CN105801593A

CN105801593A - Decursitin preparing method

Info

Publication number: CN105801593A
Application number: CN201610196268.6A
Authority: CN
Inventors: 王维娜; 邓力
Original assignee: SHANGHAI TAUTO BIOTECH CO Ltd
Current assignee: SHANGHAI TAUTO BIOTECH CO Ltd
Priority date: 2016-03-31
Filing date: 2016-03-31
Publication date: 2016-07-27

Abstract

The invention relates to a decursitin preparing method.The method comprises the steps of 1, smashing dried peucedani decursivi radix, adding ethanol solution, conducting ultrasonic extraction, and concentrating extracting solution to obtain an extractum; 2, conducting gradient elution with silica gel as filler by means of column chromatography and normal hexane-ethyl acetate two-phase solvent, collecting target eluant, and conducting concentration and drying to obtain a crude extract; 3, separating the crude extract with the high-speed counter-current chromatography method, conducting online monitoring with an ultraviolet detector, collecting distillate, and conducting vacuum concentration, crystallization and drying to obtain decursitin.Decursitin prepared with the method is high in purity and quality and suitable for high-speed counter-current chromatographic instruments of various types, and industrialized amplification can be achieved easily.

Description

A kind of preparation method of decursin

Technical field

The invention belongs to natural drug and extract separation field, particularly to the preparation method of a kind of decursin.

Background technology

RADIX PEUCEDANI Angelica decursivum (Miq.) Maxim system Umbelliferae Peucedanum plant, has scattered wind heat clearing away, lowering the adverse-rising QI to resolve phlegm Effect.It is mainly used in cough due to pathogenic wind-heat, the treatment of the diseases such as expectorant dyspnea of heat type is full.In recent years pharmacological research shows, the alcohol extraction of RADIX PEUCEDANI Thing has spasmolytic of eliminating the phlegm, antiplatelet aggregation and an antiinflammatory action, and the growth of anticancer and metabolism；Decursin There is antibacterial, anticancer, antitumor, anti-senile dementia, antioxidation, antithrombotic and platelet aggregation effect.As can be seen here, mesh The exploitation of mark composition has commercial value widely.Structural formula is as follows:

At present, the exploitation of the domestic method for separating and preparing rarely having decursin, and use column chromatography, separation cycle Long, consume solvent many, operation complexity.High speed adverse current chromatogram (High-speed countercurrent chromatography, HSCCC) As a kind of novel separating and purifying technology, easy to operate, fast, separating degree is higher, overcomes the sample brought by solid phase carrier In conjunction with, inactivate, the shortcoming such as pollution, be widely used in the analysis mirror separating preparation and total medical herbs of natural product active ingredient Fixed.

At present, there are no the document isolated and purified decursin of report high speed adverse current chromatogram.

Summary of the invention

The technical problem to be solved is to provide the preparation method of a kind of decursin, and the method prepares decursin, Product purity is high, quality better, it is adaptable to various model high-speed counter-current chromatographs, it is easy to industrialization is amplified.

A kind of preparation method of the decursin of the present invention, including:

(1) dried RADIX PEUCEDANI is pulverized, add ethanol solution, supersound extraction, concentrated extracting solution with solid-liquid mass ratio 1:5-10 Obtain extractum；

(2) use column chromatography, with silica gel as filler, with the n-hexane-ethyl acetate above-mentioned extractum of two-phase solvent gradient elution, collect mesh Mark eluent, concentrates, and is dried, obtains crude extract；

(3) to above-mentioned crude extract use high-speed countercurrent chromatography separate, the solvent system of high speed adverse current chromatogram by normal hexane, ethyl acetate, First alcohol and water 2-6:3:6:2-4 by volume forms；UV-detector on-line monitoring, collects stream part concentrating under reduced pressure, crystallization, is dried, Obtain decursin.

The volume fraction of the ethanol solution in described step (1) is 30-90%.

Supersound extraction number of times in described step (1) is 2～3 times.

Silica gel mesh number in described step (2) is 100-300 mesh.

Gradient elution in described step (2) is that the normal hexane using volume ratio to be 5:1 to 1:1 successively is washed with ethyl acetate gradient De-.

The separation condition of the high speed adverse current chromatogram in described step (3) is: engine speed 750-1000rpm/min, and flow phase 2.5-15.0ml/min, bath temperature 10-35 DEG C.

The upper phase of the solvent system in described step (3) is fixing phase, and lower phase is flowing phase.

Crystallization in described step (3) uses methanol crystallization.

Beneficial effect

Sample is carried out supersound extraction by the present invention, and extraction time is short, and extraction efficiency is high, nontoxic, tasteless, cheap and easy to get；Use Silica gel column chromatography enriching and purifying, can remove plurality of impurities, alleviates subsequent operation amount；Use high speed adverse current chromatogram good separating effect, Product purity is high.

Accompanying drawing explanation

Fig. 1 is the high speed adverse current chromatogram figure of decursin of the present invention.

Detailed description of the invention

Below in conjunction with specific embodiment, the present invention is expanded on further.Should be understood that these embodiments be merely to illustrate the present invention and not For limiting the scope of the present invention.In addition, it is to be understood that after having read the content that the present invention lectures, those skilled in the art can To make various changes or modifications the present invention, these equivalent form of values fall within the application appended claims limited range equally.

Embodiment 1

Learn from else's experience dried RADIX PEUCEDANI raw material pulverizing, claim 2000g, add 10L 80% ethanol solution supersound extraction 60 every time Minute, extracting 3 times, concentrating under reduced pressure extracting solution obtains extractum 89g.It is the silicagel column of 100-200 by mesh number on extractum, adopts successively With the normal hexane that volume ratio is 5:1 to 1:1 and ethyl acetate gradient, collecting eluent, concentrate drying obtains crude extract, is used for High-speed counter-current separates.

Normal hexane, ethyl acetate, methanol, water two phase solvent system, by volume 2:3:6:2 mixing is prepared in separatory funnel, Stratification after shaking, ultrasonic degassing 30min.Take phase and fill high-speed counter-current chromatograph as fixing phase, rotate main frame, turn Speed 800rpm, pumping into lower phase with 5ml/min does flowing phase simultaneously, and bath temperature is 20 DEG C, after setting up dynamic equilibrium, with flowing Phased soln crude extract, by injection valve sample introduction, UV-detector is on-line monitoring under 254nm wavelength, collects target component, continuously Preparation, reclaims reagent corresponding merging, methanol crystallization after concentration, from 1000mg crude extract isolated highly purified pale reddish brown before Element 200mg recklessly.Detect through HPLC, content 98.0%, characterize its physics through UV, IR, MS, HNMR, CNMR etc. The data of character are consistent with prior art.

Embodiment 2

Normal hexane, ethyl acetate, methanol, water two phase solvent system, by volume 6:3:6:4 mixing is prepared in separatory funnel, Stratification after shaking, ultrasonic degassing 30min.Take phase and fill high-speed counter-current chromatograph as fixing phase, rotate main frame, turn Speed 800rpm, pumping into lower phase with 5ml/min does flowing phase simultaneously, and bath temperature is 25 DEG C, after setting up dynamic equilibrium, with flowing Phased soln crude extract, by injection valve sample introduction, UV-detector is on-line monitoring under 254nm wavelength, collects target component, continuously Preparation, reclaims reagent corresponding merging, methanol crystallization after concentration, from 1000mg crude extract isolated highly purified pale reddish brown before Element 100mg recklessly.Detect through HPLC, content 98.8%, characterize its physics through UV, IR, MS, HNMR, CNMR etc. The data of character are consistent with prior art.

Embodiment 3

Learn from else's experience dried RADIX PEUCEDANI raw material pulverizing, claim 2000g, add 10L 80% ethanol solution supersound extraction 60 points every time Clock, extracts 3 times, and concentrating under reduced pressure extracting solution obtains extractum 89g.It is the silicagel column of 100-200 by mesh number on extractum, uses successively Volume ratio is normal hexane and the ethyl acetate gradient of 5:1 to 1:1, collects eluent, and concentrate drying obtains crude extract, for height Speed counter-current separation.

Normal hexane, ethyl acetate, methanol, water two phase solvent system, by volume 4:3:6:3 mixing is prepared in separatory funnel, Stratification after shaking, ultrasonic degassing 30min.Take phase and fill high-speed counter-current chromatograph as fixing phase, rotate main frame, turn Speed 800rpm, pumping into lower phase with 5ml/min does flowing phase simultaneously, and bath temperature is 30 DEG C, after setting up dynamic equilibrium, with flowing Phased soln crude extract, by injection valve sample introduction, UV-detector is on-line monitoring under 254nm wavelength, collects target component, continuously Preparation, reclaims reagent corresponding merging, methanol crystallization after concentration, from 1000mg crude extract isolated highly purified pale reddish brown before Element 150mg recklessly.Detect through HPLC, content 98.2%, characterize its physics through UV, IR, MS, HNMR, CNMR etc. The data of character are consistent with prior art.

Claims

1. a preparation method for decursin, including:

The preparation method of a kind of decursin the most according to claim 1, it is characterised in that: the second in described step (1) The volume fraction of alcoholic solution is 30-90%.

The preparation method of a kind of decursin the most according to claim 1, it is characterised in that: surpassing in described step (1) Sound extraction time is 2～3 times.

The preparation method of a kind of decursin the most according to claim 1, it is characterised in that: the silicon in described step (2) Glue mesh number is 100-300 mesh.

The preparation method of a kind of decursin the most according to claim 1, it is characterised in that: the ladder in described step (2) Degree eluting is normal hexane and the ethyl acetate gradient using volume ratio to be 5:1 to 1:1 successively.

The preparation method of a kind of decursin the most according to claim 1, it is characterised in that: the height in described step (3) The separation condition of speed adverse current chromatogram is: engine speed 750-1000rpm/min, and flow phase 2.5-15.0ml/min, bath temperature 10-35℃。

The preparation method of a kind of decursin the most according to claim 1, it is characterised in that: molten in described step (3) The upper phase of agent system is fixing phase, and lower phase is flowing phase.

The preparation method of a kind of decursin the most according to claim 1, it is characterised in that: the knot in described step (3) Brilliant employing methanol crystallization.