JPS63119409A - Skin cosmetic - Google Patents

Abstract

PURPOSE:To obtain a skin cosmetic having excellent practical use characteristics and improving effects on roughened skin, by adding oleic acid and rapeseed fatty acid to a culture medium prepared by aerobically cultivating a protozoan euglena, keeping the culture medium in an anaerobic state and blending the resultant fat or oil. CONSTITUTION:A skin cosmetic produced by aerobically cultivating a protozoan euglena in a Koren . Hutner culture medium, etc., at 20-33 deg.C and initial pH 3.0-7.0, adding oleic acid and rapeseed fatty acid at 60:40-90:10 weight ratio in the total amount of 0.5-2%, keeping the culture medium in an anaerobic state for 1-5 days by passing N2, etc., to provided an euglena fat or oil and blending the resultant fat or oil. The above mentioned euglena fat or oil in an amount of 2.0-5.0wt% is blended with skin cosmetic based, e.g. creams, milky lotions, lipsticks, foundations, etc. The afore-mentioned cosmetic exhibits characteristics excellent in extensibility, compatibility with the skin, refreshing feeling, humectant property, etc.

Description

DETAILED DESCRIPTION OF THE INVENTION (Field of the Invention) The present invention is directed to a method for culturing a protozoan, Euglena, in which oleic acid and rapeseed fatty acid are added to a medium, and then the medium is placed in an anaerobic state. Regarding skin cosmetics containing improved unsaturated oils and fats,
More specifically, the present invention relates to a skin cosmetic that has excellent practical properties and an effect on improving rough skin.

(Prior Art) It has been known that wax esters can be obtained from cultures of the protozoan Euglena (Japanese Patent Laid-Open No. 59-118
Publication No. 090). Furthermore, it has been announced that unsaturated wax esters obtained by enriching Euglena with oleic acid can be used for cosmetics and lubricating oils (published in the Nippon Kogyo Shimbun on July 16, 1985).

However, skin cosmetics containing these wax esters and unsaturated wax esters have poor practical properties (spreadiness, skin affinity, light feel, moisturizing properties, oily feel) and roughness when applied to the skin. The skin improvement effect was not fully satisfactory.

That is, the wax ester is a solid ester consisting of a saturated fatty acid and a saturated alcohol, and has poor characteristics as an oily substance for skin cosmetics.

Further, the unsaturated wax ester is a liquid or paste ester mainly having a total carbon number of C, L, to C3,000, and skin cosmetics containing this are blended with olive oil or macadamian nut oil. It exhibited almost the same practical properties and rough skin improvement effect as the skin cosmetics formulated with it, and had the disadvantage that no superior advantages could be found.

(Disclosure of the Invention) Therefore, as a result of intensive research on improving the oils and fats obtained by culturing Euglena, the present inventors added oleic acid and rapeseed fatty acids to the medium in which Euglena was cultured, and then added this medium to the medium. It was confirmed that the fats and oils obtained by holding the oil in an anaerobic state for 1 to 5 days mainly consist of wax esters having a total carbon number of 32 to 3 days. Since this composition is extremely similar to that of wax ester derived from human sebum, skin cosmetics containing this Euglena oil have much superior characteristics in terms of practical properties and rough skin improvement effect. This discovery led to the completion of the present invention.

(Objective of the Invention) That is, the object of the present invention is to provide a skin cosmetic that is excellent in practical properties (spreadability, affinity with the skin, light feeling, moisturizing property, oily feeling) and improvement effect on rough skin. .

(Structure of the Invention) The present invention involves culturing the protozoan Euglena aerobically, then adding oleic acid and rapeseed fatty acid to the medium, and then blending the oil and fat obtained by maintaining the medium in an anaerobic state. It is a skin cosmetic product.

(Specific description of the structure) Euglena used in the present invention is a protozoan that belongs to Euglena genus (genus Euglena) according to the zoological classification, and includes species, varieties, and mutant strains belonging to this genus.

Typical examples are Eugle's Gracilis 2 (
- Euglena gracilis bacillis (Eu Iena)
LL Tsuyu ε1” Yu±s var, bac i
1 jar is), Euglena viridis (Eu
1ena virid±1), Astasia longa (Astasia 4), and the like.

Euglena also naturally inhabits natural water systems such as ponds and swamps, and these can also be collected and used. Furthermore, various mutant strains obtained by treating these with known methods such as ultraviolet light treatment, heat treatment, antibiotic treatment, and nitrosoguanidine treatment can also be used.

The medium used for culturing Euklena is Koren-Hallertner medium (Journal op Protozoology (Jo)).
urnal of Protozoology)1
Volume 4 (1967, supplemented page 17) and Hallatner medium (Journal of Protodology Volume 6 (1959)).
Known media such as those described on page 23) can be used. In addition, glucose, starch hydrolyzate, molasses hydrolyzate, glutamic acid, acetic acid, ethanol, etc. are used as carbon sources,
As a nitrogen source, an inorganic nitrogen source such as ammonium nitrate, diammonium phosphate, ammonium sulfate, etc., an amino acid such as glutamic acid, aspartic acid, or an organic nitrogen source such as peptone, casamino acid, yeast extract, corn steep liquor, etc. are combined as appropriate. calcium, magnesium,
It is also possible to use a medium to which inorganic salts such as manganese and iron, and trace amounts of vitamin supplements and Inn are added. Alternatively, photosynthesis of Euglena can be used without using a carbon source.

The appropriate culture temperature for Euglena is 20 to 33°C, and the appropriate initial pH is 3.0 to 7.0. Cultivation may be performed either under light irradiation or in the dark.

Further, during culturing, it is preferable to perform shaking 50 to 250 times per minute and moderate aeration.

By culturing under such aerobic conditions, Euglena reaches the stationary phase of growth at approximately 3 to 60 mL, and the cell yield is
The dry weight per 12 culture fluids is 10 to 20 g.

The oleic acid used in the present invention is obtained from various animal fats and vegetable fats, and the content of oleic acid is 70% by weight (
Hereinafter, it will be abbreviated as wt%. ) or more are preferred.

The rapeseed fatty acid referred to in the present invention is a raw material widely used industrially, and its fatty acid composition is generally within the following range.
(Margin below) (Note) In this specification, for example, octadecanoic acid is expressed as C+6-Q, and octadecenoic acid is expressed as C+9=l.

In the present invention, any of those within these ranges can be used, but among them, those with a high content of C22=1 (erucic acid) are preferably used.

The weight ratio of oleic acid and rapeseed fatty acid added in the present invention is preferably in the range of 60:40 to 90:10. If the weight ratio of oleic acid is higher than this, the moisture retention and oily feel will be poor, and if the weight ratio of rapeseed fatty acid is higher than this, the spreadability and lightness will tend to be poor.

In addition, the fatty acid used in the present invention can be added in the form of various amine salts, metal salts, etc. in addition to the free acid form.

The total amount of oleic acid and rapeseed fatty acid added is about 0.1 to 5%, particularly about 0.5 to 2%, based on the medium.

Also, regarding the timing of addition to the culture medium, it is possible to add it to the culture medium from the beginning, but if there is a high concentration of fatty acids at the beginning of culture, it will delay the growth of bacteria, so it should be added after the bacteria have grown to a certain extent. is preferable.

However, if it is added at the early stage of culture, it is best to add it at as low a concentration as possible, for example, 1% or less, especially 0.2 to 0.5%.

Next, in order to shift the culture medium to an anaerobic state and maintain it, it is sufficient to pass nitrogen into the culture medium. After shifting the medium to anaerobic conditions,
By maintaining this state for 1 to 5 days, the Euglena oil and fat used in the present invention is accumulated in Euglena cells.

Here, the mechanism of oil production by Euglena under anaerobic conditions is considered to be as follows.

That is, when Euglena that has been cultured under aerobic conditions is placed under anaerobic conditions, paramin, which is a polysaccharide stored under aerobic conditions, is decomposed into glucose units and becomes pyruvic acid through glycolysis. The generated pyruvic acid becomes acetyl coenzyme A, and fatty acids are further synthesized from acetyl coenzyme A.

The fatty acids synthesized through this process have 14 carbon atoms.
11 to 17 saturated straight chain fatty acids, mainly
A part of it is reduced to alcohol with almost the same carbon number distribution, and an esterification reaction occurs between the two components, producing fats and oils.

Therefore, in normal culture media, both fatty acids and alcohols are saturated, and the fats and oils obtained from them are also only saturated.

However, when unsaturated fatty acids are present in the cells during the anaerobic oil production process by Euglena,
Some of it forms ester bonds with saturated long-chain alcohols derived from glucose, producing esters of unsaturated fatty acids and saturated alcohols, and the other part is reduced to unsaturated long-chain alcohols. After that, an ester bond is formed between the saturated fatty acid derived from glucose and the unsaturated fatty acid within the cell.

Once oils and fats have been accumulated within the cells in this way, the bacterial cells are separated from the culture medium by a method such as centrifugation, and the oils and fats are extracted and purified by conventional methods to obtain the Euglena oils used in the present invention. .

To summarize the constituent components of the Euglena oil and fat, the ester composition is: (1) Mainly monoesters with a total carbon number of 32 to 38 65 to a
owt% (2) Consists of 20 to 35 wt% of triglycerides whose fatty acid moiety mainly has carbon atoms of CI4 to C+s.

The skin cosmetic of the present invention contains Euglena oil obtained as described above in a skin cosmetic base such as creams, emulsions, lipsticks, foundations, etc. based on the total amount of 2.
What is necessary is just to mix 0-5.0 Wt% and manufacture it by a normal method.

(Example) Hereinafter, the present invention will be specifically explained using experimental examples and examples.

Test methods for practical properties and rough skin improvement effects described in Examples are as follows.

(1) Practical property test After 20 female subjects used the sample for 10 days, they evaluated the sample's ``spreadiness'', ``compatibility with the skin'', ``light feeling'', ``moisturizing ability'', and ``oily feel''. did.

The test results were ``spreads well when applied to the skin'', ``fits well to the skin'', ``feels light with less stickiness'', and ``leads to moderately moist skin''. It is shown by the number of people who answered "It has a pleasant oily feel" and "It has a desirable oily feel."

(2) Effect test on improving rough skin The effect of continuous application for 4 weeks was investigated on 20 middle-aged and elderly subjects who had rough skin on their lower legs. Approximately 1 g of the sample was applied to the test site of the left lower leg of the subject once a day, and the condition of the skin before and after the test was judged according to the following criteria.

No sample was applied to the right lower leg, which served as a control.

Compare the judgment results of the test site and control site before and after the test, and if the skin dryness has improved by two or more levels (for example, +-1,
+10-±) is valid, one level improvement is slightly valid,
If there was no change, it was considered invalid. The test results are shown as the number of subjects who found the drug to be effective or somewhat effective.

Experimental Examples 1 to 4 2 g of glucose 1 0.5 g of polypeptone, 25 mg of ammonium sulfate, 25 mg of ammonium hydrogen carbonate, 25 mg of diammonium phosphate, 60 mg of magnesium carbonate,
12 mg of calcium carbonate, 5 mg of Mohr's salt, 1.8 mg of manganese sulfate hydrate, 2. zinc sulfate heptahydrate.
5 mg g, 0.25 mg of vitamin BJ and 0.0005 mg of vitamin B+z O in 100 ml of deionized water.
and the pH was adjusted to 4.5.

This was put into a 500ml long neck shake flask and used as a medium, and Euglena gracilis 2 strains were inoculated into it.
The cells were cultured aerobically for 3 days at 28° C. in the dark with shaking 120 times per minute.

Subsequently, beef tallow oleic acid (oleic acid content so
After adding mixed fatty acids with varying mixing ratios of (wt%) and rapeseed fatty acids (the composition of which is shown in Table 1) as shown in Table 2, aerobic culture was continued for 3 days.

Table 1 [Composition of rapeseed fatty acids used in Experimental Examples 1 to 4] Next,
Transfer all the contents of the flask to a 200 ml three-necked round bottom flask, adjust the pH to 6.5, and stir gently at 20°C while bubbling nitrogen gas under room light.
Anaerobic culture was continued for an additional 3 days.

Next, the bacterial cells were collected by centrifugation and diluted with 0.85% saline.
After washing twice, the bacterial cells were washed with chloroform/methanol (
1: 1) It was immersed in a mixed solution and homogenized with ultrasonic waves to crush the bacterial cells, and the oil and fat were extracted, and decolorized, deodorized and purified using activated clay and activated carbon to obtain Euglena oil and fat.

Table 2 shows the composition of the monoester component (composition ratio by total carbon number) in the Euglena oil and fat obtained here.

Examples 1 to 2, Comparative Examples 1 to 3 (Two-layered skin lotion) A two-layered skin lotion with the composition shown below, containing olive oil, macadamian nut oil, Euglena oil, etc. of the above experimental example, as shown in Table 3. Each skin lotion was prepared by blending as described above, and the score test was conducted.

(2) Preparation method After each of the components (A) and (B) is uniformly dissolved, the components (A) and (B) are mixed, stirred and dispersed, and then filled into a container. When using, shake and disperse the contents uniformly.

(3) Characteristics The results of various tests conducted on each two-layer skin lotion are listed in the right column of Table 3.

As shown in Table 3, skin lotions containing olive oil and macadamian nut oil of Comparative Examples 1 and 2, and skin lotions containing Euglena oil and fat of Experimental Example 1 containing only oleic acid of Comparative Example 3. No good results were obtained in terms of practical properties and rough skin improvement effect. On the other hand, the skin lotions of Examples 1 and 2, which are skin cosmetics of the present invention, exhibited excellent properties in all tests. (Left below) Example 3, Comparative Examples 4 to 5 [Skin cream] In the same manner as in Example 1, each skin cream was prepared by blending oils and fats as shown in Table 3, and the various tests described above were carried out. carried out.

(2) Preparation method Components (A) and (B) were each heated and melted at 80°C.

Next, mix ingredients (A) and (B) and stir while stirring.
Each skin cream was prepared by cooling to 0°C.

(3) Properties As shown in Table 3, the skin cream of Example 3, which is the skin cosmetic of the present invention, had superior properties in all tests compared to the skin creams of Comparative Examples 4 and 5. showed that.

Example 4, Comparative Example 6 [Makeup 7 Soap Base] Similarly to Example 1, each makeup base was prepared with the following composition and various tests were conducted.

(Left below) (2) Characteristics The makeup base of Example 4 showed clearly superior properties compared to Comparative Example 6.

Example 5, Comparative Example 7 1 m of each lip balm having the following composition was manufactured and various tests were conducted.

The rough skin improvement effect test was conducted on 20 female subjects complaining of chapped lips, and after using the lip balm continuously for 10 days, the number of people who responded that their chapped lips were improved was shown.

(2) Preparation method Heat all ingredients to a temperature of 80°C, mix and dissolve them uniformly, pour into a mold, cool and mold each solid lipstick.
Manufactured.

(3) Characteristics The lip balm of Example 5, which is the skin cosmetic of the present invention, exhibited superior characteristics in all tests compared to Comparative Example 7.

(Effects of the Invention) As described above, the skin cosmetic of the present invention passed all practical property tests (spreadiness, skin affinity, light feel, heat retention, oily feel) and rough skin improvement effect tests. It is clear that these products exhibit excellent properties and provide useful skin cosmetics.

(Blank below) Voluntary amendment to the procedure) November 11, 1985 Relationship with the nature of the patent application submitted on November 8, 1986 Patent applicant address 671-4 Mizuashi, Noguchi-cho, Kakogawa-shi, Hyogo Name Harima Kasei Kogyo Co., Ltd. Representative: Mikichi Hase 4, Agent: Postal code: 530 Address: 3-21-5, Suehiro-cho, Kita-ku, Osaka City, Date of amendment order: Voluntary amendment: 6, Subject of amendment + 11 Details of the invention in the specification An explanation column.

7. Contents of the amendment +11 On page 6 of the specification, line 10, character 11, the word "mouth" is amended to read "日".

(2) Insert the two characters "Shio no" in the 16th character of the second line on page 8 of the specification.

(3) r from page 10, line 19, character 12 of the specification
"s, o w t%" should be corrected to "rsowt%".

(4) Page 19 of the specification, 4th line, 1st O character, 1
Insert “(Table 3)” and correct it as “・Euglenoid oil, etc. (Table 3).”

(4) On page 20 of the specification, line 10, character 10, “
(Table 3)" is inserted.

(5) On page 21 of the specification, line 5, change the second character “warm” to “warm”.
Corrected as "humid."

(6) On page 22 of the specification, in the left column of Table 2, the phrase "total carbon number constituting monoester" is corrected to "composition by total carbon number of monoester".

Claims (1)

[Scope of Claims] 1) The protozoan Euglena is cultivated aerobically, then oleic acid and rapeseed fatty acids are added to the medium, and then oil and fat obtained by maintaining this medium in an anaerobic state is blended. skin cosmetics. 2) Addition weight ratio of oleic acid and rapeseed fatty acid is 60:
The skin cosmetic composition according to claim 1, which has a ratio of 40 to 90:10.