CN103030567A - Propranolol medicine enantiomer resolution method - Google Patents
Propranolol medicine enantiomer resolution method Download PDFInfo
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- CN103030567A CN103030567A CN2012102595296A CN201210259529A CN103030567A CN 103030567 A CN103030567 A CN 103030567A CN 2012102595296 A CN2012102595296 A CN 2012102595296A CN 201210259529 A CN201210259529 A CN 201210259529A CN 103030567 A CN103030567 A CN 103030567A
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Abstract
The invention discloses a propranolol medicine enantiomer resolution method comprising the following steps of: (1) uniformly mixing an organic solvent containing L-tartrate and acetic acid/triethylamine buffer solution containing boric acid according to volume ratio of 1:1, standing and layering, and separating to obtain an organic phase and an aqueous phase; (2) transferring propranolol hydrochloride into free propranolol, and dissolving by the aqueous phase obtained in the step (1) to prepare a sample for standby; (3) resolving propranolol by a high-speed countercurrent chromatography method and respectively collecting prepeak eluate and postpeak eluate; and (4) respectively recycling from the collected prepeak eluate and postpeak eluate of the step (3) to obtain monomers of laevo-propranolol and dextro-propranolol. When the propranolol medicine enantiomer resolution method disclosed by the invention is used for resolving chiral medicine propranolol, high resolution degree can be obtained; and the propranolol medicine enantiomer resolution method is suitable for various types of preparative countercurrent chromatograph and the monomers of laevo-propranolol and dextro-propranolol can be obtained.
Description
Technical field
The present invention relates to a kind of method that from the chiral drug Propranolol Enantiomers, splits out left-handed Proprasylyte and D-propranolol.
Background technology
Different enantiomorphs demonstrate different biological activitys in vivo in the chiral drug, this so that seek efficiently, method for splitting seems particularly important easily.Proprasylyte belongs to β-suprarenal gland retarding agent class medicine, is widely used in clinically treatment hypertension, the diseases such as myocardial ischemia and arrhythmia, and wherein the drug effect of S type Proprasylyte approximately is 40 times of R type, and R type Proprasylyte also has very strong antifertility action.At present, the method for Separation of propranolol enantiomers mainly contains high performance liquid chromatography, capillary electrophoresis, paired ion chromatography, tlc etc.
The exploitation of chiral drug tends to develop single enantiomer over past ten years, world market the increasing with 20% above speed in every year of single enantiomer medicine.Food and drug administration is regulation just as far back as 1992, and from now on all exploitations have the medicine of asymmetric center, must provide the result of chiral separation.Therefore, carry out the fractionation of chiral drug and obtain single chiral drug, become an important branch in separation science field.
High-speed countercurrent chromatography (high-speed countercurrent chromatography, HSCCC) be that grow up the eighties in 20th century a kind of need not solid-state supporter or the liquid liquid distribution technique of carrier, in sepn process, eliminated irreversible adsorption phenomenon common in gas, the liquid chromatography fully, can the saboteur, be applicable to the large chipal compounds of separating polar and biomacromolecule; Because the separation mechanism of its uniqueness makes it can be used for preparation and separates, and therefore is applicable to very much preparation property separating chiral compound.The in recent years research of relevant adverse current chromatogram chiral separation aspect is accelerated to some extent.
Summary of the invention
The purpose of this invention is to provide a kind of method that adopts high-speed countercurrent chromatography to split the Proprasylyte drug enantiomer.
For achieving the above object, the present invention is take propranolol hydrochloride as splitting object, first propranolol hydrochloride is converted into the Proprasylyte episome, with L-TARTARIC ACID ester class and boric acid as resolution reagent, L-TARTARIC ACID ester class is added in the organic phase, boric acid adds aqueous phase to, adopts high-speed countercurrent chromatography to obtain left-handed Proprasylyte and D-propranolol.
The concrete technical scheme that the present invention adopts is as follows:
A kind of Proprasylyte drug enantiomer method for splitting comprises the steps:
Acetic acid/the triethylamine buffer solution that (1) will contain the organic solvent of L-TARTARIC ACID ester and contain boric acid mixes according to volume ratio 1:1, and standing demix separates obtaining organic phase and water; Described organic solvent is halogenated alkane, substituted arene, the alkane of C1 ~ C8 or the ether of C2 ~ C8 of C1 ~ C8, the substituting group of described substituted arene is one or more, described substituting group independently is selected from the alkyl of halogen or C1 ~ C4 separately, and the concentration of L-TARTARIC ACID ester is 0.02 ~ 0.5mol/L in the described organic solvent; The pH of the described acetic acid/triethylamine buffer solution that contains boric acid is 3.0 ~ 5.0, and the concentration of its mesoboric acid is 0.02 ~ 0.30mol/L;
(2) propranolol hydrochloride is converted into the Proprasylyte episome, and with step (1) gained aqueous phase dissolved, makes sample stand-by, sample concentration is 0.1 ~ 15.0mg/ml;
(3) adopt high-speed countercurrent chromatography to split Proprasylyte: the organic phase that obtains take step (1) respectively and water are as stationary phase and moving phase, countercurrent chromatography separation column is filled up stationary phase, column temperature is 5 ~ 30 ℃, the opening speed controller, rotating speed is 500 ~ 2000rpm, flow velocity with 0.2 ~ 3.0ml/min pumps into moving phase in the post, after two phase solvent system reaches fluid dynamic equilibrium, (when moving phase flows out from the chromatographic column exit, just think and reach balance), by the sampling valve sample introduction, UV-detector with wavelength 190 ~ 400nm detects, according to the ultraviolet detection spectrogram, collect respectively leading peak elutriant and postpeak elutriant;
(4) reclaim the monomer that obtains left-handed Proprasylyte and D-propranolol in the leading peak elutriant that collection obtains from step (3) respectively and the postpeak elutriant.
Organic solvent of the present invention is halogenated alkane, substituted arene, the alkane of C1 ~ C8 or the ether of C2 ~ C8 of C1 ~ C8, the halogenated alkane of described C1 ~ C8 can be chloroform, methylene dichloride etc., the preferred substituted benzene of described substituted arene, such as toluene, chlorobenzene etc., the alkane of described C1 ~ C8 can be normal hexane, normal heptane, Skellysolve A, hexanaphthene, sherwood oil etc., and the ether of described C2 ~ C8 can be ether, methyl tertiary butyl ether etc.Preferred organic solvent is chloroform or methylene dichloride, and most preferred organic solvent is chloroform.
L-TARTARIC ACID ester of the present invention can be the just own ester of L-TARTARIC ACID, the positive butyl ester of L-TARTARIC ACID, L-TARTARIC ACID n-octyl, L-TARTARIC ACID n-pentyl ester, L-TARTARIC ACID isobutyl ester or L-TARTARIC ACID-2-ethylhexyl.
The preferred described pH=4.0 that contains the acetic acid/triethylamine buffer solution of boric acid of the present invention ~ 4.4, wherein boric acid concentration is preferably 0.05 ~ 0.1mol/L; The described pH=4.4 that contains the acetic acid/triethylamine buffer solution of boric acid most preferably, wherein boric acid concentration is 0.1mol/L.
The concentration of L-TARTARIC ACID ester is 0.1 ~ 0.2mol/L in the preferred described organic solvent of the present invention, most preferably is 0.1mol/L.
In the step of the present invention (2), can propranolol hydrochloride be converted into the Proprasylyte episome by existing method, for example can adopt following method: get propranolol hydrochloride and join in 1 ~ 2mol/L sodium hydroxide solution, be heated to 40 ~ 50 ℃ and stir 2～3min, then use dichloromethane extraction, the gained organic phase is washed to neutrality, is namely got crude product with anhydrous sodium sulfate drying, filtration, filtrate steaming removal solvent with ultrapure water, crude product is with the methylene dichloride dissolving and add several normal hexane recrystallizations, obtains the Proprasylyte episome.
In the step of the present invention (3), after high-speed countercurrent chromatography split, the elutriant that collection is obtained carried out the high performance liquid phase detection, and the purity of two monomer elutriants all can be greater than 95%.In split process, the column temperature of preferred countercurrent chromatography separation column is 5 ~ 15 ℃; Preferably the flow velocity with 0.5 ~ 2.0ml/min pumps into moving phase in the post.
In the step of the present invention (4), can from leading peak elutriant and postpeak elutriant, reclaim respectively by existing method the monomer of left-handed Proprasylyte and D-propranolol, for example can adopt following method: with leading peak elutriant or postpeak elutriant alkalize to pH be 9 ~ 14, directly use dichloromethane extraction after separating out a large amount of white powders, dichloromethane layer is extremely neutral with the saturated common salt water washing, anhydrous sodium sulfate drying, filter, filtrate steaming removal solvent namely gets crude product, and crude product is with the methylene dichloride dissolving and add left-handed Proprasylyte monomer or the D-propranolol monomer that several normal hexane recrystallizations can obtain purifying.Wherein leading peak elutriant or postpeak elutriant available hydrogen sodium hydroxide solution alkalization, concentration of sodium hydroxide solution can be 1 ~ 2mol/L.
The present invention is concrete to recommend described method for splitting to carry out in accordance with the following steps:
Acetic acid/the triethylamine buffer solution that (1) will contain the organic solvent of L-TARTARIC ACID ester and contain boric acid mixes according to volume ratio 1:1, and standing demix separates obtaining organic phase and water; Described organic solvent is halogenated alkane, substituted arene, the alkane of C1 ~ C8 or the ether of C2 ~ C8 of C1 ~ C8, the substituting group of described substituted arene is one or more, described substituting group independently is selected from the alkyl of halogen or C1 ~ C4 separately, and the concentration of L-TARTARIC ACID ester is 0.02 ~ 0.5mol/L in the described organic solvent; The pH of the described acetic acid/triethylamine buffer solution that contains boric acid is 3.0 ~ 5.0, and the concentration of its mesoboric acid is 0.02 ~ 0.30mol/L;
(2) propranolol hydrochloride is converted into the Proprasylyte episome, and with step (1) gained aqueous phase dissolved, makes sample stand-by, Proprasylyte episome concentration is 0.1 ~ 15.0mg/ml in the sample;
(3) adopt high-speed countercurrent chromatography to split Proprasylyte: the organic phase that obtains take step (1) respectively and water are as stationary phase and moving phase, countercurrent chromatography separation column is filled up stationary phase, column temperature is 5 ~ 30 ℃, the opening speed controller, rotating speed is 500 ~ 2000rpm, flow velocity with 0.2 ~ 3.0ml/min pumps into moving phase in the post, after two phase solvent system reaches fluid dynamic equilibrium, (when moving phase flows out from the chromatographic column exit, just think and reach balance), by the sampling valve sample introduction, UV-detector with wavelength 190 ~ 400nm detects, according to the ultraviolet detection spectrogram, collect respectively leading peak elutriant and postpeak elutriant;
(4) reclaim the monomer that obtains left-handed Proprasylyte and D-propranolol in the leading peak elutriant that collection obtains from step (3) respectively and the postpeak elutriant.
Further preferred method for splitting of the present invention carries out in accordance with the following steps:
Acetic acid/the triethylamine buffer solution that (1) will contain the organic solvent of L-TARTARIC ACID ester and contain boric acid mixes according to volume ratio 1:1, and standing demix separates obtaining organic phase and water; Described organic solvent is chloroform or methylene dichloride, and the concentration of L-TARTARIC ACID ester is 0.1 ~ 0.2mol/L in the described organic solvent; The pH of the described acetic acid/triethylamine buffer solution that contains boric acid is 4.0 ~ 4.4, and the concentration of its mesoboric acid is 0.05 ~ 0.1mol/L;
(2) propranolol hydrochloride is converted into the Proprasylyte episome, and with step (1) gained aqueous phase dissolved, makes sample stand-by, Proprasylyte episome concentration is 0.1 ~ 15.0mg/ml in the sample;
(3) adopt high-speed countercurrent chromatography to split Proprasylyte: the organic phase that obtains take step (1) respectively and water are as stationary phase and moving phase, countercurrent chromatography separation column is filled up stationary phase, column temperature is 5 ~ 15 ℃, the opening speed controller, rotating speed is 500 ~ 2000rpm, flow velocity with 0.5 ~ 2.0ml/min pumps into moving phase in the post, after two phase solvent system reaches fluid dynamic equilibrium, by the sampling valve sample introduction, UV-detector with wavelength 190 ~ 400nm detects, according to the ultraviolet detection spectrogram, collect respectively leading peak elutriant and postpeak elutriant;
(4) reclaim the monomer that obtains left-handed Proprasylyte and D-propranolol in the leading peak elutriant that collection obtains from step (3) respectively and the postpeak elutriant.
The present invention can adopt analysis mode and half preparation type counter current chromatograph (analysis mode separator column column volume is 20ml, and half preparation type separator column column volume is 300ml), and counter current chromatograph is comprised of constant flow pump, main frame (separator column), UV-detector, registering instrument etc.
Compared with prior art, advantage of the present invention is: adopt present method resolving chiral medicine Proprasylyte can reach higher resolution, and the preparation type counter current chromatograph that the method is applicable to various models can separate the monomer that obtains left-handed Proprasylyte and D-propranolol.
Description of drawings
Analysis mode high speed adverse current chromatogram figure under Fig. 1: embodiment 1 experiment condition;
Analysis mode high speed adverse current chromatogram figure under Fig. 2: embodiment 2 experiment conditions;
Analysis mode high speed adverse current chromatogram figure under Fig. 3: embodiment 5 experiment conditions;
Analysis mode high speed adverse current chromatogram figure under Fig. 4: embodiment 7 experiment conditions;
Analysis mode high speed adverse current chromatogram figure under Fig. 5: embodiment 10 experiment conditions;
Half countercurrent chromatography figure under Fig. 6: embodiment 11 experiment conditions;
Fig. 7: (110min ~ 200min) high performance liquid chromatography of elutriant detects spectrogram, i.e. R type Proprasylyte to A part among Fig. 6;
Fig. 8: (235min ~ 540min) high performance liquid chromatography of elutriant detects spectrogram, i.e. S type Proprasylyte to B part among Fig. 6.
Embodiment
The present invention will be further described with specific embodiment for the below, but protection scope of the present invention is not limited to this.
Propranolol hydrochloride is converted into the Proprasylyte episome: get the 610mg propranolol hydrochloride, join in the sodium hydroxide solution of 5mL1mol/L, be heated to 50 ℃ and stir 2～3min, then be transferred in the separating funnel with on a small quantity in batches repeatedly extraction of methylene dichloride, merge organic phase and extremely neutral with the ultrapure water washing, anhydrous sodium sulfate drying, steam to desolventize after filtering and namely get the Proprasylyte episome, crude product adds several normal hexane recrystallizations with the methylene dichloride dissolving, obtain colourless acicular crystal 488mg, be free Proprasylyte.
With chloroform: water (organic phase contains the just own ester of 0.1mol/L L-TARTARIC ACID, and water is the acetic acid/triethylamine buffered soln pH=4.4 of 0.05mol/L and contains 0.1mol/L boric acid) is disposed in the separating funnel according to the volume ratio of 1:1, shakes up rear standing demix.Ready to balance will separate up and down after for some time mutually, and organic phase is as stationary phase, and water is as moving phase.Take by weighing the 10mg Propranolol Enantiomers with the dissolving of 5ml moving phase, make sample solution after the dissolving stand-by.
Adopt analysis mode high-speed counter-current chromatograph Separation of propranolol enantiomers, the separator column volume is 20ml.Before the sample introduction, countercurrent chromatography separation column is filled up stationary phase, column temperature is 15 ℃, and opening speed controller, rotating speed are 1800rpm, with the flow velocity of 0.50ml/min moving phase is pumped in the post, after two phase solvent system reaches fluid dynamic equilibrium, by the sampling valve sample introduction.Then the UV-detector with wavelength 280nm detects, and according to the ultraviolet detection spectrogram, calculating resolution is 1.08.Collect respectively leading peak elutriant and postpeak elutriant, the elutriant that collection is obtained carries out the high performance liquid phase detection, and the purity of two monomer elutriants is all greater than 95%.The testing conditions of high performance liquid chromatography is: Chiralcel OD-R column (4.6 * 250mm); Column temperature: 20 ℃; Moving phase: 0.1mol/L KPF6: acetonitrile (60:40, v/v) isocratic elution; Flow velocity 0.5ml/min; Detect wavelength 215nm; Sample size: 10 μ l.
With chloroform: water (organic phase contains 0.1mol/L L-TARTARIC ACID isobutyl ester, and water is the acetic acid/triethylamine buffered soln pH=4.4 of 0.05mol/L and contains 0.1mol/L boric acid) is disposed in the separating funnel according to the volume ratio of 1:1, shakes up rear standing demix.Ready to balance will separate up and down after for some time mutually, and organic phase is as stationary phase, and water is as moving phase.Take by weighing the 10mg Propranolol Enantiomers that obtains among the embodiment 1 and dissolve with 5ml moving phase, make sample solution after the dissolving stand-by.
Adopt analysis mode high-speed counter-current chromatograph Separation of propranolol enantiomers, the separator column volume is 20ml.Before the sample introduction, countercurrent chromatography separation column is filled up stationary phase, column temperature is 15 ℃, and opening speed controller, rotating speed are 1800rpm, with the flow velocity of 0.50ml/min moving phase is pumped in the post, after two phase solvent system reaches fluid dynamic equilibrium, by the sampling valve sample introduction.Then the UV-detector with wavelength 280nm detects, and according to the ultraviolet detection spectrogram, calculating resolution is 1.10.Collect respectively leading peak elutriant and postpeak elutriant, the elutriant that collection is obtained carries out the high performance liquid phase detection, and the purity of two monomer elutriants is all greater than 95%.The testing conditions of high performance liquid chromatography is with embodiment 1.
Embodiment 3
With hexanaphthene: water (organic phase contains the just own ester of 0.1mol/L L-TARTARIC ACID, and water is the acetic acid/triethylamine buffered soln pH=4.4 of 0.05mol/L and contains 0.1mol/L boric acid) is disposed in the separating funnel according to the volume ratio of 1:1, shakes up rear standing demix.Ready to balance will separate up and down after for some time mutually, and organic phase is as stationary phase, and water is as moving phase.Take by weighing the 10mg Propranolol Enantiomers that obtains among the embodiment 1 and dissolve with 5ml moving phase, make sample solution after the dissolving stand-by.
Adopt analysis mode high-speed counter-current chromatograph Separation of propranolol enantiomers, the separator column volume is 20ml.Before the sample introduction, countercurrent chromatography separation column is filled up stationary phase, column temperature is 15 ℃, and opening speed controller, rotating speed are 1800rpm, with the flow velocity of 0.50ml/min moving phase is pumped in the post, after two phase solvent system reaches fluid dynamic equilibrium, by the sampling valve sample introduction.Then the UV-detector with wavelength 280nm detects, and according to the ultraviolet detection spectrogram, calculating resolution is 0.34.Collect respectively leading peak elutriant and postpeak elutriant, the elutriant that collection is obtained carries out the high performance liquid phase detection, and the purity of two monomer elutriants is all greater than 95%.The testing conditions of high performance liquid chromatography is with embodiment 1.
Embodiment 4
With methylene dichloride: water (organic phase contains the just own ester of 0.1mol/L L-TARTARIC ACID, and water is the acetic acid/triethylamine buffered soln pH=4.4 of 0.05mol/L and contains 0.1mol/L boric acid) is disposed in the separating funnel according to the volume ratio of 1:1, shakes up rear standing demix.Ready to balance will separate up and down after for some time mutually, and organic phase is as stationary phase, and water is as moving phase.Take by weighing the 10mg Propranolol Enantiomers that obtains among the embodiment 1 and dissolve with 5ml moving phase, make sample solution after the dissolving stand-by.
Adopt analysis mode high-speed counter-current chromatograph Separation of propranolol enantiomers, the separator column volume is 20ml.Before the sample introduction, countercurrent chromatography separation column is filled up stationary phase, column temperature is 15 ℃, and opening speed controller, rotating speed are 1800rpm, with the flow velocity of 0.50ml/min moving phase is pumped in the post, after two phase solvent system reaches fluid dynamic equilibrium, by the sampling valve sample introduction.Then the UV-detector with wavelength 280nm detects, and according to the ultraviolet detection spectrogram, calculating resolution is 1.10.Collect respectively leading peak elutriant and postpeak elutriant, the elutriant that collection is obtained carries out the high performance liquid phase detection, and the purity of two monomer elutriants is all greater than 95%.The testing conditions of high performance liquid chromatography is with embodiment 1.
With chloroform: water (organic phase contains the just own ester of 0.1mol/L L-TARTARIC ACID, and water is the acetic acid/triethylamine buffered soln pH=4.4 of 0.05mol/L and contains 0.1mol/L boric acid) is disposed in the separating funnel according to the volume ratio of 1:1, shakes up rear standing demix.Ready to balance will separate up and down after for some time mutually, and organic phase is as stationary phase, and water is as moving phase.Take by weighing the 10mg Propranolol Enantiomers that obtains among the embodiment 1 and dissolve with 5ml moving phase, make sample solution after the dissolving stand-by.
Adopt analysis mode high-speed counter-current chromatograph Separation of propranolol enantiomers, the separator column volume is 20ml.Before the sample introduction, countercurrent chromatography separation column is filled up stationary phase, column temperature is 5 ℃, and opening speed controller, rotating speed are 1800rpm, with the flow velocity of 0.50ml/min moving phase is pumped in the post, after two phase solvent system reaches fluid dynamic equilibrium, by the sampling valve sample introduction.Then the UV-detector with wavelength 280nm detects, and according to the ultraviolet detection spectrogram, calculating resolution is 1.13.Collect respectively leading peak elutriant and postpeak elutriant, the elutriant that collection is obtained carries out the high performance liquid phase detection, and the purity of two monomer elutriants is all greater than 95%.The testing conditions of high performance liquid chromatography is with embodiment 1.
Embodiment 6
With chloroform: water (organic phase contains the just own ester of 0.1mol/L L-TARTARIC ACID, and water is the acetic acid/triethylamine buffered soln pH=4.4 of 0.05mol/L and contains 0.1mol/L boric acid) is disposed in the separating funnel according to the volume ratio of 1:1, shakes up rear standing demix.Ready to balance will separate up and down after for some time mutually, and organic phase is as stationary phase, and water is as moving phase.Take by weighing the 10mg Propranolol Enantiomers that obtains among the embodiment 1 and dissolve with 5ml moving phase, make sample solution after the dissolving stand-by.
Adopt analysis mode high-speed counter-current chromatograph Separation of propranolol enantiomers, the separator column volume is 20ml.Before the sample introduction, countercurrent chromatography separation column is filled up stationary phase, column temperature is 15 ℃, and opening speed controller, rotating speed are 1800rpm, with the flow velocity of 0.80ml/min moving phase is pumped in the post, after two phase solvent system reaches fluid dynamic equilibrium, by the sampling valve sample introduction.Then the UV-detector with wavelength 280nm detects, and according to the ultraviolet detection spectrogram, calculating resolution is 1.01.Collect respectively leading peak elutriant and postpeak elutriant, the elutriant that collection is obtained carries out the high performance liquid phase detection, and the purity of two monomer elutriants is all greater than 95%.The testing conditions of high performance liquid chromatography is with embodiment 1.
Embodiment 7
With chloroform: water (organic phase contains the just own ester of 0.2mol/L L-TARTARIC ACID, and water is the acetic acid/triethylamine buffered soln pH=4.4 of 0.05mol/L and contains 0.1mol/L boric acid) is disposed in the separating funnel according to the volume ratio of 1:1, shakes up rear standing demix.Ready to balance will separate up and down after for some time mutually, and organic phase is as stationary phase, and water is as moving phase.Take by weighing the 10mg Propranolol Enantiomers that obtains among the embodiment 1 and dissolve with 5ml moving phase, make sample solution after the dissolving stand-by.
Adopt analysis mode high-speed counter-current chromatograph Separation of propranolol enantiomers, the separator column volume is 20ml.Before the sample introduction, countercurrent chromatography separation column is filled up stationary phase, column temperature is 15 ℃, and opening speed controller, rotating speed are 1800rpm, with the flow velocity of 0.50ml/min moving phase is pumped in the post, after two phase solvent system reaches fluid dynamic equilibrium, by the sampling valve sample introduction.Then the UV-detector with wavelength 280nm detects, and according to the ultraviolet detection spectrogram, calculating resolution is 1.21.Collect respectively leading peak elutriant and postpeak elutriant, the elutriant that collection is obtained carries out the high performance liquid phase detection, and the purity of two monomer elutriants is all greater than 95%.The testing conditions of high performance liquid chromatography is with embodiment 1.
With chloroform: water (organic phase contains the just own ester of 0.1mol/L L-TARTARIC ACID, and water is the acetic acid/triethylamine buffered soln pH=4.4 of 0.05mol/L and contains 0.1mol/L boric acid) is disposed in the separating funnel according to the volume ratio of 1:1, shakes up rear standing demix.Ready to balance will separate up and down after for some time mutually, and organic phase is as stationary phase, and water is as moving phase.Take by weighing the 10mg Propranolol Enantiomers that obtains among the embodiment 1 and dissolve with 5ml moving phase, make sample solution after the dissolving stand-by.
Adopt analysis mode high-speed counter-current chromatograph Separation of propranolol enantiomers, the separator column volume is 20ml.Before the sample introduction, countercurrent chromatography separation column is filled up stationary phase, column temperature is 15 ℃, and opening speed controller, rotating speed are 1500, with the flow velocity of 0.50ml/min moving phase is pumped in the post, after two phase solvent system reaches fluid dynamic equilibrium, by the sampling valve sample introduction.Then the UV-detector with wavelength 280nm detects, and according to the ultraviolet detection spectrogram, calculating resolution is 1.01.Collect respectively leading peak elutriant and postpeak elutriant, the elutriant that collection is obtained carries out the high performance liquid phase detection, and the purity of two monomer elutriants is all greater than 95%.The testing conditions of high performance liquid chromatography is with embodiment 1.
With chloroform: water (organic phase contains the just own ester of 0.1mol/L L-TARTARIC ACID, and water is the acetic acid/triethylamine buffered soln pH=4.4 of 0.05mol/L and contains 0.05mol/L boric acid) is disposed in the separating funnel according to the volume ratio of 1:1, shakes up rear standing demix.Ready to balance will separate up and down after for some time mutually, and organic phase is as stationary phase, and water is as moving phase.Take by weighing the 10mg Propranolol Enantiomers that obtains among the embodiment 1 and dissolve with 5ml moving phase, make sample solution after the dissolving stand-by.
Adopt analysis mode high-speed counter-current chromatograph Separation of propranolol enantiomers, the separator column volume is 20ml.Before the sample introduction, countercurrent chromatography separation column is filled up stationary phase, column temperature is 15 ℃, and opening speed controller, rotating speed are 1800rpm, with the flow velocity of 0.50ml/min moving phase is pumped in the post, after two phase solvent system reaches fluid dynamic equilibrium, by the sampling valve sample introduction.Then the UV-detector with wavelength 280nm detects, and according to the ultraviolet detection spectrogram, calculating resolution is 1.02.Collect respectively leading peak elutriant and postpeak elutriant, the elutriant that collection is obtained carries out the high performance liquid phase detection, and the purity of two monomer elutriants is all greater than 95%.The testing conditions of high performance liquid chromatography is with embodiment 1.
With chloroform: water (organic phase contains the just own ester of 0.1mol/L L-TARTARIC ACID, and water is the acetic acid/triethylamine buffered soln pH=4.0 of 0.05mol/L and contains 0.1mol/L boric acid) is disposed in the separating funnel according to the volume ratio of 1:1, shakes up rear standing demix.Ready to balance will separate up and down after for some time mutually, and organic phase is as stationary phase, and water is as moving phase.Take by weighing the 10mg Propranolol Enantiomers that obtains among the embodiment 1 and dissolve with 5ml moving phase, make sample solution after the dissolving stand-by.
Adopt analysis mode high-speed counter-current chromatograph Separation of propranolol enantiomers, the separator column volume is 20ml.Before the sample introduction, countercurrent chromatography separation column is filled up stationary phase, column temperature is 15 ℃, and opening speed controller, rotating speed are 1800rpm, with the flow velocity of 0.50ml/min moving phase is pumped in the post, after two phase solvent system reaches fluid dynamic equilibrium, by the sampling valve sample introduction.Then the UV-detector with wavelength 280nm detects, and according to the ultraviolet detection spectrogram, calculating resolution is 1.07.Collect respectively leading peak elutriant and postpeak elutriant, the elutriant that collection is obtained carries out the high performance liquid phase detection, and the purity of two monomer elutriants is all greater than 95%.The testing conditions of high performance liquid chromatography is with embodiment 1.
Embodiment 11
With chloroform: water (organic phase contains the just own ester of 0.1mol/L L-TARTARIC ACID, and water is the acetic acid/triethylamine buffered soln pH=4.4 of 0.05mol/L and contains 0.1mol/L boric acid) is disposed in the separating funnel according to the volume ratio of 1:1, shakes up rear standing demix.Ready to balance will separate up and down after for some time mutually, and organic phase is as stationary phase, and water is as moving phase.Take by weighing the 108mg Propranolol Enantiomers that obtains among the embodiment 1 and dissolve with 20ml moving phase, make sample solution after the dissolving stand-by.
Adopt half countercurrent chromatography instrument Separation of propranolol enantiomers, the separator column volume is 300ml.Before the sample introduction, countercurrent chromatography separation column is filled up stationary phase, column temperature is 15 ℃, and opening speed controller, rotating speed are 800rpm, with the flow velocity of 2.00ml/min moving phase is pumped in the post, after two phase solvent system reaches fluid dynamic equilibrium, by the sampling valve sample introduction.Then the UV-detector with wavelength 280nm detects, and according to the ultraviolet detection spectrogram, receives leading peak elutriant and postpeak elutriant, and calculating resolution is 1.05.The elutriant that collection is obtained carries out the high performance liquid phase detection, and the purity of two monomer elutriants is all greater than 95%.The testing conditions of high performance liquid chromatography is with embodiment 1.
Recovery sample from elutriant: leading peak and postpeak elutriant are alkalized to the pH11 with the 1mol/L sodium hydroxide solution respectively, directly repeatedly extract on a small quantity the combined dichloromethane layer with methylene dichloride after separating out a large amount of white powders, extremely neutral with saturated common salt water washing dichloromethane layer, anhydrous sodium sulfate drying, steam to desolventize after filtering and namely get left-handed Proprasylyte and D-propranolol, add several normal hexane recrystallizations with the methylene dichloride dissolving at last, obtain the left-handed Proprasylyte of 22.9mg and the 30.1mg D-propranolol of purifying, enantiomeric purity is greater than 95%.
Claims (9)
1. a Proprasylyte drug enantiomer method for splitting comprises the steps:
Acetic acid/the triethylamine buffer solution that (1) will contain the organic solvent of L-TARTARIC ACID ester and contain boric acid mixes according to volume ratio 1:1, and standing demix separates obtaining organic phase and water; Described organic solvent is halogenated alkane, substituted arene, the alkane of C1 ~ C8 or the ether of C2 ~ C8 of C1 ~ C8, the substituting group of described substituted arene is one or more, described substituting group independently is selected from the alkyl of halogen or C1 ~ C4 separately, and the concentration of L-TARTARIC ACID ester is 0.02 ~ 0.5mol/L in the described organic solvent; The pH of the described acetic acid/triethylamine buffer solution that contains boric acid is 3.0 ~ 5.0, and the concentration of its mesoboric acid is 0.02 ~ 0.30mol/L;
(2) propranolol hydrochloride is converted into the Proprasylyte episome, and with step (1) gained aqueous phase dissolved, makes sample stand-by, Proprasylyte episome concentration is 0.1 ~ 15.0mg/ml in the sample;
(3) adopt high-speed countercurrent chromatography to split Proprasylyte: the organic phase that obtains take step (1) respectively and water are as stationary phase and moving phase, countercurrent chromatography separation column is filled up stationary phase, column temperature is 5 ~ 30 ℃, the opening speed controller, rotating speed is 500 ~ 2000rpm, flow velocity with 0.2 ~ 3.0ml/min pumps into moving phase in the post, after two phase solvent system reaches fluid dynamic equilibrium, by the sampling valve sample introduction, UV-detector with wavelength 190 ~ 400nm detects, according to the ultraviolet detection spectrogram, collect respectively leading peak elutriant and postpeak elutriant;
(4) reclaim the monomer that obtains left-handed Proprasylyte and D-propranolol in the leading peak elutriant that collection obtains from step (3) respectively and the postpeak elutriant.
2. Proprasylyte drug enantiomer method for splitting as claimed in claim 1 is characterized in that: described organic solvent is selected from one of following: chloroform, methylene dichloride, toluene, chlorobenzene, normal hexane, normal heptane, Skellysolve A, hexanaphthene, ether, sherwood oil or methyl tertiary butyl ether.
3. Proprasylyte drug enantiomer method for splitting as claimed in claim 1, it is characterized in that: described organic solvent is chloroform or methylene dichloride.
4. Proprasylyte drug enantiomer method for splitting as claimed in claim 1, it is characterized in that: described organic solvent is chloroform.
5. such as the described Proprasylyte drug enantiomer of one of claim 1 ~ 4 method for splitting, it is characterized in that: described L-TARTARIC ACID ester is the just own ester of L-TARTARIC ACID, the positive butyl ester of L-TARTARIC ACID, L-TARTARIC ACID n-octyl, L-TARTARIC ACID n-pentyl ester, L-TARTARIC ACID isobutyl ester or L-TARTARIC ACID-2-ethylhexyl.
6. Proprasylyte drug enantiomer method for splitting as claimed in claim 5 is characterized in that: the described pH=4.0 that contains the acetic acid/triethylamine buffer solution of boric acid ~ 4.4, and wherein boric acid concentration is 0.05 ~ 0.1mol/L; The concentration of L-TARTARIC ACID ester is 0.1 ~ 0.2mol/L in the described organic solvent.
7. Proprasylyte drug enantiomer method for splitting as claimed in claim 5 is characterized in that: the described pH=4.4 that contains the acetic acid/triethylamine buffer solution of boric acid, and wherein boric acid concentration is 0.1mol/L; The concentration of L-TARTARIC ACID ester is 0.1mol/L in the described organic solvent.
8. Proprasylyte drug enantiomer method for splitting as claimed in claim 5 is characterized in that: the column temperature of control countercurrent chromatography separation column is 5 ~ 15 ℃, with the flow velocity of 0.5 ~ 2.0ml/min moving phase is pumped in the post.
9. Proprasylyte drug enantiomer method for splitting as claimed in claim 1 is characterized in that described method carries out in accordance with the following steps:
Acetic acid/the triethylamine buffer solution that (1) will contain the organic solvent of L-TARTARIC ACID ester and contain boric acid mixes according to volume ratio 1:1, and standing demix separates obtaining organic phase and water; Described organic solvent is chloroform or methylene dichloride, and the concentration of L-TARTARIC ACID ester is 0.1 ~ 0.2mol/L in the described organic solvent; The pH of the described acetic acid/triethylamine buffer solution that contains boric acid is 4.0 ~ 4.4, and the concentration of its mesoboric acid is 0.05 ~ 0.1mol/L;
(2) propranolol hydrochloride is converted into the Proprasylyte episome, and with step (1) gained aqueous phase dissolved, makes sample stand-by;
(3) adopt high-speed countercurrent chromatography to split Proprasylyte: the organic phase that obtains take step (1) respectively and water are as stationary phase and moving phase, countercurrent chromatography separation column is filled up stationary phase, column temperature is 5 ~ 15 ℃, the opening speed controller, rotating speed is 500 ~ 2000rpm, flow velocity with 0.5 ~ 2.0ml/min pumps into moving phase in the post, after two phase solvent system reaches fluid dynamic equilibrium, by the sampling valve sample introduction, UV-detector with wavelength 190 ~ 400nm detects, according to the ultraviolet detection spectrogram, collect respectively leading peak elutriant and postpeak elutriant;
(4) reclaim the monomer that obtains left-handed Proprasylyte and D-propranolol in the leading peak elutriant that collection obtains from step (3) respectively and the postpeak elutriant.
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| CN104237401A (en) * | 2014-09-02 | 2014-12-24 | 浙江工业大学 | Chiral resolution method of racemic tropine |
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| CN109839444A (en) * | 2017-11-27 | 2019-06-04 | 武汉科福新药有限责任公司 | The separation method of naphthol derivative and its application |
| CN109839444B (en) * | 2017-11-27 | 2022-03-15 | 武汉科福新药有限责任公司 | Process for separating naphthol derivatives and their use |
| CN110386879A (en) * | 2018-04-20 | 2019-10-29 | 北京睿创康泰医药研究院有限公司 | The novel crystal forms of Propranolol free alkali |
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