).

4. Results of the experiment

4.1 determination of skin damage repair Rate in Experimental mice

The animal model of the epidermal barrier damage of the skin on the back of the mouse is prepared by adopting a tape repeated adhesion stripping method, the ability of promoting epidermal barrier repair of the examples and the comparative examples is detected by measuring TWEL values of the skin damage area on the back at different time points, and the skin damage repair rate of each group of mice at the corresponding time points is obtained by formula calculation.

4.2 determination of OD value of HaCaT cells at 370nm

Keratinocytes are the main cells that make up the epidermal structure. To observe the effect of each example and comparative example on HaCaT cell proliferation, the OD (optical density) value of HaCAT cells at 370nm after 24 hours of treatment in each example and comparative example was determined by BrdU method (refer to the prior art, Queensland, Wenxingxin, Huangle pine, etc.. BrdU method for observing the effect of a normally-open oral liquid on proliferation of adherent peritoneal fibroblasts [ J ]. Chinese medicinal material, 2009,32(008): 1279-.

Barrier repair rates calculated from TEWL values measured at 12, 24, 36, 48, 60, and 72 hours after the peony oil prepared in each of the above examples and comparative examples was used for the treatment of skin lesions in mice are shown in table 2 below.

The OD of HaCaT cells at 370nm after 24h treatment with the peony oil prepared in each of the above examples and comparative examples measured by BrdU method is shown in Table 3 below.

TABLE 2

TABLE 3

As can be seen from the data in table 2: the repair rates after 60 hours for examples 1-3 were 78.8%, 79.2%, and 80.0%, while the repair rates for the blank and comparative examples 1-8 were only 39.8%, 57.5%, 61.1%, 55.5%, 53.1%, 39.0%, 56.3%, 40.4%, and 55.9%. Therefore, the peony oil prepared in the embodiments 1-3 of the present invention can well promote the repair of damaged epidermal barrier.

From the data in table 3, it can be seen that: the peony flower oil prepared in examples 1-3 induced HaCaT cells to proliferate for 24h, and the HaCaT cells had OD values of 1.8 or more at 370nm, while the peony flower oil prepared in comparative examples 1-8 induced HaCaT cells to proliferate for 24h, and the HaCaT cells had OD values of less than 1.8 at 370 nm. Therefore, the peony oil prepared in the embodiments 1 to 3 of the present invention can significantly induce keratinocyte proliferation.

In addition, it is also apparent from FIG. 1 that (in the figure, ". X" represents: P < 0.05, ". X" represents: P < 0.01, the smaller the P value, the more significant the difference), the OD values at 370nm of HaCaT cells were significantly increased in examples 1-3 compared to the blank group, as high as 1.8 or more, while the OD values in comparative examples 1, 2, 3, 4, 6, 8 compared to the blank group were slightly increased, but all were less than 1.8. Comparative examples 5 and 7 were not significantly different from the blank. It was also confirmed that the peony oil prepared in examples 1 to 3 of the present invention significantly induced keratinocyte proliferation.

Effect example 2 appearance stability test

1. Test method

The color, clarity and smell of the product were examined after 30 days when the product was placed at 4 deg.C, 48 deg.C, protected from light at normal temperature and illuminated at normal temperature (illumination intensity: 300 Lux).

2. Results of the experiment

The results of the appearance stability test of the above examples and comparative examples are shown in table 4.

TABLE 4 appearance stability test results

As can be seen from Table 4, the color, clarity and smell did not change after the rest of the examples and comparative examples for 30 days except for comparative examples 1 and 3; comparative example 1 the color deepened at 48 ℃ and under normal temperature illumination; the clarity of comparative example 3 was unstable under 4 conditions, and the color deepened at 48 ℃ and under ambient temperature light conditions. Therefore, the peony oil prepared in the embodiments 1 to 3 of the present invention has stable appearance.

Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications may be made to the embodiments or portions thereof without departing from the spirit and scope of the invention.

Claims

1. The preparation method of the peony oil is characterized by comprising the following steps: grinding peony petals and peony seeds, standing and squeezing;

2. The method of claim 1, wherein the temperature of said resting is between 50 ℃ and 60 ℃, such as 55 ℃;

and/or the standing time is 14h to 18h, such as 18 h.

3. The method for preparing peony oil according to claim 1, wherein the weight portion of peony petals is 1-99 parts, preferably 5-20 parts;

and/or, the weight portion of the peony seeds can be 1-99 parts, preferably 80-95 parts;

preferably, the dosage ratio of the peony petals to the peony seeds is (1-4) in parts by weight: (16-19), more preferably (1-2): (8-9), for example, 1: 9;

and/or the peony petals are cleaned and dried peony petals; wherein the cleaning condition is cleaning with normal temperature water; the drying condition is wiping;

and/or the peony seeds are cleaned and dried peony seeds; wherein the cleaning condition is cleaning with normal temperature water; the drying condition is airing.

4. The method for preparing peony oil according to claim 1, wherein during the grinding, peony petals and peony seeds are ground separately, or a mixture of peony petals and peony seeds are directly ground;

the grinding condition is grinding by adopting a colloid mill.

5. The method for preparing peony oil according to claim 1, wherein the pressing conditions are pressing with a static pressure type cold press;

the pressing pressure is preferably 45MPa to 65MPa, for example 55 MPa.

6. The method for preparing peony oil according to claim 1, further comprising the step of separating oil from water after said pressing.

7. The method for preparing peony oil according to claim 1, wherein no solvent is added during the preparation of peony oil; the solvent is preferably water and/or an oily organic solvent.

8. A peony oil prepared by the preparation method of the peony oil as claimed in any one of claims 1-7.

9. Use of the peony oil according to claim 8 as an antioxidant active ingredient or an active ingredient for repairing skin barrier in the preparation of an external preparation for skin.

10. The use according to claim 9, wherein the peony oil is used in an amount of 0.5 to 10 wt%, preferably 1 to 10 wt%, based on the total weight of the raw materials of the skin external preparation;

and/or, the peony oil is used as a skin barrier repair agent or a keratinocyte proliferation-inducing agent when preparing a skin external preparation.