阅读说明：本技术 一种粗糙凤尾蕨中提取黄酮苷的制备方法 (The preparation method of flavonoid glycoside is extracted in a kind of coarse brake fern ) 是由 李杨 吴娇 尹瀚林 石玉生 李春斌 于 2019-09-20 设计创作，主要内容包括：本发明属于化合物提取应用领域,公开了一种粗糙凤尾蕨中提取黄酮苷的制备方法。具体步骤为：(1)用有机溶剂对粗糙凤尾蕨茎叶进行提取及萃取；(2)聚酰胺树脂柱色谱分离及中压制备；(3)高效液相色谱检测及制备。本发明制备的黄酮苷纯度较高,经高效液相色谱法检测黄酮苷的含量可达到95％以上。该提取制备方法操作简单、获得的纯度较高,生产成本低、便于产业化生产。(The invention belongs to compounds to extract application field, disclose the preparation method that flavonoid glycoside is extracted in a kind of coarse brake fern.Specific steps are as follows: (1) coarse brake fern cauline leaf is extracted and extracted with organic solvent；(2) polyamide resin column chromatography separation and middle compacting are standby；(3) high performance liquid chromatography detection and preparation.Flavonoid glycoside purity prepared by the present invention is higher, and the content through high performance liquid chromatography detection flavonoid glycoside can reach 95% or more.The purity that the extraction preparation method is easy to operate, obtains is higher, and production cost is low, is convenient for industrialization production.)

1. extracting the preparation method of flavonoid glycoside in a kind of coarse brake fern, characterized in that specific steps are as follows:

(1) coarse brake fern cauline leaf is extracted and is extracted with organic solvent；

(2) polyamide resin column chromatography separation and middle compacting are standby；

(3) high performance liquid chromatography detection and preparation；

It is tested and analyzed, is determined specific with all-wave length high performance liquid chromatography after 60 standby components of middle compacting are recovered under reduced pressure Flavonoid glycoside ingredient may be contained in component, preferable ingredient is prepared with half preparative high-performance liquid chromatographic, preparation condition: chromatography Column is half preparation ODS chromatographic column, A phase :+0.1% trifluoroacetic acid of water, B phase: methanol, elution requirement: 45% methanol or 46% methanol Or 47% methanol isocratic elution, flow velocity 3.0mL/min, ultraviolet detection wavelength are 210nm and 254nm.

2. extracting the preparation method of flavonoid glycoside in a kind of coarse brake fern as described in claim 1, characterized in that the step (1) specifically: dry coarse brake fern cauline leaf 25kg crushes, the raw materials particles crushed are placed in the extractor of 200L In, after being 1:5 immersion 6-8 hours with solid-liquid ratio with 95% ethyl alcohol, heating and refluxing extraction 3-5 times, 1 hour every time, extracting solution subtracted Receipts are pushed back, coarse brake fern cauline leaf crude extract medicinal extract is concentrated to get；Medicinal extract uses petroleum ether, ethyl acetate, extracting n-butyl alcohol respectively, Obtain petroleum ether, four ethyl acetate, n-butanol and water positions.

3. extracting the preparation method of flavonoid glycoside in a kind of coarse brake fern as described in claim 1, characterized in that the step (2) specifically: the wherein polyamide resin column chromatography elution of ethyl acetate extract -30 mesh of 14 mesh of model, respectively with water and The ethyl alcohol of various concentration: 30%, 60%, 95% carrying out gradient elution, is recovered under reduced pressure to obtain water, 30% eluate, 60% elution Four positions of object and 95% eluate；By pressure preparative liquid chromatography separation in the progress of 60% eluate medicinal extract, it is with volumetric concentration 10%-80% methanol elution gradient obtains 60 components, chromatographic column filler ODS.

4. extracting the preparation method of flavonoid glycoside in a kind of coarse brake fern as described in claim 1, characterized in that the step (1) solid-liquid ratio unit is kg/L.

Technical field

The invention belongs to compounds to extract application field, and present invention relates particularly to extract flavonoid glycoside in a kind of coarse brake fern Preparation method.

Background technique

Flavone compound is a kind of Polyphenols antioxidant, is pueraria lobata, psoralea corylifolia, Huang Cutters, ginkgo, sea-buckthorn, sophora bud etc. The main active of clinical parts of generic medicinal plants.Such compound amounts is numerous, and structure is complicated, living with many important physiology Property, such as it is antitumor, anti-diabetic, hepatoprotective effect, prevention and treatment cardiovascular and cerebrovascular disease, anti-oxidant, antibacterial, anti-inflammatory.Modern pharmacology Studies have shown that flavone compound is used as a kind of important native compound, in chemical damage, immunological liver injury, drug Property hepatic injury and alcoholic liver injury etc. is significant in efficacy.

Pteris plant majority has detumescence, removing toxic substances, stop dysentery and other effects, civil often to be used for the platymiscium to treat abdomen It rushes down, enteritis, lung hemoptysis, traumatic hemorrhage, abscess of throat, dysentery, ephritis, the diseases such as rheumatism.Pteris plant chemical ingredient is more Sample mainly has the compound of the types such as flavones, diterpene, sequiterpene, volatile oil, polysaccharide.The category Activities of Some Plants extract has anti- The bioactivity such as oxidation, antibacterial, anti-inflammatory, antiviral and antitumor.

In recent years, many researchers study growing interest to extraction, purifying of effective components from natural materials etc..But to phoenix Tail Cyclosorus plant research is less, and existing flavonoid glycoside extracting method or purifying technique are relatively complicated, time-consuming bothersome, production At high cost or yield is lower, there is no large-scale production.

Summary of the invention

For overcome the deficiencies in the prior art, the present invention provides the preparation sides that flavonoid glycoside is extracted in a kind of coarse brake fern Method, the extraction preparation method is easy to operate, the purity of acquisition is higher, and production cost is low, is convenient for industrialization production.The coarse phoenix Flavonoid glycoside is extracted in tail fern has 1 structure of following formula:

The extraction preparation method of the flavonoid glycoside extracted in above-mentioned coarse brake fern is claimed in the present invention simultaneously, and the present invention adopts Technical solution are as follows:

The preparation method of flavonoid glycoside is extracted in a kind of coarse brake fern；Specific steps are as follows:

(1) coarse brake fern cauline leaf is extracted and is extracted with organic solvent；

Dry coarse brake fern cauline leaf 25kg, crushes, the raw materials particles crushed is placed in the extractor of 200L, After being 1:5 immersion 6-8 hours with solid-liquid ratio with 95% ethyl alcohol, heating and refluxing extraction 3-5 times, 1 hour every time, extracting solution was depressurized back It receives, is concentrated to get coarse brake fern cauline leaf crude extract medicinal extract；Medicinal extract uses petroleum ether, ethyl acetate, extracting n-butyl alcohol respectively, obtains Four petroleum ether, ethyl acetate, n-butanol and water positions；

(2) polyamide resin column chromatography separation and middle compacting are standby；

The wherein polyamide resin column chromatography elution of ethyl acetate extract -30 mesh of 14 mesh of model, respectively with water and not With the ethyl alcohol of concentration: 30%, 60%, 95% carrying out gradient elution, be recovered under reduced pressure to obtain water, 30% eluate, 60% eluate With 95% eluate, four positions；By pressure preparative liquid chromatography separation in the progress of 60% eluate medicinal extract, it is with volumetric concentration 10%-80% methanol elution gradient obtains 60 components, chromatographic column filler ODS；

(3) high performance liquid chromatography detection and preparation；

It is tested and analyzed, is determined with all-wave length high performance liquid chromatography after 60 standby components of middle compacting are recovered under reduced pressure Flavonoid glycoside ingredient may be contained in concrete component, preferable ingredient is prepared with half preparative high-performance liquid chromatographic, preparation condition: Chromatographic column is half preparation ODS chromatographic column, A phase :+0.1% trifluoroacetic acid of water, B phase: methanol, elution requirement: 45% methanol or 46% Methanol or 47% methanol isocratic elution, flow velocity 3.0mL/min, ultraviolet detection wavelength are 210nm and 254nm.

Further, the solid-liquid ratio unit in step (1) is kg/L.

The Structural Identification of monomeric compound after preparation: by nuclear magnetic resonance (¹H-NMR、¹³) etc. C-NMR means are to singulation It closes object and carries out Structural Identification.

The present invention compared with prior art the utility model has the advantages that

The flavonoid glycoside purity that the present invention extracts preparation method preparation is higher, through the pure of high performance liquid chromatography detection flavonoid glycoside Degree can reach 95% or more.Raw material of the invention is coarse brake fern, is widely distributed in south China and southwest, but at present to phoenix The research of tail Cyclosorus plant is less, and the present invention can make full use of the resources of medicinal plant in China.And it is mentioned from coarse brake fern Pure to be not easy to mention the active constituent of purity is high because component, the flavonoid glycoside purity that the preparation method of the application obtains is higher.And this The invention coarse brake fern of raw material used has 25kg, and large-scale in this way to extract in contrast, experimental data is more accurate, mentions It takes and purification technique has biggish reference value.And a large amount of extract uses application suitable for enterprise's industrialization promotion.This hair Bright isolated flavonoid glycoside be it is isolated in coarse brake fern for the first time, the present invention is that the medical value of flavonoid glycoside is made full use of to mention The preparation method of high-purity is supplied.

Detailed description of the invention

Fig. 1 is the flavonoid glycoside high performance liquid chromatography detection figure that preparation is extracted in embodiment 1.

Fig. 2 is the flavonoid glycoside hydrogen nuclear magnetic resonance spectrogram that preparation is extracted in embodiment 1.

Fig. 3 is the flavonoid glycoside carbon-13 nmr spectra figure that preparation is extracted in embodiment 1.

Specific embodiment

The present invention is described in detail below by specific embodiment, but is not limited the scope of the invention.Unless otherwise specified, originally Experimental method used by inventing is conventional method, and experiment equipment used, material, reagent etc. commercially obtain.