阅读说明：本技术 一种桑葚提取物、提取分离方法及其应用 (Mulberry extract, extraction and separation method and application thereof ) 是由 白乃生 彭赛男 高兵 郭森 张姗姗 郭雅欣 岳文平 于 2019-09-17 设计创作，主要内容包括：本发明公开了一种桑葚提取物、提前分离方法和应用,该提取物包含以下组分：花青素3-O-β-吡喃葡萄糖苷、槲皮素-3-O-β-D-吡喃葡萄糖苷、β-谷甾醇亚油酸酯、(3S,4R,5R)-N-(3,5-二羟基哌啶-4-基)乙酰胺、二氢山萘酚7-O-β-D-吡喃葡萄糖苷、山奈酚3-O-β-D-芸香糖苷、山奈酚3-O-芸香苷、N-乙酰基-L-丝氨酸、叶黄素、5,7-二羟基原色酮、谷甾醇-3-O-β-D-4’-O-辛烷-吡喃葡萄糖苷、槲皮素、芦丁、邻苯二酚等。本发明所得的桑葚提取物在制备皮肤增白药物、治疗II型糖尿病药物和认知性障碍疾病药物中具有美好的应用前景。(The invention discloses a mulberry extract, a pre-separation method and application, wherein the extract comprises the following components: anthocyanidin 3-O-beta-glucopyranoside, quercetin-3-O-beta-D-glucopyranoside, beta-sitosterol linoleate, (3S,4R,5R) -N- (3, 5-dihydroxypiperidin-4-yl) acetamide, dihydrokaempferol 7-O-beta-D-glucopyranoside, kaempferol 3-O-beta-D-rutinoside, kaempferol 3-O-rutinoside, N-acetyl-L-serine, lutein, 5, 7-dihydroxychromone, sitosterol-3-O-beta-D-4 '-O-octane-glucopyranoside, quercetin, rutin, beta-D-4' -O-octane-glucopyranoside, Catechol, and the like. The mulberry extract obtained by the invention has good application prospect in preparing skin whitening drugs, drugs for treating type II diabetes and cognitive disorder diseases.)

1. A mulberry extract, characterized in that the extract comprises the following components:

anthocyanin 3-O-beta-glucopyranoside shown in formula I, quercetin 3-O-beta-D-glucopyranoside shown in formula II, beta-sitosterol linoleate shown in formula III, (3S,4R,5R) -N- (3, 5-dihydroxypiperidine-4-yl) acetamide shown in formula IV, (dihydrokaempferol 7-O-beta-D-glucopyranoside shown in formula IV, (kaempferol 3-O-beta-D-rutinoside shown in formula IV), kaempferol 3-O-rutin shown in formula IV, (N-acetyl-L-serine shown in formula IV, (xanthophyll) shown in formula III, (5, 7-dihydroxychromone) shown in formula IV, Formula (II)sitosterol-3-O-beta-D-4' -O-octane-glucopyranoside shown in the formulaQuercetin, formulaRutin shown in the formulaCatechol of the formulaN-hexos-5, 8, 11-trienic acid shown in the formulaMorachalcone A shown in the formulaP-hydroxybenzoic acid shown in the formulaKaempferol 3-O-beta-glucopyranoside of the formulaThe isobavachalone shown;

2. the extraction and separation method of the mulberry extract as set forth in claim 1, comprising the steps of:

s1, carrying out leakage extraction on the dried mulberry by using an ethanol solution with the volume concentration of 90% for 3 times, soaking for 48 hours each time, then starting leakage, combining three extracting solutions, and carrying out reduced pressure concentration to obtain an ethanol extract;

wherein the feed-liquid ratio of the dried mulberry to the ethanol solution is 1g:1.5 mL;

s2, completely dissolving the ethanol extract obtained in the step S1 in water to obtain an extract solution, sequentially and respectively extracting with petroleum ether, ethyl acetate and n-butanol which have the same volume as the extract solution for 2-3 times, and then concentrating under reduced pressure to obtain petroleum ether, ethyl acetate and n-butanol extracts;

s3, subjecting the ethyl acetate extract obtained in S2 to polyamide gradient elution with water-ethanol solution to obtain 7 fractions, performing TLC and HPLC detection on each fraction, selecting chromatographic separation mode and corresponding eluent according to the detection result to obtain formula (I) -formula (VII), formula (III) and formula (III)8 compounds as shown;

s4, subjecting the n-butanol extract obtained in S2 to polyamide, performing gradient elution with water-ethanol solution to obtain 7 fractions, performing TLC and HPLC detection on each fraction, and selecting a chromatographic separation mode and corresponding eluents according to the detection results to obtain formulas (VI), (VIII), (Nc) and (VII)And11 compounds are shown.

3. The extraction and separation method of the mulberry extract according to claim 2, wherein the chromatographic separation manner in S3 and S4 is as follows: repeatedly performing chromatographic separation by using one or more of silica gel column, polyamide column, MCI CHP-20P reverse phase column, and Sephadex LH-20 reverse phase column;

the eluents of the chromatographic separation process in S3 and S4 are: one or more of dichloromethane-methanol solution, water-methanol solution and water-ethanol solution.

4. The extraction and separation method of the mulberry extract according to claim 2, wherein the conditions of the reduced pressure concentration in the S1 and S2 are 30-40 ℃ and 0.09-0.1 MPa of vacuum degree.

5. The extraction and separation method of the mulberry extract according to claim 2, wherein the water-ethanol solution in S3 and S4 comprises water, ethanol, and a mixture of water and ethanol at a volume ratio of 10:90-90: 10.

6. The use of a mulberry extract as claimed in claim 1, in the preparation of a medicament or health product for skin whitening, treatment of type II diabetes and cognitive impairment.

7. The use of a mulberry extract according to claim 6, wherein the mulberry extract comprises anthocyanidin 3-O- β -glucopyranoside, quercetin 3-O- β -D-glucopyranoside, β -sitosterol linoleate, kaempferol 3-O- β -D-rutinoside, kaempferol 3-O-rutin, N-acetyl-L-serine, lutein, 5, 7-dihydroxychromone, quercetin, N-hexocos-5, 8, 11-trienic acid, morachalcoA, and isobavachalone in the preparation of drugs for inhibiting α -glucosidase, tyrosinase, and acetylcholinesterase activities.

Technical Field

The invention relates to the technical field of plant extraction and separation, and particularly relates to a mulberry extract, an extraction and separation method and application thereof.

Background

The mulberry (Morus alba L.) plants of Moraceae (Moraceae) Morus (Morus L.) are distributed in various places of China, and due to the difference of growing environments of various places, under the influence of natural selection and artificial selection, abundant mulberry resources are formed, wherein the mulberry resources comprise 4 cultivars, 11 wild mulberry varieties, 3 varieties of white mulberry and varieties of Mongolian mulberry. The diameter of small and medium-sized trees or shrubs with the height of 3-10 m or more can reach 50cm, and the bark is gray and thick; the branch buds are oval, the color is reddish brown, and the small branches have fine hair; the leaf is egg-shaped or wide egg-shaped, the surface is bright green, has no hair, and the leaf stalk has soft hair; the florescence is 4-5 months, the fruit period is 5-8 months, the mulberry is in a round shape, and the length is generally 2-3 cm. The mulberry leaves have the functions of dispelling wind and heat, clearing lung-heat, moistening dryness, clearing liver and improving vision; the dried tender branches have the functions of expelling wind-damp and benefiting joints; cortex Mori has effects in purging lung fire, relieving asthma, inducing diuresis, and relieving swelling.

Mulberry (Fructus Mori) is the fruit ear of mulberry, also called Morus plant of Moraceae such as Mulberry jujube, Mulberry and Mulberry fruit, and it is first recorded in Tang. New repair materia Medica, it has the functions of nourishing yin, tonifying blood, promoting the production of body fluid and moistening dryness, and can be used for treating yin deficiency of liver and kidney, dark tinnitus of eye, deficiency of body fluid, senile constipation and diabetes. Li Shizhen is mentioned in Ben Cao gang mu, the mulberry is a sweet and slightly cold medicine with the functions of nourishing yin, enriching blood, promoting the production of body fluid and tranquilizing the mind. Modern pharmaceutical research shows that the mulberry has the pharmacological effects of regulating immunity, supplementing deficient gastric juice and improving gastrointestinal digestion capacity, and also has the effects of relieving eyestrain and dryness of eyes, removing oxygen free radicals, resisting oxidation and aging, preventing arteriosclerosis, promoting metabolism, reducing blood sugar and blood fat, resisting mutagenesis and the like.

The main chemical components of the plants in the genus of Morus include flavonoids, stilbenes, alkaloids, phenols, etc., wherein polyphenol is one of the most important active substances in the plants in the genus of Morus, and is a secondary metabolite of the plants. The polyphenol compounds are a wide range of compound types, and have diversity of biological activities due to the diversity of structures. Among them, 47 phenolic compounds, mostly flavonols, mainly rutin, quercetin, myricetin, kaempferol and derivatives thereof, have been found from mulberries.

Disclosure of Invention

In order to solve the problems, the invention provides a mulberry extract, an extraction and separation method and application thereof, the extraction rate of the main components of mulberry is high, the main compounds of the mulberry can be effectively extracted, 19 monomer compounds can be separated at one time, and part of the compounds have application prospects.

In order to achieve the purpose, the invention adopts the technical scheme that:

a mulberry extract comprising the following components:

anthocyanin 3-O-beta-glucopyranoside shown in formula I, quercetin 3-O-beta-D-glucopyranoside shown in formula II, beta-sitosterol linoleate shown in formula III, (3S,4R,5R) -N- (3, 5-dihydroxypiperidine-4-yl) acetamide shown in formula IV, (dihydrokaempferol 7-O-beta-D-glucopyranoside shown in formula IV, (kaempferol 3-O-beta-D-rutinoside shown in formula IV), kaempferol 3-O-rutin shown in formula IV, (N-acetyl-L-serine shown in formula IV, (xanthophyll) shown in formula III, and (5, 7-dihydroxyketone) primary color shown in formula III, Formula (II)sitosterol-3-O-beta-D-4' -O-octane-glucopyranoside shown in the formulaQuercetin, formulaRutin shown in the formulaCatechol of the formulaN-hexos-5, 8, 11-trienic acid shown in the formulaMorachalcone A shown in the formulaP-hydroxybenzoic acid shown in the formulaKaempferol 3-O-beta-glucopyranoside of the formulaThe isobavachalone shown;

the invention also provides an extraction and separation method of the mulberry extract, which comprises the following steps:

s1, carrying out leakage extraction on the dried mulberry by using an ethanol solution with the volume concentration of 90% for 3 times, soaking for 48 hours each time, then starting leakage, combining three extracting solutions, and carrying out reduced pressure concentration to obtain an ethanol extract;

s2, completely dissolving the ethanol extract obtained in the step S1 in water to obtain an extract solution, sequentially and respectively extracting with petroleum ether, ethyl acetate and n-butanol which have the same volume as the extract solution for 2-3 times, and then concentrating under reduced pressure to obtain petroleum ether, ethyl acetate and n-butanol extracts;

s3, processing the ethyl acetate extract obtained in the S2 by polyamide,gradient eluting with water-ethanol solution to obtain 7 fractions, detecting each fraction by TLC and HPLC, selecting chromatographic separation mode and corresponding eluent according to detection result to obtain formula (I) -formula (V), formula (III) and formula (III)8 compounds as shown;

s4, subjecting the n-butanol layer extract obtained in S2 to polyamide, performing gradient elution with water-ethanol solution to obtain 7 fractions, performing TLC and HPLC detection on each fraction, selecting a chromatographic separation mode and corresponding eluent according to the detection result to obtain formulas (VI), (VII), (ninC) and (VII)And formula-formula (la)11 compounds are shown.

Preferably, the separation mode in S3 and S4 is: repeatedly performing chromatographic separation by using one or more of silica gel column, polyamide column, MCI CHP-20P reverse phase column, and Sephadex LH-20 reverse phase column;

the eluents of the chromatographic separation process in S3 and S4 are: one or more of dichloromethane-methanol solution, water-methanol solution and water-ethanol solution.

Preferably, the conditions of the reduced pressure concentration in S1 and S2 are that the temperature is 30-40 ℃ and the vacuum degree is 0.09-0.1 MPa.

Preferably, the water-ethanol solution in S3 and S4 comprises water, ethanol and a mixed solution of water and ethanol according to a volume ratio of 10:90-90: 10.

The invention also protects the application of the mulberry extract, and the application of the extract in preparing medicines or health-care products for whitening skin and treating type II diabetes and cognitive disorder diseases.

Preferably, the mulberry extract is the application of anthocyanin 3-O-beta-glucopyranoside, quercetin-3-O-beta-D-glucopyranoside, beta-sitosterol linoleate, kaempferol 3-O-beta-D-rutinoside, kaempferol 3-O-rutinoside, N-acetyl-L-serine, lutein, 5, 7-dihydroxychromone, quercetin, N-hexos-5, 8, 11-trienic acid, morachalone A and isobavachalone in the preparation of drugs for inhibiting alpha-glucosidase, tyrosinase and acetylcholinesterase activities.

The invention has the following beneficial effects:

the extraction rate of the main components of the mulberry is high, the main compounds of the mulberry can be effectively extracted, and 19 monomer compounds can be separated at one time; part of the compounds have good tyrosinase inhibitory activity, acetylcholinesterase inhibitory activity and alpha-glucosidase inhibitory activity, and have great application prospects.

Drawings

FIG. 1 is an HPLC chromatogram of a portion of ethyl acetate in example 1 of the present invention;

FIG. 2 is an HPLC chromatogram of a part of an n-butanol layer in example 1 of the present invention;

FIG. 3 is a diagram showing a partial separation process of an n-butanol layer in example 1 of the present invention;

FIG. 4 is a diagram showing a partial separation process of an ethyl acetate layer in example 1 of the present invention;

FIG. 5 shows the corresponding positions of 19 compounds of the present invention in the HPLC chromatogram of 90% ethanol extract.

Detailed Description

In order that the objects and advantages of the invention will be more clearly understood, the invention is described in detail below by way of specific embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.