阅读说明：本技术 一种藤茶中提取黄酮苷的方法及其应用 (The method and its application of flavonoid glycoside are extracted in a kind of vine tea ) 是由 辛文锋 蔡群虎 刘石磊 张文生 胡会泽 袁敏惠 王芬 张芳盛 郭霞 于 2019-08-16 设计创作，主要内容包括：本发明涉及植物活性成分提取领域,具体涉及一种从藤茶中提取黄酮苷的方法及其应用,包括以下步骤：(1)将藤茶粉碎、粗提得到提取液；(2)冷藏沉淀除杂；(3)树脂柱脱糖脱酸；(4)水相萃取精制；(5)脱盐、干燥。本发明所得到的产物黄酮苷含量较高、纯度较高、杂质较少。本发明所述方法得到的黄酮苷制成饮品后口感较好,味淡回甘极强。(The present invention relates to active components of plants to extract field, and in particular to a kind of method and its application that flavonoid glycoside is extracted from vine tea, comprising the following steps: (1) crushes vine tea, slightly raises extracting solution；(2) removing impurity by means of precipitation is refrigerated；(3) resin column desugar depickling；(4) water phase extracting and refining；(5) desalination, drying.The obtained product flavonoid glycoside content of the present invention is higher, purity is higher, impurity is less.The flavonoid glycoside that the method for the invention obtains is made good in taste after drink, and lightly seasoned time sweet extremely strong.)

1. extracting the method for flavonoid glycoside in a kind of vine tea, which comprises the following steps:

(1) vine tea crushed, slightly raise extracting solution；

(2) it refrigerates removing impurity by means of precipitation: extracting solution being stood, centrifugal filtration obtains filtrate；

(3) resin column desugar depickling: by filtrate after resin column, discarding efflux, then carries out solvent elution, collects elution Liquid, low temperature are spin-dried for obtaining water-soluble vine tea flavone glycosides crude product；

(4) water phase extracting and refining: crude product with water is dissolved, and adds polyethylene glycol, and ultrasound is stood, and retains lower layer's product；

(5) desalination, drying: obtained lower layer's product low temperature is spin-dried for, ethyl alcohol dissolution, filters after standing, filtrate is freeze-dried Obtain vine tea flavone glycosides.

2. extracting the method for flavonoid glycoside in a kind of vine tea, which comprises the following steps:

(1) vine tea crushed, slightly raise extracting solution；

(2) it refrigerates removing impurity by means of precipitation: extracting solution being stood, centrifugal filtration obtains filtrate；

(3) resin column desugar depickling: by filtrate after resin column, discarding efflux, then carries out solvent elution, collects elution Liquid, low temperature are spin-dried for obtaining water-soluble vine tea flavone glycosides crude product；

(4) it refines: crude product with water being dissolved, oversized particles polyamide resin column, eluent, concentrate drying obtains vine tea Flavonoid glycoside.

3. method according to claim 1 to 2, which is characterized in that vine tea is crushed to 60-80 by the step (1) Mesh.

4. method according to claim 1 to 2, which is characterized in that the step (1), slightly proposing process is, after crushing Crude drug in plus 15-20 times of quality of pure water, refluxing extraction, filter to obtain extracting solution, the dregs of a decoction add 5 times of crude drug quality measure it is pure Water refluxing extraction, filters to obtain secondary raffinate, and combined extract is concentrated in vacuo to 5 times of crude drug amount aqueous extracts.

5. according to the method described in claim 4, it is characterized in that, in smashed crude drug plus 15-20 times of quality of pure water, time Stream extracts 1.5-4h；The dregs of a decoction add the pure water refluxing extraction 1-1.5h of 5 times of crude drug quality amounts；Temperature concentrated in vacuo is 50-60 ℃。

6. method according to claim 1 to 2, which is characterized in that the step (2), by 2-5 DEG C of condition of extracting solution Lower standing 12-16h.

7. method according to claim 1 to 2, which is characterized in that the step (3), resin column are nonpolar macroporous One of resin XAD-1600, XAD-7, D101-C, weight resin: crude drug=1-2:3；Eluting solvent is 70%-100% second Alcohol or 70%-100% methanol.

8. the method according to claim 1, wherein the step (4), the polyethylene glycol is polyethylene glycol 2000 or cetomacrogol 1000, by weight, crude product: water=1:10-15, polyethylene glycol: water=0.07-0.11:1.

9. the method according to claim 1, wherein the step (4), when ultrasonic, be added potassium dihydrogen phosphate, press Parts by weight meter, water: potassium dihydrogen phosphate=1:0.07-0.09；The step (5), stands 2-4h under the conditions of -10--15 DEG C.

10. application of the flavonoid glycoside that method described in claim 1-9 obtains in preparation drink containing flavonoid glycoside.

Technical field

The present invention relates to active components of plants extract field, and in particular to it is a kind of from vine tea extract flavonoid glycoside method and It is applied.

Background technique

Vine tea also known as Maoyanmei tea, long-lived rattan, vine tea is not belonging to Theaceae plant, but ampelopsis grossdentata kind is perennial Liana, scientific name are ampelopsis grossdentata.

Vine tea is decaffeinated, non-stimulated to gastrointestinal tract, has the bitter taste of tea and returns sugariness, it is more lasting especially to return sugariness It is aromatic.Traditional tea has some acerbities without acerbity, vine tea.

Vine tea also belongs to Chinese medicine scope, and vine tea slightly sweet flavor is cool in nature, have drop heat promotes the production of body fluid, anti-inflammatory diuresis, the anti-rhythm of the heart is unbalance, Resist myocardial ischemia and other effects.All to skin tinea leprosy, icterepatitis, cold anemopyretic, abscess of throat, acute conjunctivitis, insomnia etc. Play the role of it is fabulous, be medical and edible dual purpose plant.

Vine tea mainly contains flavones ingredient, and between general flavone content about 7-10%, main component is dihydromyricetin, red bayberry Element, Quercetin, kaempferol etc..Wherein the highest Ampeloptin of content has distinguished heat-clearing toxin expelling, sharp larynx detumescence, and antibacterial disappears Inflammation, the effect of anti influenza hepatitis B and auxiliary drop " three is high " prevention and treatment cardio-cerebrovascular diseases.

Vine tea return sugariness it is splendid, than traditional tea return sweet effect it is stronger, persistently, mouthfeel is in sweet flavor, surpass Yunnan olive return it is sweet, Sweet and oiliness, without the sweet taste of apparent olive, however after vine tea comparison traditional tea brews, aqueous is muddy, band acerbity, so that Some people is difficult to receive, and the Main Flavonoids ingredient as contained by vine tea, dihydromyricetin, myricetin, Quercetin etc. is bitter, And it is water-soluble poor, muddiness is caused, acerbity also leads to mouthfeel elementary errors.So the vine tea of taste and sweet mouthfeel to be developed, need to use up can The removal dihydromyricetin of energy, myricetin, the bitter principles such as Quercetin.

Application for a patent for invention CN 106942436A discloses a kind of high flavonoid glycoside vine tea, is original with fresh vine tea young stem and leaf Expect by spreading for cooling, high temperature stir-fry is killed, spreading for cooling gets damp again, rubs, deblocking, high temperature is baked, the process of high temperature fried dry is made.The present invention also public affairs The processing technology of high flavonoid glycoside vine tea is opened, as follows: using fresh vine tea young stem and leaf as material, spreading for cooling, cauline leaf is put into 260-280 DEG C high temperature pot, fried using throwing with the stewing frying mode fried and combined；Vine tea spreading for cooling resurgence, is put into rolling machine and rubs, make into Item rate reaches 50% or more；The deblocking of vine tea cauline leaf；Obtained vine tea cauline leaf is put into dryer, 95-110 DEG C of baking, until cauline leaf Water content is 40-50%；Being fried on 90-100 DEG C of roasting machine again to the water content of cauline leaf is 5-6%.The present invention is pressed down using high temperature The activity of oxygenerating enzyme and hydrolase reduces the conversion and loss of flavonoid glycoside compound in manufacturing process, and fragrance object Matter is obviously increased than fresh leaf.By the method treated vine tea, flavonoid glycoside content only has 20-40%, dihydromyricetin cellulose content Higher, up to 16-34%, mouthfeel is bitter.

Application for a patent for invention CN 104644816A disclose it is a kind of simple, practical, can be in the Huang that laboratory is synthesized Ketoside preparation method.Flavonoid glycoside preparation method of the invention, it is characterised in that: precise Fructus Aurantii medicinal material powder 5-30g, with 60- 80% ethyl alcohol 200mL ultrasonic extraction 2-5 times, each 30min.Ethanol extract is filtered, merging filtrate.Reduced-pressure backflow flings to second Alcohol is steamed to no alcohol taste.It accurately weighs 100g macroreticular resin and loads 20-40mm internal diameter glass column, by the pharmaceutical aqueous solution portion of recycling Divide and be splined on resin column, absorption cleans cylinders overnight, with 3 times of column volume ultrapure waters, then molten with 3 times of column volume 60-80% ethyl alcohol Liquid elutes cylinder, collects eluent.60 DEG C of reduced-pressure backflows of eluent fling to liquid, and solid portion is molten with 50% acetonitrile-water of 20mL Liquid dissolution, after 0.22 μm of membrane filtration, obtains flavonoid glycoside solution.But pass through the method treated flavonoid glycoside solution, ingredient It include the bitter tastes impurity components such as flavones, dihydromyricetin glucoside, though general flavone content is high, mouthfeel is bitter.

Currently, there are no a kind of methods for extracting flavonoid glycoside in good taste, sweet in flavor in vine tea, to obtain good in taste Ampelopsis grossedentata extrat, it is necessary to remove bitter taste flavone component in vine tea, only retain sweet sweet ingredient.It is mentioned by obtained by this patent preparation method The flavonoid glycoside purity for taking object to obtain is higher, content is high, is free of bitter taste flavones impurity component.

Summary of the invention

In order to solve the problems such as flavonoid glycoside content is low, impurity content is high obtained in the prior art, the present invention provides one kind The method of flavonoid glycoside is extracted in vine tea, comprising the following steps:

(1) vine tea crushed, slightly raise extracting solution；

(2) it refrigerates removing impurity by means of precipitation: extracting solution being stood, centrifugal filtration obtains filtrate；

(3) resin column desugar depickling: by filtrate after resin column, discarding efflux, then carries out solvent elution, collects Eluent, low temperature are spin-dried for obtaining water-soluble vine tea flavone glycosides crude product；

(4) water phase extracting and refining: crude product with water is dissolved, and adds polyethylene glycol, and ultrasound is stood, and is retained lower layer and is produced Object；

(5) desalination, drying: obtained lower layer's product low temperature is spin-dried for, ethyl alcohol dissolution, filters after standing, filtrate is freezed It is dried to obtain vine tea flavone glycosides.

Further, the step (1), is crushed to 60-80 mesh for vine tea.

Further, the step (1), slightly proposing process is, 15-20 times of quality of pure water is added in smashed crude drug, is returned Stream extracts, and filters to obtain extracting solution, and the dregs of a decoction add the pure water refluxing extraction of 5 times of crude drug quality amounts, filter to obtain secondary raffinate, closes And extracting solution, it is concentrated in vacuo to 5 times of crude drug amount aqueous extracts.

Further, 15-20 times of quality of pure water, refluxing extraction 1.5-4h are added in smashed crude drug；The dregs of a decoction add again Enter the pure water refluxing extraction 1-1.5h of 5 times of crude drug quality amounts；Temperature concentrated in vacuo is 50-60 DEG C.

The sour and astringent slight bitter of crude extract mouthfeel obtained after step (1) is returned sweet.

Further, the step (2) will stand 12-16h under the conditions of 2-5 DEG C of extracting solution.

Step (2) is to remove myricetin, dihydromyricetin, the bitter tastes impurity such as Quercetin.It will be extracted obtained by step 1 Liquid is put in 2-5 DEG C of low temperature of refrigerator-freezer and stands overnight, and yellow crystal precipitating occurs, and centrifugal filtration is precipitated as the production of low temperature yellow crystal Object, bitter, product are mainly myricetin, dihydromyricetin, and Quercetin etc. accounts for 4-5% in crude drug.It precipitates individually placed, takes Centrifugal filtering liquid is spare, and the liquid time is sweet, slightly sour puckery, not bitter, and returning sweet ingredient is vine tea water-soluble flavone glycosides, mainly returns for vine tea Sweet constituents, sour and astringent ingredient are mainly vine tea tannic acid, meat pectin class impurity.

Further, the step (3), resin column are in non-polar macroporous resin XAD-1600, XAD-7, D101-C It is a kind of.

Further, the step (3), weight resin: crude drug=1-2:3.

Further, the step (3), eluting solvent are 70%-100% methanol or 70%-100% ethyl alcohol.

In step (3), non-polar macroporous resin is taken, the filtrate that step 2 is obtained crosses macroreticular resin, and efflux is that vine tea is more Sugar, amino acid, tannin, the impurity such as pectin (the part efflux takes a small amount of freeze-drying, deutostoma taste without obvious sweet taste or return it is sweet, Show that vine tea sweet taste returns the sweet non-polysaccharide of ingredient), efflux abandons, and pure water washes column to colourless rear 70% ethanol elution vine tea Huang Ketones component, ferric chloride reaction differentiate that no flavones ingredient stops elution, collect eluent low temperature and are spin-dried for obtaining water-soluble rattan Tea flavones, flavonoid glycoside, vine tea phenols component, lightly seasoned time sweet, without sour and astringent, the peculiar smell such as hardship.

Further, the step (4), polyethylene glycol be polyethylene glycol-1000 or polyethylene glycol 2000, by weight, Crude product: water=1:10-15, polyethylene glycol: water=0.07-0.11:1.

Further, the step (4), when ultrasonic, addition potassium dihydrogen phosphate, by weight, water: potassium dihydrogen phosphate= 1:0.07-0.09。

After step (4), the upper layer discarded is the slightly larger water-soluble flavone and phenols of polarity, accounts for desciccate 20-30%, Lower layer is flavonoid glycoside (lower layer's product trifluoroacetic acid hydrolysis, thin layer identify, and show that lower layer is flavonoid glycoside), and gained flavonoid glycoside is light Yellow, comparison step (3) products therefrom lightly seasoned time sweet extremely strong.

Further, the step (5), stands 2-4h under the conditions of -10--15 DEG C.

Step 5 gained lower layer solution low temperature is spin-dried for, dehydrated alcohol dissolution is placed in -10-15 DEG C of refrigerator and refrigerates 3 hours, Ethanol is filtered out, insoluble matter is potassium dihydrogen phosphate, and recycling ethyl alcohol is freeze-dried to obtain vine tea flavone glycoside, is mainly returned for vine tea sweet Taste effective component.

The present invention also provides a kind of methods that flavonoid glycoside is extracted in vine tea, comprising the following steps:

(1) vine tea crushed, slightly raise extracting solution；

(2) it refrigerates removing impurity by means of precipitation: extracting solution being stood, centrifugal filtration obtains filtrate；

(3) resin column desugar depickling: by filtrate after resin column, discarding efflux, then carries out solvent elution, collects Eluent, low temperature are spin-dried for obtaining water-soluble vine tea flavone glycosides crude product；

(4) it refines: crude product with water being dissolved, oversized particles polyamide resin column, eluent, concentrate drying obtains Vine tea flavone glycosides.

Further, the step (1), is crushed to 60-80 mesh for vine tea.

Further, the step (1), slightly proposing process is, 15-20 times of quality of pure water is added in smashed crude drug, is returned Stream extracts, and filters to obtain extracting solution, and the dregs of a decoction add the pure water refluxing extraction of 5 times of crude drug quality amounts, filter to obtain secondary raffinate, closes And extracting solution, it is concentrated in vacuo to 5 times of crude drug amount aqueous extracts.

Further, 15-20 times of quality of pure water, refluxing extraction 1.5-4h are added in smashed crude drug；The dregs of a decoction add again Enter the pure water refluxing extraction 1-1.5h of 5 times of crude drug quality amounts；Temperature concentrated in vacuo is 50-60 DEG C.

The sour and astringent slight bitter of crude extract mouthfeel obtained after step (1) is returned sweet.

Further, the step (2) will stand 12-16h under the conditions of 2-5 DEG C of extracting solution.

Step (2) is to remove myricetin, dihydromyricetin, the bitter tastes impurity such as Quercetin.It will be extracted obtained by step 1 Liquid is put in 2-5 DEG C of low temperature of refrigerator-freezer and stands overnight, and yellow crystal precipitating occurs, and centrifugal filtration is precipitated as the production of low temperature yellow crystal Object, bitter, product are mainly myricetin, dihydromyricetin, and Quercetin etc. accounts for 4-5% in crude drug.It precipitates individually placed, takes Centrifugal filtering liquid is spare, and the liquid time is sweet, slightly sour puckery, not bitter, and returning sweet ingredient is vine tea water-soluble flavone glycosides, mainly returns for vine tea Sweet constituents, sour and astringent ingredient are mainly vine tea tannic acid, meat pectin class impurity.

Further, the step (3), resin column are in non-polar macroporous resin XAD-1600, XAD-7, D101-C It is a kind of.

Further, the step (3), weight resin: crude drug=1-2:3.

Further, the step (3), eluting solvent are 70%-100% methanol or 70%-100% ethyl alcohol

Further, the step (4), by weight, crude product: water=1:10-15.

Further, the step (4), by weight, polyamide: crude product=20-25:1.

Further, the step (4), the method for eluent are first with 40% ethanol elution, 3 column volumes, elution Impurity, then with 70% ethanol elution, 5 column volumes, recycle eluent.

The present invention also provides application of the flavonoid glycoside obtained by the above method in preparation drink containing flavonoid glycoside.

This patent returns sugariness ingredient to vine tea and carries out Phytochemical Analysis, find vine tea polysaccharide by basic research early period Without obvious sweet taste, without sweet property is obviously returned, separation analysis is extracted to vine tea total flavonoid ingredient, finds general flavone ingredient Bitterness takes back sweet, crystallized method, and silica gel column chromatography separation is found in general flavone, the several higher flavones of content such as dihydros Myricetin, myricetin, 4,5 ingredients such as Quercetin all have bitter taste, sweet without obviously returning, then separate to big polarity flavone component It is found when analysis, time sugariness ingredient is flavonoid glycoside, and flavonoid glycoside is lightly seasoned, faint sweet taste, and it is sweet extremely strong lasting to return.It is logical It crosses experimental technique to study to obtain a kind of sweet ingredient process of preparing of vine tea flavone time, taste good can be drink top grade, removal Sour and astringent, peculiar smell such as hardship existing for vine tea, extract enter that water is instant, and clarification is returned sweet.

It is compared with the prior art, the beneficial effects of the present invention are:

1. the obtained product flavonoid glycoside content of the present invention is higher, purity is higher, impurity is less.

2. the flavonoid glycoside that the method for the invention obtains is made good in taste after drink, lightly seasoned time sweet extremely strong.

3. the method for the present invention is simple, toxic solvent is free of, environmentally protective, safety is beneficial to industrialized production.

Specific embodiment

The invention will now be further described with reference to specific embodiments, the advantages and features of the present invention will be with description and It is apparent, but these examples are merely exemplary, it is not intended to limit the scope of the present invention in any way.Those skilled in the art Member it should be understood that without departing from the spirit and scope of the invention can details to technical solution of the present invention and form into Row modifications or substitutions, but these modifications and replacement are fallen within the protection scope of the present invention.