CN108276375A - A method of isolating Stilbene effective constituents from acanthopanax leucorrhizus Harms stem skin - Google Patents
A method of isolating Stilbene effective constituents from acanthopanax leucorrhizus Harms stem skin Download PDFInfo
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- C07D313/02—Seven-membered rings
- C07D313/06—Seven-membered rings condensed with carbocyclic rings or ring systems
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- C07C37/68—Purification; separation; Use of additives, e.g. for stabilisation
- C07C37/70—Purification; separation; Use of additives, e.g. for stabilisation by physical treatment
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- C07D307/78—Benzo [b] furans; Hydrogenated benzo [b] furans
- C07D307/82—Benzo [b] furans; Hydrogenated benzo [b] furans with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to carbon atoms of the hetero ring
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Abstract
The present invention relates to a kind of methods that Stilbene effective constituents are isolated in stem skin from acanthopanax leucorrhizus Harms, and this approach includes the following steps：(1) after fresh acanthopanax leucorrhizus Harms stem skin being dried, crushed, being sieved, through Microwave Extraction, it is concentrated under reduced pressure, obtains crude extract；(2) slightly take object to be extracted to respectively after extract liquor becomes colorless with petroleum ether, ethyl acetate successively, acetic acid ethyl acetate extract is merged, sample to be separated is obtained through evaporated under reduced pressure；(3) sample solution will be obtained after sample to be separated mixed solvent ultrasonic dissolution；(4) preparative HSCCC two phase solvent systems are determined；(5), by after stationary phase and mobile phase respectively ultrasound degassing, it is pumped into successively in the multi-layer helix-tube of preparative counter-current chromatograph, reinjects sample solution and carry out purifies and separates；(6) it is collected into 7 chromatographic peak components successively by chromatogram, 7 components is obtained into 7 kinds of stilbenes compounds after being concentrated under reduced pressure, being dried in vacuo respectively.Operation is simple by the present invention, extracted amount is big, favorable reproducibility, and the purity of 7 kinds of stilbenes compounds of gained is above 98.0%.
Description
Technical field
The present invention relates to active ingredients from traditional Chinese medicinal extractive technique fields, more particularly to one kind to be isolated from acanthopanax leucorrhizus Harms stem skin
The method of Stilbene effective constituents.
Background technology
Acanthopanax leucorrhizus Harms（Acanthopanax leucorrhizus）Belong to the perennial machaka of Acanthopanax, is that China is peculiar
Folk medicinal plants, be distributed mainly on the ground such as Shaanxi, Gansu, Yunnan-Guizhou, the middle and lower reach of Yangtze River.Its medicinal part be mainly root skin and
Stem skin has wind-damp dispelling, degrading the channel, strong boat bone and other effects, cure mainly arthralgia pain due to rheumatism, contraction, soreness and weakness of waist and knees, hemiplegia,
Traumatic injury, oedema, skin temperature are itched, the wet diseases such as swollen of scrotum.The main pharmacodynamics ingredient of acanthopanax leucorrhizus Harms stem skin is Stilbene class, flavones, phenylpropyl alcohol
The constituents such as element, terpene, lignanoid, cumarin, polysaccharide.The study found that the part Stilbene class isolated from acanthopanax leucorrhizus Harms stem skin at
Divide to SMMC-7721（Human liver cancer cell）、HL-60（People in loop）And MCF-7（Human breast cancer cell）Deng tool
There are different degrees of selective inhibitory, only a few Stilbene constituents to can be used as the anti-swollen of development prospect after structural modification
The primer of tumor medicine.But content is low in nature due to most of stilbenes compounds, is difficult to detach and Structural Identification, limitation
To its system research and development.
Currently, being mainly that traditional solvent assists various chromatographies for the separation method of Stilbene constituents（Such as silicagel column color
Spectrum, polyamide column chromatography the methods of prepare thin-layer chromatography, ODS, Sephadex LH-20 column chromatographys and HPLC）, but all different journeys
Degree ground there is complex process, the period is long, solvent-oil ratio is big, recovery rate is low, purity is not high the shortcomings of.
Invention content
Technical problem to be solved by the invention is to provide a kind of fast, efficiently Stilbene class being isolated from acanthopanax leucorrhizus Harms stem skin
The method of active ingredient.
To solve the above problems, isolating the side of Stilbene effective constituents in a kind of stem skin from acanthopanax leucorrhizus Harms of the present invention
Method includes the following steps：
(1) fresh acanthopanax leucorrhizus Harms stem skin dried, crushed, after the sieve of 40 ~ 60 mesh excessively, through Microwave Extraction, be concentrated under reduced pressure, slightly carried
Object；
(2) slightly object is taken to be extracted respectively 3 ~ 4 times with the petroleum ether of its 3 ~ 4 times of volume of quality, ethyl acetate successively by described, until extraction
After taking liquid to become colorless, acetic acid ethyl acetate extract is merged, sample to be separated is obtained through evaporated under reduced pressure；
(3) the sample to be separated mixed solvent is pressed into 1000 mg:Sample solution is obtained after the ratio ultrasonic dissolution of 50 mL；
(4) preferred through TLC screenings, analytic type HSCCC, by n-hexane, ethyl acetate, first alcohol and water according to 5:4:7:5 volume ratio
It is added in separatory funnel, is stood overnight after abundant shaken well, after two-phase system balance, upper phase is stationary phase, and lower phase is
Mobile phase is to get preparative HSCCC two phase solvent systems；
(5), by after the stationary phase and the mobile phase respectively 15 min of ultrasound degassing, preparative counter-current chromatograph is pumped into successively
Multi-layer helix-tube in, reinject the sample solution and carry out purifies and separates；
(6) it is collected into 7 chromatographic peak components successively by chromatogram, 7 components is obtained after being concentrated under reduced pressure, being dried in vacuo respectively
7 kinds of stilbenes compounds；7 kinds of stilbenes compounds be respectively 4- hydroxyl -11,12,13- trimethoxies dibenzo [b,f] Evil heptan
English, isorhapontigenin, 3,4 ', 5- trimethoxies-(E)-talan, 2 ', 3,4 ', 5- tetrahydroxys-(E)-talan,
3 '-isopentene groups-(E)-resveratrol, 3,4 ', 5- trihydroxies -2 '-methoxyl group-(E)-talan and 2- (3 '-hydroxyls -5 ' -
Methoxybenzene) -4- hydroxyl benzofurans.
The step (1) in the condition of Microwave Extraction refer to Extraction solvent be volumetric concentration be 75% ~ 95% ethyl alcohol, liquid-solid ratio
For 10 ~ 20 mL/g, speed of agitator be 600 ~ 700 rpm, Extracting temperature is 55 ~ 65 DEG C, microwave power is 650 ~ 750 W, is carried
It is 15 ~ 20 min to take the time, extraction time is 2 ~ 3 times.
The step (1) in reduced pressure condition be finger pressure be 0.05 ~ 0.06 MPa, temperature be 40 ~ 50 DEG C, rotating speed
For 100 ~ 150 rpm.
The step (2) in evaporated under reduced pressure condition be finger pressure be 0.04 ~ 0.05 MPa, temperature be 30 ~ 40 DEG C, rotating speed
For 80 ~ 100 rpm.
The step (3) in mixed solvent refer to by n-hexane, ethyl acetate, first alcohol and water according to 5:4:7:5 volume
Proportioning is added in separatory funnel, is stood overnight after abundant shaken well, after two-phase system balance, is divided into two-phase, then will
Upper phase solvent and lower phase solvent are obtained by mixing in equal volume.
The step (3) in ultrasonic dissolution condition refer to supersonic frequency be 40 kHz, temperature is 40 DEG C, and ultrasonic time is
5 min。
The step (5) in purifies and separates condition refer to temperature be 25 DEG C, helix tube rotating speed be 800 ~ 900 rpm, inspection
Survey wavelength is 325 nm, and note sample flow velocity is 1.5 ~ 2.0 mL/min, and single injected sampling amount is 160 mg.
The step (6) in reduced pressure condition be finger pressure be 0.03 ~ 0.05 MPa, temperature be 30 ~ 50 DEG C, rotating speed
For 80 ~ 150 rpm.
The step (6) in vacuum drying condition be finger pressure be 0.05 ~ 0.06 MPa, temperature is 30 ~ 50 DEG C, dry
2 ~ 10 h of time.
Compared with the prior art, the present invention has the following advantages：
1, the present invention is compared with traditional solvent extraction techniques：Under microwave action, Stilbene constituents can be selectively heated, and
It is oozed out from plant matrix, so as to quickly and efficiently realize the separation of Stilbene constituents and matrix, is effectively improved recovery rate
And purity.
2, the present invention is compared with traditional chromatographic separation technology：Using HSCCC method of purification, same dicyandiamide solution is repeatable to be made
With fractional dose is big, favorable reproducibility；Solid carrier is not needed simultaneously, avoids the separated object matter because of caused by Irreversible Adsorption
The problems such as loss, inactivation, denaturation, hangover, the rate of recovery of active ingredient is improved, is very suitable for Chinese herbal medicine effective ingredients control
The preparation of product.
3, the present invention realizes the extraction separation of Stilbene constituents in acanthopanax leucorrhizus Harms stem skin, purifying preparation integration, MAE crude extracts
It can be directly entered HSCCC after concentration to isolate and purify, greatly reduce intermediary operation link, reduce labor intensity and active ingredient
Loss, shorten disengaging time, improve separative efficiency.
4, present invention collection Microwave Extraction and high performance countercurrent chromatography purifying prepare integration, operation is simple, extracted amount is big,
The purity of favorable reproducibility, 7 kinds of stilbenes compounds of gained is above 98.0%.
Specific implementation mode
The method that Stilbene effective constituents are isolated in a kind of stem skin from acanthopanax leucorrhizus Harms of embodiment 1, includes the following steps：
(1) the fresh acanthopanax leucorrhizus Harms stem skins of 300 g dried, crushed, after the sieve of 40 mesh excessively, obtaining acanthopanax leucorrhizus Harms stem skin powder, the acanthopanax leucorrhizus Harms stem
It is 75% ethyl alcohol that Pi Fenzhong, which is 10 mL/g addition volumetric concentrations by liquid-solid ratio, is put into stirrer magneton, is placed in Microwave Extraction instrument,
Speed of agitator is 600 rpm, Extracting temperature is 60 DEG C, microwave power is 700 W, extraction time is 15 min, extraction time
Into Microwave Extraction of passing through under conditions of being 2 times, extracting solution is obtained；Extracting solution is in pressure is 0.05 MPa, temperature is 40 DEG C, turns
Speed is that 100 rpm are concentrated under reduced pressure, and obtains crude extract.
(2) it will slightly take object to be extracted respectively 3 times with the petroleum ether of its 3 times of volume of quality, ethyl acetate successively, until extract liquor
After becoming colorless, acetic acid ethyl acetate extract is merged, in the item that pressure is 0.04 MPa, temperature is 30 DEG C, rotating speed is 80 rpm
4.20 g of sample to be separated is obtained through evaporated under reduced pressure under part, yield is that acanthopanax leucorrhizus Harms dry the 1.40% of stem cortex amount.
(3) sample to be separated mixed solvent is pressed into 1000 mg:The ratio of 50 mL is 40 kHz, temperature in supersonic frequency
Ultrasonic dissolution under conditions of being 40 DEG C obtains the sample solution of 20 mg/mL after 5 min.
Wherein：Mixed solvent refers to by n-hexane, ethyl acetate, first alcohol and water according to 5:4:7:5 volume proportion is added
It into separatory funnel, is stood overnight after abundant shaken well, after two-phase system balance, is divided into two-phase, then by upper phase solvent
It is obtained by mixing in equal volume with lower phase solvent.
(4) preferred through TLC screenings, analytic type HSCCC, by n-hexane, ethyl acetate, first alcohol and water according to 5:4:7:5 body
Product ratio is added in separatory funnel, is stood overnight after abundant shaken well, and after two-phase system balance, upper phase is stationary phase, under
Phase is mobile phase to get preparative HSCCC two phase solvent systems.
(5), by after stationary phase and mobile phase respectively 15 min of ultrasound degassing, the more of preparative counter-current chromatograph are pumped into successively
In helical layer pipe, reinjects sample solution and carry out purifies and separates.
Detailed process：Stationary phase is pumped into the multi-layer helix-tube of preparative counter-current chromatograph with the flow velocity of 6.0 mL/min
In, after borded pile is completely filled with, opens constant-temperature circulating device and keep the temperature at 25 DEG C, start-up detector and HSCCC master
Machine slowly rotates in the forward direction（FWD）Adjusting helix tube rotating speed is 900 rpm, then enters mobile phase extremely with the flow pump of 2.0 mL/min
Until helix tube tail end has mobile phase outflow.After system reaches kinetic balance（After baseline stability）, then with 2.0 mL/min
Flow velocity by sample solution by sampling valve inject helix tube in detach.Detection wavelength is 325 nm, and single injected sampling amount is
160 mg。
(6) after about 45 min of sample solution injection separation helix tube, chromatographic peak starts to occur, and is collected successively by chromatogram
To 7 chromatographic peak components, 3.0 h are taken.By 7 components pressure is 0.03 MPa, temperature is 30 DEG C, rotating speed is 80 rpm
Under conditions of through being concentrated under reduced pressure to give concentrate, the concentrate is under conditions of pressure is 0.05 MPa, temperature is 30 DEG C through true
After dry 10 h of sky, 6 kinds of solid powders and a kind of yellow oil totally 7 kinds of stilbenes compounds are obtained.
It is parsed through NMR, MS, IR and UV, 7 kinds of stilbenes compounds of confirmation are respectively 4- hydroxyls -11,12,13- trimethoxies two
Benzo [b,f] Evil English in heptan（4-hydroxy-11,12,13-trimethoxyldibenz[b,f] oxepin）, different red leaf rheum officinale
Element（isorhapontigenin）, 3,4 ', 5- trimethoxies-(E)-talan（3,4′,5 -trimethoxy-(E)-
stilbene）, 2 ', 3,4 ', 5- tetrahydroxys-(E)-talan（2′,3,4′,5-tetrahydroxy- (E)-
stilbene）, 3 '-isopentene groups-(E)-resveratrol（3′-isopentenyl-(E)-resveratrol）, 3,4 ', 5- tri-
Hydroxyl -2 '-methoxyl group-(E)-talan（3,4′,5-trihydroxy-2′-methoxy-(E)-stilbene）And 2-
(3 '--5 '-methoxybenzene of hydroxyl) -4- hydroxyl benzofurans（2-(3′-hydroxy-5′-methoxyphenyl)-4-
hydroxybenzofuran）, and it is named as ZC I, ZC II, ZC III, ZC IV, ZC V, ZC VI and ZC VII successively.
By to 4 continuous sample introductions of HSCCC splitters, adding up sample size 640 mg, 15.5 h of total time-consuming.
Pass through HPLC purity assays：The quality and purity of 7 components are followed successively by ZC I（9.23 mg, 98.6%）、ZCⅡ
（23.01 mg, 99.2%）、ZCⅢ（13.62 mg, 99.5%）、ZCⅣ（6.18 mg, 99.3%）、ZCⅤ（7.14 mg,
98.1%）、ZCⅥ（14.25 mg, 99.4%）With ZC VII（8.61 mg, 98.7%）, purity is 98.0% or more.
The method that Stilbene effective constituents are isolated in a kind of stem skin from acanthopanax leucorrhizus Harms of embodiment 2, includes the following steps：
(1) the fresh acanthopanax leucorrhizus Harms stem skins of 500 g dried, crushed, after the sieve of 50 mesh excessively, obtaining acanthopanax leucorrhizus Harms stem skin powder, the acanthopanax leucorrhizus Harms stem
It is 80% ethyl alcohol that Pi Fenzhong, which is 15 mL/g addition volumetric concentrations by liquid-solid ratio, is put into stirrer magneton, is placed in Microwave Extraction instrument,
Speed of agitator is 650 rpm, Extracting temperature is 60 DEG C, microwave power is 650 W, extraction time is 20 min, extraction time
Into Microwave Extraction of passing through under conditions of being 3 times, extracting solution is obtained；Extracting solution is in pressure is 0.06 MPa, temperature is 50 DEG C, turns
Speed is that 150 rpm are concentrated under reduced pressure, and obtains crude extract.
(2) it will slightly take object to be extracted respectively 3 times with the petroleum ether of its 4 times of volume of quality, ethyl acetate successively, until extract liquor
After becoming colorless, acetic acid ethyl acetate extract is merged, in pressure is 0.05 MPa, temperature is 40 DEG C, rotating speed is 100 rpm
Under the conditions of through evaporated under reduced pressure obtain 7.10 g of sample to be separated, yield is that acanthopanax leucorrhizus Harms dry the 1.42% of stem cortex amount.
(3) sample to be separated mixed solvent is pressed into 1000 mg:The ratio of 50 mL is 40 kHz, temperature in supersonic frequency
Ultrasonic dissolution under conditions of being 40 DEG C obtains sample solution after 5 min.
Wherein：Mixed solvent is the same as embodiment 1.
(4) preparative HSCCC two phase solvent systems are the same as embodiment 1.
(5), by after stationary phase and mobile phase respectively 15 min of ultrasound degassing, the more of preparative counter-current chromatograph are pumped into successively
In helical layer pipe, reinjects sample solution and carry out purifies and separates.
Detailed process：Stationary phase is pumped into the multi-layer helix-tube of preparative counter-current chromatograph with the flow velocity of 6.0 mL/min
In, after borded pile is completely filled with, opens constant-temperature circulating device and keep the temperature at 25 DEG C, start-up detector and HSCCC master
Machine slowly rotates in the forward direction（FWD）Adjusting helix tube rotating speed is 900 rpm, then enters mobile phase extremely with the flow pump of 2.0 mL/min
Until helix tube tail end has mobile phase outflow.After system reaches kinetic balance（After baseline stability）, then with 1.5 mL/min
Flow velocity by sample solution by sampling valve inject helix tube in detach.Detection wavelength is 325 nm, and single injected sampling amount is
160 mg。
(6) after about 45 min of sample solution injection separation helix tube, chromatographic peak starts to occur, and is collected successively by chromatogram
To 7 chromatographic peak components, 3.5 h are taken.By 7 components pressure is 0.05 MPa, temperature is 50 DEG C, rotating speed is 150 rpm
Under conditions of through being concentrated under reduced pressure to give concentrate, the concentrate is under conditions of pressure is 0.06 MPa, temperature is 50 DEG C through true
After dry 2 h of sky, obtains 6 kinds of solid powders and a kind of yellow oil amounts to 7 kinds of stilbenes compounds.
By to 4 continuous sample introductions of HSCCC splitters, adding up sample size 640 mg, 17.5 h of total time-consuming.
Pass through HPLC purity assays：The quality and purity of 7 components are followed successively by ZC I（9.47 mg, 98.3%）、ZCⅡ
（23.46 mg, 98.6%）、ZCⅢ（13.10 mg, 99.3%）、ZCⅣ（6.25 mg, 99.6%）、ZCⅤ（7.31 mg,
98.5%）、ZCⅥ（13.43 mg, 98.7%）With ZC VII（9.45 mg, 98.1%）, purity is 98.0% or more.
The method that Stilbene effective constituents are isolated in a kind of stem skin from acanthopanax leucorrhizus Harms of embodiment 3, includes the following steps：
(1) the fresh acanthopanax leucorrhizus Harms stem skins of 400 g dried, crushed, after the sieve of 60 mesh excessively, obtaining acanthopanax leucorrhizus Harms stem skin powder, the acanthopanax leucorrhizus Harms stem
It is 95% ethyl alcohol that Pi Fenzhong, which is 20 mL/g addition volumetric concentrations by liquid-solid ratio, is put into stirrer magneton, is placed in Microwave Extraction instrument,
Speed of agitator is 700 rpm, Extracting temperature is 65 DEG C, microwave power is 750 W, extraction time is 20 min, extraction time
Into Microwave Extraction of passing through under conditions of being 3 times, extracting solution is obtained；Extracting solution is in pressure is 0.055 MPa, temperature is 45 DEG C, turns
Speed is that 125 rpm are concentrated under reduced pressure, and obtains crude extract.
(2) it will slightly take object to be extracted respectively 4 times with the petroleum ether of its 3 times of volume of quality, ethyl acetate successively, until extract liquor
After becoming colorless, acetic acid ethyl acetate extract is merged, in pressure is 0.045 MPa, temperature is 35 DEG C, rotating speed is 90 rpm
Under the conditions of through evaporated under reduced pressure obtain 5.50 g of sample to be separated, yield is that acanthopanax leucorrhizus Harms dry the 1.38% of stem cortex amount.
(3) sample to be separated mixed solvent is pressed into 1000 mg:The ratio of 50 mL is 40 kHz, temperature in supersonic frequency
Ultrasonic dissolution under conditions of being 40 DEG C obtains sample solution after 5 min.
Wherein：Mixed solvent is the same as embodiment 1.
(4) preparative HSCCC two phase solvent systems are the same as embodiment 1.
(5), by after stationary phase and mobile phase respectively 15 min of ultrasound degassing, the more of preparative counter-current chromatograph are pumped into successively
In helical layer pipe, reinjects sample solution and carry out purifies and separates.
Detailed process：Stationary phase is pumped into the multi-layer helix-tube of preparative counter-current chromatograph with the flow velocity of 6.0 mL/min
In, after borded pile is completely filled with, opens constant-temperature circulating device and keep the temperature at 25 DEG C, start-up detector and HSCCC master
Machine slowly rotates in the forward direction（FWD）Adjusting helix tube rotating speed is 800 rpm, then enters mobile phase extremely with the flow pump of 2.0 mL/min
Until helix tube tail end has mobile phase outflow.After system reaches kinetic balance（After baseline stability）, then with 2.0 mL/min
Flow velocity by sample solution by sampling valve inject helix tube in detach.Detection wavelength is 325 nm, and single injected sampling amount is
160 mg。
(6) after about 45 min of sample solution injection separation helix tube, chromatographic peak starts to occur, and is collected successively by chromatogram
To 7 chromatographic peak components, 3.2 h are taken.By 7 components pressure is 0.04 MPa, temperature is 40 DEG C, rotating speed is 100 rpm
Under conditions of through being concentrated under reduced pressure to give concentrate, which passes through under conditions of pressure is 0.055 MPa, temperature is 40 DEG C
After being dried in vacuo 6 h, obtains 6 kinds of solid powders and a kind of yellow oil amounts to 7 kinds of stilbenes compounds.
By to 4 continuous sample introductions of HSCCC splitters, adding up sample size 640 mg, 16.0 h of total time-consuming.
Pass through HPLC purity assays：The quality and purity of 7 components are followed successively by ZC I（9.21 mg, 99.1%）、ZCⅡ
（22.65 mg, 98.4%）、ZCⅢ（14.02 mg, 98.6%）、ZCⅣ（6.36 mg, 98.9%）、ZCⅤ（7.05 mg,
98.5%）、ZCⅥ（14.32 mg, 98.6%）With ZC VII（8.51 mg, 99.2%）, purity is 98.0% or more.
The method that Stilbene effective constituents are isolated in a kind of stem skin from acanthopanax leucorrhizus Harms of embodiment 4, includes the following steps：
(1) the fresh acanthopanax leucorrhizus Harms stem skins of 350 g dried, crushed, after the sieve of 40 mesh excessively, obtaining acanthopanax leucorrhizus Harms stem skin powder, the acanthopanax leucorrhizus Harms stem
It is 90% ethyl alcohol that Pi Fenzhong, which is 10 mL/g addition volumetric concentrations by liquid-solid ratio, is put into stirrer magneton, is placed in Microwave Extraction instrument,
Speed of agitator is 650 rpm, Extracting temperature is 55 DEG C, microwave power is 750 W, extraction time is 15 min, extraction time
Into Microwave Extraction of passing through under conditions of being 2 times, extracting solution is obtained；Extracting solution is in pressure is 0.05 MPa, temperature is 40 DEG C, turns
Speed is that 100 rpm are concentrated under reduced pressure, and obtains crude extract.
(2) it will slightly take object to be extracted respectively 4 times with the petroleum ether of its 4 times of volume of quality, ethyl acetate successively, until extract liquor
After becoming colorless, acetic acid ethyl acetate extract is merged, in the item that pressure is 0.04 MPa, temperature is 30 DEG C, rotating speed is 85 rpm
4.69 g of sample to be separated is obtained through evaporated under reduced pressure under part, yield is that acanthopanax leucorrhizus Harms dry the 1.34% of stem cortex amount.
(3) sample to be separated mixed solvent is pressed into 1000 mg:The ratio of 50 mL is 40 kHz, temperature in supersonic frequency
Ultrasonic dissolution under conditions of being 40 DEG C obtains sample solution after 5 min.
Wherein：Mixed solvent is the same as embodiment 1.
(4) preparative HSCCC two phase solvent systems are the same as embodiment 1.
(5), by after stationary phase and mobile phase respectively 15 min of ultrasound degassing, the more of preparative counter-current chromatograph are pumped into successively
In helical layer pipe, reinjects sample solution and carry out purifies and separates.
Detailed process：Stationary phase is pumped into the multi-layer helix-tube of preparative counter-current chromatograph with the flow velocity of 6.0 mL/min
In, after borded pile is completely filled with, opens constant-temperature circulating device and keep the temperature at 25 DEG C, start-up detector and HSCCC master
Machine slowly rotates in the forward direction（FWD）Adjusting helix tube rotating speed is 850 rpm, then enters mobile phase extremely with the flow pump of 2.0 mL/min
Until helix tube tail end has mobile phase outflow.After system reaches kinetic balance（After baseline stability）, then with 1.8 mL/min
Flow velocity by sample solution by sampling valve inject helix tube in detach.Detection wavelength is 325 nm, and single injected sampling amount is
160 mg。
(6) after about 45 min of sample solution injection separation helix tube, chromatographic peak starts to occur, and is collected successively by chromatogram
To 7 chromatographic peak components, 3.6 h are taken.By 7 components pressure is 0.03 MPa, temperature is 30 DEG C, rotating speed is 90 rpm
Under conditions of through being concentrated under reduced pressure to give concentrate, the concentrate is under conditions of pressure is 0.05 MPa, temperature is 30 DEG C through true
After dry 10 h of sky, obtains 6 kinds of solid powders and a kind of yellow oil amounts to 7 kinds of stilbenes compounds.
By to 4 continuous sample introductions of HSCCC splitters, adding up sample size 640 mg, 17.2 h of total time-consuming.
Pass through HPLC purity assays：The quality and purity of 7 components are followed successively by ZC I（9.18 mg, 98.6%）、ZCⅡ
（22.37 mg, 98.9%）、ZCⅢ（13.96 mg, 98.5%）、ZCⅣ（6.14 mg, 99.2%）、ZCⅤ（7.08 mg,
98.3%）、ZCⅥ（14.22 mg, 98.7%）With ZC VII（8.24 mg, 99.1%）, purity is 98.0% or more.
The method that Stilbene effective constituents are isolated in a kind of stem skin from acanthopanax leucorrhizus Harms of embodiment 5, includes the following steps：
(1) the fresh acanthopanax leucorrhizus Harms stem skins of 300 g dried, crushed, after the sieve of 40 mesh excessively, obtaining acanthopanax leucorrhizus Harms stem skin powder, the acanthopanax leucorrhizus Harms stem
It is 80% ethyl alcohol that Pi Fenzhong, which is 15 mL/g addition volumetric concentrations by liquid-solid ratio, is put into stirrer magneton, is placed in Microwave Extraction instrument,
Speed of agitator is 700 rpm, Extracting temperature is 65 DEG C, microwave power is 650 W, extraction time is 15 min, extraction time
Into Microwave Extraction of passing through under conditions of being 3 times, extracting solution is obtained；Extracting solution is in pressure is 0.06 MPa, temperature is 50 DEG C, turns
Speed is that 150 rpm are concentrated under reduced pressure, and obtains crude extract.
(2) it will slightly take object to be extracted respectively 4 times with the petroleum ether of its 3 times of volume of quality, ethyl acetate successively, until extract liquor
After becoming colorless, acetic acid ethyl acetate extract is merged, in pressure is 0.05 MPa, temperature is 40 DEG C, rotating speed is 100 rpm
Under the conditions of through evaporated under reduced pressure obtain 4.23 g of sample to be separated, yield is that acanthopanax leucorrhizus Harms dry the 1.41% of stem cortex amount.
(3) sample to be separated mixed solvent is pressed into 1000 mg:The ratio of 50 mL is 40 kHz, temperature in supersonic frequency
Ultrasonic dissolution under conditions of being 40 DEG C obtains sample solution after 5 min.
Wherein：Mixed solvent is the same as embodiment 1.
(4) preparative HSCCC two phase solvent systems are the same as embodiment 1.
(5), by after stationary phase and mobile phase respectively 15 min of ultrasound degassing, the more of preparative counter-current chromatograph are pumped into successively
In helical layer pipe, reinjects sample solution and carry out purifies and separates.
Detailed process：Stationary phase is pumped into the multi-layer helix-tube of preparative counter-current chromatograph with the flow velocity of 6.0 mL/min
In, after borded pile is completely filled with, opens constant-temperature circulating device and keep the temperature at 25 DEG C, start-up detector and HSCCC master
Machine slowly rotates in the forward direction（FWD）Adjusting helix tube rotating speed is 900 rpm, then enters mobile phase extremely with the flow pump of 2.0 mL/min
Until helix tube tail end has mobile phase outflow.After system reaches kinetic balance（After baseline stability）, then with 1.5 mL/min
Flow velocity by sample solution by sampling valve inject helix tube in detach.Detection wavelength is 325 nm, and single injected sampling amount is
160 mg。
(6) after about 45 min of sample solution injection separation helix tube, chromatographic peak starts to occur, and is collected successively by chromatogram
To 7 chromatographic peak components, 3.5 h are taken.By 7 components pressure is 0.05 MPa, temperature is 50 DEG C, rotating speed is 95 rpm
Under conditions of through being concentrated under reduced pressure to give concentrate, the concentrate is under conditions of pressure is 0.06 MPa, temperature is 50 DEG C through true
After dry 2 h of sky, obtains 6 kinds of solid powders and a kind of yellow oil amounts to 7 kinds of stilbenes compounds.
By to 4 continuous sample introductions of HSCCC splitters, adding up sample size 640 mg, 17.0 h of total time-consuming.
Pass through HPLC purity assays：The quality and purity of 7 components are followed successively by ZC I（9.15 mg, 98.5%）、ZCⅡ
（22.41 mg, 99.2%）、ZCⅢ（13.84 mg, 98.4%）、ZCⅣ（6.09 mg, 99.0%）、ZCⅤ（7.12 mg,
98.5%）、ZCⅥ（14.51 mg, 98.8%）With ZC VII（8.13 mg, 99.4%）, purity is 98.0% or more.
The method that Stilbene effective constituents are isolated in a kind of stem skin from acanthopanax leucorrhizus Harms of embodiment 6, includes the following steps：
(1) the fresh acanthopanax leucorrhizus Harms stem skins of 400 g dried, crushed, after the sieve of 50 mesh excessively, obtaining acanthopanax leucorrhizus Harms stem skin powder, the acanthopanax leucorrhizus Harms stem
It is 80% ethyl alcohol that Pi Fenzhong, which is 20 mL/g addition volumetric concentrations by liquid-solid ratio, is put into stirrer magneton, is placed in Microwave Extraction instrument,
Speed of agitator is 700 rpm, Extracting temperature is 60 DEG C, microwave power is 650 W, extraction time is 20 min, extraction time
Into Microwave Extraction of passing through under conditions of being 3 times, extracting solution is obtained；Extracting solution is in pressure is 0.055 MPa, temperature is 45 DEG C, turns
Speed is that 130 rpm are concentrated under reduced pressure, and obtains crude extract.
(2) it will slightly take object to be extracted respectively 3 times with the petroleum ether of its 3 times of volume of quality, ethyl acetate successively, until extract liquor
After becoming colorless, acetic acid ethyl acetate extract is merged, in pressure is 0.045 MPa, temperature is 35 DEG C, rotating speed is 90 rpm
Under the conditions of through evaporated under reduced pressure obtain 5.54 g of sample to be separated, yield is that acanthopanax leucorrhizus Harms dry the 1.39% of stem cortex amount.
(3) sample to be separated mixed solvent is pressed into 1000 mg:The ratio of 50 mL is 40 kHz, temperature in supersonic frequency
Ultrasonic dissolution under conditions of being 40 DEG C obtains sample solution after 5 min.
Wherein：Mixed solvent is the same as embodiment 1.
(4) preparative HSCCC two phase solvent systems are the same as embodiment 1.
(5), by after stationary phase and mobile phase respectively 15 min of ultrasound degassing, the more of preparative counter-current chromatograph are pumped into successively
In helical layer pipe, reinjects sample solution and carry out purifies and separates.
Detailed process：Stationary phase is pumped into the multi-layer helix-tube of preparative counter-current chromatograph with the flow velocity of 6.0 mL/min
In, after borded pile is completely filled with, opens constant-temperature circulating device and keep the temperature at 25 DEG C, start-up detector and HSCCC master
Machine slowly rotates in the forward direction（FWD）Adjusting helix tube rotating speed is 900 rpm, then enters mobile phase extremely with the flow pump of 2.0 mL/min
Until helix tube tail end has mobile phase outflow.After system reaches kinetic balance（After baseline stability）, then with 2.0 mL/min
Flow velocity by sample solution by sampling valve inject helix tube in detach.Detection wavelength is 325 nm, and single injected sampling amount is
160 mg。
(6) after about 45 min of sample solution injection separation helix tube, chromatographic peak starts to occur, and is collected successively by chromatogram
To 7 chromatographic peak components, 3.1 h are taken.By 7 components pressure is 0.04 MPa, temperature is 40 DEG C, rotating speed is 100 rpm
Under conditions of through being concentrated under reduced pressure to give concentrate, which passes through under conditions of pressure is 0.055 MPa, temperature is 40 DEG C
After being dried in vacuo 6 h, obtains 6 kinds of solid powders and a kind of yellow oil amounts to 7 kinds of stilbenes compounds.
By to 4 continuous sample introductions of HSCCC splitters, adding up sample size 640 mg, 15.0 h of total time-consuming.
Pass through HPLC purity assays：The quality and purity of 7 components are followed successively by ZC I（9.32 mg, 98.3%）、ZCⅡ
（22.45 mg, 99.0%）、ZCⅢ（13.67 mg, 98.2%）、ZCⅣ（6.05 mg, 98.9%）、ZCⅤ（7.24 mg,
98.7%）、ZCⅥ（14.38 mg, 98.7%）With ZC VII（8.24 mg, 99.2%）, purity is 98.0% or more.
The method that Stilbene effective constituents are isolated in a kind of stem skin from acanthopanax leucorrhizus Harms of embodiment 7, includes the following steps：
(1) the fresh acanthopanax leucorrhizus Harms stem skins of 500 g dried, crushed, after the sieve of 60 mesh excessively, obtaining acanthopanax leucorrhizus Harms stem skin powder, the acanthopanax leucorrhizus Harms stem
It is 95% ethyl alcohol that Pi Fenzhong, which is 20 mL/g addition volumetric concentrations by liquid-solid ratio, is put into stirrer magneton, is placed in Microwave Extraction instrument,
Speed of agitator is 700 rpm, Extracting temperature is 55 DEG C, microwave power is 650 W, extraction time is 15 min, extraction time
Into Microwave Extraction of passing through under conditions of being 3 times, extracting solution is obtained；Extracting solution is in pressure is 0.05 MPa, temperature is 40 DEG C, turns
Speed is that 100 rpm are concentrated under reduced pressure, and obtains crude extract.
(2) it will slightly take object to be extracted respectively 4 times with the petroleum ether of its 4 times of volume of quality, ethyl acetate successively, until extract liquor
After becoming colorless, acetic acid ethyl acetate extract is merged, in the item that pressure is 0.04 MPa, temperature is 30 DEG C, rotating speed is 80 rpm
7.14 g of sample to be separated is obtained through evaporated under reduced pressure under part, yield is that acanthopanax leucorrhizus Harms dry the 1.43% of stem cortex amount.
(3) sample to be separated mixed solvent is pressed into 1000 mg:The ratio of 50 mL is 40 kHz, temperature in supersonic frequency
Ultrasonic dissolution under conditions of being 40 DEG C obtains sample solution after 5 min.
Wherein：Mixed solvent is the same as embodiment 1.
(4) preparative HSCCC two phase solvent systems are the same as embodiment 1.
(5), by after stationary phase and mobile phase respectively 15 min of ultrasound degassing, the more of preparative counter-current chromatograph are pumped into successively
In helical layer pipe, reinjects sample solution and carry out purifies and separates.
Detailed process：Stationary phase is pumped into the multi-layer helix-tube of preparative counter-current chromatograph with the flow velocity of 6.0 mL/min
In, after borded pile is completely filled with, opens constant-temperature circulating device and keep the temperature at 25 DEG C, start-up detector and HSCCC master
Machine slowly rotates in the forward direction（FWD）Adjusting helix tube rotating speed is 900 rpm, then enters mobile phase extremely with the flow pump of 2.0 mL/min
Until helix tube tail end has mobile phase outflow.After system reaches kinetic balance（After baseline stability）, then with 2.0 mL/min
Flow velocity by sample solution by sampling valve inject helix tube in detach.Detection wavelength is 325 nm, and single injected sampling amount is
160 mg。
(6) after about 45 min of sample solution injection separation helix tube, chromatographic peak starts to occur, and is collected successively by chromatogram
To 7 chromatographic peak components, 3.2 h are taken.By 7 components pressure is 0.03 MPa, temperature is 30 DEG C, rotating speed is 80 rpm
Under conditions of through being concentrated under reduced pressure to give concentrate, the concentrate is under conditions of pressure is 0.05 MPa, temperature is 30 DEG C through true
After dry 10 h of sky, obtains 6 kinds of solid powders and a kind of yellow oil amounts to 7 kinds of stilbenes compounds.
By to 4 continuous sample introductions of HSCCC splitters, adding up sample size 640 mg, 15.5 h of total time-consuming.
Pass through HPLC purity assays：The quality and purity of 7 components are followed successively by ZC I（9.45 mg, 98.5%）、ZCⅡ
（22.48 mg, 98.6%）、ZCⅢ（13.71 mg, 98.1%）、ZCⅣ（6.01 mg, 99.1%）、ZCⅤ（7.23 mg,
98.5%）、ZCⅥ（14.24 mg, 99.0%）With ZC VII（8.17 mg, 99.3%）, purity is 98.0% or more.
The method that Stilbene effective constituents are isolated in a kind of stem skin from acanthopanax leucorrhizus Harms of embodiment 8, includes the following steps：
(1) the fresh acanthopanax leucorrhizus Harms stem skins of 300 g dried, crushed, after the sieve of 60 mesh excessively, obtaining acanthopanax leucorrhizus Harms stem skin powder, the acanthopanax leucorrhizus Harms stem
Pi Fenzhong by liquid-solid ratio be 20 mL/g be added volumetric concentration be 75% ethyl alcohol, speed of agitator be 700 rpm, Extracting temperature 60
DEG C, microwave power be 700 W, under conditions of extraction time is 18 min, extraction time is 23 times into Microwave Extraction of passing through, obtain
Extracting solution；Extracting solution is slightly carried in pressure is 0.06 MPa, temperature is 50 DEG C, rotating speed is that 150 rpm are concentrated under reduced pressure
Object.
(2) it will slightly take object to be extracted respectively 4 times with the petroleum ether of its 3.5 times of volume of quality, ethyl acetate successively, until extraction
After liquid becomes colorless, acetic acid ethyl acetate extract is merged, in pressure is 0.05 MPa, temperature is 40 DEG C, rotating speed is 100 rpm
Under conditions of through evaporated under reduced pressure obtain 4.35 g of sample to be separated, yield is that acanthopanax leucorrhizus Harms dry the 1.45% of stem cortex amount.
(3) sample to be separated mixed solvent is pressed into 1000 mg:The ratio of 50 mL is 40 kHz, temperature in supersonic frequency
Ultrasonic dissolution under conditions of being 40 DEG C obtains sample solution after 5 min.
Wherein：Mixed solvent is the same as embodiment 1.
(4) preparative HSCCC two phase solvent systems are the same as embodiment 1.
(5), by after stationary phase and mobile phase respectively 15 min of ultrasound degassing, the more of preparative counter-current chromatograph are pumped into successively
In helical layer pipe, reinjects sample solution and carry out purifies and separates.
Detailed process：Stationary phase is pumped into the multi-layer helix-tube of preparative counter-current chromatograph with the flow velocity of 6.0 mL/min
In, after borded pile is completely filled with, opens constant-temperature circulating device and keep the temperature at 25 DEG C, start-up detector and HSCCC master
Machine slowly rotates in the forward direction（FWD）Adjusting helix tube rotating speed is 900 rpm, then enters mobile phase extremely with the flow pump of 2.0 mL/min
Until helix tube tail end has mobile phase outflow.After system reaches kinetic balance（After baseline stability）, then with 2.0 mL/min
Flow velocity by sample solution by sampling valve inject helix tube in detach.Detection wavelength is 325 nm, and single injected sampling amount is
160 mg。
(6) after about 45 min of sample solution injection separation helix tube, chromatographic peak starts to occur, and is collected successively by chromatogram
To 7 chromatographic peak components, 3.0 h are taken.By 7 components pressure is 0.05 MPa, temperature is 50 DEG C, rotating speed is 150 rpm
Under conditions of through being concentrated under reduced pressure to give concentrate, the concentrate is under conditions of pressure is 0.06 MPa, temperature is 50 DEG C through true
After dry 2 h of sky, obtains 6 kinds of solid powders and a kind of yellow oil amounts to 7 kinds of stilbenes compounds.
By to 4 continuous sample introductions of HSCCC splitters, adding up sample size 640 mg, 15.2 h of total time-consuming.
Pass through HPLC purity assays：The quality and purity of 7 components are followed successively by ZC I（9.44 mg, 98.6%）、ZCⅡ
（22.45 mg, 98.7%）、ZCⅢ（13.68 mg, 98.3%）、ZCⅣ（6.11 mg, 99.0%）、ZCⅤ（7.18 mg,
98.6%）、ZCⅥ（14.27 mg, 98.9%）With ZC VII（8.22 mg, 99.8%）, purity is 98.0% or more.
Claims (9)
1. a kind of method for isolating Stilbene effective constituents in stem skin from acanthopanax leucorrhizus Harms, includes the following steps：
(1) fresh acanthopanax leucorrhizus Harms stem skin dried, crushed, after the sieve of 40 ~ 60 mesh excessively, through Microwave Extraction, be concentrated under reduced pressure, slightly carried
Object；
(2) slightly object is taken to be extracted respectively 3 ~ 4 times with the petroleum ether of its 3 ~ 4 times of volume of quality, ethyl acetate successively by described, until extraction
After taking liquid to become colorless, acetic acid ethyl acetate extract is merged, sample to be separated is obtained through evaporated under reduced pressure；
(3) the sample to be separated mixed solvent is pressed into 1000 mg:Sample solution is obtained after the ratio ultrasonic dissolution of 50 mL；
(4) preferred through TLC screenings, analytic type HSCCC, by n-hexane, ethyl acetate, first alcohol and water according to 5:4:7:5 volume ratio
It is added in separatory funnel, is stood overnight after abundant shaken well, after two-phase system balance, upper phase is stationary phase, and lower phase is
Mobile phase is to get preparative HSCCC two phase solvent systems；
(5), by after the stationary phase and the mobile phase respectively 15 min of ultrasound degassing, preparative counter-current chromatograph is pumped into successively
Multi-layer helix-tube in, reinject the sample solution and carry out purifies and separates；
(6) it is collected into 7 chromatographic peak components successively by chromatogram, 7 components is obtained after being concentrated under reduced pressure, being dried in vacuo respectively
7 kinds of stilbenes compounds；7 kinds of stilbenes compounds be respectively 4- hydroxyl -11,12,13- trimethoxies dibenzo [b,f] Evil heptan
English, isorhapontigenin, 3,4 ', 5- trimethoxies-(E)-talan, 2 ', 3,4 ', 5- tetrahydroxys-(E)-talan,
3 '-isopentene groups-(E)-resveratrol, 3,4 ', 5- trihydroxies -2 '-methoxyl group-(E)-talan and 2- (3 '-hydroxyls -5 ' -
Methoxybenzene) -4- hydroxyl benzofurans.
2. the method for isolating Stilbene effective constituents in a kind of stem skin from acanthopanax leucorrhizus Harms as described in claim 1, it is characterised in that：
The step (1) in Microwave Extraction condition refer to Extraction solvent be volumetric concentration be 75% ~ 95% ethyl alcohol, liquid-solid ratio be 10 ~ 20
ML/g, speed of agitator are 600 ~ 700 rpm, Extracting temperature is 55 ~ 65 DEG C, microwave power is 650 ~ 750 W, extraction time is
15 ~ 20 min, extraction time are 2 ~ 3 times.
3. the method for isolating Stilbene effective constituents in a kind of stem skin from acanthopanax leucorrhizus Harms as described in claim 1, it is characterised in that：
The step (1) in reduced pressure condition be finger pressure be 0.05 ~ 0.06 MPa, temperature be 40 ~ 50 DEG C, rotating speed be 100 ~
150 rpm。
4. the method for isolating Stilbene effective constituents in a kind of stem skin from acanthopanax leucorrhizus Harms as described in claim 1, it is characterised in that：
The step (2) in evaporated under reduced pressure condition be finger pressure be 0.04 ~ 0.05 MPa, temperature be 30 ~ 40 DEG C, rotating speed be 80 ~ 100
rpm。
5. the method for isolating Stilbene effective constituents in a kind of stem skin from acanthopanax leucorrhizus Harms as described in claim 1, it is characterised in that：
The step (3) in mixed solvent refer to by n-hexane, ethyl acetate, first alcohol and water according to 5:4:7:5 volume proportion is added
It into separatory funnel, is stood overnight after abundant shaken well, after two-phase system balance, is divided into two-phase, then by upper phase solvent
It is obtained by mixing in equal volume with lower phase solvent.
6. the method for isolating Stilbene effective constituents in a kind of stem skin from acanthopanax leucorrhizus Harms as described in claim 1, it is characterised in that：
The step (3) in ultrasonic dissolution condition refer to supersonic frequency be 40 kHz, temperature be 40 DEG C, ultrasonic time be 5 min.
7. the method for isolating Stilbene effective constituents in a kind of stem skin from acanthopanax leucorrhizus Harms as described in claim 1, it is characterised in that：
The step (5) in purifies and separates condition refer to temperature be 25 DEG C, helix tube rotating speed is 800 ~ 900 rpm, Detection wavelength is
325 nm, note sample flow velocity are 1.5 ~ 2.0 mL/min, and single injected sampling amount is 160 mg.
8. the method for isolating Stilbene effective constituents in a kind of stem skin from acanthopanax leucorrhizus Harms as described in claim 1, it is characterised in that：
The step (6) in reduced pressure condition be finger pressure be 0.03 ~ 0.05 MPa, temperature be 30 ~ 50 DEG C, rotating speed be 80 ~ 150
rpm。
9. the method for isolating Stilbene effective constituents in a kind of stem skin from acanthopanax leucorrhizus Harms as described in claim 1, it is characterised in that：
The step (6) in vacuum drying condition be finger pressure be 0.05 ~ 0.06 MPa, temperature be 30 ~ 50 DEG C, drying time 2 ~ 10
h。
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1566054A (en) * | 2003-06-27 | 2005-01-19 | 中国医学科学院药物研究所 | Resveratrol oligo cattail compounds, its manufacturing process, pharmaceutical combination and uses thereof |
| CN106309523A (en) * | 2015-11-01 | 2017-01-11 | 长沙博海生物科技有限公司 | Total flavonoid extract of acanthopanax senticosus harms, as well as preparation method and application thereof |
-
2018
- 2018-02-02 CN CN201810105960.2A patent/CN108276375A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1566054A (en) * | 2003-06-27 | 2005-01-19 | 中国医学科学院药物研究所 | Resveratrol oligo cattail compounds, its manufacturing process, pharmaceutical combination and uses thereof |
| CN106309523A (en) * | 2015-11-01 | 2017-01-11 | 长沙博海生物科技有限公司 | Total flavonoid extract of acanthopanax senticosus harms, as well as preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| HAO-BIN HUA等: "《Isolation, modification and cytotoxic evaluation of stilbenoids from Acanthopanax leucorrhizus》", 《FITOTERAPIA》 * |
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