CN111616985B - Oil body cleansing product and preparation method thereof - Google Patents

Abstract

The invention discloses a washing and caring product of oil body and a preparation method thereof, and discloses a washing and caring product, which comprises vegetable oil body emulsion, wherein the vegetable oil body emulsion is prepared by the following method: adding the xanthan gum into deionized water, uniformly stirring to avoid bubbles, fully swelling, and slowly adding the vegetable oil body suspension under the action of a magnetic stirrer; the plant oil body is a plant oil body which over-expresses oleosin; the final concentration of the oil body is 10%; the concentration of xanthan gum is 0.05-0.5%. The washing and caring product prepared by the invention can reduce the oil content and increase the water content of skin, so that the skin elasticity is increased, the moisturizing and water-locking properties are good, and the safety is high.

Description

Oil body care product and preparation method thereof

Technical Field

The invention belongs to the field of biotechnology and cosmetics technology, and in particular relates to an oil body washing and care product and a preparation method thereof.

Background Art

Oil bodies contain highly unsaturated fatty acids, with linoleic acid content of more than 80%. Their solution can be regarded as an oil-water (O/W) emulsion. They are an important additive in food processing. Conventional cream foods, salad dressings, mayonnaise, meat and meat products are widely used in the food industry. The stability of the gel is the most important performance among its many physical and chemical properties. Only under the condition of reasonable control of system stability can the product ensure its functional characteristics.

Plant oil bodies are originally very unstable components in the in vitro state after extraction. Plant oil bodies exist in vitro and are easily changed in quality due to isomerization, oxidative degradation, etc. caused by many factors such as air, heat and metal ions. Therefore, the stability of oil bodies has attracted much attention. The addition of gel matrix plays an indispensable role in oil-water food industry products, which can cover the oil bodies and then generate repulsive force to prevent the oil bodies from agglomerating to maintain their stability.

Nowadays, many researchers at home and abroad have begun to study oil body structured systems by combining plant oil bodies with the structural materials of gels to form a viscoelastic "gel-like" structure. The gel can effectively reduce trans fatty acids and saturated fatty acids. Oil body gels are mainly divided into two categories, namely low molecular weight organic gel systems, which are mainly composed of small molecules that form self-assembled or crystalline networks. The gelling agents used are mainly saturated fatty acids or alcohols, beeswax, plant sterols and peptides, or lecithin; another method is to use hydrophobic biomacromolecules and their derivatives, such as ethyl cellulose, soy protein. Biomacromolecules have very good hydrophilicity, and the molecules are entangled with each other and form a dense network structure through hydrogen bonds and electrostatic interactions.

At present, there are many chemically synthesized cleaning products on the market, but the ingredients they contain are worrying. Although the chemicals contained in the products can clean the skin, they will also damage the skin, even cause allergic reactions, and lead to skin aging. With the development of the times, people are paying more and more attention to health, and the country's supervision of the environment is also increasing day by day. Therefore, a method for making skin care products using pure natural and non-irritating plant oil raw materials is of great significance. Natural raw materials can not only avoid damage to the skin, but also the natural ingredients of pure natural plant oils (safflower oil, soybean oil, sunflower oil, etc.) can balance sebum, regulate skin quality, nourish and moisturize, and have antibacterial and anti-inflammatory effects; the addition of active ingredients can better clean the dirt deep in the skin and deeply clean the skin. At the same time, natural ingredients will not pollute the environment. This is an inevitable trend in the development of the cosmetics industry in the future.

Summary of the invention

The present invention aims to provide an oil body care product and a preparation method thereof.

A washing product, comprising a plant oil emulsion, wherein the plant oil emulsion is prepared by the following method: adding xanthan gum to deionized water, stirring evenly without bubbles to fully swell the xanthan gum, and slowly adding the plant oil suspension under the action of a magnetic stirrer;

The plant oil body is a plant oil body that overexpresses oleosin;

The final concentration of the oil bodies is 10%;

The concentration of the xanthan gum is 0.05-0.5%.

The cleaning product is an oil emulsion, which is composed of the following components in mass fraction: 10-55% recombinant plant oil emulsion, 5-10% glycerol, 10-20% carboxyethyl cellulose, 1-5% PEG800, 5-20% vitamin C, 0.1-0.5% essence, with a total weight of 100%.

The washing product is an oil-based cold-processed soap, which is composed of the following components in weight fractions: 2-20% reconstituted vegetable oil emulsion, 15-40% coconut oil, 15-40% palm oil, 1-10% castor oil, 2-10% rosehip oil, 2-10% olive oil, 2-10% shea butter, 5-20% alkali, and the balance distilled water, with a total weight of 100%.

The cleaning product is an oil-based hand cream, which is composed of the following components in mass fractions: 5-40% recombinant plant oil suspension, 0.05-0.5% xanthan gum, 5-10% polyvinylpyrrolidone PVP, 3-8% triglyceride, 10-20% cetyl alcohol, 15-25% cetostearyl alcohol, 15-20% monoglyceride, 1-10% xanthan gum, 5-10% German BASF, 10-15% glycerol, and the balance is water, with a total weight of 100%.

The cleaning product is an oil foot cream, which is composed of the following components in mass fractions: 5-40% recombinant plant oil suspension, 0.05-0.5% xanthan gum, 15-20% glycerol, 10-20% cetyl alcohol, 10-15% cetostearyl alcohol, 15-20% monoglyceride, and the balance is water, with a total weight of 100%.

The present invention provides a washing product, which includes a plant oil body emulsion, wherein the plant oil body emulsion is prepared by the following method: adding xanthan gum to deionized water, stirring evenly without bubbles, making it fully swollen, and slowly adding the plant oil body suspension under the action of a magnetic stirrer; the plant oil body is a plant oil body overexpressing oleosin; the final concentration of the oil body is 10%; and the concentration of the xanthan gum is 0.05-0.5%. The washing product prepared by the present invention can reduce the oil content of the skin, increase the moisture content, increase the elasticity of the skin, have good moisturizing and water-locking properties, and have high safety.

BRIEF DESCRIPTION OF THE DRAWINGS

Fig. 1 Appearance of recombinant plant oil bodies;

Figure 2 Comparison of the microstructures of recombinant plant oil bodies and wild oil bodies; A, wild oil bodies; B, recombinant plant oil bodies; C, wild oil body emulsion; D, wild oil body emulsion;

Figure 3 Appearance of recombinant plant oil body suspension;

Fig. 4 is a graph showing the particle size of the recombinant plant oil body suspension;

Figure 5: Appearance of reconstituted vegetable oil body emulsion;

Figure 6 Particle size detection diagram of recombinant plant oil body emulsion;

Figure 7: Finished product of cold-process soap made from reconstituted vegetable oil;

Figure 8 Appearance of reconstituted vegetable oil hand cream.

DETAILED DESCRIPTION

Example 1 Preparation of recombinant plant oil bodies

1. Obtaining plants overexpressing oleosin

The pOBT expression vector was constructed, and the T-DNA region was modified on the basis of the vector skeleton pCAMBIA1301. The plasmid also had NcoI and HindIII restriction sites. The safflower oleosin gene (base sequence shown in the sequence table SEQ ID NO.1) was inserted into the pOBT expression vector to obtain the recombinant pOTB-Oleosin vector, which was transfected into Agrobacterium competent cells, and PCR identification was performed using the bacterial solution as a template. The transfected Agrobacterium was used to transform safflower by using the Agrobacterium-mediated method: when the germinated seeds had not yet grown true leaves, the growth point below the cotyledon was cross-cut, and the surface wound of the explant was contacted with the prepared Agrobacterium bacterial solution for infection for 10 minutes, and then cultured in a dark box at 25°C for 4 days. After 4 days of dark culture, the cotyledon node was transferred to a bud proliferation medium, and after 1 to 3 green buds grew out of the wound of the cotyledon node, it was transferred to an elongation medium, cultured for about 4 weeks, and hardened and transplanted; after harvesting, T1 generation safflower plants overexpressing oleosin were obtained.

2. Detection of gene and protein levels in seeds of plants overexpressing oleosin

Extract oleosin-overexpressing safflower genomic DNA using specific primers:

F:3’-5’:ATGGGTACGGTTGAAACGACA

R:3’-5’:AGCTCCCGACAACCGACCAA

PCR detection was performed to amplify the target band with the same size as the oleosin protein, preliminarily confirming that it had been inserted into the plant genome; recombinant oil bodies were extracted and SDS-PAGE detection was performed, and finally T3 generation safflower seeds overexpressing oleosin were obtained.

3. Preparation of recombinant plant oil bodies

Take 20g of T3 generation safflower seeds overexpressing oleosin, wash them 2-3 times, absorb water and dry them; use an ultra-high-speed grinder to crush them for 10s; add 200mL PBS buffer solution, grind them with a juicer, filter them with gauze, place the filtrate in a 50mL centrifuge tube, and use an ultracentrifuge at 4℃ 12000 r/min for 15 min; resuspend the upper solid matter with PBS buffer solution, stir with a glass rod until there are no particles, and ultracentrifuge at 4℃ 12000 r/min for 15min, repeat once; resuspend the upper solid matter with PBS buffer solution, stir with a glass rod until there are no particles, and ultracentrifuge at 4℃ 12000 r/min for 15min, repeat twice; the upper white solid is the recombinant plant oil body (oleosin overexpressing oil body), which is stored at 4℃ for future use.

Example 2 Preparation of recombinant plant oil bodies

1. Obtaining plants overexpressing oleosin

The pOBT expression vector is constructed, and the T-DNA region is modified on the basis of the vector skeleton pCAMBIA1301, and the plasmid also has NcoI and HindIII restriction sites; the Arabidopsis oleosin gene (base sequence is shown in the sequence table SEQ ID NO.2) is inserted into the pOBT expression vector to obtain the recombinant pOTB-Oleosin vector, which is transfected into Agrobacterium competent cells, and PCR identification is performed using the bacterial solution as a template; the transfected Agrobacterium is used to transform Arabidopsis explants using the Flora Dip method: when the Arabidopsis is in the state of white flowers and clips, it is inverted in the prepared Agrobacterium bacterial solution for infection for 8-15 minutes, and is cultured in the dark for 2-3 days. After harvesting, resistance screening is performed to obtain T1 generation Arabidopsis plants overexpressing oleosin.

2. Detection of gene and protein levels in seeds of plants overexpressing oleosin

Extract genomic DNA of Arabidopsis thaliana overexpressing oleosin with specific primers:

F:3’-5’:ATGGCGGATACAGC TAGAGGAA

R:3’-5’:CACCGGGTGGAGTAGTGTGCTGG;

PCR detection was performed to amplify the target band with the same size as the oleosin protein, preliminarily confirming that it had been inserted into the Arabidopsis plant genome; recombinant oil bodies were extracted for SDS-PAGE detection, and finally T3 generation Arabidopsis seeds overexpressing oleosin were obtained.

3. Preparation of recombinant plant oil bodies

Take 20g of T3 generation Arabidopsis seeds overexpressing oleosin, wash 2-3 times, absorb water and dry; add 200mL PBS buffer solution, grind with a mortar, filter with gauze, place the filtrate in a 50mL centrifuge tube, and use ultracentrifuge at 4℃ 12000 r/min for 15 min; resuspend the upper solid matter with PBS buffer solution, stir with a glass rod until there are no particles, and ultracentrifuge at 4℃ 12000 r/min for 15min, repeat once; resuspend the upper solid matter with PBS buffer solution, stir with a glass rod until there are no particles, and ultracentrifuge at 4℃ 12000 r/min for 15min, repeat twice; the upper white solid is the recombinant plant oil body (oleosin overexpressing oil body), which is stored at 4℃ for future use.

Example 3 Preparation of recombinant oil body suspension

The pH of PBS buffer was adjusted to 6-9 and the ion concentration was 10 mM using 1 mol of HCl or NaOH. The recombinant plant oil bodies were added to the PBS buffer and stirred evenly with a magnetic stirrer to prepare a recombinant oil body suspension without particles. The concentration of the recombinant oil body suspension was 10%, and the appearance was shown in Figure 3. The particle size of the recombinant oil body suspension was 400-900 nm, and the particle size of the wild-type oil body suspension was 700-1500 nm (Table 1, Figure 4).

Example 4 Preparation of reconstituted oil body emulsion

Select a suitable stabilizer from carbomer, sodium carboxymethyl cellulose, Tween-80, Tween-20, xanthan gum, dextran and other stabilizers; weigh an appropriate amount of stabilizer and add it to deionized water, stir evenly without bubbles, and make it fully swollen. Slowly add the plant oil body suspension under a magnetic stirrer to finally prepare a reconstituted plant oil body emulsion, the stabilizer concentration range is 0.3%, and the final oil body concentration is 10%; by measuring the microstructure, particle size and potential indicators, xanthan gum was finally determined to be the best stabilizer (Table 2), and the appearance of the reconstituted plant oil body emulsion is shown in Figure 5.

Example 5 Detection of the stability of reconstituted oil body emulsion

1) The prepared vegetable oil emulsion (xanthan gum as stabilizer) was placed in a constant temperature water bath and treated at 30℃, 60℃, and 90℃ for 30min, and at 50w-250w high ultrasound for 30min, 60min, and 90min. The results of microstructure, particle size, and potential index determination showed that the vegetable oil emulsion had good stability to temperature and ultrasound.

2) Alkali resistance test: When 5% sodium hydroxide is added to the vegetable oil emulsion (xanthan gum as stabilizer), it still appears as a milky white oil emulsion with no change in appearance.

Example 6 Preparation of reconstituted oil body skin care lotion

Weigh the raw materials according to their mass percentage: 10-55% recombinant vegetable oil body emulsion (recombinant oil body suspension plus xanthan gum), 5-10% glycerol, 10-20% carboxyethyl cellulose, 1-5% PEG800, 5-20% vitamin C, 0.1-0.5% flavor, and the final percentage content of all the above ingredients is 100%; first add glycerol, carboxyethyl cellulose, PEG800, and vitamin C into a stirring cup, and emulsify under a shear force of 1000rpm-12000rpm for 30 minutes; then add 10-55% recombinant vegetable oil body emulsion, stir evenly, and finally add 0.1-0.5% flavor, mix well and bottle.

Example 7 Preparation method of vegetable oil cold-process soap

According to the mass ratio of raw materials, 15-40% coconut oil, 15-40% palm oil, 0-10% castor oil, 2-10% rosehip oil, 2-10% olive oil, 2-10% shea butter, 5-20% alkali, 20-30% distilled water are added to the mixing cup, fully mixed, poured into the alkali solution that has been restored to 45±5℃ in advance, and placed in a 45±5℃ water bath for heating and stirring for 3-4 hours until the soap solution is uniform, and then 2-20% recombinant plant oil suspension is added thereto, mixed, poured into a mold and dried for 1-2 days to obtain a handmade soap product; the obtained handmade soap product is demoulded, and saponification is continued in a cool and ventilated place for 8-12 weeks, and the soap is repaired and packaged to obtain the finished handmade soap (Figure 6). 10 volunteers used the above cold-processed soap and measured the skin indicators before and after use. The results are shown in Table 5.

Example 8 Preparation method of vegetable oil cold-process soap

The weight ratio of raw materials is 2-20% reconstituted vegetable oil emulsion, 15-40% coconut oil, 15-40% palm oil, 1-10% castor oil, 2-10% rosehip oil, 2-10% olive oil, 2-10% shea butter, 5-20% alkali, and the balance is distilled water. The final percentage content of all the above ingredients is 100%;

Add coconut oil, palm oil, castor oil, rosehip oil, olive oil, and shea butter to a mixing cup in proportion, mix thoroughly, pour in the pre-prepared alkali solution that has been restored to 45±5℃, and place in a 45±5℃ water bath for heating and stirring for 3-4 hours until the soap solution is uniform; then add 2-20% of the total mass of the soap solution reconstituted vegetable oil body emulsion (reconstituted oil body suspension plus xanthan gum), mix, pour into a mold and dry for 1~2 days to obtain the first handmade soap; demould the obtained first handmade soap, continue saponification in a cool and ventilated place for 8~12 weeks, repair the soap, and package to obtain the finished handmade soap. 10 volunteers used the above cold-processed soap and measured the skin indicators before and after use. The results are shown in Table 6.

The cold-processed soap of the present invention is made of pure natural plant oil raw materials, has the effects of efficient cleaning, moisturizing the skin, improving skin quality and antibacterial and anti-inflammatory, is mild and non-irritating, natural and pollution-free, and is suitable for men and women, young and old. The plant oil components make the product of the present invention have the effects of moisturizing, locking water, cleaning, etc. (see Table 7).

Example 9 Preparation of recombinant vegetable oil hand cream

The mass fractions of each substance are: 5-40% recombinant plant oil suspension, 0.05-0.5% xanthan gum, 5-10% polyvinyl pyrrolidone PVP, 3-8% triglyceride, 10-20% cetyl alcohol, 15-25% cetostearyl alcohol, 15-20% monoglyceride, 1-10% xanthan gum, 5-10% BASF, 10-15% glycerol, the balance is water, and the total weight is 100%;

The specific steps are as follows:

1) Weigh 5-10% polyvinyl pyrrolidone (PVP), 3-8% triglyceride, 10-20% cetyl alcohol, 15-25% cetostearyl alcohol, 15-20% monoglyceride, 1-10% xanthan gum, 5-10% BASF, and 10-15% glycerol, add water, place in a 75°C water bath, and stir with a glass rod until all solid particles are dissolved; 2) Dissolve xanthan gum in 0.05 mol/L phosphate buffer at pH 7.0, heat to 60°C, stir magnetically for 2 hours to absorb water and swell, then mix with the recombinant plant oil body suspension, and stir for another 2 hours to form a recombinant plant oil body emulsion;

3) The liquids in step 1) and 2) were mixed and stirred, and intermittently emulsified for 30 minutes using an emulsifier until the emulsion was milky white with uniform color and no particles; a small amount of pure natural plant essential oil was added, stirred, mixed, and packaged to obtain a reconstituted plant oil hand cream; 10 volunteers were selected to use the hand cream, and skin indicators before and after use were measured. The results are shown in Table 8. After using the hand cream, the oil content in the hands of the volunteers decreased and the water content increased.

Example 10 Preparation of recombinant plant oil foot cream

The mass fractions of the substances are: 5-40% recombinant plant oil suspension, 0.05-0.5% xanthan gum, 15-20% glycerol, 10-20% cetyl alcohol, 10-15% cetostearyl alcohol, 15-20% monoglyceride, the balance is water, and the total weight is 100%;

The specific steps are as follows:

1) Weigh 15-20% glycerin, 10-20% cetyl alcohol, 10-15% cetostearyl alcohol, and 15-20% monoglyceride as basic hand cream matrix, add 420mL water, place in a 75℃ water bath, and stir with a glass rod until all solid particles are dissolved;

2) Xanthan gum was first dissolved in 0.05 mol/L phosphate buffer at pH 7.0, heated to 60°C, magnetically stirred for 2 h to absorb water and swell, then mixed with the recombinant plant oil body suspension, and stirred for another 2 h to form a recombinant plant oil body emulsion;

3) The liquids in step 1) and 2) were mixed and stirred, and emulsified intermittently for 30 minutes using an emulsifier, and then packaged. 10 volunteers used the above foot cream, and the skin indicators before and after use were measured. The results are shown in Table 9. The oil content of the foot cream increased, the skin elasticity was enhanced, and the moisture content was increased.

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Sequence Listing

<110> Jilin Agricultural University

<120> Oil body care product and preparation method thereof

<160> 2

<170> SIPOSequenceListing 1.0

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<213> Safflower (Carthamus tinctorius L.)

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aaagctcagg atctgaaaga cagagctcag tactacggac agcaacatac tggtggggaa 720

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Claims (3)

1. A cleaning and care article, characterized in that: the recombinant vegetable oil body emulsion is prepared by the following method: adding the xanthan gum into deionized water, uniformly stirring to avoid bubbles, fully swelling, and slowly adding the recombinant vegetable oil suspension under the action of a magnetic stirrer;

the recombinant plant oil body is a plant oil body which over-expresses oleosin;

the final concentration of the recombinant vegetable oil body suspension is 10%;

the concentration of the xanthan gum is 0.3-0.5%.

2. The washing and caring product according to claim 1, wherein the washing and caring product is an oil body skin care emulsion, and comprises the following components in percentage by mass: 10-55% of recombinant vegetable oil emulsion, 5-10% of glycerol, 10-20% of carboxyethyl cellulose, 1-5% of PEG800,5-20% of vitamin C and 0.1-0.5% of essence, wherein the total weight is 100%.

3. The washing and caring product according to claim 1, wherein the washing and caring product is an oil body cold soap, and comprises the following components in percentage by mass: 2-20% of recombinant vegetable oil emulsion, 15-40% of coconut oil, 15-40% of palm oil, 1-10% of castor oil, 2-10% of rose hip oil, 2-10% of olive oil, 2-10% of shea butter, 5-20% of alkali and the balance of distilled water, wherein the total weight is 100%.

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