JPH03227909A - Cosmetic containing natural red pigment - Google Patents

Abstract

PURPOSE:To obtain a non-toxic and safe cosmetic suitable for cosmetics such as lipstick, rouge and nail enamel by using a natural red pigment obtained from a callus derived from the tissue of a plant of genus Malus as a colorant. CONSTITUTION:A tissue piece (e.g. shoot apex, cotyledon or hypocotyl) of a plant of genus Malus is planted on a medium and a callus is induced under irradiation. A callus capable of stably producing a red pigment is selected. The cell is proliferated in dark place and the proliferated cells are collected as incipient cells. The cells are cultured in a medium to collect a callus having red or dark red color. The callus is extracted with a proper solvent and the solvent is removed to obtain a red pigment to be used as a raw material for a cosmetic. The non-toxic and safe natural red pigment produced by this process is used as a component of cosmetics such as lipstick, rouge and nail enamel.

Description

[Detailed Description of the Invention] [Industrial Application Field] The present invention relates to cosmetics such as lipsticks, blushers, and nail enamels containing a red natural pigment obtained from callus derived from tissues of plants of the genus Malus. .

[Prior art and problems to be solved by the present invention] Cosmetics such as blushers, blushers, and nail polishes usually contain coloring agents such as oils and fats.
An oily base such as wax, hydrocarbon, or higher alcohol, and if desired, other components such as antioxidants, surfactants, etc.
It is manufactured by blending it with plasticizers, solvents, etc.

Conventionally, synthetic colorants have been used as such colorants, but in recent years, the use of such synthetic colorants has been discouraged due to drawbacks such as roughening of the skin and nails, and negative effects on the human body as lipsticks are swallowed with food. Avoid non-toxic colorants (especially red colorants commonly used in lipsticks, blushers, and nail enamels)
It is desired that safe cosmetics containing . Although various studies have been made for this purpose, a cosmetic containing the apple red natural pigment of the present invention is not yet known.

[Means for Solving the Problems 1] In order to solve the above problems, the present inventors focused on the red pigment contained in apples. However, conventional apples (Malus pumila Mill, va.
r.

domestica C, 5chn) has anthocyanin pigment (cyanidin-3-galac) in the pericarp.
Although it is attracting attention as a raw material for red pigments, it has been difficult to use it industrially as a coloring agent because the pigments are only localized in several layers of epidermal cells.

As a result of intensive studies, the present inventors succeeded in creating an industrial method for producing apple red natural pigment by tissue culture of a specific apple, as described below. It has been found that this compound is not only non-toxic and safe, but also has physical properties that are extremely suitable for the production of cosmetics such as lipstick, blusher, and nail enamel.

That is, the present invention is: 14 Cosmetics 2 containing apple red pigment, Lipstick 3 containing apple red pigment, and Blush containing apple red pigment.

Hereinafter, a method for producing a red natural pigment, which is a raw material of the present invention, and a method for producing a cosmetic using the red natural pigment as a raw material will be explained in detail.

The red natural pigment used as a raw material for the cosmetic of the present invention consists of four steps: callus induction, selection, dispersed cell proliferation, and pigment production.

The steps will be explained below in order.

Callus induction step Callus is induced and cultured from apple plant tissue by a method known per se. As the apple plant tissue, for example, a tissue piece of an apple branch, leaf, fruit, root, etc., preferably a growing part, such as a shoot tip, cotyledon, hypocotyl, etc., is used. This tissue piece is placed on a medium, for example, a liquid medium or a solid medium, and a callus is induced under illumination.

It is best to sterilize apple plants with disinfectant alcohol, sodium hypochlorite solution, bleaching powder suspension filtrate, etc., wash them with sterile water, cut them into small pieces, and place them on a culture medium.

As a medium, in addition to normal solid media, liquid media used for plant tissue culture, such as Murashige-Skoog's (hereinafter abbreviated as MS) medium, Ganpaug's B5 (hereinafter referred to as B
5), Linsmeyer-Skoog's medium, White's medium, Hella's medium, Sienck-Hildebrand's medium, Nilachie-Nirachie's medium, and modified media thereof. Alternatively, a solid medium obtained by adding a solidifying agent (agar, agarose, gelrite, etc.) to these liquid media may be used.

Such a medium may contain a carbon source, a nitrogen source, inorganic salts, organic substances, etc. as appropriate.

Examples of carbon sources that can be used include sugars such as sucrose, glucose, galactose, fructose, and maltose, and soluble starch. As the nitrogen source, for example, nitrates, ammonium salts, etc. are used. Examples of inorganic salts used include those containing elements such as phosphorus, potassium, calcium, magnesium, manganese, copper, zinc, molybdenum, boron, iron, cobalt, and nickel. Examples of organic substances include inositol, nicotine, pyridoxine hydrochloride, thiamine hydrochloride, calcium pantothenate, folic acid, p-aminobenzoic acid, biotin, choline chloride, riboflavin, ascorbic acid, vitamins such as vitamin A1, vitamin D3, and vitamin BI2. Organic acids such as sodium pyruvate, citric acid, malic acid, fumaric acid, natural substances such as coconut milk, casein hydrolyzate, yeast extract, etc. are used.

Further, plant growth regulators such as auxins and cytokinins, agar, and the like may be appropriately added to the medium. Examples of such auxins include 2,4-dichlorophenoxyacetic acid, 2,4-dichlorophenylacetic acid, indole-3-acetic acid, and indole-3-acetic acid.
Butyric acid, l-naphthaleneacetic acid (hereinafter abbreviated as NAA), 2
-Naphthoxyacetic acid, parachlorophenoxyacetic acid, 2.4
5-Trichloro[7-phenoxyacetic acid, 1-naphthaleneacetamide, etc. are used. In addition, examples of cytokinins include 6-benzyladenine (hereinafter referred to as BAP).
), 2-isopentyladenine, 2-isopentenyladenine, kinetin, zeatin, dihydrozeatin, zeatin riboside, diphenylurea, etc. are used. Among these, NAA and the like are frequently used as auxins, and BAP and the like are frequently used as cytokinins.

Normally, auxins are about 0.01 to 20 ppm, and cytokinins are about 0.01 to 20 ppm. It is added to the medium at a rate of 1 to 15 ppm. When cultured in the light at approximately 15-35°C, approximately 1
Callus is induced from the tissue piece after 0 to 40 days.

Of course, inorganic or organic acids, alkalis, buffers, etc. may be added for the purpose of adjusting the pH of the medium, or an appropriate amount of fats and oils 9 surfactants may be added for the purpose of defoaming.

The culturing method may be static culture, shaking culture, or aeration/agitation culture. For large-scale processing, it is desirable to use so-called shaking culture. Although the culture conditions naturally vary depending on the state and composition of the medium, the means of culture, etc., it is usually preferable to select a temperature between 10° C. and 45° C. and an initial pH around neutrality. In particular, the temperature during the middle stage of culture is 20°C to 37°C1, and the initial pH is 5.
．． Conditions of 0 to 8.5 are desirable. The culture time may be about 1 day to 3 months, but 1 week to 4 weeks is particularly preferable.

Cell selection process The formed reddish callus is separated from the tissue piece and transplanted into a solid medium or liquid medium similar to that used for inducing calf callus. Cultivation can be performed under the same conditions as in the callus induction step.

After culturing, the dark red callus portion is cultured again to obtain a red pigment culture with high pigment concentration. If this operation is repeated every 5 to 60 days, preferably every 14 days, and subplant selection is performed, calli that stably produce red pigment can usually be obtained after 1 to 5 months.

This is selected and used in the next step.

Dispersed Cell Proliferation Step The obtained callus that stably produces red pigment is transferred to the liquid medium described above, preferably in the dark, and grown every 5 to 14 days.
The cells are expanded by repeating the subculture 10 times. When this cell proliferation is carried out for about 3 weeks, a brittle and homogeneous dispersed cell mass is obtained. This cell mass is crushed into fine cell particles in a liquid and then cultured in suspension for further cell proliferation. As the medium, a liquid medium similar to that used for callus induction, preferably,
B5 medium is used. Culture can be performed under the same conditions as those used for callus induction.

This culture allows the cells to proliferate rapidly, and this 8 Cultivation can be carried out in large quantities.

The resulting cell mass is subcultured every 5 to 14 days.

At this time, it is preferable to use the cell mass after passing it through a sieve.

After about 2 to 3 weeks, the proliferated cells are collected as initial cells and used in the next step of producing red pigment.

If this step is performed in the light, the pectin-like substance will gradually increase and the proportion of red cells will decrease, so it is preferable to perform it in the dark.

The initial cells obtained in the previous step of the red pigment production process are cultured in the same solid medium or liquid medium as used for callus induction. A preferred medium is a liquid medium, such as MS medium.

The culture period is 3 to 14 days, preferably 5 to 10 days. The culture temperature is about 10-30°C.

Cultivation is performed under illumination, preferably under blue light irradiation with a wavelength of 220 to 500 nm. This culture yields red to dark red callus. These calli contain large amounts of red pigment. The callus is collected, squeezed or pulverized if necessary, extracted with an appropriate solvent, and the solvent removed to obtain a red pigment that can be used as a raw material for the cosmetic composition of the present invention. The pigment may be extracted from the collected callus without drying it, or it may be extracted after being dried by a conventional drying method such as vacuum drying. As the extraction solvent, alcohols such as methanol, ethanol, etc. are preferred, and these preferably contain hydrochloric acid. Particularly preferred is 0.05-2, Ow/w
% hydrochloric acid in methanol. The preferred temperature for extraction is about θ to 20°C and the time is about 16 hours to 1 week. This extraction is preferably carried out in the dark.

The red natural pigment produced in this way may optionally be
For example, dissolution, concentration (preferably vacuum concentration).

Further purification may be performed by means known per se such as chromatography and crystallization.

Method for producing cosmetics containing apple red pigment A lipstick and blusher containing the red natural pigment of the present invention.

Cosmetics such as nail enamel and lipstick are made from red pigment powder obtained from apple callus using the method described above.

It can be easily manufactured by incorporating it into a diluent for manufacturing blushers, nail enamels, etc. As diluents, those known for the production of such cosmetics are conveniently used in the present invention. Such diluents include, for example, oils, waxes, and hydrocarbons.

Oily bases such as higher alcohols are preferred. The oil bases used here vary depending on the type of cosmetic, but in the case of lipstick, castor oil, butyl stearate, tetrahydrofurfuryl esters, polyethylene glycol, propylene glycol monoester, cacao butter, hydrogenated oil , Vaseline.

Lanolin, lecithin, carnauba wax, candelilla wax, beeswax, oshkelite, paraffin.

Examples include hydrogenated castor oil.

For blushing, use beeswax or carnauba wax.

Candelilla wax, ceresin, paraffin, spermaceti, lanolin, lanolin absorption base, petrolatum, liquid paraffin, castor oil, cocoa butter, butyl stearate, tetrafurfuryl alcohol ester, cetanol, stearyl alcohol, hydrogenated vegetable oil, etc. Examples include fats and oils, waxes, hydrocarbons, and higher alcohols. In the case of nail enamel, butyl acetate, toluene, etc. can be used as an oil base instead.

The cosmetic composition of the present invention can be easily produced by mixing the above-mentioned components by means known per se, and if desired, other components such as antioxidants may be added.

Surfactants, plasticizers, solvents, humectants, thickeners, preservatives,
Fragrances, other non-toxic or low-toxic colorants or pigments, etc. may be added.

In addition, when producing nail enamel as a cosmetic,
It is preferable to add and mix a film-forming resin such as nitrocellulose or polyacrylic acid ester.

Although the amount of red natural pigment used in the cosmetics of the present invention cannot be definitively stated, it is usually about 0.001% to 0.001% based on the finished cosmetics.
It is about 20%.

[Example] The present invention will be further explained below in the form of reference examples and examples.

In each example, % indicates weight % unless otherwise specified.

Reference example (callus induction process) The apical bud or axillary bud of an apple (variety Nistar King) was
% ethanol for 5 minutes, then immersed in 1% sodium hypochlorite solution for 10 minutes, and then washed five times with sterile water. Next, the shoot apex was removed under a stereomicroscope and placed on MS agar medium (
NAA2. Omg/12.

BAP2.5mg/ff, agar 9g/Q, pH5,
8). Under lighting (white fluorescent light, 2000Lu)
x.

After culturing at 25°C for 3 weeks (16 hour photoperiod), the reddish callus formed on the cross section of the shoot apex was separated and transplanted onto the above MS agar medium.

(Cell selection process) After that, the dark red callus parts were transplanted every 7E1, and
After several months, callus that stably produced red pigment was obtained.

(Dispersed cell growth step) The callus obtained above was grown in a growth medium (NAA2.Omg/
(2, BAP2.5mg/(2, B5 with 3% sucrose added)
The cells were cultured in the dark (60 rpm) using a liquid medium). Cells were repeatedly subcultured every 7 days, and uniformly dispersed cells were obtained after one month.

(Red pigment production process) The dispersed cells were collected by filtration on a nylon mesh, and the pigment production medium (NAA 2.0 mg 10.0, BAP 2.5 mg /
Q + MS medium supplemented with 3% sucrose) 9c containing 20 cells
The cells were transplanted into a plastic petri dish of m.

The initial cell amount was 20 mg/m12. Culture was carried out under blue fluorescent lamp (product name: FL-40SB, NEC) lighting.
The cells were cultured at 25° for 0.8 days (60 rpm).

After filtering the culture solution containing the callus obtained above using a 50 μm nylon mesh and freeze-drying it,
The same weight of methanol containing 1% hydrochloric acid was added, and the mixture was left in the dark at 4°C for 1 day to extract the red pigment. After extraction, the callus was filtered off using a filter paper, and the solution was dried under reduced pressure using an evaporator while keeping the temperature below 35° C. to obtain a red pigment powder.

This red pigment powder was dissolved in water, and a cation exchange resin (Dowex 50W-X2, 200-400 mesh, Dow Chemical Company) was added to this solution and stirred for 16 hours to adsorb the pigment to the resin. This was washed successively with water and methanol to remove impurities such as sugar and lipids, and then the dye was eluted with methanol containing 1% hydrochloric acid. The obtained dye fraction was further subjected to preparative cellulose TLC (/i opening solvent: A
cOH:HC(1:H20=15+3:82
), scrape off the pigment component band, elute with methanol containing 1% hydrochloric acid, concentrate under reduced pressure until a small amount of methanol is present, and then add 20 times the amount of ether to precipitate the pigment to obtain purified pigment powder. Ta. In the above method, 2 mg of red pigment powder was obtained from 3.34 g (fresh weight) of callus, and 1 mg of purified pigment powder was obtained from this.

Example 1 3g of red pigment powder obtained from apple callus was added to Nilacol H.
CO-20 (manufactured by Nikko Shokai) 0-5gs castor oil 68.
0g and sufficiently mixed and dispersed. Separate carnauba wax 9.5
14.5 g of beeswax and 4.5 g of Lano 5 phosphorus were heated and melted together, and the red pigment powder dispersion was added thereto and thoroughly mixed. Finally, 10 mg of fragrance was added and further mixed, poured into a mold while stirring at room temperature, and rapidly cooled to obtain a red lipstick.

Example 2 4.0 g of apple red pigment powder, 6.0 gs of Carbowax 400, 10 g of castor oil. A liquid blush was prepared by mixing 80.0 g of Og, ethyl alcohol and lomg of fragrance.

Example 3 Nitrocellulose 15.0g, Santorite 745g5
Nail enamel was produced by mixing 3.75 g of diptylphthalate, 29.35 g of butyl acetate, 6.4 g of ethyl alcohol, 1.1 g of butyl alcohol, 36.9 g of doluol, and 4.0 g of apple red pigment powder.

Claims (3)

[Claims] (1) Cosmetics containing apple red pigment. (2) Lipstick containing apple red pigment. (3) A blusher containing apple red pigment.