阅读说明：本技术 一种蔷薇花活性提取物及其应用 (Active rose extract and application thereof ) 是由 李甫 王明奎 王伦 陈斌 于 2020-12-31 设计创作，主要内容包括：本发明涉及护肤品技术领域,具体涉及一种蔷薇花活性提取物及其应用。具体技术方案为：一种蔷薇花活性提取物,所述活性提取物中含有的活性成分为GeminA,和/或,GeminB,所述Gemin A的质量百分含量≥4.0％,和/或,所述Gemin B的质量百分含量≥0.8％。其活性成分具有良好的酪氨酸酶抑制活性,可以在美白护肤品和果蔬保鲜剂中作为酪氨酸酶抑制剂使用；也可以在酪氨酸酶抑制剂中加入Gemin A,和/或,Gemin B或多苞蔷薇花作为其活性成分使用。(The invention relates to the technical field of skin care products, in particular to a rose active extract and application thereof. The specific technical scheme is as follows: the active extract of the rose flowers comprises Gemin A and/or Gemin B as active ingredients, wherein the Gemin A accounts for more than or equal to 4.0% by mass, and/or the Gemin B accounts for more than or equal to 0.8% by mass. The active ingredients of the compound have good tyrosinase inhibitory activity, and can be used as a tyrosinase inhibitor in whitening skin care products and fruit and vegetable preservatives; gemin A and/or Gemin B or Rosa multiflora can also be added into tyrosinase inhibitor as active ingredients.)

1. An active rose flower extract, which is characterized in that: the active ingredients contained in the active extract are GeminA and/or GeminB, wherein the mass percentage of the GeminA is more than or equal to 4.0 percent, and/or the mass percentage of the GeminB is more than or equal to 0.8 percent.

2. An active rose flower extract, which is characterized in that: the structural formula of an active ingredient GeminA contained in the active extract is as follows:

3. an active rose flower extract, which is characterized in that: the structural formula of an active ingredient GeminB contained in the active extract is as follows:

4. an active rose flower extract as claimed in any one of claims 1 to 3, wherein: the flos Rosae Multiflorae is flos Rosae Multiflorae.

5. A tyrosinase inhibitor, characterized by: contains active ingredients of Rosa multiflora or its active extract.

6. A fruit and vegetable fresh-keeping agent is characterized in that: contains active ingredients of Rosa multiflora or its active extract.

7. A method for extracting an active rose flower extract is characterized by comprising the following steps: extracting with ethanol and concentrating under reduced pressure to obtain flos Rosae Multiflorae extract; then eluting the rose extract by macroporous adsorption resin, and concentrating under reduced pressure to obtain an active extract containing active ingredients; then carrying out reversed phase preparative chromatography separation on the active extract to obtain the active component of the rose flower.

8. The method for extracting an active rose flower extract as claimed in claim 7, wherein the method comprises the steps of: dissolving rose extract in water, passing through macroporous adsorption resin, eluting by pure water for 5-10 column volumes, eluting by ethanol solution with volume fraction of 20-30% for 5-10 column volumes, and then carrying out reduced pressure concentration on the eluent at 40-50 ℃ to obtain the active extract.

9. The application of the rose active extract is characterized in that: the GeminA and/or the GeminB are/is applied to whitening skin care products as tyrosinase inhibitors.

10. The application of the rose active extract is characterized in that: the GeminA and/or GeminB is applied to the fruit and vegetable preservative as a tyrosinase inhibitor.

Technical Field

The invention relates to the technical field of skin care products, in particular to a rose active extract and application thereof.

Background

It is known that excessive ultraviolet radiation causes a large amount of free radicals and active oxygen to be generated by the human body, and further causes oxidative damage to the body. The existence of excessive active oxygen in human body can promote tyrosinase to catalyze L-tyrosine to generate a series of complex reactions to finally form melanin, and the proper amount of melanin can protect the personal safety, but a large amount of melanin accumulated in human body can cause a series of diseases such as pigmented spots, malignant melanoma and the like. Therefore, the tyrosinase inhibitor has good application potential in the fields of medicines, whitening cosmetics and the like.

Currently, with the continuous improvement of the living standard of substances, people pay more attention to skin care, and put forward higher requirements on the efficacy of cosmetics, so that how to safely whiten skin becomes the basic skin care of people to meet the requirements of the people on deeper layers. Currently, the melanin production inhibitors widely used in the market include gentisic acid, arbutin, hydroquinone, and the like, but all show certain side effects. Such as: long-term use of hydroquinone may cause brown yellow disease and dermal elastic fibrosis, while improper use of arbutin will not inhibit the production of melanin, but will increase the production of melanin. Therefore, the search for efficient and safe tyrosinase inhibitors from natural sources as active additives of whitening skin care products has become a development trend of the cosmetic industry.

Meanwhile, tyrosinase is also called polyphenol oxidase, is widely present in various fruits and vegetables, is a main enzyme causing enzymatic browning of the fruits and vegetables, and seriously affects the flavor, nutrition and appearance of the fruits and vegetables. The tyrosinase inhibitor can effectively prevent the browning of fruits and vegetables, and is one of important means for keeping the fruits fresh. Tyrosinase is an essential enzyme for the life activities of various microorganisms, and the inhibition of the activity of tyrosinase can also inhibit the growth of putrefying bacteria, so that the purposes of fresh preservation and corrosion prevention are achieved, and the flavor and the taste of food are not influenced. In the early days, sulfur-containing compounds were widely used in the field of food preservation, but were limited in use due to the negative effects of sulfur residues on human health. At present, the tyrosinase inhibitors widely used in food preservation comprise cysteine, ascorbic acid, citric acid and the like, and have high safety.

Therefore, the search for new plant raw materials with potential value and the extraction of active ingredients with tyrosinase inhibition effect from the plant raw materials have important significance for developing new safe and efficient whitening skin care products and fruit and vegetable preservatives.

Disclosure of Invention

Aiming at the defects of the prior art, the invention provides a rose active extract which has excellent tyrosinase inhibitory activity and can be used for whitening skin care products and/or fruit and vegetable preservative additives.

In order to achieve the purpose, the invention is realized by the following technical scheme:

the invention discloses a rose active extract, wherein the active component in the rose active extract is Gemin A and/or Gemin B, the Gemin A content by mass is more than or equal to 4.0%, and/or the Gemin B content by mass is more than or equal to 0.8%.

Correspondingly, the active extract of the rose flowers contains an active ingredient Gemin A with the following structural formula:

correspondingly, the active extract of the rose flowers contains an active ingredient Gemin B with the following structural formula:

preferably, the rose is a multiflora rose flower.

Accordingly, a tyrosinase inhibitor comprising an active ingredient of Rosa multiflora or an active extract thereof.

Correspondingly, the fruit and vegetable fresh-keeping agent contains active ingredients in the rosa multiflora thunb or the active extract thereof.

Correspondingly, the extraction method of the rose active extract adopts the modes of ethanol extraction and reduced pressure concentration to extract rose extract; then eluting the rose extract by macroporous adsorption resin, and concentrating under reduced pressure to obtain an active extract containing active ingredients; then carrying out reversed phase preparative chromatography separation on the active extract to obtain the active component of the rose flower.

Preferably, the preparation process of the rose extract comprises the following steps: soaking the crushed rose in 5-12 times of 30-50% ethanol solution for 7-15 h, and then concentrating the extracting solution at 40-50 ℃ under reduced pressure to obtain rose extract.

Preferably, the rose flower extract is dissolved in water, the rose flower extract is subjected to macroporous adsorption resin, pure water is firstly used for eluting for 5-10 column volumes, then 20-30% of ethanol solution in volume fraction is used for eluting for 5-10 column volumes, and then the eluent is subjected to reduced pressure concentration at 40-50 ℃ to remove ethanol, so that the active extract is obtained.

Correspondingly, the application of the rose active extract and the application of GeminA and/or GeminB as tyrosinase inhibitors in whitening skin care products.

Correspondingly, the application of the rose active extract and the application of GeminA and/or GeminB as tyrosinase inhibitors in fruit and vegetable preservatives.

The invention has the following beneficial effects:

the invention extracts an active extract of a multi-bud rose flower from the multi-bud rose flower, wherein the mass fractions of active components Gemin A and Gemin B are not less than 4.0% and 0.8% respectively. Meanwhile, the active component of the rosa multiflora has good tyrosinase inhibitory activity and can be used as a tyrosinase inhibitor in whitening skin care products and fruit and vegetable preservatives; gemin A and/or Gemin B or Rosa multiflora can also be added into tyrosinase inhibitor as active ingredients. The multi-bud rose flower active extract prepared by the invention is a natural source, has the advantages of safety, no toxicity, no teratogenicity, no carcinogenicity, no sensitization and the like, and has an inhibition rate of more than 50% on tyrosinase under the test concentration of 60 mug/mL in a tyrosinase inhibition activity test.

Detailed Description

The following will clearly and completely describe the technical solutions in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Unless otherwise indicated, the technical means used in the examples are conventional means well known to those skilled in the art.

Example 1 preparation and chemical composition analysis of crude extracts of different roses

Taking 50g of each dry petal of each rose (wild collected Sichuan Yunnan rose, common anthriscus, northwest rose, common emetic rose and yellow rose), crushing, putting into different triangular bottles, respectively adding 500mL of 50% ethanol water solution, extracting for 10h at 50 ℃, filtering to obtain each rose flower extract, respectively concentrating the extract at 50 ℃ under reduced pressure to obtain each rose flower QWH crude extract, which is sequentially named as CDQWH-1A, JMQWH-1A, EMQWH-1A, XBQWH-1A, DBQWH-1A and HQWH-1A. And analyzing the 6 rose crude extracts by using Gemin A and Gemin B obtained by separation as reference substances through reversed phase liquid chromatography, wherein the Gemin A and Gemin B in the multi-bud rose crude extract are 1.1% and 0.2% in mass percentage in sequence, and Gemin A and Gemin B are not detected in other 5 rose flowers.

Example 2

Pulverizing 50g of Rosa multiflora Thunb, soaking in 10 times of 50% ethanol water solution at 50 deg.C for 10 hr for 3 times, mixing extractive solutions, concentrating under reduced pressure at 50 deg.C, and drying to obtain 16.8g of extract. Suspending the extract in water, extracting with petroleum ether-ethyl acetate (volume ratio 1: 1) mixed solvent for 3 times to remove liposoluble components, extracting with n-butanol for 3 times, mixing n-butanol extractive solutions, and concentrating under reduced pressure at 50 deg.C to obtain n-butanol extract 5.1 g. Dissolving the n-butanol extract in water, passing through HPD-100A macroporous adsorbent resin, eluting with pure water for 10 column volumes, eluting with 25% ethanol water for 10 column volumes, mixing eluates of 25% ethanol water, and concentrating under reduced pressure at 50 deg.C to obtain active extract rich in target active ingredient 2.3 g; subjecting the active extract to reverse phase C₁₈Separating by preparative chromatography (column size of 5 × 25cm, acetonitrile: water 14: 86 (volume ratio), flow rate of 50mL/min, detection wavelength of 280nm) to obtain Gemin A (153mg) and Gemin B (38mg) with chemical structures shown below：

Nuclear magnetic resonance spectrogram data of Gemin A and Gemin B (¹H-NMR and¹³C-NMR data) as follows:

Gemin A：¹H-NMR(400MHz，Acetone-d₆:D₂o) δ: 7.29, 7.26, 6.76 (each 1H, brs, DHDG-H), 7.01, 6.96 (each 2H, s, gal-H), 6.66, 6.63, 6.61, 6.48, 6.47, 6.35 (each 1H, s, HHDP × 3), 6.48(1H, overlap, Glu-1-H), 6.10(1H, d, J ═ 7.8Hz, Glu-1 '-H), 4.43-5.50(Glu-H-2-5), 3.76((1H, d, J ═ 13.2Hz, Glu-6-H), 3.60((1H, d, J ═ 12.8Hz, Glu-6' -H)).

¹³C-NMR(100MHz，Acetone-d₆:D₂O) δ: 169.5, 168.4, 168.3, 168.2, 168.0, 167.6, 166.5, 165.8, 165.1, 162.9(10 ester-based carbons), 119.6, 119.5 (toll-C-1), 109.9, 109.7 (each 2C, toll-C-2, 2 ', 6, 6'), 145.7, 145.5 (each 2C, toll-C-3, 3 ', 5, 5'), 139.3, 139.2 (toll-C-4, 4 '), 115.9, 115.7, 115.5, 115.4, 114.7, 114.2(HHDP-C-1, 1'), 125.8, 125.7, 125.7, 125.6, 125.2, 125.1 (HH-C-2, 2 '), 107.9(2C), 107.6(2C), 107.3, 107.1 (HH-C-3, 107.1, 3, 144, 136.3, 136.2', 136.9 (HHDP-C-1, 144, 136, 136.3, 136.4, 136.2 ', 136, 136.4, 136.2',136,136,136,136,136,136,136,136,136,136,136,136,144, 5 ') 118.7(115.9) (DHDG-C-1, 1 '), 112.0(136.5) (DHDG-C-2, 2 '), 147.9(140.7) (DHDG-C-3, 3 '), 141.0(139.8) (DHDG-C-4, 4 '), 146.4(143.2) (DHDG-C-5, 5 '), 107.0(109.9) (DHDG-C-6, 6 '), 93.5(90.4) (Glu-1, 1 '), 74.0(71.2) (Glu-2, 2 '), 75.7(73.0) (Glu-3, 3 '), 68.7 (70.3) (Glu-4, 4 '), 71.0(72.6) (Glu-5, 5 '), 62.9(62.7) (Glu-6, 6 ').

Gemin B：¹H-NMR(400MHz，Acetone-d₆:D₂O) δ: 7.26, 7.23, 6.73 (each 1H, brs, DHDG-H), 7.01, 6.97 (each 2H, s, toll-H), 6.71, 6.63, 6.48, 6.47,(each of 1H, s, HHDP × 2), 6.40(1H, overlap, Glu-1-H), 6.12(1H, d, J ═ 8.0Hz, Glu-1 '-H) 4.40-5.80(Glu-H-2-5), 3.79 ((1H, overlap, Glu-6-H), 3.75((1H, overlap, Glu-6' -H)).

¹³C-NMR(100MHz，Acetone-d₆:D₂O) δ: 170.5, 169.0, 168.6, 168.3, 166.6, 165.7, 165.6, 163.0(8 ester carbons), 119.5, 119.2 (gal-C-1), 110.0, 109.9 (each 2C, gal-C-2, 2 ', 6, 6 '), 145.8, 145.7 (each 2C, gal-C-3, 3 ', 5, 5 '), 139.4, 139.2 (gal-C-4, 4 '), 116.1, 115.7, 115.8, 114.4(HHDP-C-1, 1 '), 125.7, 125.6, 125.3, 125.2(HHDP-C-2, 2 '), 107.8, 107.7, 107.4, 107.0(HHDP-C-3, 3 '), 145.3(2C), 145.1(2C), 144.4, 144.3 (2C-3), 2.3 (HHDP-C-1, 144.3, 144, 136.6, 136.9.6, 136.9 (each 2C, 1, 9-C-3, 136, 136.2 (HHDP-C-1, 1 ',136,136,136,136,136,136,136,136,136,2,2,2,136,136,2,2, 1 '), 112.1(136.3) (DHDG-C-2, 2'), 148.1(141.1) (DHDG-C-3, 3 '), 140.9(139.7) (DHDG-C-4, 4'), 146.5(143.4) (DHDG-C-5, 5 '), 107.3(110.3) (DHDG-C-6, 6'), 94.2(91.0) (Glu-1, 1 '), 74.3(71.9) (Glu-2, 2'), 79.0(74.0) (Glu-3, 3 '), 67.7 (71.5) (Glu-4, 4'), 76.2(73.8) (Glu-5, 5 '), 63.6(61.1) (Glu-6, 6').

Example 3

Pulverizing 200g flos Rosae Multiflorae, extracting with 10 times of 35% ethanol water solution at 50 deg.C for 10 hr for 3 times, mixing extractive solutions, and concentrating under reduced pressure at 40 deg.C to obtain 62g extract. Dissolving the extract in 2L of pure water, passing through HPD-100A macroporous adsorbent resin, eluting with pure water for 8 column volumes, eluting with 30% ethanol water solution for 10 column volumes, mixing 30% ethanol eluates, concentrating under reduced pressure at 45 deg.C until the solution has no obvious alcohol smell, freezing the solution, and vacuum drying at low temperature to obtain 9.1g active extract (number DBQWH-1). Gemin A and Gemin B obtained by separation are used as reference substances, and the active extract is analyzed by reversed phase liquid chromatography, wherein the mass fraction of Gemin A is 5.2%, and the mass fraction of Gemin B is 1.1%.

Example 4

Pulverizing 200g flos Rosae Multiflorae, extracting with 8 times of 50% ethanol water solution at 40 deg.C for 10 hr for 3 times, mixing extractive solutions, and concentrating under reduced pressure at 50 deg.C to obtain 58g extract. Dissolving the extract in 2L of pure water, passing through HPD-100A macroporous adsorbent resin, eluting with pure water for 5 column volumes, eluting with 20% ethanol water solution for 10 column volumes, mixing 20% ethanol eluates, concentrating under reduced pressure at 40 deg.C until the solution has no obvious alcohol smell, freezing the solution, and vacuum drying at low temperature to obtain 8.8g active extract (number DBQWH-2). Gemin A and Gemin B obtained by separation are used as reference substances, and the active extract is analyzed by reversed phase liquid chromatography, wherein the mass fraction of Gemin A is 4.8%, and the mass fraction of Gemin B is 1.0%.

Example 5 tyrosinase inhibiting Activity of different Rosa extracts

Experimental materials: crude rose extract prepared in example 1, active extracts prepared in example 3 and example 4.

The experimental method comprises the following steps: mixing a sample to be tested with levodopa (the final concentration is 1.25mM), adding tyrosinase (the final concentration is 25U/mL) to start reaction, setting 3 repeated wells, setting a positive control (alpha-arbutin), reacting for 5 minutes at room temperature, and measuring the OD value at the wavelength of 490nm by using an enzyme labeling instrument. The tyrosinase inhibition rate was calculated as follows:

tyrosinase inhibition (%) (1-sample OD)_490nmExperimental control well OD_490nm)×100％

Specific results are shown in the following table 1, and the results show that the activity of the rosa multiflora extract is excellent, the activity of the rosa multiflora extract is better than that of the positive control alpha-arbutin under the same mass concentration, and the activities of other 5 rosa multiflora extracts are general or poor. Further separation analysis shows that the active extract of the rosa multiflora is rich in tannin compounds, while other rosa multiflora extracts mainly contain flavonoid components and almost do not contain tannin compounds. The activity experiment shows that the tannin compound tyrosinase in the rosa multiflora flowers has good inhibitory activity and is an active ingredient.

TABLE 1 tyrosinase inhibition of active extracts of different roses

Sample numbering	Concentration (μ g/mL)	Tyrosinase inhibition (%)	IC50 value (μ g/mL)
				Alpha-arbutin	60	25.33	-
CDQWH-1A	60	12.37	-
				JMQWH-1A	60	9.86	-
EMQWH-1A	60	11.22	-
				XBQWH-1A	60	7.81	-
DBQWH-1A	60	39.29	-
				HQWH-1A	60	14.16	-
DBQWH-1	60	60.45	39.58±1.21
				DBQWH-2	60	58.33	45.02±0.88
Gemin A	60	54.69	53.44±1.43
				Gemin B	60	45.22	71.35±1.25

The above-described embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various modifications and improvements of the technical solutions of the present invention can be made by those skilled in the art without departing from the spirit of the present invention, and the technical solutions of the present invention are within the scope of the present invention defined by the claims.