CN102295629A - Preparation method of ligustilide - Google Patents
Preparation method of ligustilide Download PDFInfo
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- CN102295629A CN102295629A CN2011101842751A CN201110184275A CN102295629A CN 102295629 A CN102295629 A CN 102295629A CN 2011101842751 A CN2011101842751 A CN 2011101842751A CN 201110184275 A CN201110184275 A CN 201110184275A CN 102295629 A CN102295629 A CN 102295629A
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Abstract
The invention relates to the field of chemistry, and specifically relates to a preparation method of a ligustilide. The method employs vacuum fractionation and molecule distillation to separate and prepare the ligustilide from Chinese angelica oil and ligusticum wallichii oil, so as to increase a ligustilide purity to 93.2-98.8%. Besides, no separating medium or organic solvent is used during the preparation process, and no pollution is caused on the environment.
Description
Technical field
The present invention relates to chemical field, particularly a kind of preparation method of Z-ligustilide.
Background technology
Z-ligustilide (Ligustilide) is the main effective constituent in traditional Chinese medicine Radix Angelicae Sinensis, the Ligusticum wallichii, content reaches more than 1% in medicinal material, content is up to more than 30% in the volatile oil of Radix Angelicae Sinensis, Ligusticum wallichii, leading indicator composition for Radix Angelicae Sinensis, Ligusticum wallichii medicinal material and medicine preparation quality control, be the oily liquids of yellowish colour band fragrance, boiling point is 168-169 ℃/6mmHg, and is water insoluble, dissolve in organic solvents such as ethanol, methyl alcohol, ether, ethyl acetate, sherwood oil, normal hexane, molecular formula is C
12H
14O
2, molecular weight is 190.24, chemical structural formula is:
Z-ligustilide is special unsaturated phthalide, and the conjugated double bond and the exocyclic double bond of six-ring are arranged, and has undersaturated five-ring lactone, at C
3Also have active butenyl on the position, these active structure groups make Z-ligustilide have very strong pharmacologically active, have also increased chemical instability simultaneously.Because the singularity of Z-ligustilide structure, its chemical total synthesis method is not overripened at present, thereby is the effective ways of preparation Z-ligustilide from the Chinese medicine extraction separation.Because Z-ligustilide is water-fast oily liquids, boiling point is higher under the normal temperature, and Z-ligustilide less stable at high temperature, thereby can not use under the normal pressure distillatory method to carry out separation and purification.
In recent years, Z-ligustilide silica gel column chromatography method commonly used is carried out separation and purification, uses a large amount of organic solvents such as sherwood oil, ethyl acetate, acetone, normal hexane and silica gel etc. to separate medium in this method.Patent CN1583735A discloses " a kind of technology of enrichment Z-ligustilide and preparation ", obtain Rhizoma Chuanxiong oil with the Ligusticum wallichii medicinal material extract, after the sherwood oil dilution, last silicagel column, wash post with 1～2 times of sherwood oil after, again with 12～16 times of sherwood oils flushings and collect elutriant, concentrate promptly, the Z-ligustilide purity that makes is 50～60%, but this method velocity of separation is slow and Z-ligustilide purity that obtain is lower, is unsuitable for suitability for industrialized production.Patent CN1552702A disclose the Ligusticum wallichii medicinal material with organic solvents such as ethanol, ethyl acetate in the high-shear instrument, shear extract behind the volatile oil, directly be splined on silicagel column, with the mixed solution wash-out of sherwood oil and ethyl acetate, collect second colour band, concentrate and obtain cis form ligustilide.This method is separated the purity that obtains Z-ligustilide can reach 90%, but this method is separated Z-ligustilide with conventional normal pressure or pressurized column chromatography, velocity of separation is slow, complicated operation, and much can not separate fully with Z-ligustilide in the Rhizoma Chuanxiong oil with the composition of Z-ligustilide similar performance, the purity of Z-ligustilide is only judged by the colour band of observing chromatography column, obtains very difficulty of highly purified Z-ligustilide, can not be used on a large scale producing.Add the use of organic solvents such as sherwood oil and ethyl acetate, cause environmental pollution easily.Patent CN1778799 discloses with Ligusticum wallichii or Radix Angelicae Sinensis volatile oil, with silica gel is parting material, but prepare Z-ligustilide with decompression column chromatography sharp separation, be applicable to large-scale industrialization production, but to be elutriant also with organic solvents such as a large amount of sherwood oils and ethyl acetate, easily contaminate environment causes actual bodily harm to the production operation personnel simultaneously.Therefore, provide and do not use separating medium and organic solvent in a kind of preparation process, from Radix Angelicae Sinensis, Ligusticum wallichii, separate the method for preparing Z-ligustilide, have realistic meaning.
Summary of the invention
In view of this, the invention provides a kind of preparation method of Z-ligustilide.This method adopts vacuum fractionation and molecular distillation to separate from Radix Angelicae Sinensis oil and Rhizoma Chuanxiong oil to prepare Z-ligustilide, can improve the purity of Z-ligustilide, and not use separating medium and organic solvent in the preparation process, can not pollute environment.
In order to realize the foregoing invention purpose, the invention provides following technical scheme:
A kind of preparation method of Z-ligustilide comprises:
Step 1: under the vacuum state of 5～100Pa, Radix Angelicae Sinensis oil or Rhizoma Chuanxiong oil heating are carried out vacuum fractionation, collect 100～110 ℃ cut;
Step 2: the described cut of step 1 is carried out molecular distillation under 70～90 ℃, the condition of 0.1～10Pa vacuum tightness, collect and steam thing, obtain Z-ligustilide; The rotating speed of described molecular distillation rotor knifing is that the charging flow velocity of 50～500r/min, described cut is 0.5～5.0mL/min.
Vacuum fractionation is meant fractionating system under vacuum state, by the successive heat supply of heating unit, mixed solution is carried out repeatedly the partly process of vaporization and partial condensation, and this process can make mixed solution be separated more completely.The vacuum tightness of fractionating system is high more, and the separation temperature of mixed solution is then low more, and lower boiling component is separated fully, has kept the physicochemical property of cut simultaneously.Because mixed solution mostly is multi-component material, the vaporization temperature difference of each component, therefore, in the vacuum fractionation process, when differing temps and mutual unbalanced gas-liquid two-phase contact, will inevitably produce the dual function of biography and mass transfer simultaneously, when the liquid-phase reflux of upper level directly contacts with the gas phase of next stage, the repeatedly part vaporization of low boiling component and the partial condensation of higher component just can take place.Repeatedly carry out by so repeating, reach the purpose that fractionation is purified.Wherein, vacuum tightness and cut point are the important factors of decision fractionating effect.Because Z-ligustilide is water-fast oily liquids, boiling point is higher under the normal temperature, less stable at high temperature, therefore among the preparation method provided by the invention, under the vacuum state of 5～100Pa, Radix Angelicae Sinensis oil or Rhizoma Chuanxiong oil heating are carried out vacuum fractionation, collect 100～110 ℃ cut, do not change to guarantee Z-ligustilide character.
As preferably, preparation method provided by the invention, the vacuum tightness in the step 1 are 5～50Pa.
As preferably, preparation method provided by the invention collects 105～110 ℃ cut with Radix Angelicae Sinensis oil or Rhizoma Chuanxiong oil heating in the described step 1, can access the higher Z-ligustilide of purity.
Molecular distillation is a kind of special liquid liquid isolation technique, is different from the tradition distillation and relies on the boiling-point difference separation principle, but realize separating by the difference of different substances molecular motion mean free path.Its principle of work is under the extra-high vacuum degree, according to the difference of mixture molecular motion mean free path, under far below the temperature of its boiling point with mixture separation.The molecule of distilling material is not subjected to the influence of intermolecular resistance to impact pi by generating surface to the stroke of condensing surface, and the distance between the two sides is less than the molecular motion mean free path of distilling material molecule.Molecular distillation apparatus can be divided into scraped film type molecular distillation, falling film type molecular distillation and centrifugal molecular distillating according to the design difference of stroke evaporation liquid film, and wherein the scraped film type molecular distillation is the most commonly used.
The scraped film type molecular distillation is a kind of non-equilibrium still-process under high vacuum state, the rotor of high speed rotating is swiped continuously to the liquid on the wall in the vaporizer, be that liquid is dispersed in the evaporation wall, utilize the difference of molecular free path between different substances again, under far below the temperature of boiling point of liquid with its separation, higher to boiling point under the normal temperature, the relatively poor Z-ligustilide of high temperatures can effectively separate.Therefore, among the preparation method provided by the invention, method and the coupling of vacuum distilling method with molecular distillation separate Z-ligustilide.
The molecular distillation common instrument is a rotary film evaporator, and it partly is made up of the drive part on top and the evaporation concentration of bottom, and inner-cooled rotary film evaporator bottom also has internal condensation device part.The evaporation concentration part is made up of the evaporation cylindrical shell and the rotor of strap clamp cover.Structures such as rotor banding sparger, mist eliminator, main shaft, groove scraper plate and support.When having the scraper plate of certain interval to rotate with cylinder inner wall, promote material and form a fluid bow wave in the above, because of scraper plate general knowledge tilts, or have the groove of certain angle, bow wave inner fluid curl flows downward.Form the towing eddy current in the scraper plate back, then form storehouse tower stream between scraper plate and the wall, and the elsewhere of heating surface, because action of gravity, material is bonded at slowly creeps mobilely vertically downward on the wall, form film stream.Because the rotation and the stirring of scraper plate, promote bow wave constantly and film stream carry out the exchange of matter and energy, help the evaporation of material at heating surface.
When the material that is preheating to certain temperature enters vaporizer from the luwa evaporator upper inlet, the liquid distributor that is rotated is divided into four bursts of logistics and flows into cylinder inner walls.Per share logistics is dispersed on the evaporation cylinder heating inwall by the agitaion of corresponding clamping plate and forms even liquid film.Heating medium (heating steam or thermal oil) is passed to the heat of water surface of evaporation (cylinder inner wall) in the liquid film absorption chuck, carries out the rapid evaporation of material on its surface.The molecule effusion liquid level that energy is enough, the mean free path of light molecule is big, the mean free path of weight molecule is little, if, make light molecule constantly be captured, thereby destroyed the transient equilibrium of light molecule and the light molecule in the mixed solution is constantly overflowed greater than weight molecule mean free path place one trap being set less than the mean free path of light molecule from liquid level, and weight molecule is not caught device and is tending towards running balance very soon because of reaching, no longer overflow from mixed solution, like this, liquid mixture has just reached isolating purpose.
The main operation controlled variable of knifing thin-film evaporator has: feeding temperature, inlet amount, heat transfer temperature difference, vacuum tightness, scraper plate rotating speed etc.
When considering feeding temperature, satisfy logistics and after entering vaporizer, reach the boiling temperature of operating under the vacuum tightness.Feeding temperature is higher than evaporated components during pairing boiling temperature, will cause splashing of liquid under this vacuum tightness, be entrained with a large amount of liquid by in the gas that steams, and is difficult to control this moment.When temperature is lower than the boiling temperature of component, just needs a part of sensible heat of water surface of evaporation to add thermal material and make it to reach boiling temperature, so just can not effectively utilize heating surface, cause energy loss.As preferably, among the preparation method provided by the invention, also comprise before the distillation of the conducting molecule of cut described in the step 2 being preheated to 40～45 ℃ step.
In the evaporation operation process, the selection of the temperature difference is very important.When the temperature difference was too small, resulting heat was not enough in heat transfer process, and evaporation can not be carried out well; But during excessive temperature differentials, heat transfer coefficient is descended.Therefore, select for use appropriate temperature difference extremely important to guarantee having bigger heat transfer coefficient to improve evaporation capacity.As preferably, among the preparation method provided by the invention, molecular distillation is to carry out under 80～85 ℃ the condition in temperature in the step 2.
The size of inlet amount is subjected to the influence of the wetting ratio of vaporizer.In evaporative process, flow rate of fluid constantly diminishes on the short transverse of heating entire body, and wetting ratio is also constantly reduced.Minimum wetting ratio can guarantee to form the pairing wetting ratio of fluid film exactly in the squeegee fields of bottom.When wetting ratio is lower than minimum value, can occur doing the surface, cause the logistics coking, strengthen the wearing and tearing of scraper plate, increase power consumption at water surface of evaporation.Maximum wetting ratio is the liquid flooding rate again, flows to not keeping film at water surface of evaporation to the fluidic flow is high, and bottom or whole scraper plate are all by soaked with liquid at this moment, and heat transfer efficiency and vaporization efficiency descend, and vaporator rate reduces to the same level of falling film evaporation.Thin-film evaporator should be maximum wetting ratio that causes the top liquid flooding and minimum wettingly operating between than these two limit of causing the withered coking in bottom.As preferably, among the preparation method provided by the invention, molecular distillation is to carry out under the condition of 1.0～2.0mL/min at the cut flow velocity in the step 2.
When handling heat-sensitive material, molecular distillation need be operated under certain vacuum degree condition, to guarantee material when the water surface of evaporation, can thermolysis not take place because of temperature is high or thermopolymerization is gone bad.So the operation vacuum tightness of selecting will satisfy the pairing boiling temperature of material under this vacuum tightness, this temperature is lower than pairing thermolysis of material generation chemical reaction or thermopolymerization temperature.As preferably, among the preparation method provided by the invention, molecular distillation is to carry out under the condition of 0.1～5Pa in vacuum tightness in the step 2.
Rotor rotation speed should make in the evaporation cylinder and form even liquid film, finishes the arc exchange fully that involves the matter and energy of liquid film.Along with rotating speed increases, rate of heat transfer also can increase, but rotating speed is too high, can increase kinetic energy consumption.Therefore, select suitable rotor speed, the effect of molecular distillation is also had material impact.As preferably, among the preparation method provided by the invention, molecular distillation is to carry out under the condition of 80～350r/min at the rotating speed of rotor knifing in the step 2.
The invention provides a kind of preparation method of Z-ligustilide.This method adopts vacuum fractionation and molecular distillation to separate the preparation Z-ligustilide from Radix Angelicae Sinensis oil and Rhizoma Chuanxiong oil, purity to 93.2～98.8% of raising Z-ligustilide, and do not use separating medium and organic solvent in the preparation process, can not pollute environment.
Description of drawings
Fig. 1 shows the Z-ligustilide HPLC color atlas that embodiment 2 makes.
Fig. 2 shows the total ion collection of illustrative plates of Z-ligustilide GC-MS purity detecting that embodiment 2 makes.
Fig. 3 shows the MS collection of illustrative plates of the Z-ligustilide GC-MS purity detecting principal constituent that embodiment 2 makes.
Embodiment
The invention discloses a kind of preparation method of Z-ligustilide, those skilled in the art can use for reference this paper content, suitably improve processing parameter and realize.Special needs to be pointed out is that all similarly replace and change apparent to those skilled in the art, they all are regarded as being included in the present invention.Method of the present invention and application are described by preferred embodiment, the related personnel obviously can change or suitably change and combination methods and applications as herein described in not breaking away from content of the present invention, spirit and scope, realizes and use the technology of the present invention.
Among the preparation method provided by the invention, Radix Angelicae Sinensis oil and Rhizoma Chuanxiong oil can be buied by market, also can extract voluntarily.Each reagent all can be buied by market.
Below in conjunction with embodiment, further set forth the present invention:
The selection of vacuum fractionation condition among embodiment 1 preparation method of the present invention
The extraction of Radix Angelicae Sinensis oil: after taking by weighing the suitable chopping of Radix Angelicae Sinensis crude drug 200kg, add 8 times of amount 95% ethanol in extractor after the soaked overnight, heating and refluxing extraction 2h, filter, the dregs of a decoction add 6 times, 6 times amount 95% ethanol, heating and refluxing extraction 2h respectively more respectively, filter and merging filtrate, filtrate decompression concentrates and reclaims ethanol to the greatest extent, obtains the Radix Angelicae Sinensis oil of the about 8000mL of the dense thick liquid of Vandyke brown, and is standby.
Test group 1: get the 1000mL Radix Angelicae Sinensis oil in the 3000mL round-bottomed flask, install the vacuum fractionation device, charge into all air in the argon gas remover, vacuumize vacuum state with the vacuum oil pump into 20Pa, begin heating with oil bath, after cut point reaches 80 ℃, has a large amount of yellow cuts to steam, use special-purpose run tank to collect the first cut 215mL; Temperature continues to rise, and when cut point reaches 110 ℃, has a large amount of yellow cuts to go out again, collects the second cut 348mL; When cut point rises to 120 ℃, have iota fraction to steam, continue to be warming up to steam without any liquid till, collect the 3rd cut 76mL.
Test group 2: get the 1500mL Radix Angelicae Sinensis oil in the 5000mL round-bottomed flask, install the vacuum fractionation device, charge into all air in the argon gas remover, vacuumize vacuum state with the vacuum oil pump into 10Pa, begin heating with oil bath, after cut point reaches 75 ℃, has a large amount of yellow cuts to steam, use special-purpose run tank to collect the first cut 320mL; Temperature continues to rise, and when cut point reaches 105 ℃, has a large amount of yellow cuts to go out again, collects the second cut 480mL; When the fractionation calibration rises to 115 ℃, have iota fraction to steam, continue to be warming up to steam without any liquid till, collect the 3rd cut 120mL.
Test group 3: get the 2000mL Rhizoma Chuanxiong oil in the 5000mL round-bottomed flask, install the vacuum fractionation device, charge into all air in the argon gas remover, vacuumize vacuum state with the vacuum oil pump into 5Pa, begin heating with oil bath, after cut point reaches 90 ℃, has a large amount of yellow cuts to steam, use special-purpose run tank to collect the first cut 256mL; Temperature continues to rise, when cut point reaches: 100 ℃, there are a large amount of yellow cuts to go out again, collect the second cut 539mL; When the fractionation calibration rises to 120 ℃, have iota fraction to steam, continue to be warming up to steam without any liquid till, collect the 3rd cut 192mL.
First cut, second cut and the 3rd cut 1mL that get collection respectively add methanol constant volume to 10mL, centrifugal 5min (10000RPM), and supernatant liquor is sample introduction 5 μ L respectively, measure.
HPLC chromatogram testing conditions is as follows: instrument: Agilent 1260 type liquid chromatographs (quaternary pump, automatic sampler, DAD detector); Chromatographic column: Hypersil ODS (4.6 * 150mm, 5 μ m); Moving phase: methyl alcohol-5% Virahol (60: 40); Detect wavelength: 280nm; Flow velocity: 1mL/min; Column temperature: 25 ℃.Detected result sees Table 1.
Table 1HPLC detects the concentration of Z-ligustilide
| Group | First cut (%) | Second cut (%) | The 3rd cut (%) |
| Test group 1 | 5 | 88 | 11 |
| Test group 2 | 5 | 89 | 12 |
| Test group 3 | 3 | 88 | 12 |
As shown in Table 1, in the test group 1 to 3 in second cut content utmost point of Z-ligustilide be significantly higher than with first cut in the group and the 3rd cut (P＜0.01), therefore, cut that select to collect the shunting temperature and be 100～110 ℃ carries out follow-up molecular distillation.
Embodiment 2 Z-ligustilide preparation methods provided by the invention
Get the 1500mL Radix Angelicae Sinensis oil in the 5000mL round-bottomed flask, install the vacuum fractionation device, charge into all air in the argon gas remover, vacuumize vacuum state with the vacuum oil pump, begin heating with oil bath, when cut point reaches 105 ℃ into 10Pa, there are a large amount of yellow cuts to go out again, collect cut 480mL.
Molecular distillation apparatus is charged into all air in the argon gas remover, be heated to 80 ℃, opening vacuum pump vacuumizes when making vacuum tightness reach 1Pa, after being heated to 45 ℃, the cut 400mL that vacuum fractionation is collected adds from opening for feed, the rotating speed of regulating the rotor knifing is 50r/min, and the reinforced flow velocity 1.5mL/min of cut that vacuum fractionation is collected must steam thing respectively and reserve thing from two discharge ports, collection steams thing 290mL, is light yellow liquid.
Use respectively proton nmr spectra (
1H-NMR) and carbon-13 nmr spectra (
13C-NMR) thing that steams of present embodiment molecular distillation is identified that concrete outcome is as follows:
EI-MS?m/z：190(M
+)，161(M
+-C
2H
5)，148，133(M
+-C
2H
5-CO)，106，76，55，27。
1H-NMR(CDCl
3)δ：0.96(t，J＝8，3H)，1.52(m，J＝8，2H)，2.38-2.63(m，6H)，5.24(t，J＝8，1H)，6.01(m，J＝8，1H)，6.29(d，J＝8，1H)。
13C-NMR(CDCl
3)δ：13.67(-CH
3)，18.45(-CH
2-)，22.32(-CH
2-)，22.36(-CH
2-)，28.05(-CH
2-)，112.73(C＝)，117.01(C＝)，123.92(C＝)，129.83(C＝)，146.99(C＝)，148.54(O-C＝)，167.46(O-C＝O)。
In sum: can learn that the material that present embodiment makes contains :-CH
3,-CH
2-, main functional group such as C=, O-C=, O-C=O, and above data are consistent with the Z-ligustilide of bibliographical information, show that steaming thing is Z-ligustilide (1igustilide), its chemical structure, molecular formula and molecular weight are as follows:
Z-ligustilide to the present embodiment preparation is tested: gets the Z-ligustilide 1mL that makes and adds methanol constant volume to 10mL, and centrifugal 5min (10000RPM), supernatant liquor is sample introduction 5 μ L respectively, measure.
HPLC chromatogram testing conditions is as follows: instrument: Agilent 1260 type liquid chromatographs (quaternary pump, automatic sampler, DAD detector); Chromatographic column: Hypersil ODS (4.6 * 150mm, 5 μ m); Moving phase: methyl alcohol-5% Virahol (60: 40); Detect wavelength: 280nm; Flow velocity: 1mL/min; Column temperature: 25 ℃.
The purity that detects Z-ligustilide with HPLC is 98.6%, and color atlas is seen Fig. 1.
GC-MS purity detecting condition: instrument: Agilent6890/5973 type GC-MS combined instrument; Chromatographic column: HP-5ms (5% phenyl methyl polysiloxane, 30m * 320um * 0.25um); Carrier gas: He; Injector temperature: 250 ℃; Column temperature: keep 6min for 210 ℃, (20 ℃/mim) be worth 300 ℃ of temperature programmings again; Input mode: split stream sampling, splitting ratio 50: 1.
Measure: get the Z-ligustilide behind the purifying, sample introduction is measured by chromatographic condition, respectively the MS collection of illustrative plates (Fig. 3) of total ion collection of illustrative plates (Fig. 2) and principal constituent, be 98.4% by peak area normalization method calculating Z-ligustilide purity.
Embodiment 3 Z-ligustilide preparation methods provided by the invention
Get the 1000mL Radix Angelicae Sinensis oil in the 3000mL round-bottomed flask, install the vacuum fractionation device, charge into all air in the argon gas remover, vacuumize vacuum state with the vacuum oil pump, begin heating with oil bath, when cut point reaches 110 ℃ into 50Pa, there are a large amount of yellow cuts to go out, collect cut 348mL.
Molecular distillation apparatus is charged into all air in the argon gas remover, be heated to 80 ℃, opening vacuum pump vacuumizes when making vacuum tightness reach 7.5Pa, add from opening for feed after the cut 300g that vacuum fractionation is collected is heated to 40 ℃, the rotating speed of regulating the rotor knifing is 300r/min, and the collected reinforced flow velocity of cut of vacuum fractionation is 1.8mL/min, must steam thing respectively and reserve thing from two discharge ports, collection steams thing 240mL, obtains steaming thing, is light yellow liquid.
Carry out structure detection and purity check to steaming thing:
Use respectively proton nmr spectra (
1H-NMR) and carbon-13 nmr spectra (
13C-NMR) thing that steams of present embodiment molecular distillation is identified that the result shows that the thing that steams that present embodiment makes is a Z-ligustilide.
Get the Z-ligustilide 1mL that makes and add methanol constant volume to 10mL, centrifugal 5min (10000RPM), supernatant liquor is sample introduction 5 μ L respectively, measure.
HPLC chromatogram testing conditions is as follows: instrument: Agilent 1260 type liquid chromatographs (quaternary pump, automatic sampler, DAD detector); Chromatographic column: Hypersil ODS (4.6 * 150mm, 5 μ m); Moving phase: methyl alcohol-5% Virahol (60: 40); Detect wavelength: 280nm; Flow velocity: 1mL/min; Column temperature: 25 ℃.
The purity that detects Z-ligustilide with HPLC is 95.8%.
GC-MS purity detecting condition: instrument: Agilent6890/5973 type GC-MS combined instrument; Chromatographic column: HP-5ms (5% phenyl methyl polysiloxane, 30m * 320um * 0.25um); Carrier gas: He; Injector temperature: 250 ℃; Column temperature: keep 6min for 210 ℃, (20 ℃/mim) be worth 300 ℃ of temperature programmings again; Input mode: split stream sampling, splitting ratio 50: 1.The result shows that the Z-ligustilide purity that present embodiment makes is 95.8%.
Embodiment 4 Z-ligustilide preparation methods provided by the invention
Rhizoma Chuanxiong oil extracts: after taking by weighing the suitable chopping of Ligusticum wallichii crude drug 100kg, add 10 times of amount 95% ethanol in extractor after the soaked overnight, heating and refluxing extraction 2h, filter, the dregs of a decoction add 6 times, 4 times amount 95% ethanol, heating and refluxing extraction 2h respectively more respectively, filter and merging filtrate, filtrate decompression concentrates and reclaims ethanol to the greatest extent, obtains the Rhizoma Chuanxiong oil of the dense magma 4200mL of tawny, and is standby.
Get the 2000mL Rhizoma Chuanxiong oil in the 5000mL round-bottomed flask, install the vacuum fractionation device, charge into all air in the argon gas remover, vacuumize vacuum state with the vacuum oil pump, begin heating with oil bath, when cut point reaches 104 ℃ into 9Pa, there are a large amount of yellow cuts to go out, collect cut 653mL.
Molecular distillation apparatus is charged into all air in the argon gas remover, be heated to 80 ℃, opening vacuum pump vacuumizes when making vacuum tightness reach 0.1Pa, after being heated to 40 ℃, the cut 600mL that vacuum fractionation is collected adds from opening for feed, the rotating speed of regulating the rotor knifing is 80r/min, and the reinforced flow velocity 2.0mL/min of cut that vacuum fractionation is collected must steam thing respectively and reserve thing from two discharge ports, collection steams thing 420mL, is light yellow liquid.
Carry out structure detection and purity check to steaming thing:
Use respectively proton nmr spectra (
1H-NMR) and carbon-13 nmr spectra (
13C-NMR) thing that steams of present embodiment molecular distillation is identified that the result shows that the thing that steams that present embodiment makes is a Z-ligustilide.
Get the Z-ligustilide 1mL that makes and add methanol constant volume to 10mL, centrifugal 5min (10000RPM), supernatant liquor is sample introduction 5 μ L respectively, measure.
HPLC chromatogram testing conditions is as follows: instrument: Agilent 1260 type liquid chromatographs (quaternary pump, automatic sampler, DAD detector); Chromatographic column: Hypersil ODS (4.6 * 150mm, 5 μ m); Moving phase: methyl alcohol-5% Virahol (60: 40); Detect wavelength: 280nm; Flow velocity: 1mL/min; Column temperature: 25 ℃.
The purity that detects Z-ligustilide with HPLC is 98.8%.
Embodiment 5 Z-ligustilide preparation methods provided by the invention
Get the 1000mL Radix Angelicae Sinensis oil in the 3000mL round-bottomed flask, install the vacuum fractionation device, charge into all air in the argon gas remover, vacuumize vacuum state with the vacuum oil pump, begin heating with oil bath, when cut point reaches 100 ℃ into 40Pa, there are a large amount of yellow cuts to go out, collect cut 431mL.
Molecular distillation apparatus is charged into all air in the argon gas remover, be heated to 70 ℃, opening vacuum pump vacuumizes when making vacuum tightness reach 5Pa, after being heated to 40 ℃, the cut 400mL that vacuum fractionation is collected adds from opening for feed, the rotating speed of regulating the rotor knifing is 200r/min, and the reinforced flow velocity of Z-ligustilide is 0.5mL/min, must steam thing respectively and reserve thing from two discharge ports, collection steams thing 286mL, is light yellow liquid.
Carry out structure detection and purity check to steaming thing:
Use respectively proton nmr spectra (
1H-NMR) and carbon-13 nmr spectra (
13C-NMR) thing that steams of present embodiment molecular distillation is identified that the result shows that the thing that steams that present embodiment makes is a Z-ligustilide.
Get the Z-ligustilide 1mL that makes and add methanol constant volume to 10mL, centrifugal 5min (10000RPM), supernatant liquor is sample introduction 5 μ L respectively, measure.
HPLC chromatogram testing conditions is as follows: instrument: Agilent 1260 type liquid chromatographs (quaternary pump, automatic sampler, DAD detector); Chromatographic column: Hypersil ODS (4.6 * 150mm, 5 μ m); Moving phase: methyl alcohol-5% Virahol (60: 40); Detect wavelength: 280nm; Flow velocity: 1mL/min; Column temperature: 25 ℃.
The purity that detects Z-ligustilide with HPLC is 96.7%.
Embodiment 6 Z-ligustilide preparation methods provided by the invention
Get the 2000mL Rhizoma Chuanxiong oil in the 5000mL round-bottomed flask, install the vacuum fractionation device, charge into all air in the argon gas remover, vacuumize vacuum state with the vacuum oil pump, begin heating with oil bath, when cut point reaches 108 ℃ into 70Pa, there are a large amount of yellow cuts to go out, collect cut 905mL.
Molecular distillation apparatus is charged into all air in the argon gas remover, be heated to 90 ℃, opening vacuum pump vacuumizes when making vacuum tightness reach 2Pa, after being heated to 45 ℃, the cut 500mL that vacuum fractionation is collected adds from opening for feed, the rotating speed of regulating the rotor knifing is 500r/min, and the reinforced flow velocity of Z-ligustilide is 5.0mL/min, must steam thing respectively and reserve thing from two discharge ports, collection steams thing 369mL, is light yellow liquid.
Carry out structure detection and purity check to steaming thing:
Use respectively proton nmr spectra (
1H-NMR) and carbon-13 nmr spectra (
13C-NMR) thing that steams of present embodiment molecular distillation is identified that the result shows that the thing that steams that present embodiment makes is a Z-ligustilide.
Get the Z-ligustilide 1mL that makes and add methanol constant volume to 10mL, centrifugal 5min (10000RPM), supernatant liquor is sample introduction 5 μ L respectively, measure.
HPLC chromatogram testing conditions is as follows: instrument: Agilent 1260 type liquid chromatographs (quaternary pump, automatic sampler, DAD detector); Chromatographic column: Hypersil ODS (4.6 * 150mm, 5 μ m); Moving phase: methyl alcohol-5% Virahol (60: 40); Detect wavelength: 280nm; Flow velocity: 1mL/min; Column temperature: 25 ℃.
The purity that detects Z-ligustilide with HPLC is 94.1%.
Embodiment 7 Z-ligustilide preparation methods provided by the invention
Get the 1000mL Radix Angelicae Sinensis oil in the 3000mL round-bottomed flask, install the vacuum fractionation device, charge into all air in the argon gas remover, vacuumize vacuum state with the vacuum oil pump, begin heating with oil bath, when cut point reaches 110 ℃ into 85Pa, there are a large amount of yellow cuts to go out, collect cut 550mL.
Molecular distillation apparatus is charged into all air in the argon gas remover, be heated to 75 ℃, opening vacuum pump vacuumizes when making vacuum tightness reach 10Pa, after being heated to 42 ℃, the cut 400mL that vacuum fractionation is collected adds from opening for feed, the rotating speed of regulating the rotor knifing is 350r/min, and the reinforced flow velocity of Z-ligustilide is 3.0mL/min, must steam thing respectively and reserve thing from two discharge ports, collection steams thing 218mL, is light yellow liquid.
Carry out structure detection and purity check to steaming thing:
Use respectively proton nmr spectra (
1H-NMR) and carbon-13 nmr spectra (
13C-NMR) thing that steams of present embodiment molecular distillation is identified that the result shows that the thing that steams that present embodiment makes is a Z-ligustilide.
Get the Z-ligustilide 1mL that makes and add methanol constant volume to 10mL, centrifugal 5min (10000RPM), supernatant liquor is sample introduction 5 μ L respectively, measure.
HPLC chromatogram testing conditions is as follows: instrument: Agilent 1260 type liquid chromatographs (quaternary pump, automatic sampler, DAD detector); Chromatographic column: Hypersil ODS (4.6 * 150mm, 5 μ m); Moving phase: methyl alcohol-5% Virahol (60: 40); Detect wavelength: 280nm; Flow velocity: 1mL/min; Column temperature: 25 ℃.
The purity that detects Z-ligustilide with HPLC is 93.2%.
Embodiment 8 Z-ligustilide preparation methods provided by the invention
Get the 2000mL Radix Angelicae Sinensis oil in the 5000mL round-bottomed flask, install the vacuum fractionation device, charge into all air in the argon gas remover, vacuumize vacuum state with the vacuum oil pump, begin heating with oil bath, when cut point reaches 106 ℃ into 30Pa, there are a large amount of yellow cuts to go out, collect cut 961mL.Molecular distillation apparatus is charged into all air in the argon gas remover, be heated to 85 ℃, opening vacuum pump vacuumizes when making vacuum tightness reach 1Pa, after being heated to 45 ℃, the cut 800mL that vacuum fractionation is collected adds from opening for feed, the rotating speed of regulating the rotor knifing is 250r/min, and the reinforced flow velocity of Z-ligustilide is 1.0mL/min, must steam thing respectively and reserve thing from two discharge ports, collection steams thing 579mL, is light yellow liquid.
Carry out structure detection and purity check to steaming thing:
Use respectively proton nmr spectra (
1H-NMR) and carbon-13 nmr spectra (
13C-NMR) thing that steams of present embodiment molecular distillation is identified that the result shows that the thing that steams that present embodiment makes is a Z-ligustilide.
Get the Z-ligustilide 1mL that makes and add methanol constant volume to 10mL, centrifugal 5min (10000RPM), supernatant liquor is sample introduction 5 μ L respectively, measure.HPLC chromatogram testing conditions is as follows: instrument: Agilent 1260 type liquid chromatographs (quaternary pump, automatic sampler, DAD detector); Chromatographic column: Hypersil ODS (4.6 * 150mm, 5 μ m); Moving phase: methyl alcohol-5% Virahol (60: 40); Detect wavelength: 280nm; Flow velocity: 1mL/min; Column temperature: 25 ℃.
The purity that detects Z-ligustilide with HPLC is 98.7%.
The above only is a preferred implementation of the present invention; should be pointed out that for those skilled in the art, under the prerequisite that does not break away from the principle of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (8)
1. the preparation method of a Z-ligustilide is characterized in that, comprising:
Step 1: under the vacuum state of 5～100Pa, Radix Angelicae Sinensis oil or Rhizoma Chuanxiong oil heating are carried out vacuum fractionation, collect 100～110 ℃ cut;
Step 2: the described cut of step 1 is carried out molecular distillation under 70～90 ℃, the condition of 0.1～10Pa vacuum tightness, collect and steam thing, obtain Z-ligustilide; The rotating speed of described molecular distillation rotor knifing is that the charging flow velocity of 50～500r/min, described cut is 0.5～5.0mL/min.
2. preparation method as claimed in claim 1 is characterized in that, vacuum tightness is 5～50Pa in the described step 1.
3. preparation method as claimed in claim 1 is characterized in that, in the described step 1 Radix Angelicae Sinensis oil or Rhizoma Chuanxiong oil heating is collected 105～110 ℃ cut.
4. preparation method as claimed in claim 1 is characterized in that, also comprises before the distillation of cut conducting molecule described in the described step 2 being preheated to 40～45 ℃ step.
5. preparation method as claimed in claim 1 is characterized in that, molecular distillation is to carry out under 80～85 ℃ the condition in temperature in the described step 2.
6. preparation method as claimed in claim 1 is characterized in that, molecular distillation is to carry out under the condition of 0.1～5Pa in vacuum tightness in the described step 2.
7. preparation method as claimed in claim 1 is characterized in that, molecular distillation is to carry out under the condition of 80～350r/min at the rotating speed of rotor knifing in the described step 2.
8. preparation method as claimed in claim 1 is characterized in that, molecular distillation is to carry out under the condition of 1.0～2.0mL/min at the flow velocity of the described cut of step 1 in the described step 2.
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| CN106138135A (en) * | 2016-08-18 | 2016-11-23 | 四川省中医药科学院 | Medicine for treatment of vascular cognitive disorder and preparation method thereof |
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