WO2025121819A1 - Cosmetic composition for improving skin condition of body, comprising makgeolli lees and beer lees, and preparation method therefor - Google Patents

Abstract

The present invention relates to a cosmetic composition for improving skin condition of the body, comprising Makgeolli lees and beer lees, and a preparation method therefor. The composition of the present invention exhibits excellent moisturizing effects and itchiness reducing effects when applied to the human body, and thus can be usefully employed for the purpose of improving skin condition.

Description

Cosmetic composition for improving body skin condition including makgeolli lees and beer lees and method for producing same

The present invention relates to a cosmetic composition for improving body skin condition including makgeolli lees and beer lees, and a method for producing the same.

The skin protects the organs in the body from external stimuli such as changes in temperature and humidity, ultraviolet rays, and pollutants, and acts as a barrier to prevent moisture from evaporating from the skin.

The skin above gradually ages over time. Internal factors of skin aging include genetic mutations and tissue changes, and external factors include the influence of ultraviolet rays and humidity. As aging progresses, itching occurs due to dryness of the skin or weakened immunity. This itchiness takes away peace from daily life, and especially for seniors who live alone, it can become a problem because it can become a switch that leads to depression, such as having no one to scratch your back.

Meanwhile, demand for eco-friendly products is increasing due to increasing awareness of the harmful effects of synthetic chemicals and increasing interest in sustainability and ethical consumption.

TechSci Research, an international market research firm headquartered in Burnaby, British Columbia, Canada, predicts that the global organic cosmetics market will continue to grow due to the increasing demand for chemical-free personal care products and changing consumer lifestyles regarding organic care. The eco-friendly (organic) body products market is expected to reach USD 9.83 billion in 2021, USD 10.49 billion in 2022, and USD 21.16 billion in 2030, registering a CAGR of 8.9% during the forecast period of 2023-2030.

Jigemi refers to the dregs left after filtering alcohol, which are the residue left after water is added to the dregs and the mother liquor is squeezed out. It is also called suljimi, jubak, juja, jujae, and duljeongbak.

The above-mentioned waste is usually disposed of, which causes environmental pollution and causes disputes between local residents due to landfills, etc. However, since the above-mentioned waste contains useful components for the human body, it is necessary to give a different value to these discarded resources (Re-value).

Although there have been various attempts to utilize the lees in the past, the extracts produced from the lees have limitations in extracting useful components, and as a result, it is still difficult to find products developed with the lees as the main ingredient.

Therefore, creative reuse (upcycling) is important to transform waste materials, which are discarded resources, into products that are environmentally friendly, provide products at good prices by reducing production costs, and create new high added value products that are rich in ingredients useful to the human body by efficiently extracting various ingredients contained in the waste materials.

The present inventors recognized the problems of the above-mentioned prior art and confirmed that a cosmetic composition including a mixed extract of makgeolli lees and beer lees extracted by a method including specific steps can exhibit an excellent moisturizing effect and an itchiness-reducing effect when applied to the human body, thereby improving the skin condition, and thus completed the present invention.

The present invention relates to a cosmetic composition for improving skin condition, which comprises a mixed extract of makgeolli lees and beer lees, wherein the makgeolli lees and beer lees extracts are prepared by a method comprising the following steps.

S1) A step of drying, crushing and mixing makgeolli lees and beer lees;

S2) A step of immersing the above mixture in a citric acid aqueous solution;

S3) A step of cooling the above immersed mixture;

S4) A step of applying ultrasonic waves to the cooled mixture;

S5) Step of adding a strain to the sonicated mixture and fermenting it;

S6) a step of preparing an extract from the fermented mixture; and

S7) Step of filtering and concentrating the above extract.

The present invention relates to a method for producing a cosmetic composition for improving skin condition.

Hereinafter, this will be described in detail. All combinations of various elements disclosed in the present invention fall within the scope of the present invention. In addition, the scope of the present invention cannot be considered limited by the following specific description.

Additionally, unless otherwise defined herein, all terms used in the specification should be understood to have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention belongs.

Also, singular expressions include plural expressions unless the context clearly dictates otherwise.

In this specification, the terms first, second, etc. may be used to distinguish or describe one component from another, but the components should not be construed as being limited by these terms. For example, within the scope of the present invention, the first component may be named the second component, or vice versa.

Additionally, the term "degree" combined with a number in this specification represents degrees Celsius ( ^o C) and may be used interchangeably with " ^o C" (for example, "100 degrees" is used with the same meaning as "100 ^o C").

The present invention provides a cosmetic composition for improving skin condition comprising a mixed extract of makgeolli lees and beer lees.

In the present invention, 'Makgeolli' is a type of Korean traditional liquor, and is also called Takju, Dokdong, Jaeju, Hoeju, or Baekju. According to an embodiment of the present invention, it is preferable that the Makgeolli is rice Makgeolli fermented by adding nuruk to rice, but is not limited thereto.

In the present invention, 'beer' refers to an alcoholic beverage made by fermenting malt made from grains and adding hops, a spice, to the malt as the main ingredient to give it flavor.

In the present invention, 'jjimi' refers to the dregs remaining after filtering alcohol, which are the dregs, and are also referred to as liquor dregs, liquor residue, juja, jujae, and liquor residue. It is known that the dregs contain various nutrients such as starch, protein, fiber, minerals, vitamins, enzymes, and yeast.

In the present invention, 'mixed extract' means an extract manufactured from a mixture of raw materials rather than from each raw material.

In the present invention, 'extract' means a result of extraction using an appropriate extraction solvent, and the extract includes the extract itself and all formulations that can be formed using the extract, such as an extract liquid obtained by extraction, a diluted or concentrated extract liquid, a dried product obtained by drying the extract liquid, a controlled or purified product thereof, a fraction thereof, or a mixture thereof.

The extract of the present invention may be manufactured and used in the form of a dried powder after extraction, but is not limited thereto.

In the present invention, the extract may be prepared by an extraction method commonly used in the art. For example, methods such as hot water extraction, hot water extraction, cold immersion extraction, reflux cooling extraction, ultrasonic extraction, and high-temperature pressure extraction may be used.

In preparing the extract of the present invention, water; lower alcohols having 1 to 4 carbon atoms such as methanol, ethanol, propyl alcohol, and butyl alcohol; polyhydric alcohols such as glycerin, butylene glycol, and propylene glycol; hydrocarbon solvents such as chloroform, methyl acetate, ethyl acetate, benzene, hexane, diethyl ether, petroleum ether, and dichloromethane; or mixed forms thereof can be used as the extraction solvent, but is not limited thereto.

In the present invention, the mixed extract of makgeolli lees and beer lees can be produced by a method comprising the following steps:

S1) A step of drying, crushing and mixing makgeolli lees and beer lees;

S2) A step of immersing the above mixture in a citric acid aqueous solution;

S3) A step of cooling the above immersed mixture;

S4) A step of applying ultrasonic waves to the cooled mixture;

S5) Step of adding a strain to the sonicated mixture and fermenting it;

S6) a step of preparing an extract from the fermented mixture; and

S7) Step of filtering and concentrating the above extract.

In the present invention, steps S1 to S4 are pretreatment steps prior to fermentation, which are performed to increase the activity of useful components contained in makgeolli lees and beer lees, and to obtain an extract with a high extraction yield during the subsequent fermentation process.

In the above step S1, the drying of the makgeolli lees and beer lees is not limited to any commonly known method such as hot air drying, freeze drying, or room temperature drying. However, according to an embodiment of the present invention, it may be preferable to perform room temperature drying, but is not limited thereto.

The pulverization in the above step S1 is performed to secure a large surface area to facilitate reaction or extraction in subsequent steps, and can be performed using a commonly known method.

In the above step S1, the makgeolli lees and the beer lees can be mixed in a weight ratio of 2-1:1, and according to an embodiment of the present invention, the makgeolli lees and the beer lees can be mixed in a weight ratio of 1:1, but is not limited thereto.

In the above step S2, the concentration of the citric acid aqueous solution may be 0.01 to 1% (v/v), preferably 0.01 to 0.3% (v/v), but is not limited thereto. In addition, according to an embodiment of the present invention, the concentration of the citric acid aqueous solution may be preferably 0.05% (v/v), but is not limited thereto.

In the above step S2, the citric acid aqueous solution may be added in an amount of 100 to 500 parts by weight relative to 100 parts by weight of the mixture of the makgeolli lees and the beer lees, and according to an embodiment of the present invention, the citric acid aqueous solution may be added in an amount of 200 to 400 parts by weight, or 150 to 350 parts by weight relative to 100 parts by weight of the mixture of the makgeolli lees and the beer lees, but is not limited thereto.

In the above step S2, according to an embodiment of the present invention, the immersion using the citric acid aqueous solution may be performed at a temperature of 50 to 80 ^o C for 1 to 10 minutes, but is not limited thereto.

In the above step S3, the cooling may be lowering the temperature to 15 to 25 ^o C, preferably lowering the temperature to 20 to 25 ^o C, but is not limited thereto.

In the above step S4, the ultrasonic waves may be low frequency ultrasonic waves, and the low frequency may have a frequency of 5 to 30 kHz, and according to an embodiment of the present invention, the frequency may be 23 kHz, but is not limited thereto.

In the above step S4, the ultrasonic irradiation time may be 3 to 7 hours, and according to an embodiment of the present invention, the ultrasonic irradiation time may be 4 to 6 hours, but is not limited thereto.

According to an embodiment of the present invention, after performing step S4 and before performing step S5, a step of maturing the mixture of the pretreated makgeolli lees and beer lees at a low temperature may be further included, and the maturing may be performed at a temperature of 1 to 5 ^o C for 5 to 15 hours, but is not limited thereto.

In the present invention, the step S5 is a step of performing fermentation on the pretreated makgeolli lees and beer lees.

The strain used in the above step S5 may be a lactic acid bacterium, specifically Lactobacillus, and more specifically Lactobacillus sakei, Lactobacillus brevis or Lactobacillus plantarum, and according to an embodiment of the present invention, the lactic acid bacterium may be Lactobacillus brevis, but is not limited thereto.

The above S6 step is a step of performing extraction from a fermented mixture, and according to an embodiment of the present invention, the extraction can be performed by a method including the following steps:

S6-1) A step of first extracting the fermented mixture at a temperature of 45 to 60 ^o C; and

S6-2) A step of adding ethanol to the first extracted mixture and performing a second extraction at a temperature of 45 to 60 ^o C.

According to an embodiment of the present invention, steps S6-1 and S6-2 may be performed for 1 to 5 hours, preferably 1 to 3 hours, but are not limited thereto.

According to an embodiment of the present invention, in the step S6-2, ethanol may have a concentration of 60% (v/v) and may be introduced in an amount of 30% of the volume of the resultant obtained in the step S5, but is not limited thereto.

The above step S7 is a step of finally filtering and concentrating the extract. The filtration and concentration can be performed using a method conventional in the art, and the method is not limited.

According to an embodiment of the present invention, a freeze-drying step may be further included after the S7 step, but is not limited thereto.

In the present invention, the mixed extract of makgeolli lees and beer lees may be included in an amount of 0.1 to 20 wt% relative to the total weight of the cosmetic composition, preferably 1 to 10 wt%, and more preferably 3 to 7 wt%, but is not limited thereto.

In the present invention, the cosmetic composition may be for the body, and in the present invention, 'for the body' means use for a part other than the face.

In the present invention, 'improvement in skin condition' may refer to one or more of the effects of improving skin moisture content, alleviating itchiness, alleviating inflammation, and improving skin tone. According to an embodiment of the present invention, it may refer to, but is not limited to, improving skin moisture content and alleviating itchiness.

The ingredients included in the cosmetic composition of the present invention may include ingredients commonly used in cosmetic compositions, and include, for example, conventional auxiliary agents such as preservatives, antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances, and carriers.

In the present invention, the composition may further include a preservative. Examples of the preservative include, but are not limited to, parabens, alcohols, organic acids and their salts, isothiazole compounds, urea, and triclosan.

The cosmetic composition of the present invention may be formulated in the form of, for example, a solution, a suspension, an emulsion, a paste, a gel, a powder, a cream, a lotion, a toner, a water-soluble liquid, an essence, a surfactant-containing cleansing, an oil, a powder foundation, an emulsion foundation, a wax foundation, a spray, a shampoo, a rinse, a body wash, a soap, etc., but is not limited thereto, and may be manufactured in any form of a formulation commonly manufactured in the cosmetic industry.

When the cosmetic composition of the present invention is formulated as a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide can be used as a carrier component.

When the cosmetic composition of the present invention is formulated as a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and particularly in the case of a spray, a propellant such as chlorofluorohydrocarbon, propane/butane or dimethyl ether may be additionally included.

When the cosmetic composition of the present invention is formulated as a solution or emulsion, a solvent, a solubilizer or an emulsifier is used as a carrier component, and may include, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan.

When the cosmetic composition of the present invention is formulated as a suspension, liquid diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth, etc. can be used as carrier components.

When the cosmetic composition of the present invention is formulated as a surfactant-containing cleansing agent, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkyl amidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester may be used as a carrier component.

In addition to the various moisturizing ingredients mentioned above, those skilled in the art can easily select and mix other ingredients appropriately depending on the formulation or intended use.

Another aspect of the present invention provides a method for producing a cosmetic composition comprising a mixed extract of makgeolli lees and beer lees, comprising the following steps:

M1) A step for producing a mixed extract of makgeolli lees and beer lees; and

M2) A step of mixing the mixed extract of the above-mentioned makgeolli lees and beer lees so as to be 0.1 to 20 wt% of the total composition.

In the present invention, the mixed extract of makgeolli lees and beer lees can be mixed in an amount of 0.1 to 20 wt% relative to the total weight of the cosmetic composition, preferably 1 to 10 wt%, and more preferably 3 to 7 wt%, but is not limited thereto.

In the present invention, the mixed extract of makgeolli lees and beer lees may be produced by a method comprising the following steps:

S1) A step of drying, crushing and mixing makgeolli lees and beer lees;

S2) A step of immersing the above mixture in a citric acid aqueous solution;

S3) A step of cooling the above immersed mixture;

S4) A step of applying ultrasonic waves to the cooled mixture;

S5) Step of adding a strain to the sonicated mixture and fermenting it;

S6) a step of preparing an extract from the fermented mixture; and

S7) Step of filtering and concentrating the above extract.

In this specification, specific details related to the method for manufacturing a cosmetic composition are substantially the same as those described in the cosmetic composition unless otherwise stated, and are omitted to avoid unnecessary repetition.

The cosmetic composition for improving the body skin condition of the present invention exhibits excellent moisturizing effects and itchiness-reducing effects when applied to the human body, and thus can be usefully used for the purpose of improving the skin condition.

In addition, the cosmetic composition for improving the body skin condition of the present invention is made by recycling discarded waste resources, such as lees, and is environmentally friendly and allows for the production of a product at a good price with reduced production costs.

Hereinafter, the present invention will be described in more detail by way of examples. However, these examples are intended to exemplify the present invention, and the scope of the present invention is not limited by these examples. In addition, each component or feature may be considered optional unless otherwise explicitly stated. Each component or feature may be implemented in a form that is not combined with other components or features. In addition, various embodiments may be configured by combining some components and/or features. Some components or features of one embodiment may be included in another embodiment, or may be replaced with corresponding components or features of another embodiment. In addition, terms not specifically defined in this specification should be understood to have the meaning commonly used in the technical field to which the present invention belongs.

Example

First, the makgeolli (main ingredient: rice) lees and beer (main ingredient: malt) lees were completely dried using a room temperature desiccator and then finely ground.

Then, the crushed makgeolli lees and beer lees were evenly mixed at a weight ratio of about 1:1.

A 0.05% (w/v) citric acid aqueous solution was added to the above mixture of seaweeds in an amount about 3 times its weight, and left to stand (immerse) at a temperature of 50 to 80 ^o C for about 5 minutes.

Next, the immersed squid mixture was cooled to 20 to 25 ^o C, and while maintaining the temperature, ultrasonic waves (frequency: 23 kHz) were applied using an ultrasonic device for about 5 hours. During this time, the container was continuously stirred.

Then, the sonicated mixture was cooled to a temperature of 1 to 5 ^o C and aged for about 10 hours while maintaining the temperature.

Thereafter, Lactobacillus brevis, a lactic acid bacterium, was added to the sonicated mixture and fermentation was performed. Specifically, Lactobacillus brevis B3-18, cultured in MRS broth for 20 hours, was inoculated to 10 ⁸ cfu/g and cultured at 27 ^o C for 24 hours.

Next, the fermented lees mixture was stirred at a temperature of 50±5 ^o C for about 2 hours to perform a first extraction, and 60% (v/v) ethanol corresponding to 30% of the volume of the contents of the container was additionally added, and the second extraction was performed by stirring for about 2 hours, followed by filtration, concentration under reduced pressure, and freeze-drying to obtain an extract.

Next, a cosmetic composition of the example was prepared using 5.00% of the above-mentioned prepared extract, 6.00% of propanediol, 5.00% of glycerin, 0.50% of betaine, 0.40% of polyglyceryl-10 laurate, 0.35% of carbomer, 0.30% of arginine, 0.5% of butylene glycol, and the remainder of purified water.

Comparative example

First, the makgeolli lees and beer lees were completely dried using a room temperature desiccator and then finely ground.

Then, the crushed makgeolli lees and beer lees were evenly mixed at a weight ratio of about 1:1.

Next, Lactobacillus brevis, a lactic acid bacterium, was added to the mixture and fermentation was performed. Specifically, Lactobacillus brevis B3-18, cultured in MRS broth for 18 hours, was inoculated to 10 ⁸ cfu/g and cultured at 27 ^o C for 24 hours.

Next, the fermented lees mixture was stirred at a temperature of 50±5 ^o C for about 4 hours to proceed with extraction, and the extract was obtained by filtering and concentrating.

Next, a cosmetic composition of a comparative example was prepared using 5.00% of the above-mentioned prepared extract, 6.00% of propanediol, 5.00% of glycerin, 0.50% of betaine, 0.40% of polyglyceryl-10 laurate, 0.35% of carbomer, 0.30% of arginine, 0.5% of butylene glycol, and the remainder of purified water.

Experimental example

1. Selection criteria for test subjects

1) A person who has received a sufficient explanation of the matters that the test manager or a person authorized by the test manager must inform the test subject of and voluntarily completed and signed a consent form.

2) People who usually have itchiness due to dryness in the test area

3) Adult men and women aged 65 years or older (n=20) who are healthy and do not have any acute or chronic physical illnesses, including skin diseases.

4) People who can be monitored during the test period

2. Exclusion criteria

1) Pregnant or lactating women and women of childbearing potential

2) People who have used topical skin care products containing steroids for more than 1 month to treat skin diseases

3) People who have not passed 6 months since participating in the same test

4) People with sensitive or hypersensitive skin

5) People with skin abnormalities such as spots, acne, erythema, or telangiectasia on the test area

6) People who received treatment on the test area within 6 months prior to the start of the study

7) Any other person deemed unsuitable for the test by the test manager

3. Criteria for dropping out

1) If adverse effects such as burning sensation or erythema occur at the test site

2) If the test subject exposes the test area to excessive ultraviolet rays during the test process or consumes excessive alcohol or smoking, which hinders the evaluation of the results.

3) If the test taker has difficulty in follow-up observation due to personal reasons during the test process

4) If the test subject violates the usage method or schedule without any special reason.

4. Test method

^{Corneometer®} is a device that indirectly measures moisture content by utilizing the property that the capacitance of the skin increases in proportion to the moisture content. The unit of measurement is expressed as Arbitrary Unit (AU), which is a relative, arbitrary value of the skin moisture content.

The subjects washed their upper arms with the same body cleanser, gently patted dry with a paper towel, and rested for 30 minutes in a constant temperature and humidity room (temperature: 22±2 ^o C, humidity: 50±5%). Then, measurements were taken five times each using a corneometer, and the average value of the measurements was recorded as the measurement value.

The subjects were instructed on precautions and usage, and the compositions of examples (n=10) and comparative examples (n=10) were distributed. The subjects were instructed to use them twice a day, in the morning and evening, for 4 weeks.

Device measurements were performed on the same area before use of the test substance, immediately after use, after two weeks of use, and after four weeks of use.

5. Test Results (1)

The test results related to the skin moisture improvement effect are shown in the table below.

division Before use Immediately after first use After 2 weeks of use After 4 weeks of use Example 35.23±5.11 53.33±5.10 55.12±5.27 57.12±5.11 Comparative example 35.25±5.10 45.72±6.06 47.26±5.45 48.03±5.27

As can be seen from the above results, in the case of the example, the moisture content of the skin increased by 51.38% immediately after one use, 56.46% after two weeks of use, and 62.13% after four weeks of use compared to before use of the test substance.

In contrast, in the comparative example, moisture content increased by 29.70% immediately after one use, 34.07% after two weeks of use, and 36.26% after four weeks of use.

Looking at the above results, it can be confirmed that the cosmetic composition of the present invention can exhibit an excellent moisturizing effect. In addition, it can be confirmed that it can exhibit a more excellent moisturizing effect compared to the comparative example in which some steps were omitted in the preparation of the extract.

6. Test Results (2)

A tickling relief assessment (itching assessment) was conducted on the study subjects before the above test, 2 weeks after the test, and 4 weeks after the test.

The evaluation criteria were classified from 0 to 4 points according to the degree of tickling, as shown in Table 2 below.

<Evaluation criteria>

4: Very ticklish

3: Severe ticklishness

2: The tickling is severe

1: I feel a little ticklish

0: No tickling

division Before use After 2 weeks of use After 4 weeks of use Example 2.21 1.12 0.09

As can be seen from the above results, the average value related to the degree of itching relief of the test subjects significantly decreased in score compared to before use, confirming that the composition of the present invention can have a significant improvement effect on itching when applied to the skin.

The various embodiments described above may be embodied in other specific forms without departing from the technical idea and essential characteristics thereof. Therefore, the above detailed description should not be construed as limiting in all aspects but should be considered as illustrative. The scope of the various embodiments should be determined by a reasonable interpretation of the appended claims, and all changes within the equivalent scope of the various embodiments are included in the scope of the various embodiments. In addition, claims that do not have an explicit citation relationship in the patent claims may be combined to form an embodiment or included as a new claim by post-application amendment.

Claims (5)

Contains a mixed extract of makgeolli lees and beer lees, The above makgeolli lees and beer lees extracts are prepared by a method comprising the following steps, a cosmetic composition for improving skin condition: S1) A step of drying, crushing and mixing makgeolli lees and beer lees; S2) A step of immersing the above mixture in a citric acid aqueous solution; S3) A step of cooling the above immersed mixture; S4) A step of applying ultrasonic waves to the cooled mixture; S5) Step of adding a strain to the sonicated mixture and fermenting it; S6) a step of preparing an extract from the fermented mixture; and S7) Step of filtering and concentrating the above extract. In the first paragraph, A cosmetic composition for improving skin condition, wherein, in the above step S1, makgeolli lees and beer lees are mixed in a weight ratio of 1:1. In the first paragraph, A cosmetic composition for improving skin condition, wherein in the step S2, the citric acid aqueous solution has a concentration of 0.01 to 1% (v/v). In the first paragraph, A cosmetic composition for improving skin condition, wherein the ultrasound in the above step S4 is performed at a frequency of 5 to 30 kHz for 3 to 7 hours. In the first paragraph, In the above step S6, the extract is a cosmetic composition for improving skin condition, which is manufactured by a method comprising the following steps: S6-1) A step of first extracting the fermented mixture at a temperature of 45 to 60 ^o C; and S6-2) A step of adding ethanol to the first extracted mixture and performing a second extraction at a temperature of 45 to 60 ^o C.