KR20200073015A - Pretreatment technology for enhancing physiological activity of plant extracts of paeonia - Google Patents

Abstract

The present disclosure provides a method for manufacturing a Magnolia sieboldii extract which pre-treats Magnolia sieboldii with chilling stress or heat treatment stress. The Magnolia sieboldii extract undergone the pretreatment of the present disclosure increases the content of total polyphenol, paeoniflorin, which is an index material, and total saponin, thereby exhibiting excellent antioxidant activity.

Description

Pretreatment technology to enhance the physiological activity of Paeonia plant extracts {PRETREATMENT TECHNOLOGY FOR ENHANCING PHYSIOLOGICAL ACTIVITY OF PLANT EXTRACTS OF PAEONIA}

The present disclosure relates to a pre-treatment technique for enhancing the physiological activity of a plant extract of the genus Paeonia . More specifically, it relates to a technique of heat treatment prior to extraction in order to enhance the physiological activity of the paeonia genus extract.

For skin function, the conventional peony extract was mainly limited to the root portion. In particular, functional substances have been extracted by drying the root skin. As a conventional method, the roots of peonies grown for more than 6 years have been collected, and the roots and necks have been removed. This method requires a lot of labor and time, and has been limited to use in industrial mass production. In addition, there has been a demand to promote the utilization of the remaining parts by using only the root part of the roots, and among them, only the bark part of the root.

Korean Patent Publication No. 2016-0124478

Accordingly, the present inventors tried to develop a cosmetic composition having enhanced bioactive functions using the whole plant of Paeonia genus. In addition, the purpose of the present invention was to provide a cosmetic composition having enhanced bioactive functions using leaves, stems, and flowers, which are parts other than the roots of the Paeonia plant outpost.

This disclosure par Pioneer (paeonia) to provide a method of producing in a plant a chilling stress, heat treated or stress par undergoes a combination of both the pre-treatment step Pioneer (paeonia) in plant extracts.

In addition, the present disclosure is to provide a cosmetic composition comprising a plant extract of the genus Paeonia ( paeonia ) prepared by the above manufacturing method.

In addition, the present disclosure has a total polyphenol content of 12.5% or more as measured by a colorimetric method using a Folin-Denis reagent, and a fionifluorine content of 2.5% or more when analyzed using an HPLC/UV method, and a colorimetric method. Using the analysis to provide a plant extract of the genus Paeonia, the total saponin content is 3.8% or more.

In addition, the present disclosure intends to provide a plant extract of the genus Paeonia , wherein the minimum concentration IC50 for 50% elimination of DPPH is 0.51 μg/ml or less.

According to the present disclosure, extracts of plant roots of the genus Paeonia extracted through a pre-treatment process have increased content of total polyphenols, indicators and total saponins. In addition, it was confirmed that the extract of the present disclosure has enhanced DPPH scavenging activity and enhanced physiological function.

The technology disclosed herein is a method for producing a plant extract of the genus Paeonia , the method comprising a pre-treatment step of pre-treating a plant of the genus Paeonia ; The pre-treatment step is to provide a method for preparing a plant extract of the genus Paeonia , which is a step of treating chilling stress, heat treatment stress, or a combination thereof.

In one embodiment, the paeonia ( paeonia ) genus plant may be at least one selected from the group consisting of outpost, root, leaf, stem and flowers. More preferably, the plant of the genus Paeonia may be a root.

In one embodiment, the pre-treatment step may be heat treatment stress.

In one embodiment, the pre-treatment step may be to apply heat treatment stress to the root of a plant in the genus Paeonia .

In one embodiment, the manufacturing method comprises the steps of powdering the pre-treated paeonia genus plants by hot air drying after pretreatment; Extracting the powdered plant using a solvent; And concentrating the extract under reduced pressure.

In one embodiment, the solvent is not particularly limited and may be, for example, water, hot water, an organic solvent, or a mixture thereof. Specifically, the solvent may be hot water.

In one embodiment, the step of separating the plant of the genus Paeonia ( paeonia ) into the root, leaf, stem, flower parts before the pre-treatment step; Washing the separated three parts; And cutting the washed portion.

In one embodiment, the cutting may be a step of cutting the washed root, leaf, or stem into 1 to 4 cm. Specifically, 1 cm or more, 1.3 cm or more, 1.6 cm or more, 1.9 cm or more, 2.2 cm or more, 2.3 cm or more, or 2.4 cm or more, 4 cm or less, 3.7 cm or less, 3.4 cm or less, 3.1, cm or less , 2.8 cm or less, 2.7 cm or less, or 2.6 cm or less. More specifically, the cut size may be 2.5 cm.

In one embodiment, the chilling stress may be to refrigerate the paeonia genus plant at a temperature of -5 to 9°C for 36 to 60 hours.

Specifically, the chilling stress time is 36 hours or more, 38 hours or more, 40 hours or more, 42 hours or more, 44 hours or more, 46 hours or more, or 47 hours or more, 60 hours or less, 58 hours or less, 56 hours or less, It may be 54 hours or less, 52 hours or less, 50 hours or less, or 49 hours or less. More specifically, the chilling stress time may be 48 hours.

Specifically, the chilling stress temperature is -5°C or higher, -4°C or higher, -3°C or higher, -2°C or higher, -1°C or higher, 0°C or higher, 0.3°C or higher, 0.6°C or higher, 0.9°C or higher, 1.2 °C or higher, 1.5 °C or higher, 1.8 °C or higher, or 1.9 °C or higher, 9 °C or lower, 8 °C or lower, 7 °C or lower, 6 °C or lower, 5 °C or lower, 4 °C or lower, 3.7 °C or lower, 3.4 °C or lower, 3.1 ℃ or less, 2.8 ℃ or less, 2.5 ℃ or less, 2.2 ℃ or less, or 2.1 ℃ or less. More specifically, the chilling stress temperature may be 2°C.

In the field of extracting herbal medicines, there is a formulation method, which is a process of heat treatment after drying to change the medicinal properties of the herbal medicine. In the heat treatment used in the preparation method, there is a method of boiling or roasting. In the present disclosure, the preparation method can be simulated and boiled by heat treatment after plant collection.

In one embodiment, the heat treatment stress may be that the plants of the genus Paeonia are waterproofed and heated for 0.5 to 1.5 hours in a water bath at 80 to 110°C.

Specifically, the heat treatment temperature is 80°C or higher, 83°C or higher, 86°C or higher, 89°C or higher, 92°C or higher, 93°C or higher, or 94°C or higher, 110°C or lower, 107°C or lower, 104°C or lower, 101 ℃ or less, 98 ℃ or less, 97 ℃ or less, or 96 ℃ or less. More specifically, it may be 95 ℃.

Specifically, the heating time of the heat treatment stress is 0.5 hours or more, 0.6 hours or more, 0.7 hours or more, 0.8 hours or more, or 0.9 hours or more, 1.5 hours or less, 1.4 hours or less, 1.3 hours or less, 1.2 hours or less, or 1.1 It can be less than an hour. More specifically, it may be 1 hour.

In one embodiment, the waterproofing treatment may be to seal plants of the genus Paeonia in a waterproof packaging. This is to prevent the active ingredient of the paeonia genus during the pretreatment process from being dissolved by hot water.

In one embodiment, the par Pioneer (paeonia) in the plant is not particularly limited, par Pioneer (paeonia), but as long as in a plant, e.g. Tibet hwangmo is (Paeonia ludlowii), salted peony (Paeonia rockii ), Dravemoran ( Paeonia delavayi ), wind protein ( Paeonia ostii ), Paeonia jishanensis ), and may be one or more of the group consisting of Paeonia suffruticosa . Specifically, it may be, the par Pioneer (paeonia) in plants sieboldii (Paeonia suffruticosa).

The technology disclosed herein provides a paeonia genus plant extract produced by any one of the above manufacturing methods.

The technology disclosed herein provides a cosmetic composition comprising the paeonia genus plant extract.

The formulation of the cosmetic composition is not particularly limited, and may be appropriately selected according to the purpose. For example, skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, foundation, essence, nutrition essence, pack, soap, cleansing Foam, cleansing lotion, cleansing cream, body lotion and body cleanser may be prepared from one or more formulations selected from the group consisting of, but is not limited thereto.

The technology disclosed herein is a plant extract of the genus Paeonia , a total polyphenol content of at least 12% based on the total weight of the extract, and at least 2.4% of the total weight of the extract based on the total weight of the extract (paeoniflorin), and the extract It is a plant extract of the genus Paeonia , which contains a total saponin content of 3.5% or more based on the total weight, and has a minimum concentration IC50 of 0.51 μg/ml or less to eliminate 50% of DPPH.

In one embodiment, the par Pioneer (paeonia) in the plant is not particularly limited, par Pioneer (paeonia), but as long as in a plant, e.g. Tibet hwangmo is (Paeonia ludlowii), salted peony (Paeonia rockii ), Dravemoran ( Paeonia delavayi ), wind protein ( Paeonia ostii ), Paeonia jishanensis ), and may be one or more of the group consisting of Paeonia suffruticosa . Specifically, the par Pioneer (paeonia) in plants sieboldii (Paeonia suffruticosa ).

In one embodiment, the total polyphenol content may be 12% or more to 15% or less. Specifically, 12% or more, 12.2% or more, 12.4% or more, 12.6% or more, 12.8% or more, 13% or more, 13.1% or more, 13.2% or more, 13.3% or more, or 13.4% or more, 15% or less, 14.8 % Or less, 14.6% or less, 14.4% or less, 14.2% or less, 14% or less, 13.9% or less, 13.8% or less, 13.7% or less, or 13.6% or less.

In one embodiment, the content of the paeoniflorin (paeoniflorin) may be 2.4% or more to 4.2% or less. Specifically, 2.4% or more, 2.6% or more, 2.8% or more, 2.9% or more, 3.0% or more, 3.1% or more, or 3.2% or more, 4.2% or less, 4% or less, 3.8% or less, 3.6% or less , 3.5% or less, 3.4% or less, or 3.3% or less.

In one embodiment, the total saponin content may be 3.5% or more to 5.5%. Specifically, 3.5% or more, 3.6% or more, 3.7% or more, 3.8% or more, 3.9% or more, 4.0% or more, 4.1% or more, 4.2% or more, 4.3% or more, or 4.4% or more, and 5.5% or less, 5.4% or less, 5.3% or less, 5.2% or less, 5.1% or less, 5.0% or less, 4.9% or less, 4.8% or less, 4.7% or less, 4.6% or less, or 4.5% or less.

In one embodiment, the IC50 may be 0.01 μg/ml or more to 0.051 μg/ml or less. Specifically, 0.01 µg/ml or more, 0.015 µg/ml or more, 0.02 µg/ml or more, 0.025 µg/ml or more, 0.026 µg/ml or more, 0.027 µg/ml or more, 0.028 µg/ml or more, or 0.029 µg/or more ml or more, 0.051 µg/ml or less, 0.045 µg/ml or less, 0.04 µg/ml or less, 0.035 µg/ml or less, 0.034 µg/ml or less, 0.033 µg/ml or less, 0.032 µg/ml or less, 0.031 µg/ml or less Or less, or 0.03 μg/ml or less.

In addition, the mock (Paeonia suffruticosa) referred to in the present disclosure may be referred to as a mock, peony, or mod, but in the present disclosure, it is unified as a mock.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as limited by these examples.

One. Moran Preparation of extract

(1) Domestic Moran Outpost Collection and sample preparation

In the Gangjin region of Jeollanam-do, a tree that has been cultivated for more than 5 years was collected and washed with roots, leaves, stems and flowers. After washing, the roots, leaves, and stems were prepared by cutting to a size of about 2 to 3 cm for easy pretreatment.

(2) Pre-treatment step using temperature stress

The collected eggs were subjected to a pre-treatment step of chilling stress or heat treatment stress described in more detail below.

1) chilling stress

In the chilling stress stage, chilling stress was applied to the collected eggs samples for 48 hours using a facility refrigerated at 0 to 4°C, and then dried after 24 hours at room temperature.

2) Heat treatment stress

In the field of extracting herbal medicines, there is a formulation method, which is a process of heat treatment after drying to change the medicinal properties of the herbal medicine. In the heat treatment used in the preparation method, there is a method of boiling or roasting. In this embodiment, heat treatment stress was performed to boil the plants collected by simulating the preparation method.

In the heat treatment stress stage, the collected eggs were heated in a water bath at 95° C. for 1 hour. At this time, heat treatment was performed using a packaging material capable of waterproofing to prevent direct contact with water. This is to prevent the active ingredient of the eggs from being dissolved by hot water during the pre-treatment process.

The control group was the untreated group, which did not undergo any pre-treatment with the Molan extract.

(3) Extract manufacturing step

After the pre-treatment, the sample of each peony outpost was dried by hot air to lower the moisture to a level of 5% and powdered. After extraction with hot water, it was concentrated under reduced pressure to prepare an extract. In the case of the prepared extract, there was no significant difference in yield according to the pretreatment method, and the yield for each part was 24.35%±1.2% by weight for the flower part, 27.95%±0.922% by weight for the leaf and stem parts, and 29.0%± for the root part. 0.28% by weight. Leaves and stems were extracted together. The yield of the extract represents the weight of the extract relative to the dry weight of the raw material before extraction.

2. Moran Functional evaluation of extract

(1) Total polyphenol content according to pretreatment

The total polyphenol content was measured by a colorimetric method using a Folin-Denis reagent. Weigh 100 mg of the sample, put it in a 100 ml volumetric flask, add distilled water to the mark, and dissolve by ultrasonic treatment. 7.5 ml of distilled water was taken into a new test tube, and 1 ml of sample solution was taken, followed by adding 0.5 ml of polylinide reagent and 1 ml of 35% sodium carbonate in order. After standing in the cow for 1 hour, absorbance was measured at 760 nm. As a standard solution, 10 mg of tannic acid was precisely weighed, contained in a 100 ml volumetric flask, filled with distilled water to the mark, dissolved, and diluted to an appropriate concentration.

Table 1 shows the total polyphenol content measured.

Moran root extract Total polyphenol content (% by weight) No treatment 11.9 Chilling stress 10.1 Heat treatment stress 13.3

It was confirmed that the total polyphenol content was increased in the extract of the root part of the orchid, which was subjected to heat treatment stress by pretreatment.

(2) Content of indicators (Piononiflorin)

The analysis of the indicator material, fioniflorine, was performed using an HPLC/UV method. Using a C18 analytical column (250X4.60 mm, 5 μm), mobile phase A (acetonitrile) and mobile phase B (0.005% formic acid-containing distilled water) were analyzed by adjusting the concentration over time in the following compositions. Hourly mobile phase A: The volume ratio of mobile phase B is as follows: 0 to 10:90, 20 to 20:80, 30 to 40:60, 50 to 40:60, 60 to 100:0, 70 10:90 in minutes. The analysis was conducted in the 230 nm region. Fioniflorin used as a standard was purchased from Sigma, and a standard curve was drawn to obtain the index content. The results are shown in Table 2 below.

Moran root extract Index (paeoniflorin) content (% by weight) No treatment 2.38 Chilling stress 2.73 Heat treatment stress 3.27

It was confirmed that the content of the indicator material, paeoniflorin, was increased in both the group subjected to chilling stress and the group subjected to heat treatment stress.

(3) Total saponin content

A colorimetric method was used to analyze the total saponin content. After adding 0.3 ml of 8% vanillin solution to 100 μl of the obtained extract, 4 ml of 75% sulfuric acid solution was added in a cold water bath. Then, the mixture was warmed at 60° C. for 10 minutes to develop color, and absorbance was measured at 545 nm to obtain the total saponin content from the standard curve. The standard curve was prepared by sequentially preparing ginsenoside Re (purchased from Sigma) at a concentration of 0.2 to 1.0 mg/ml, reacting in the same manner as the sample solution, and measuring absorbance. The results are shown in Table 3 below.

Moran root extract Total saponin content (% by weight) No treatment 3.4 Chilling stress 4.2 Heat treatment stress 4.5

It was confirmed that the total saponin content increased in the heat treatment stress group. It was thought that a higher amount of saponin was extracted as the tissue softened as it was heated during the heat treatment stress process.

(4) Free radical scavenging activity (DPPH scavenging activity)

In the free radical scavenging activity experiment, 2,2-diphenyl-1-pyrylhydrazyl (2,2-diphenyl-1-picrylhydrazyl, DPPH) dissolved in methanol solution at 0.002% (w/v) was used for each concentration. The amount of erase was compared by mixing with the prepared extract. After mixing, the cows were left at room temperature for 30 minutes, and the absorbance was measured at 515 nm using an optical spectrometer and compared. Tables 4 to 6 below are the results, respectively, showing the results of the root extract, leaf and stem extract, and flower extract.

Moran root extract IC50 (µg/ml) No treatment 0.052 Chilling stress 0.051 Heat treatment stress 0.036

It was confirmed that the DPPH scavenging activity was increased in the extract of the root part of the fever treated with heat treatment stress.

Magnolia leaf and stem part extract IC50 (µg/ml) No treatment 0.055 Chilling stress 0.045

Magnolia flower part extract IC50 (µg/ml) No treatment 0.045 Chilling stress 0.041

In addition, it was confirmed that the DPPH scavenging activity was also increased in other parts of leaves, stems, and flowers of the outpost.

From the above results, when pre-treatment such as chilling stress or heat treatment stress is applied to the mock sample, the content of the total polyphenol is increased, the content of the indicator material, fioniflorine, and the total saponin, than that of extracting and using mock-neck in the conventional method. It was confirmed that the content of is increased to show superior antioxidant capacity than the extract by the conventional method. In addition, it was confirmed that when applying the chilling stress pre-treatment, the extracts of the leaves, stems, and flowers other than the roots of the orchids have better antioxidant activity than the existing untreated extracts. Accordingly, it was confirmed that the mock outpost can be used in a cosmetic composition for skin improvement.

Claims (17)

As a method of producing a plant extract of the genus Paeonia ,
The manufacturing method includes a pre-treatment step of pre-treatment of plants of the genus Paeonia ;
The pre-treatment step is a process of treating chilling stress, heat treatment stress, or a combination thereof, a method for preparing a plant extract of the genus Paeonia . The method of claim 1 wherein the par Pioneer (paeonia) in plant sentinel, roots, leaves, stems and the not less than one selected from the group consisting of floral, par Pioneer (paeonia) in method for manufacturing a plant extract. The method of claim 1, wherein the plant of the genus Paeonia is the root, the method of manufacturing a plant extract of the genus Paeonia . The method of claim 1, wherein the pre-treatment step is stress of heat treatment, a plant extract of the genus Paeonia . The method of claim 1, wherein the pre-treatment step is on par Pioneer A method for producing a, par Pioneer (paeonia) in plant extracts to apply a heat treatment of the root stress (paeonia) in plants. According to claim 1, The manufacturing method after the pre-treatment,
Pulverizing the pre-treated paeonia genus plants by hot air drying;
Extracting the powdered plant using a solvent; And
Further comprising the step of concentrating the extract under reduced pressure, Paeonia ( paeonia ) method of producing a plant extract. The method of claim 6, wherein the solvent is water, hot water, an organic solvent, or a mixture thereof, a method for producing a plant extract of the genus Paeonia . The method of claim 6, wherein the solvent is hot water, a method for producing a plant extract of the genus Paeonia . The method of claim 1, wherein the sludge pre-treatment stage before par ah (paeonia) in a step of cutting the plant with 1 to 4 cm more, par Pioneer (paeonia) in method for manufacturing a plant extract. The method of claim 1, wherein the method for producing the chilling stress par Pioneer (paeonia) in the plant 36 to 60 to the refrigerating treatment at a temperature of -5 to 9 ℃ time for, par Pioneer (paeonia) in plant extracts . The method of claim 1, wherein the heat treatment stress is to prepare a plant extract of Paeonia ( paeonia ), waterproofing the plant of Paeonia and heating it in a water bath at 80 to 110° C. for 0.5 to 1.5 hours. Way. The method of claim 1 wherein the par Pioneer (paeonia) in plants sieboldii (Paeonia suffruticosa) would of, par Pioneer (paeonia) in method for manufacturing a plant extract. A plant extract of the genus Paeonia produced by the method of any one of claims 1 to 12. Claim 13 paeonia ( paeonia ) cosmetic composition comprising a plant extract. As a plant extract of the genus Paeonia ,
Total polyphenol content of at least 12% based on the total weight of the extract,
More than 2.4% of the extract by weight based on the total weight of pioniflorin (paeoniflorin), and
Plant extract of the genus paeonia , comprising a total saponin content of at least 3.5% based on the total weight of the extract As a plant extract of the genus Paeonia,
A plant extract of the genus Paeonia, with a minimum concentration IC50 of 0.51 μg/ml or less to eliminate 50% of DPPH. 16. The method of claim 15 or 16 in the par Pioneer (paeonia) in the plant would sieboldii (Paeonia suffruticosa), par Pioneer (paeonia) in plant extracts.