CN103450145A - Method for separating and preparing Brazilin and Protosappanin B from Sappanwood by using high-speed countercurrent chromatography - Google Patents
Method for separating and preparing Brazilin and Protosappanin B from Sappanwood by using high-speed countercurrent chromatography Download PDFInfo
- Publication number
- CN103450145A CN103450145A CN 201310393712 CN201310393712A CN103450145A CN 103450145 A CN103450145 A CN 103450145A CN 201310393712 CN201310393712 CN 201310393712 CN 201310393712 A CN201310393712 A CN 201310393712A CN 103450145 A CN103450145 A CN 103450145A
- Authority
- CN
- China
- Prior art keywords
- brazilin
- hematorylin
- former
- bush
- preparing
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Abstract
The invention discloses a method for separating and preparing Brazilin and Protosappanin B from Sappanwood by using high-speed countercurrent chromatography, which comprises the following steps: preparing a sample solution firstly, then carrying out separation and preparation on the sample solution by using a high-speed countercurrent chromatographic instrument and an ultraviolet detector, respectively collecting flow elements containing target components according to an ultraviolet map, and carrying out decompression concentration on the flow elements until the target components including Brazilin and Protosappanin B are obtained, wherein the nature determination of the target components is determined by a mass spectrum in combination with H nuclear magnetic resonance and a carbon spectrum, and the purity is determined by HPLC. According to the invention, a solid phase carrier is not required to be used, reversible adsorption does not exist, solvents used in the process of separation and preparation can be greatly reduced, the solvent reduced amount is over 80%, the separation time is saved by at least five times, and the yield is increased by more than 50%; separated target components are little in loss, high in purity, low in cost and economic and has environmental-friendly effect.
Description
Technical field
The present invention relates to the method for separating compound in natural drug, be specifically related to a kind of high speed adverse current chromatogram that utilizes and separate the method for preparing brazilin and former Hematorylin B from bush.
Background technology
Bush is traditional Chinese medicine, in China, long medicinal history is arranged, Chinese Pharmacopoeia record bush be the leguminous plants bush (
caesalpinia sappanlingum) dry duramen, have capable blood and dissolving stasis, the effect of swelling and pain relieving.The chemical composition that bush contains mainly contains the compositions such as flavonoid, homoisoflavone class, chalcone.
Brazilin is a kind of natural medium fuel, is usually used in the dyeing of silk, fur and leather and the aspects such as stamp of cotton; Brazilin and former Hematorylin B are the main a large amount of compositions in the bush medicinal material, are also main activeconstituentss.Therefore develop a kind of economy, simply, method obtains these two chemical compositions and has larger production application and be worth efficiently.
The separation of natural product adopts the method for column chromatography to be separated mostly, if adopt column chromatography to separate from bush, prepares brazilin and former Hematorylin B, needs repeatedly repeatedly to separate, and its deficiency has: (1) is consuming time long, often will be over one month; (2) need to use a large amount of solvents; (3) have dead adsorption phenomena, sample more or less all can have loss; (4) labor capacity is larger; (5) sample size that single obtains is less, if obtain the above sample of 10mg, and be through separating for several times.
Summary of the invention
The present invention separates for solving traditional column chromatography problems such as preparing in the target compound process length consuming time, the solvent-oil ratio that run into are large, labor capacity is large, sample absorption is serious, sample loss, the invention provides a kind of high speed adverse current chromatogram that utilizes and separate the method for preparing brazilin and former Hematorylin B from bush, at first prepare sample solution, then adopt high-speed counter-current chromatograph coupling UV-detector to separate preparation, collect respectively the flow point that contains target components brazilin and former Hematorylin B according to uv-spectrogram.The present invention adopts high-speed countercurrent chromatography coupling UV-detector not need to use solid phase carrier, without irreversible adsorption, can greatly reduce and separate the solvent used in preparation process, its solvent reduction is more than 80%, and disengaging time is at least saved more than 5 times, and yield improves more than 50%, isolated target component loss is few, purity is high, and expense is low, economic environmental protection.
The high speed adverse current chromatogram that utilizes of the present invention separates the method for preparing brazilin and former Hematorylin B from bush, at first prepare sample solution, then adopt high-speed counter-current chromatograph coupling UV-detector to separate preparation, collect respectively the flow point that contains brazilin and former Hematorylin B according to uv-spectrogram, then be concentrated into dryly with Rotary Evaporators, obtain respectively final product brazilin and former Hematorylin B.Described target component brazilin and former Hematorylin B's is qualitative definite in conjunction with proton nmr spectra, carbon spectrum by mass spectrum, and its purity is utilized HPLC to analyze and obtained, and its weight obtains by weighing on analytical balance.
The structural formula of former Hematorylin B of the present invention and brazilin is as follows:
The chromatographic condition that described high speed adverse current chromatogram adopts is: solvent system is chloroform: methyl alcohol: the volume ratio of water is 4:2-4:2, and upper is stationary phase mutually, and lower is moving phase mutually; The instrument rotating speed: adopt and clockwise rotate, rotating speed is 400 ~ 550 turn/min, and flow rate of mobile phase is 5.0 ~ 8.0 mL/min; Bath temperature: 20 ℃-30 ℃; Retention rate: 30%-80%; Detect wavelength: 280nm.
Except above-mentioned chromatographic condition, other conditions are the existing installation normal condition.
Such chromatographic condition, can make the retention rate of stationary phase between 30%-80%, thereby realize separating efficiently; And under the prerequisite that guarantees separating effect, go out peak the fastest, cost is minimum.
According to the physico-chemical property of target compound brazilin and former Hematorylin B itself and obtain in conjunction with its uv scan spectrogram: the ultraviolet detection wavelength adopted during described ultraviolet detection is 280nm.
Concrete steps comprise:
(1) preparation of solvent system
By chloroform: methyl alcohol: water be take the ratio that volume ratio is 4:2-4:2 and is mixed, stratification, and upper is stationary phase mutually, lower is moving phase mutually;
(2) preparation of sample solution
Bush is extracted 1 ~ 5 time with the solvent orange 2 A thermal backflow of 2 ~ 5 times of amounts, united extraction liquid, be evaporated to the residual extractum A that obtains of solvent-free A, the extractum A water disperses fully, then uses the ethyl acetate extraction extractum A of 2 ~ 5 times of water gagings, ethyl acetate layer obtains medicinal extract B after being evaporated to and doing, upper macroporous adsorbent resin, used the aqueous ethanolic solution wash-out of 5 ~ 20 times of amounts, and elutriant is evaporated to dry, obtain medicinal extract C, i.e. bush efficient part; Then, in the solvent that the moving phase that to be dissolved in by volume ratio fully by the bush efficient part be 1:1 and stationary phase form, obtain sample solution;
(3) separate and prepare brazilin and former Hematorylin B
Extracting sample solution adopts high-speed counter-current chromatograph to carry out sample introduction, and collects respectively according to the UV-detector spectrogram flow point that contains brazilin and former Hematorylin B, then with Rotary Evaporators, is concentrated into dryly, obtains respectively final product brazilin and former Hematorylin B;
The described solvent orange 2 A of step (2) is methanol solution or ethanolic soln.
Bush of the present invention is commercially available medicinal material.
Of the present invention times of amount refered in particular to volume ratio.
The aqueous ethanolic solution of described 5 ~ 20 times of amounts of step (2), its volume refers to 5 ~ 20 times with respect to the column volume of macroporous adsorbent resin column chromatography.
The stationary phase of high speed adverse current chromatogram of the present invention and moving phase are layering after chloroform, first alcohol and water mix, upper as stationary phase, described upper be the less solvent system of proportion after layering mutually, lower to moving phase, described lower be the larger solvent system of proportion after layering mutually.
Described chloroform: methyl alcohol: the volume ratio of water is 4:2-4:2, is preferably 4:4:2.Adopt the solvent system of such volume ratio, be because the described bush efficient part of step (2) dissolves wherein, and can realize separating efficiently in high-speed counter-current chromatograph.
Further, in the solvent that the described bush efficient part of step (2) moving phase that to be dissolved in by volume ratio fully be 1:1 and stationary phase are formed, as sample solution, carry out sample introduction.Can avoid the waste of solvent like this, reduce costs.
The described solvent orange 2 A of step (2) is: the aqueous ethanolic solution that the methanol aqueous solution that volume fraction is 30%-100% or volume fraction are 30%-100%.This is because target components brazilin and former Hematorylin B are soluble in the methanol aqueous solution or aqueous ethanolic solution that volume fraction is 30%-100%.
The described macroporous adsorbent resin of step (2) is the D101 macroporous adsorbent resin, for commercially available prod, the macroporous resin of described D101 macroporous adsorbent resin and commercially available different model, macroporous resin as LX-17, LSA-40, HP-20 model, the contriver finds to adopt the D101 macroporous adsorbent resin to have good selectivity for isolate the efficient part that contains the former Hematorylin B of target components and brazilin from bush, and the separation of bush efficient part preparation is brazilin separates preparation prerequisite with former Hematorylin B.
The described eluting solvent of step (2) is the aqueous ethanolic solution that volume fraction is 5%-15%, and this is because target components brazilin and former Hematorylin B mainly concentrate in the flow point of the aqueous ethanolic solution that volume fraction is 5%-15%.
The model of the present invention's high-speed counter-current chromatograph used is TBE-1000A, is the currently available products of Shanghai Tongtian Biotechnology Co., Ltd.'s production; The UV-detector that HSCCC is equipped with is that gold reaches UV-detector, is the currently available products of Shanghai Jin Da Science and Technology Ltd. production; Rotary Evaporators is N-1000 type Rotary Evaporators, is the currently available products of Shanghai Ai Lang instrument company production; The temperature control instrument is the circulator bath system, is the currently available products of Beijing rich doctor health biological medicine company limited production; All reagent is all purchased from Chemical Reagent Co., Ltd., Sinopharm Group, and purity is analytical pure.
The temperature control instrument of described high-speed counter-current chromatograph is the circulator bath system, and the bath temperature adopted is 20 ℃-30 ℃, is preferably 25 ℃, and the resolution of target components, peak type reach effective separation like this.
The high speed adverse current chromatogram that utilizes of the present invention separates the method for preparing brazilin and former Hematorylin B from bush, at first prepare sample solution, then adopt high-speed counter-current chromatograph coupling UV-detector to separate preparation, collect respectively the target component that contains brazilin and former Hematorylin B according to uv-spectrogram.The qualitative of described target component composed and determines in conjunction with proton nmr spectra, carbon by mass spectrum, and its purity is determined by HPLC.Described HPLC analysis condition is as follows: the serial HPLC of Agilent 1260, the auxiliary work station, Agilent SB-C18 chromatographic column (column length 150mm * post footpath 4.6mm, particle diameter 5 μ m), adopt the methanol aqueous solution that volume fraction is 70% to carry out isocratic elution as moving phase, detecting wavelength is 280 nm, and the flow velocity of moving phase is 1.0 mL/min.
The present invention adopts high-speed countercurrent chromatography coupling UV-detector to separate the method for preparing brazilin and former Hematorylin B from bush, do not need to use solid phase carrier, without irreversible adsorption, can greatly reduce and separate the solvent used in preparation process, its solvent reduction is more than 80%, disengaging time is at least saved more than 5 times, yield improves more than 50%, and isolated target component loss is few, and purity is high, expense is low, economic environmental protection.
The accompanying drawing explanation
Fig. 1 is that embodiment 1 separates the HSCCC collection of illustrative plates for preparing target compound;
Fig. 2 is that embodiment 2 separates the HSCCC collection of illustrative plates for preparing target compound;
Fig. 3 is that embodiment 3 separates the HSCCC collection of illustrative plates for preparing target compound;
Fig. 4 is the HPLC purity check figure that separates the former Hematorylin B prepared;
Fig. 5 is the HPLC purity check figure that separates the brazilin prepared.
Embodiment
High-speed counter-current chromatograph model of the present invention is TBE-1000A, is the currently available products of Shanghai Tongtian Biotechnology Co., Ltd.'s production; The UV-detector that HSCCC is equipped with is that gold reaches UV-detector, is the currently available products of Shanghai Jin Da Science and Technology Ltd. production; Rotary Evaporators is N-1000 type Rotary Evaporators, is the currently available products of Shanghai Ai Lang instrument company production; The temperature control instrument is circulator bath, is the currently available products of Beijing rich doctor health biological medicine company limited production; All reagent is all purchased from Chemical Reagent Co., Ltd., Sinopharm Group, and purity is analytical pure.
Embodiment 1
A kind of high speed adverse current chromatogram that utilizes separates the method for preparing brazilin and former Hematorylin B from bush, and concrete steps comprise:
(1) preparation of solvent system
By chloroform: methyl alcohol: water be take the ratio that volume ratio is 4:4:2 and is mixed, stratification, and upper is stationary phase mutually, lower is moving phase mutually;
(2) preparation of sample solution
The aqueous ethanolic solution 5000mL thermal backflow that bush medicinal material 1 Kg is 80% by the volume fraction of 3 times of amounts is extracted 3 times, each 2h, united extraction liquid, be evaporated to after residual without aqueous ethanolic solution to obtain extractum A 10g, after extractum A is disperseed fully with 200mL water, extract extractum A 3 times with the ethyl acetate 600mL of 3 times of amounts again, ethyl acetate layer obtains medicinal extract B 5g after being evaporated to and doing, upper D101 macroporous adsorbent resin, the aqueous ethanolic solution 2000mL wash-out that the volume fraction of using 10 times of amounts (column volume of macroporous adsorbent resin chromatography post is 200mL) is 10%, elutriant is evaporated to dry, obtain 2g medicinal extract C, it is the bush efficient part, then, in the 80mL solvent that the moving phase that to be dissolved in by volume ratio fully by the bush efficient part be 1:1 and stationary phase form, obtain sample solution,
(3) separate and prepare brazilin and former Hematorylin B
The chromatographic condition of described high speed adverse current chromatogram is: solvent system is chloroform: methyl alcohol: the volume ratio of water is 4:4:2, upper as stationary phase, lower to moving phase; The instrument rotating speed: the stationary phase system clockwise rotates, 500 turn/min; Flow rate of mobile phase: 7.5 mL/min; Bath temperature: 25 ℃; Retention rate: 75%; Detect wavelength: 280 nm;
First adopt stationary phase to be full of the chromatographic column of high-speed counter-current chromatograph, then make its main frame clockwise rotate, then moving phase is pumped in chromatographic column, extracting sample solution is by the sampling valve sample introduction; And be under 280nm at the ultraviolet detection wavelength, collect respectively the flow point that contains brazilin and former Hematorylin B according to the UV-detector spectrogram, then be concentrated into dryly with Rotary Evaporators, obtain respectively the former Hematorylin B(of final product 671 mg purity 97.1%) and 865 mg brazilins (purity 92.9%).
Embodiment 2
A kind of high speed adverse current chromatogram that utilizes separates the method for preparing brazilin and former Hematorylin B from bush, and concrete steps comprise:
(1) preparation of solvent system
By chloroform: methyl alcohol: water be take the ratio that volume ratio is 4:3:2 and is mixed, stratification, and upper is stationary phase mutually, lower is moving phase mutually;
(2) preparation of sample solution
The methanol aqueous solution 5000 mL thermal backflows that bush medicinal material 1 Kg is 30% by the volume fraction of 2 times of amounts are extracted 1 time, each 2h, united extraction liquid, be evaporated to after residual without methanol-water to obtain extractum A 15g, after extractum A is disperseed fully with 300ml water, extract extractum A 3 times with ethyl acetate 600 mL of 2 times of amounts again, ethyl acetate layer obtains medicinal extract B 5g after being evaporated to and doing, upper D101 macroporous adsorbent resin, the aqueous ethanolic solution 1000 mL wash-outs that the volume fraction of using 5 times of amounts (column volume of macroporous adsorbent resin chromatography post is 200mL) is 5%, elutriant is evaporated to dry, obtain 2g medicinal extract C, it is the bush efficient part, then, in the 80mL solvent that the moving phase that to be dissolved in by volume ratio fully by the bush efficient part be 1:1 and stationary phase form, obtain sample solution,
(3) separate and prepare brazilin and former Hematorylin B
The chromatographic condition of described high speed adverse current chromatogram is: separation system is chloroform: methyl alcohol: the volume ratio of water is 4:3:2, upper as stationary phase, lower to moving phase; The instrument rotating speed: the stationary phase system clockwise rotates, 400 turn/min; Flow rate of mobile phase: 6mL/min; Bath temperature: 20 ℃; Retention rate: 30%; Detect wavelength: 280 nm;
First adopt stationary phase to be full of the chromatographic column of high-speed counter-current chromatograph, then make its main frame clockwise rotate, then moving phase is pumped in chromatographic column, extracting sample solution is by the sampling valve sample introduction; And be under 280nm at the ultraviolet detection wavelength, collect respectively the flow point that contains brazilin and former Hematorylin B according to the UV-detector spectrogram, then be concentrated into dryly with Rotary Evaporators, obtain respectively the former Hematorylin B(of final product 692 mg purity 96.4%) and 841 mg brazilins (purity 92.6%).
Embodiment 3
A kind of high speed adverse current chromatogram that utilizes separates the method for preparing brazilin and former Hematorylin B from bush, and concrete steps comprise:
(1) preparation of solvent system
By chloroform: methyl alcohol: water be take the ratio that volume ratio is 4:2:2 and is mixed, stratification, and upper is stationary phase mutually, lower is moving phase mutually;
(2) preparation of sample solution
The aqueous ethanolic solution 10000 mL thermal backflows that bush medicinal material 1 Kg is 100% by the volume fraction of 5 times of amounts are extracted 5 times, each 2h, united extraction liquid, be evaporated to after residual without aqueous ethanolic solution to obtain extractum A 8g, extractum A is disperseed fully with 160 ml pure water, extract extractum A 3 times with the ethyl acetate 800mL of 5 times of amounts again, ethyl acetate layer obtains medicinal extract B 5g after being evaporated to and doing, upper D101 macroporous adsorbent resin, the aqueous ethanolic solution 4000mL wash-out that the volume fraction of using 20 times of amounts (column volume of macroporous adsorbent resin chromatography post is 200mL) is 15%, elutriant is evaporated to dry, obtain 2g medicinal extract C, it is the bush efficient part, then, in the 80mL solvent that the moving phase that to be dissolved in by volume ratio fully by the bush efficient part be 1:1 and stationary phase form, obtain sample solution,
(3) separate and prepare brazilin and former Hematorylin B
The chromatographic condition of described high speed adverse current chromatogram is: separation system is chloroform: methyl alcohol: the volume ratio of water is 4:2:2, upper as stationary phase, lower to moving phase; The instrument rotating speed: the stationary phase system clockwise rotates, 500 turn/min; Flow rate of mobile phase: 5 mL/min; Bath temperature: 30 ℃; Retention rate: 80%; Detect wavelength: 280 nm;
First adopt stationary phase to be full of the chromatographic column of high-speed counter-current chromatograph, then make its main frame clockwise rotate, then moving phase is pumped in chromatographic column, extracting sample solution is by the sampling valve sample introduction; And be under 280nm at the ultraviolet detection wavelength, collect respectively the flow point that contains brazilin and former Hematorylin B according to the UV-detector spectrogram, then be concentrated into dryly with Rotary Evaporators, obtain respectively the former Hematorylin B(of final product 682 mg purity 96.5%) and 847 mg brazilins (purity 92.3%).
Embodiment 1-3 gained compound determines by mass spectrum, proton nmr spectra, carbon-13 nmr spectra, and data are as follows:
Former Hematorylin B: mass spectrum ESI-MS (m/z): [M-H]
-303, the hydrogen spectrum
1h-NMR (400MHz, DMSO-
d 6 ) δ: 2.42-2.58 (4H, each H J=13.2 Hz, H-8), (3.16-3.38 4H, each H J=10.8 Hz, 7-CH2OH), (3.73-4.26 4H, each H J=12 Hz, H-6), 6.35-6.36 (2H, dd, J=2.4 Hz, H-2), 6.45-6.68 (2H, d, J=8.0,2.4 Hz, H-2), 6.88-6.93 (4H, s, H-9 and 12), 6.45-6.68 (2H, d, J=8.0 Hz, H-1).The carbon spectrum
13c-NMR (100MHz, DMSO-
d 6 ) δ: 132.0/131.0 (C-1), 111.1/110.2 (C-2), 157.6/157.8 (C-3), 107.9/107.0 (C-4), 157.8/159.2 (C-4a), 76.3/74.8 (C-6), 71.4/71.3 (C-7), 42.0/ 39.5 (C-8), 123.8/122.0 (C-8a), 116.6/116.1 (C-9), 143.6/143.6 (C-10), 143.6/143.6 (C-11), 119.4/118.3 (C-12), 130.0/129.5 (C-12a), 126.8/126.3 (C-12b), 66.7/69.1 (7-CH2OH).Consistent with bibliographical information, therefore be defined as former Hematorylin B.
Brazilin: mass spectrum ESI-MS (
m/
z): [
m-H]
-285, the hydrogen spectrum
1h NMR (400MHz, DMSO-
d 6) δ: 3.82 (1H, d, J=11.2Hz, 2-H), 3.54 (1H, d, J=11.2 Hz, 2-H), 6.60 (1H, s, 2 ￠-OH), 6.51 (1H, s, 3 ￠-OH), 3.81 (1H, s, 4-H), 5.30 (1H, s, 3-OH), 8.61 (1H, s, 3 ￠-OH), 8.63 (1H, s, 4 ￠-OH), 7.11 (1H, d, J=8.4 Hz, 5-H), 6.39 (1H, s, J=2.4, J=8.4 Hz, 6-H), 9.24 (1H, s, 7-OH), 6.18 (1H, d, 8-H), 2.85 (1H, d, 8 Hz, 9-H), 2.67 (1H, d, 8 Hz, 9-H).The carbon spectrum
13c NMR (100MHz, DMSO-
d 6 ) δ: 143.9 (C-1 ￠), 69.6 (C-2), (112.1 C-2 ￠), 76.3 (C-3), (144.0 C-3 ￠), 49.5 (C-4), 144.3 (C-4 ￠), 114.4 (C-4a), 131.0 (C-5), (111.7 C-5 ￠), 108.7 (C-6), 135.6 (C-6 ￠), 156.5 (C-7), 102.8 (C-8), 154.1 (C-8a), 42.0 (C-9).Consistent with bibliographical information, therefore be defined as brazilin.
The weight of embodiment 1-3 gained compound obtains by the analytical balance weighing, and its purity is to adopt HPLC to determine, adopts the purity of the former Hematorylin B of the method for the invention gained more than 96%, and the purity of brazilin is more than 92%.Wherein said HPLC analysis condition is as follows: the serial HPLC of Agilent 1260, the auxiliary work station, Agilent SB-C18 chromatographic column (column length 150mm * post footpath 4.6mm, particle diameter 5 μ m), adopt the methanol aqueous solution that volume fraction is 70% to carry out isocratic elution as moving phase, detecting wavelength is 280 nm, and the flow velocity of moving phase is 1.0 mL/min.
The present invention's high-speed countercurrent chromatography used can greatly reduce the solvent used in the separation preparation process, and reduction is more than 80% usually, and disengaging time is at least saved more than 5 times, yield improves more than 50%, and isolated target component loss is few, and purity is high, expense is low, economic environmental protection.
Claims (10)
1. one kind is utilized high speed adverse current chromatogram to separate the method for preparing brazilin and former Hematorylin B from bush, it is characterized in that: at first prepare sample solution, then adopt high-speed counter-current chromatograph coupling UV-detector to separate preparation, collect respectively the flow point that contains brazilin and former Hematorylin B according to uv-spectrogram, be evaporated to dry target compound brazilin and former Hematorylin B;
The chromatographic condition that described high speed adverse current chromatogram adopts is: solvent system is chloroform: methyl alcohol: the volume ratio of water is 4:2-4:2, and upper is stationary phase mutually, and lower is moving phase mutually; The instrument rotating speed: adopt and clockwise rotate, rotating speed is 400 ~ 550 turn/min, and flow rate of mobile phase is 5.0 ~ 8.0 mL/min; Bath temperature: 20 ℃-30 ℃; Retention rate: 30%-80%; Detect wavelength: 280nm.
2. the high speed adverse current chromatogram that utilizes according to claim 1 separates the method for preparing brazilin and former Hematorylin B from bush, and it is characterized in that: concrete steps comprise:
(1) preparation of solvent system
By chloroform: methyl alcohol: water be take the ratio that volume ratio is 4:2-4:2 and is mixed, stratification, and upper is stationary phase mutually, lower is moving phase mutually;
(2) preparation of sample solution
Bush is extracted 1 ~ 5 time with the solvent orange 2 A thermal backflow of 2 ~ 5 times of amounts, united extraction liquid, be evaporated to the residual extractum A that obtains of solvent-free A, the extractum A water disperses fully, then uses the ethyl acetate extraction extractum A of 2 ~ 5 times of water gagings, ethyl acetate layer obtains medicinal extract B after being evaporated to and doing, upper macroporous adsorbent resin, used the aqueous ethanolic solution wash-out of 5 ~ 20 times of amounts, and elutriant is evaporated to dry, obtain medicinal extract C, i.e. bush efficient part; Then, in the solvent that the moving phase that to be dissolved in by volume ratio fully by the bush efficient part be 1:1 and stationary phase form, obtain sample solution;
(3) separate and prepare brazilin and former Hematorylin B
Extracting sample solution adopts high-speed counter-current chromatograph to carry out sample introduction, and collects respectively according to the UV-detector spectrogram flow point that contains brazilin and former Hematorylin B, then with Rotary Evaporators, is concentrated into dryly, obtains respectively final product brazilin and former Hematorylin B;
The described solvent orange 2 A of step (2) is methanol solution or ethanolic soln.
3. the high speed adverse current chromatogram that utilizes according to claim 1 separates the method for preparing brazilin and former Hematorylin B from bush, and it is characterized in that: described solvent system is chloroform: methyl alcohol: the volume ratio of water is 4:4:2.
4. the high speed adverse current chromatogram that utilizes according to claim 1 separates the method for preparing brazilin and former Hematorylin B from bush, and it is characterized in that: described bath temperature is 25 ℃.
5. the high speed adverse current chromatogram that utilizes according to claim 1 separates the method for preparing brazilin and former Hematorylin B from bush, it is characterized in that: described rotating speed is 550 turn/min.
6. the high speed adverse current chromatogram that utilizes according to claim 1 separates the method for preparing brazilin and former Hematorylin B from bush, and it is characterized in that: described flow rate of mobile phase is 7.5 mL/min.
7. the high speed adverse current chromatogram that utilizes according to claim 1 separates the method for preparing brazilin and former Hematorylin B from bush, and it is characterized in that: described retention rate is 75%.
8. the high speed adverse current chromatogram that utilizes according to claim 2 separates the method for preparing brazilin and former Hematorylin B from bush, it is characterized in that: the described solvent orange 2 A of step (2) is: the ethanolic soln that the methanol solution that volume fraction is 30%-100% or volume fraction are 30%-100%.
9. the high speed adverse current chromatogram that utilizes according to claim 2 separates the method for preparing brazilin and former Hematorylin B from bush, it is characterized in that: the described macroporous adsorbent resin of step (2) is the D101 macroporous adsorbent resin.
10. the high speed adverse current chromatogram that utilizes according to claim 2 separates the method for preparing brazilin and former Hematorylin B from bush, it is characterized in that: the described eluting solvent of step (2) is the aqueous ethanolic solution that volume fraction is 5%-15%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201310393712 CN103450145A (en) | 2013-09-03 | 2013-09-03 | Method for separating and preparing Brazilin and Protosappanin B from Sappanwood by using high-speed countercurrent chromatography |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201310393712 CN103450145A (en) | 2013-09-03 | 2013-09-03 | Method for separating and preparing Brazilin and Protosappanin B from Sappanwood by using high-speed countercurrent chromatography |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103450145A true CN103450145A (en) | 2013-12-18 |
Family
ID=49733034
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201310393712 Pending CN103450145A (en) | 2013-09-03 | 2013-09-03 | Method for separating and preparing Brazilin and Protosappanin B from Sappanwood by using high-speed countercurrent chromatography |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103450145A (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107115336A (en) * | 2017-05-22 | 2017-09-01 | 北京大学 | It is a kind of to be used to treat composition of ischemia apoplexy and preparation method thereof and medical usage |
| CN108329292A (en) * | 2018-04-16 | 2018-07-27 | 黄榜昇 | A method of preparing former haematoxylin B |
| CN108546261A (en) * | 2018-04-16 | 2018-09-18 | 黄榜昇 | A method of preparing protosappanin A |
| CN108546260A (en) * | 2018-04-16 | 2018-09-18 | 黄榜昇 | A method of preparing former haematoxylin C |
| CN109288835A (en) * | 2018-08-08 | 2019-02-01 | 北京大学 | Application of the compound in the drug of preparation treatment acute pulmonary embolism |
| CN111533726A (en) * | 2020-05-16 | 2020-08-14 | 西安都创医药科技有限公司 | Preparation method of hematoxylin derivative, product prepared by preparation method and application of product |
-
2013
- 2013-09-03 CN CN 201310393712 patent/CN103450145A/en active Pending
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107115336A (en) * | 2017-05-22 | 2017-09-01 | 北京大学 | It is a kind of to be used to treat composition of ischemia apoplexy and preparation method thereof and medical usage |
| CN107115336B (en) * | 2017-05-22 | 2019-10-22 | 北京大学 | A kind of composition for treating ischemia apoplexy and preparation method thereof and medical usage |
| CN108329292A (en) * | 2018-04-16 | 2018-07-27 | 黄榜昇 | A method of preparing former haematoxylin B |
| CN108546261A (en) * | 2018-04-16 | 2018-09-18 | 黄榜昇 | A method of preparing protosappanin A |
| CN108546260A (en) * | 2018-04-16 | 2018-09-18 | 黄榜昇 | A method of preparing former haematoxylin C |
| CN109288835A (en) * | 2018-08-08 | 2019-02-01 | 北京大学 | Application of the compound in the drug of preparation treatment acute pulmonary embolism |
| CN109288835B (en) * | 2018-08-08 | 2020-09-25 | 北京大学 | Application of compound in preparation of medicine for treating acute pulmonary embolism |
| CN111533726A (en) * | 2020-05-16 | 2020-08-14 | 西安都创医药科技有限公司 | Preparation method of hematoxylin derivative, product prepared by preparation method and application of product |
| CN111533726B (en) * | 2020-05-16 | 2022-07-26 | 西安都创医药科技有限公司 | Preparation method of hematoxylin derivative, product prepared by preparation method and application of product |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103450145A (en) | Method for separating and preparing Brazilin and Protosappanin B from Sappanwood by using high-speed countercurrent chromatography | |
| CN103145677B (en) | Method for separating active ingredients from aquilaria sinensis lamina by utilizing high-speed countercurrent chromatography | |
| CN101353363B (en) | Method for separating and purifying Momordica grosvenori leaf chromocor compound by high-speed countercurrent chromatography and products thereof | |
| CN107011308B (en) | The method of polymethoxyflavone class compound is isolated and purified from bowl mandarin orange fruit | |
| CN104031013B (en) | A kind of utilize the isolated and purified method preparing salvianolic acid B and rosmarinic acid of high speed adverse current chromatogram | |
| CN110483599A (en) | The separation method of flavones ingredient in a kind of manaca leaf | |
| CN110294673A (en) | Caffeoylquinic acids butyl ester class isomer and its preparation method and application | |
| CN101357147A (en) | Extract of dogbane leaf with finger print and preparation and analytical method thereof | |
| CN102372754A (en) | Method for preparing specnuezhenide | |
| CN106957310B (en) | The high efficiency preparation method of flavonoids monomer in a kind of leaves of Hawthorn | |
| Wang et al. | Determination and isolation of potential α-glucosidase and xanthine oxidase inhibitors from Trifolium pratense L. by ultrafiltration liquid chromatography and high-speed countercurrent chromatography | |
| CN103232427B (en) | Xanthone compound as well as preparation method and application thereof | |
| CN103145775B (en) | The preparation of high purity Herba Cleidion brevipetiolae glycosides A and quality controlling means thereof | |
| CN104974122A (en) | Coumarin compound originated from tobacco, and preparation method and application thereof | |
| CN105131063B (en) | From Meconopsis integrifolia spend in and meanwhile the method that isolates and purifies a variety of flavones ingredients | |
| CN105330710B (en) | A kind of method for extracting three kinds of iridoid glycosides chemical reference substances simultaneously from the fruit of glossy privet | |
| CN1268631C (en) | Technique for preparing general flavone of Chinese globeflower with short petal some medicinal substances in high purity | |
| CN103254165A (en) | Preparation method of atractylenolide II | |
| CN101982466A (en) | Method for extracting isoflavonoids compound in all-grass of Twining Rhynchosia with ionic liquid | |
| CN111718318B (en) | Method for separating flavone monomer in spina gleditsiae based on countercurrent chromatography | |
| CN104892620B (en) | A kind of preparation method of high-purity karanjin | |
| CN102659549B (en) | Method for extracting brevifolin from Relinqing granula raw material and method for preparation quality control | |
| CN108752403B (en) | Method for separating quercetin rhamnoside from artemisia rupestris | |
| CN100526329C (en) | Production of high-purity peiminine and fritimine | |
| CN103058859B (en) | Simultaneous preparation and detection method of gallic acid and gallicin in toona sinensis leaves |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20131218 |