CN103694110A - Chiral resolution method of racemic alpha-cyclopentyl mandelic acid - Google Patents

Abstract

The invention discloses a chiral separation method of racemic α-cyclopentylmandelic acid. The method comprises the following steps: an organic solvent A, an organic solvent B and a phosphate buffer solution containing hydroxypropyl-β-cyclodextrin are mixed in a volume ratio of 1 to 9:1 to 9:10 to form a solvent system, the mixture is evenly mixed, the mixture is allowed to stand for stratification, and an organic phase and an aqueous phase are separated; the racemic α-cyclopentylmandelic acid is dissolved in the obtained organic phase to prepare a sample; the racemic α-cyclopentylmandelic acid is separated by a high-speed countercurrent chromatography method, and the mixture is recovered from a collected front peak eluate and a rear peak eluate to obtain a right-handed α-cyclopentylmandelic acid monomer and a left-handed α-cyclopentylmandelic acid monomer respectively; the invention adopts the countercurrent chromatography method for the first time to separate the racemic α-cyclopentylmandelic acid, and the method is applicable to various types of preparative countercurrent chromatographs, and the left-handed and right-handed α-cyclopentylmandelic acid monomers separated have a purity greater than 98% and a recovery rate greater than 90%.

Description

A chiral resolution method for racemic α-cyclopentylmandelic acid

(1) Technical field

The invention relates to a method for splitting a racemate, in particular to a method for separating L-α-cyclopentylmandelic acid and D-α-cyclopentylmandelic acid from racemic α-cyclopentylmandelic acid sour method.

(2) Background technology

α-cyclopentylmandelic acid is an important class of drug intermediates, which can be used to synthesize a variety of M receptor antagonists such as phencyclonyl hydrochloride, etc. The curative effect of these drugs is closely related to the stereo configuration of the chiral carbon of the tertiary hydroxyl group. Relationship. Obtaining optically pure α-cyclopentylmandelic acid is also an inevitable requirement for drug synthesis. Therefore, it is a topic of theoretical and practical significance to study the chiral resolution of racemic α-cyclopentylmandelic acid and to establish a fast and effective resolution method.

For chiral drugs, its enantiomers have the same physical and chemical properties, but usually exhibit different biological activities, and only one isomer may be effective, while the other is ineffective or even harmful, This makes obtaining pure enantiomers all the more important.

The separation of chiral compounds by chromatography is one of the important means in the field of separation science, and high speed countercurrent chromatography (HSCCC) is a new type of liquid-liquid partition chromatography developed in the 1980s. In addition to avoiding the problems of sample loss, inactivation and denaturation caused by irreversible adsorption in gas and liquid chromatography, it also has the advantages of large amount of one-time separation and preparation, simple and easy maintenance of the instrument, and low operating cost. It has unique advantages in separation and preparation.

(3) Contents of the invention

The purpose of the present invention is to provide a method for splitting racemic α-cyclopentylmandelic acid using high-speed countercurrent chromatography.

In order to achieve the above-mentioned purpose of the invention, the present invention uses racemic α-cyclopentylmandelic acid as the resolution object, uses hydroxypropyl-β-cyclodextrin as a chiral reagent, adds it to the water phase, and adopts liquid-liquid The solvent system was screened by the method of distributing chiral extraction and resolution, and L-α-cyclopentylmandelic acid and D-α-cyclopentylmandelic acid were obtained by high-speed countercurrent chromatography.

The technical scheme adopted in the present invention is:

A chiral resolution method for racemic α-cyclopentylmandelic acid, said method is carried out as follows:

(1) Combine organic solvent A, organic solvent B and phosphate buffer containing hydroxypropyl-β-cyclodextrin in a volume ratio of 1 to 9:1 to 9:10 to form a solvent system, mix well, and let stand to separate layers , liquid separation to obtain an organic phase and an aqueous phase; the degree of substitution of the hydroxypropyl-β-cyclodextrin is 4.5 to 8.0; the pH of the phosphate buffer containing the hydroxypropyl-β-cyclodextrin is 2.0～8.0; the concentration of hydroxypropyl-β-cyclodextrin in the phosphate buffer containing hydroxypropyl-β-cyclodextrin is 0.02～0.40mol/L; the organic solvent A is C1～C8 alkane; the organic solvent B is a C2-C8 ether or a C3-C8 ester;

(2) dissolving racemic α-cyclopentylmandelic acid with the organic phase obtained in step (1) to prepare a sample;

(3) Use high-speed countercurrent chromatography to separate racemic α-cyclopentylmandelic acid: the organic phase obtained in step (1) is used as the stationary phase, and the aqueous phase is used as the mobile phase, and the countercurrent chromatographic separation column is filled with the stationary phase. The column temperature is 2-35°C, the speed controller is turned on, the rotation speed is 200-2000rpm, and the mobile phase is pumped into the column at a flow rate of 0.1-3.0mL/min. After the two-phase solvent system reaches the hydrodynamic equilibrium, the When the mobile phase flows out from the outlet of the chromatographic column, the sample prepared in step (2) is injected through the injection valve, and the ultraviolet detector detects the effluent at a wavelength of 190-400nm. Collect the eluate at the gradient time interval between 30min and 200min, and according to the time period when the enantiomer double peaks appear in the ultraviolet detection spectrum, combine the eluent collected in the corresponding time period of the previous peak to obtain the target component The eluate from the front peak of the right-handed α-cyclopentylmandelic acid monomer is combined with the eluent collected during the corresponding time period of the back peak to obtain the back peak containing the target component L-cyclopentylmandelic acid monomer Eluent; The front peak refers to the main peak that comes out earlier in the enantiomer doublet, and the described back peak refers to the main peak that comes out later in the enantiomer doublet;

(4) recovering from the pre-peak eluate collected in step (3) to obtain D-α-cyclopentylmandelic acid monomer; recovering from the post-peak eluate collected in step (3), Obtain L-α-cyclopentylmandelic acid monomer.

In step (1) of the chiral resolution method for racemic α-cyclopentylmandelic acid of the present invention, preferably the organic solvent A is n-hexane, n-heptane, n-pentane, cyclohexane or petroleum ether; preferably The organic solvent B is ether, methyl tert-butyl ether, ethyl acetate, methyl acetate or ethyl formate; preferably the pH of the phosphate buffer containing hydroxypropyl-β-cyclodextrin is 2.0～4.0; preferably the concentration of hydroxypropyl-β-cyclodextrin in the phosphate buffer containing hydroxypropyl-β-cyclodextrin is 0.05～0.40mol/L; preferably the hydroxypropyl-β-cyclodextrin The substitution degree of β-cyclodextrin is 5.5-8.0.

In step (1) of the present invention, the preferred solvent system is n-hexane, methyl tert-butyl ether and phosphate buffer solution containing hydroxypropyl-β-cyclodextrin with a pH value of 2.7 at a volume ratio of 9:1 :10 mixed and made, wherein the concentration of hydroxypropyl-β-cyclodextrin in the phosphate buffer containing hydroxypropyl-β-cyclodextrin is 0.1mol/L.

In step (2) of the present invention, preferably, the concentration of racemic α-cyclopentylmandelic acid in the sample is 1-20 mg/mL.

In step (3) of the present invention, the gradient time interval is collected every 0.5-5 minutes, preferably every 2 minutes.

In step (3) of the present invention, preferably, the column temperature is 5-25° C.; the rotational speed is 800-1800 rpm; the flow rate of the mobile phase pumped into the column is 0.5-2.5 mL/min.

In step (4) of the present invention, the recovery is carried out as follows: adjust the pH value of the front peak eluent and the rear peak eluent to 1 to 2 with hydrochloric acid, and then extract 2 ~ 3 times, combined the methyl tert-butyl ether layer, washed with water until neutral, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to remove the solvent, that is, the d-α-cyclopentylmandelic acid monomer and the L-α- Cyclopentylmandelic acid monomer.

Specifically, it is recommended that the splitting method of the present invention be carried out according to the following steps:

(1) Make n-hexane, methyl tert-butyl ether and phosphate buffer solution containing hydroxypropyl-β-cyclodextrin with a pH value of 2.7 to form a solvent system according to the volume ratio of 9:1:10, mix well, and statically Set the layers, and separate the liquid to obtain the organic phase and the aqueous phase; the degree of substitution of the hydroxypropyl-β-cyclodextrin is 5.5 to 8.0; the phosphate buffer containing the hydroxypropyl-β-cyclodextrin The concentration of hydroxypropyl-β-cyclodextrin is 0.1mol/L;

(2) Dissolving racemic α-cyclopentylmandelic acid in the organic phase obtained in step (1) to prepare a sample with a concentration of 1-20 mg/mL of racemic α-cyclopentylmandelic acid;

(3) Use high-speed countercurrent chromatography to separate racemic α-cyclopentylmandelic acid: the organic phase obtained in step (1) is used as the stationary phase, and the aqueous phase is used as the mobile phase, and the countercurrent chromatographic separation column is filled with the stationary phase. The column temperature is 5-25°C, the speed controller is turned on, the rotation speed is 800-1800rpm, and the mobile phase is pumped into the column at a flow rate of 0.5-2.5mL/min. After the two-phase solvent system reaches the hydrodynamic equilibrium, the When the mobile phase flows out from the outlet of the chromatographic column, the sample prepared in step (2) is injected through the injection valve, and the ultraviolet detector detects the effluent at a wavelength of 190-400nm. Collect the eluate at every 2min time interval between 30min and 200min, and according to the time period when the enantiomer double peaks appear in the ultraviolet detection spectrum, combine the eluate collected in the corresponding time period of the previous peak to obtain the target group Separate the eluate from the front peak of the D-α-cyclopentylmandelic acid monomer, and combine the eluents collected in the corresponding time period of the rear peak to obtain the after-peak eluate containing the target component L-α-cyclopentylmandelic acid monomer. Peak eluent;

(4) recovering from the pre-peak eluate collected in step (3) to obtain D-α-cyclopentylmandelic acid monomer; recovering from the post-peak eluate collected in step (3), Obtain the L-α-cyclopentylmandelic acid monomer; the recovery method is: adjust the pH value of the front peak eluent and the rear peak eluent to 1 to 2 with hydrochloric acid, and then use methyl tert-butyl ether to Extract 2 to 3 times, combine the methyl tert-butyl ether layers, wash with water until neutral, dry over anhydrous sodium sulfate, filter, evaporate the filtrate to remove the solvent, and obtain the dextrorotatory α-cyclopentylmandelic acid monomer and the levorotatory α-cyclopentylmandelic acid monomer respectively. α-cyclopentylmandelic acid monomer.

The present invention can adopt analytical, semi-preparative and preparative countercurrent chromatographs (the volume of the analytical separation column is 20ml, the volume of the semi-preparative separation column is 300ml, and the volume of the preparative separation column is 1000ml), and the countercurrent chromatograph consists of a constant Flow pump, host (separation column), ultraviolet detector, recorder, etc.

Compared with the prior art, the beneficial effect of the present invention is mainly reflected in: the present invention uses countercurrent chromatography to split racemic α-cyclopentylmandelic acid for the first time, and this method can basically achieve the purpose of separation, and this method is applicable to various The model preparative countercurrent chromatograph can separate the monomers of L-α-cyclopentylmandelic acid and D-α-cyclopentylmandelic acid, the purity of which is greater than 98%, and the recovery rate is greater than 90%.

(4) Description of drawings

Fig. 1: Analytical high-speed countercurrent chromatogram under the experimental conditions of embodiment 1;

Figure 2: Analytical high-speed countercurrent chromatograms under the experimental conditions of Example 2;

Fig. 3: Analytical high-speed countercurrent chromatogram under the experimental conditions of embodiment 3;

Figure 4: Analytical high-speed countercurrent chromatograms under the experimental conditions of Example 4;

Figure 5: Analytical high-speed countercurrent chromatograms under the experimental conditions of Example 5;

Figure 6: Analytical high-speed countercurrent chromatograms under the experimental conditions of Example 6;

Figure 7: Analytical high-speed countercurrent chromatograms under the experimental conditions of Example 7;

Fig. 8: Analytical high-speed countercurrent chromatogram under the experimental conditions of embodiment 8;

Figure 9: Analytical high-speed countercurrent chromatograms under the experimental conditions of Example 9;

Figure 10: Analytical high-speed countercurrent chromatograms under the experimental conditions of Example 10;

Figure 11: Analytical high-speed countercurrent chromatograms under the experimental conditions of Example 11;

Fig. 12: Semi-preparative high-speed countercurrent chromatogram under the experimental conditions of embodiment 12;

Figure 13: Semi-preparative high-speed countercurrent chromatograms under the experimental conditions of Example 13;

Figure 14: The HPLC detection spectrum of the eluate from part A (98min to 130min) in Figure 13, that is, D-α-cyclopentylmandelic acid;

Figure 15: The HPLC detection spectrum of the eluate from part B (138min-170min) in Figure 13, that is, L-α-cyclopentylmandelic acid.

(5) Specific implementation methods

The present invention is further described below in conjunction with specific embodiment, but protection scope of the present invention is not limited thereto:

Example 1

(1) Mix n-hexane: methyl tert-butyl ether: phosphate buffer containing hydroxypropyl-β-cyclodextrin (prepared with 0.1mol/L phosphate buffer solution, pH=2.7, containing 0.10mol/L Hydroxypropyl-β-cyclodextrin, with a substitution degree of 7.5) was configured in a separatory funnel at a volume ratio of 8:2:10, shaken well and allowed to stand to separate layers. After equilibrating for a period of time, the upper and lower phases were separated, the organic phase was used as the stationary phase, and the aqueous phase was used as the mobile phase.

(2) Weigh 15mg of racemic α-cyclopentylmandelic acid and dissolve it in 1mL of organic phase, and make a sample solution after dissolution for use.

(3) Use an analytical high-speed countercurrent chromatograph (Shanghai Tongtian Biotechnology Co., Ltd., instrument model TBE-20A) to resolve racemic α-cyclopentylmandelic acid: the volume of the separation column is 20 mL, and the countercurrent The chromatographic separation column is filled with the stationary phase, the column temperature is 10°C, the speed controller is turned on, the rotation speed is 1800rpm, and the mobile phase is pumped into the column at a flow rate of 0.5mL/min. After the two-phase solvent system reaches the hydrodynamic equilibrium ( That is, when the mobile phase flows out from the outlet of the chromatographic column), the sample injection is started by the injection valve. Then use a UV detector with a wavelength of 254nm (Shanghai Jinda Biochemical Instrument Co., Ltd., instrument model UVD-200UV) to detect the effluent, and according to the time gradient, use an automatic partial collector (Shanghai Huxi Analytical Instrument Factory Co., Ltd., instrument model SBS-100) respectively collect the eluate at intervals of 2 minutes between 50min and 130min, and collect the corresponding time period of the front peak according to the time period when the enantiomer double peaks appear in the ultraviolet detection spectrum, that is, within 50-86min The eluents of the eluents were combined to obtain the eluent of the front peak containing the target component D-α-cyclopentylmandelic acid monomer, and the eluents collected within the corresponding time period of the rear peak, that is, 85-130min, were combined to obtain the eluent containing The eluent of the back peak of the target component L-α-cyclopentylmandelic acid monomer. According to the ultraviolet detection spectrum (shown in Figure 1), the calculated resolution is 0.91. The pre-peak eluate containing the target component D-α-cyclopentylmandelic acid monomer and the post-peak eluate containing the target component L-α-cyclopentylmandelic acid monomer were detected by high performance liquid phase. The purity of each monomer eluate was greater than 98%. The detection conditions of high performance liquid chromatography are: J&KCHEMICAL C ₁₈ (5μm, 150mm×4.6mm) chromatographic column; column temperature: 25°C; mobile phase: 24% ethanol water (water pH2.68, phosphate buffer adjusted and containing 9.5 mmol·L ^-1 hydroxypropyl-β-cyclodextrin): acetonitrile (95:5, v/v) isocratic elution; flow rate 0.6mL/min; detection wavelength 220nm; injection volume: 20μL.

Example 2

(1) Mix n-hexane: methyl tert-butyl ether: phosphate buffer containing hydroxypropyl-β-cyclodextrin (prepared with 0.1mol/L phosphate buffer solution, pH=2.7, containing 0.10mol/L Hydroxypropyl-β-cyclodextrin, with a substitution degree of 7.5) was configured in a separatory funnel at a volume ratio of 9:1:10, shaken well and allowed to stand to separate layers. After equilibrating for a period of time, the upper and lower phases were separated, the organic phase was used as the stationary phase, and the aqueous phase was used as the mobile phase.

(2) Weigh 15mg of racemic α-cyclopentylmandelic acid and dissolve it in 1mL of the organic phase, and make a sample solution after dissolution, ready for use.

(3) Use an analytical high-speed countercurrent chromatograph (Shanghai Tongtian Biotechnology Co., Ltd., instrument model TBE-20A) to resolve racemic α-cyclopentylmandelic acid: the volume of the separation column is 20 mL, and the countercurrent The chromatographic separation column is filled with the stationary phase, the column temperature is 10°C, the speed controller is turned on, the rotation speed is 1800rpm, and the mobile phase is pumped into the column at a flow rate of 0.5mL/min. After the two-phase solvent system reaches the hydrodynamic equilibrium ( That is, when the mobile phase flows out from the outlet of the chromatographic column), the sample injection is started by the injection valve. Then use a UV detector with a wavelength of 254nm (Shanghai Jinda Biochemical Instrument Co., Ltd., instrument model UVD-200UV) to detect the effluent, and according to the time gradient, use an automatic partial collector (Shanghai Huxi Analytical Instrument Factory Co., Ltd., instrument model SBS-100) respectively collect the eluate at intervals of 2 minutes between 30 minutes and 65 minutes, and collect the corresponding time period of the front peak according to the time period when the enantiomer double peaks appear in the ultraviolet detection spectrum, that is, within 30 to 46 minutes The eluents of the eluents were combined to obtain the eluent of the front peak containing the target component D-α-cyclopentylmandelic acid monomer, and the eluents collected within the corresponding time period of the rear peak, that is, 46-65min, were combined to obtain the eluent containing The eluent of the back peak of the target component L-α-cyclopentylmandelic acid monomer. According to the ultraviolet detection spectrum (shown in Figure 2), the calculated resolution is 0.78. The pre-peak eluate containing the target component D-α-cyclopentylmandelic acid monomer and the post-peak eluate containing the target component L-α-cyclopentylmandelic acid monomer were detected by high performance liquid phase. The purity of each monomer eluate was greater than 98%. The detection condition of high performance liquid chromatography is the same as embodiment 1.

Example 3

(1) Mix n-hexane: ethyl acetate: phosphate buffer containing hydroxypropyl-β-cyclodextrin (prepared with 0.1mol/L phosphate buffer solution, pH=2.7, containing 0.10mol/L hydroxypropyl -β-cyclodextrin, the degree of substitution is 7.5) was arranged in a separatory funnel according to the volume ratio of 7:3:10, shaken well, and then stood to separate layers. After equilibrating for a period of time, the upper and lower phases were separated, the organic phase was used as the stationary phase, and the aqueous phase was used as the mobile phase.

(2) Weigh 15mg of racemic α-cyclopentylmandelic acid and dissolve it in 1mL of the organic phase, and make a sample solution after dissolution, ready for use.

(3) Use an analytical high-speed countercurrent chromatograph (Shanghai Tongtian Biotechnology Co., Ltd., instrument model TBE-20A) to resolve racemic α-cyclopentylmandelic acid: the volume of the separation column is 20 mL, and the countercurrent The chromatographic separation column is filled with the stationary phase, the column temperature is 10°C, the speed controller is turned on, the rotation speed is 1800rpm, and the mobile phase is pumped into the column at a flow rate of 0.5mL/min. After the two-phase solvent system reaches the hydrodynamic equilibrium ( That is, when the mobile phase flows out from the outlet of the chromatographic column), the sample injection is started by the injection valve. Then use a UV detector with a wavelength of 254nm (Shanghai Jinda Biochemical Instrument Co., Ltd., instrument model UVD-200UV) to detect the effluent, and according to the time gradient, use an automatic partial collector (Shanghai Huxi Analytical Instrument Factory Co., Ltd., instrument model SBS-100) respectively collect the eluate at intervals of 2 minutes between 50min and 100min, and collect the corresponding time period of the front peak according to the time period when the enantiomer double peaks appear in the ultraviolet detection spectrum, that is, within 50-75min The eluents of the eluents were combined to obtain the pre-peak eluent containing the target component D-α-cyclopentylmandelic acid monomer, and the eluents collected within the corresponding time period of the rear peak, that is, 75-100min, were combined to obtain the eluent containing The eluent of the back peak of the target component L-α-cyclopentylmandelic acid monomer. According to the ultraviolet detection spectrum (shown in Figure 3), the calculated resolution is 0.93. The pre-peak eluate containing the target component D-α-cyclopentylmandelic acid monomer and the post-peak eluate containing the target component L-α-cyclopentylmandelic acid monomer were detected by high performance liquid phase. The purity of each monomer eluate was greater than 98%. The detection condition of high performance liquid chromatography is the same as embodiment 1.

Example 4

(1) Mix n-hexane: methyl tert-butyl ether: phosphate buffer containing hydroxypropyl-β-cyclodextrin (prepared with 0.1mol/L phosphate buffer solution, pH=2.7, containing 0.10mol/L Hydroxypropyl-β-cyclodextrin, with a substitution degree of 7.5) was configured in a separatory funnel at a volume ratio of 8.5:1.5:10, shaken up and allowed to stand to separate layers. After equilibrating for a period of time, the upper and lower phases were separated, the organic phase was used as the stationary phase, and the aqueous phase was used as the mobile phase.

(2) Weigh 15mg of racemic α-cyclopentylmandelic acid and dissolve it in 1mL of the organic phase, and make a sample solution after dissolution, ready for use.

(3) Use an analytical high-speed countercurrent chromatograph (Shanghai Tongtian Biotechnology Co., Ltd., instrument model TBE-20A) to resolve racemic α-cyclopentylmandelic acid: the volume of the separation column is 20 mL, and the countercurrent The chromatographic separation column is filled with the stationary phase, the column temperature is 10°C, the speed controller is turned on, the rotation speed is 1800rpm, and the mobile phase is pumped into the column at a flow rate of 0.5mL/min. After the two-phase solvent system reaches the hydrodynamic equilibrium ( That is, when the mobile phase flows out from the outlet of the chromatographic column), the sample injection is started by the injection valve. Then use a UV detector with a wavelength of 254nm (Shanghai Jinda Biochemical Instrument Co., Ltd., instrument model UVD-200UV) to detect the effluent, and according to the time gradient, use an automatic partial collector (Shanghai Huxi Analytical Instrument Factory Co., Ltd., instrument model SBS-100) respectively collect the eluate at intervals of 2 minutes between 35min and 75min, and collect the corresponding time period of the front peak according to the time period when the enantiomer double peaks appear in the ultraviolet detection spectrum, that is, within 35-54min The eluents of the eluents were combined to obtain the eluent of the front peak containing the target component D-α-cyclopentylmandelic acid monomer, and the eluents collected within the corresponding time period of the rear peak, that is, 54 to 75 minutes, were combined to obtain the eluent containing The eluent of the back peak of the target component L-α-cyclopentylmandelic acid monomer. According to the ultraviolet detection spectrum (shown in Figure 4), the calculated resolution is 0.86. The pre-peak eluate containing the target component D-α-cyclopentylmandelic acid monomer and the post-peak eluate containing the target component L-α-cyclopentylmandelic acid monomer were detected by high performance liquid phase. The purity of each monomer eluate was greater than 98%. The detection condition of high performance liquid chromatography is the same as embodiment 1.

Example 5

(1) Mix n-hexane: methyl tert-butyl ether: phosphate buffer containing hydroxypropyl-β-cyclodextrin (prepared with 0.1mol/L phosphate buffer solution, pH=2.7, containing 0.10mol/L Hydroxypropyl-β-cyclodextrin, with a substitution degree of 7.5) was configured in a separatory funnel at a volume ratio of 8.5:1.5:10, shaken up and allowed to stand to separate layers. After equilibrating for a period of time, the upper and lower phases were separated, the organic phase was used as the stationary phase, and the aqueous phase was used as the mobile phase.

(2) Weigh 15mg of racemic α-cyclopentylmandelic acid and dissolve it in 1mL of the organic phase, and make a sample solution after dissolution, ready for use.

(3) Use an analytical high-speed countercurrent chromatograph (Shanghai Tongtian Biotechnology Co., Ltd., instrument model TBE-20A) to resolve racemic α-cyclopentylmandelic acid: the volume of the separation column is 20 mL, and the countercurrent The chromatographic separation column is filled with the stationary phase, the column temperature is 20°C, the speed controller is turned on, the rotation speed is 1800rpm, and the mobile phase is pumped into the column at a flow rate of 0.5mL/min. After the two-phase solvent system reaches the hydrodynamic equilibrium ( That is, when the mobile phase flows out from the outlet of the chromatographic column), the sample injection is started by the injection valve. Then use a UV detector with a wavelength of 254nm (Shanghai Jinda Biochemical Instrument Co., Ltd., instrument model UVD-200UV) to detect the effluent, and according to the time gradient, use an automatic partial collector (Shanghai Huxi Analytical Instrument Factory Co., Ltd., instrument model SBS-100) respectively collect the eluate at intervals of 2 minutes between 40min and 90min, and collect the corresponding time period of the front peak according to the time period when the enantiomer double peaks appear in the ultraviolet detection spectrum, that is, within 40-64min The eluents of the eluents were combined to obtain the eluent of the front peak containing the target component D-α-cyclopentylmandelic acid monomer, and the eluents collected within the corresponding time period of the rear peak, that is, 64-90min, were combined to obtain the eluent containing The eluent of the back peak of the target component L-α-cyclopentylmandelic acid monomer. According to the ultraviolet detection spectrum (shown in Figure 5), the calculated resolution is 0.87. The pre-peak eluate containing the target component D-α-cyclopentylmandelic acid monomer and the post-peak eluate containing the target component L-α-cyclopentylmandelic acid monomer were detected by high performance liquid phase. The purity of each monomer eluate was greater than 98%. The detection condition of high performance liquid chromatography is the same as embodiment 1.

Example 6

(1) Mix n-hexane: methyl tert-butyl ether: phosphate buffer solution containing hydroxypropyl-β-cyclodextrin (prepared with 0.1mol/L phosphate buffer solution, pH=2.5, containing 0.10mol/L Hydroxypropyl-β-cyclodextrin, the degree of substitution is 8.0) is arranged in a separatory funnel according to the volume ratio of 8.5:1.5:10, shake well and then stand to separate layers. After equilibrating for a period of time, the upper and lower phases were separated, the organic phase was used as the stationary phase, and the aqueous phase was used as the mobile phase.

(2) Weigh 15mg of racemic α-cyclopentylmandelic acid and dissolve it in 1mL of the organic phase, and make a sample solution after dissolution, ready for use.

(3) Use an analytical high-speed countercurrent chromatograph (Shanghai Tongtian Biotechnology Co., Ltd., instrument model TBE-20A) to resolve racemic α-cyclopentylmandelic acid: the volume of the separation column is 20 mL, and the countercurrent The chromatographic separation column is filled with the stationary phase, the column temperature is 10°C, the speed controller is turned on, the rotation speed is 1800rpm, and the mobile phase is pumped into the column at a flow rate of 0.5mL/min. After the two-phase solvent system reaches the hydrodynamic equilibrium ( That is, when the mobile phase flows out from the outlet of the chromatographic column), the sample injection is started by the injection valve. Then use a UV detector with a wavelength of 254nm (Shanghai Jinda Biochemical Instrument Co., Ltd., instrument model UVD-200UV) to detect the effluent, and according to the time gradient, use an automatic partial collector (Shanghai Huxi Analytical Instrument Factory Co., Ltd., instrument model SBS-100) respectively collect the eluate at intervals of 2 minutes between 40min and 90min, and collect the corresponding time period of the front peak according to the time period when the enantiomer double peaks appear in the ultraviolet detection spectrum, that is, within 40-62min The eluents of the eluents were combined to obtain the pre-peak eluent containing the target component D-α-cyclopentylmandelic acid monomer, and the eluents collected within the corresponding time period of the rear peak, that is, 62-90min, were combined to obtain the eluent containing The eluent of the back peak of the target component L-α-cyclopentylmandelic acid monomer. According to the ultraviolet detection spectrum (shown in Figure 6), the calculated resolution is 0.90. The pre-peak eluate containing the target component D-α-cyclopentylmandelic acid monomer and the post-peak eluate containing the target component L-α-cyclopentylmandelic acid monomer were detected by high performance liquid phase. The purity of each monomer eluate was greater than 98%. The detection condition of high performance liquid chromatography is the same as embodiment 1.

Example 7

(1) Mix n-hexane: methyl tert-butyl ether: phosphate buffer solution containing hydroxypropyl-β-cyclodextrin (prepared with 0.1mol/L phosphate buffer solution, pH=3.5, containing 0.10mol/L Hydroxypropyl-β-cyclodextrin, with a substitution degree of 7.5) was configured in a separatory funnel at a volume ratio of 8.5:1.5:10, shaken up and allowed to stand to separate layers. After equilibrating for a period of time, the upper and lower phases were separated, the organic phase was used as the stationary phase, and the aqueous phase was used as the mobile phase.

(2) Weigh 15mg of racemic α-cyclopentylmandelic acid and dissolve it in 1mL of the organic phase, and make a sample solution after dissolution, ready for use.

(3) Use an analytical high-speed countercurrent chromatograph (Shanghai Tongtian Biotechnology Co., Ltd., instrument model TBE-20A) to resolve racemic α-cyclopentylmandelic acid: the volume of the separation column is 20 mL, and the countercurrent The chromatographic separation column is filled with the stationary phase, the column temperature is 10°C, the speed controller is turned on, the rotation speed is 1800rpm, and the mobile phase is pumped into the column at a flow rate of 0.5mL/min. After the two-phase solvent system reaches the hydrodynamic equilibrium ( That is, when the mobile phase flows out from the outlet of the chromatographic column), the sample injection is started by the injection valve. Then use a UV detector with a wavelength of 254nm (Shanghai Jinda Biochemical Instrument Co., Ltd., instrument model UVD-200UV) to detect the effluent, and according to the time gradient, use an automatic partial collector (Shanghai Huxi Analytical Instrument Factory Co., Ltd., instrument model SBS-100) respectively collect the eluate at intervals of 2 minutes between 35min and 80min, and collect the corresponding time period of the front peak according to the time period when the enantiomer double peaks appear in the ultraviolet detection spectrum, that is, within 35-57min The eluents of the eluents were combined to obtain the pre-peak eluent containing the target component D-α-cyclopentylmandelic acid monomer, and the eluents collected within the corresponding time period of the rear peak, that is, 57-80min, were combined to obtain the eluent containing The eluent of the back peak of the target component L-α-cyclopentylmandelic acid monomer. According to the ultraviolet detection spectrum (shown in Figure 7), the calculated resolution is 0.83. The pre-peak eluate containing the target component D-α-cyclopentylmandelic acid monomer and the post-peak eluate containing the target component L-α-cyclopentylmandelic acid monomer were detected by high performance liquid phase. The purity of each monomer eluate was greater than 98%. The detection condition of high performance liquid chromatography is the same as embodiment 1.

Example 8

(1) Mix n-hexane: methyl tert-butyl ether: phosphate buffer containing hydroxypropyl-β-cyclodextrin (prepared with 0.1mol/L phosphate buffer solution, pH=2.7, containing 0.05mol/L Hydroxypropyl-β-cyclodextrin, with a substitution degree of 7.5) was configured in a separatory funnel at a volume ratio of 8.5:1.5:10, shaken up and allowed to stand to separate layers. After equilibrating for a period of time, the upper and lower phases were separated, the organic phase was used as the stationary phase, and the aqueous phase was used as the mobile phase.

(2) Weigh 15mg of racemic α-cyclopentylmandelic acid and dissolve it in 1mL of the organic phase, and make a sample solution after dissolution, ready for use.

(3) Use an analytical high-speed countercurrent chromatograph (Shanghai Tongtian Biotechnology Co., Ltd., instrument model TBE-20A) to resolve racemic α-cyclopentylmandelic acid: the volume of the separation column is 20 mL, and the countercurrent The chromatographic separation column is filled with the stationary phase, the column temperature is 10°C, the speed controller is turned on, the rotation speed is 1800rpm, and the mobile phase is pumped into the column at a flow rate of 0.5mL/min. After the two-phase solvent system reaches the hydrodynamic equilibrium ( That is, when the mobile phase flows out from the outlet of the chromatographic column), the sample injection is started by the injection valve. Then use a UV detector with a wavelength of 254nm (Shanghai Jinda Biochemical Instrument Co., Ltd., instrument model UVD-200UV) to detect the effluent, and according to the time gradient, use an automatic partial collector (Shanghai Huxi Analytical Instrument Factory Co., Ltd., instrument model SBS-100) respectively collect the eluate at intervals of 2 minutes between 60min and 150min, and collect the corresponding time period of the front peak according to the time period when the enantiomer double peaks appear in the ultraviolet detection spectrum, that is, within 60-101min The eluents of the eluents were combined to obtain the pre-peak eluate containing the target component D-α-cyclopentylmandelic acid monomer, and the eluents collected within the corresponding time period of the rear peak were combined to obtain the eluent containing The eluent of the back peak of the target component L-α-cyclopentylmandelic acid monomer. According to the ultraviolet detection spectrum (shown in Figure 8), the calculated resolution is 0.95. The pre-peak eluate containing the target component D-α-cyclopentylmandelic acid monomer and the post-peak eluate containing the target component L-α-cyclopentylmandelic acid monomer were detected by high performance liquid phase. The purity of each monomer eluate was greater than 98%. The detection condition of high performance liquid chromatography is the same as embodiment 1.

Example 9

(1) Mix n-hexane: methyl tert-butyl ether: phosphate buffer containing hydroxypropyl-β-cyclodextrin (prepared with 0.1mol/L phosphate buffer solution, pH=2.7, containing 0.15mol/L Hydroxypropyl-β-cyclodextrin, the degree of substitution is 6.0) is arranged in a separating funnel according to the volume ratio of 8.5:1.5:10, shake well and then stand to separate layers. After equilibrating for a period of time, the upper and lower phases were separated, the organic phase was used as the stationary phase, and the aqueous phase was used as the mobile phase.

(2) Weigh 15mg of racemic α-cyclopentylmandelic acid and dissolve it in 1mL of the organic phase, and make a sample solution after dissolution, ready for use.

(3) Use an analytical high-speed countercurrent chromatograph (Shanghai Tongtian Biotechnology Co., Ltd., instrument model TBE-20A) to resolve racemic α-cyclopentylmandelic acid: the volume of the separation column is 20 mL, and the countercurrent The chromatographic separation column is filled with the stationary phase, the column temperature is 10°C, the speed controller is turned on, the rotation speed is 1800rpm, and the mobile phase is pumped into the column at a flow rate of 0.5mL/min. After the two-phase solvent system reaches the hydrodynamic equilibrium ( That is, when the mobile phase flows out from the outlet of the chromatographic column), the sample injection is started by the injection valve. Then use a UV detector with a wavelength of 254nm (Shanghai Jinda Biochemical Instrument Co., Ltd., instrument model UVD-200UV) to detect the effluent, and according to the time gradient, use an automatic partial collector (Shanghai Huxi Analytical Instrument Factory Co., Ltd., instrument model SBS-100) respectively collect the eluate at intervals of 2 minutes between 30 minutes and 70 minutes, and collect the corresponding time period of the front peak according to the time period when the enantiomer double peaks appear in the ultraviolet detection spectrum, that is, within 30 to 50 minutes The eluents of the eluents were combined to obtain the pre-peak eluate containing the target component D-α-cyclopentylmandelic acid monomer, and the eluents collected within the corresponding time period of the rear peak, that is, 50-70min, were combined to obtain the eluent containing The eluent of the back peak of the target component L-α-cyclopentylmandelic acid monomer. According to the ultraviolet detection spectrum (shown in Figure 9), the calculated resolution is 0.78. The pre-peak eluate containing the target component D-α-cyclopentylmandelic acid monomer and the post-peak eluate containing the target component L-α-cyclopentylmandelic acid monomer were detected by high performance liquid phase. The purity of each monomer eluate was greater than 98%. The detection condition of high performance liquid chromatography is the same as embodiment 1.

Example 10

(1) Mix n-hexane: methyl tert-butyl ether: phosphate buffer containing hydroxypropyl-β-cyclodextrin (prepared with 0.1mol/L phosphate buffer solution, pH=2.7, containing 0.15mol/L Hydroxypropyl-β-cyclodextrin, with a substitution degree of 7.5) was configured in a separatory funnel at a volume ratio of 8.5:1.5:10, shaken up and allowed to stand to separate layers. After equilibrating for a period of time, the upper and lower phases were separated, the organic phase was used as the stationary phase, and the aqueous phase was used as the mobile phase.

(2) Weigh 1mg of racemic α-cyclopentylmandelic acid and dissolve it in 1mL of the organic phase, and make a sample solution after dissolution, ready for use.

(3) Use an analytical high-speed countercurrent chromatograph (Shanghai Tongtian Biotechnology Co., Ltd., instrument model TBE-20A) to resolve racemic α-cyclopentylmandelic acid: the volume of the separation column is 20 mL, and the countercurrent The chromatographic separation column is filled with the stationary phase, the column temperature is 10°C, the speed controller is turned on, the rotation speed is 1800rpm, and the mobile phase is pumped into the column at a flow rate of 0.5mL/min. After the two-phase solvent system reaches the hydrodynamic equilibrium ( That is, when the mobile phase flows out from the outlet of the chromatographic column), the sample injection is started by the injection valve. Then use a UV detector with a wavelength of 254nm (Shanghai Jinda Biochemical Instrument Co., Ltd., instrument model UVD-200UV) to detect the effluent, and according to the time gradient, use an automatic partial collector (Shanghai Huxi Analytical Instrument Factory Co., Ltd., instrument model SBS-100) respectively collect the eluate at intervals of 2 minutes between 40min and 80min, and collect the corresponding time period of the front peak according to the time period when the enantiomer double peaks appear in the ultraviolet detection spectrum, that is, within 40-60min The eluents of the eluents were combined to obtain the eluent of the front peak containing the target component D-α-cyclopentylmandelic acid monomer, and the eluents collected within the corresponding time period of the rear peak, that is, 60-80min, were combined to obtain the eluent containing The eluent of the back peak of the target component L-α-cyclopentylmandelic acid monomer. According to the ultraviolet detection spectrum (shown in Figure 10), the eluate from the front peak containing the target component D-α-cyclopentylmandelic acid monomer and the target component containing L-α-cyclopentylmandelic acid monomer The post-peak eluate was detected by high performance liquid phase, and the purity of the two monomer eluates was greater than 98%. The detection condition of high performance liquid chromatography is the same as embodiment 1.

Example 11

(1) Mix n-hexane: methyl tert-butyl ether: phosphate buffer solution containing hydroxypropyl-β-cyclodextrin (prepared with 0.1mol/L phosphate buffer solution, pH=2.7, containing 0.35mol/L Hydroxypropyl-β-cyclodextrin, with a substitution degree of 7.5) was configured in a separatory funnel at a volume ratio of 8.5:1.5:10, shaken up and allowed to stand to separate layers. After equilibrating for a period of time, the upper and lower phases were separated, the organic phase was used as the stationary phase, and the aqueous phase was used as the mobile phase.

(2) Weigh 15mg of racemic α-cyclopentylmandelic acid and dissolve it in 1mL of the organic phase, and make a sample solution after dissolution, ready for use.

(3) Use an analytical high-speed countercurrent chromatograph (Shanghai Tongtian Biotechnology Co., Ltd., instrument model TBE-20A) to resolve racemic α-cyclopentylmandelic acid: the volume of the separation column is 20 mL, and the countercurrent The chromatographic separation column is filled with the stationary phase, the column temperature is 10°C, the speed controller is turned on, the rotation speed is 1800rpm, and the mobile phase is pumped into the column at a flow rate of 0.5mL/min. After the two-phase solvent system reaches the hydrodynamic equilibrium ( That is, when the mobile phase flows out from the outlet of the chromatographic column), the sample injection is started by the injection valve. Then use a UV detector with a wavelength of 254nm (Shanghai Jinda Biochemical Instrument Co., Ltd., instrument model UVD-200UV) to detect the effluent, and according to the time gradient, use an automatic partial collector (Shanghai Huxi Analytical Instrument Factory Co., Ltd., instrument model SBS-100) respectively collect the eluate at intervals of 2 minutes between 30 minutes and 50 minutes, and collect the corresponding time period of the front peak according to the time period when the enantiomer double peaks appear in the ultraviolet detection spectrum, that is, within 30 to 38 minutes The eluents of the eluents were combined to obtain the pre-peak eluent containing the target component D-α-cyclopentylmandelic acid monomer, and the eluents collected within the corresponding time period of the rear peak, that is, 38-50min, were combined to obtain the eluent containing The eluent of the back peak of the target component L-α-cyclopentylmandelic acid monomer. According to the ultraviolet detection spectrum (shown in Figure 11), the calculated resolution is 0.56. The pre-peak eluate containing the target component D-α-cyclopentylmandelic acid monomer and the post-peak eluate containing the target component L-α-cyclopentylmandelic acid monomer were detected by high performance liquid phase. The purity of each monomer eluate was greater than 98%. The detection condition of high performance liquid chromatography is the same as embodiment 1.

Example 12

(1) Mix n-hexane: methyl tert-butyl ether: phosphate buffer containing hydroxypropyl-β-cyclodextrin (prepared with 0.1mol/L phosphate buffer solution, pH=2.7, containing 0.1mol/L Hydroxypropyl-β-cyclodextrin, with a substitution degree of 7.5) was configured in a separatory funnel at a volume ratio of 8.5:1.5:10, shaken up and allowed to stand to separate layers. After equilibrating for a period of time, the upper and lower phases were separated, the organic phase was used as the stationary phase, and the aqueous phase was used as the mobile phase.

(2) Weigh 150mg of racemic α-cyclopentylmandelic acid and dissolve it in 8mL of organic phase, and make a sample solution after dissolution, ready for use.

(3) Use semi-preparative high-speed countercurrent chromatography (Shanghai Tongtian Biotechnology Co., Ltd., instrument model TBE-200V) to separate racemic α-cyclopentylmandelic acid: the volume of the separation column is 200mL. The countercurrent chromatographic separation column is filled with stationary phase, the column temperature is 10°C, the speed controller is turned on, the rotation speed is 800rpm, and the mobile phase is pumped into the column at a flow rate of 2.0mL/min. After the two-phase solvent system reaches hydrodynamic equilibrium (that is, when the mobile phase flows out from the outlet of the chromatographic column), the injection valve starts to inject the sample. Then use a UV detector with a wavelength of 254nm (Shanghai Jinda Biochemical Instrument Co., Ltd., instrument model UVD-200UV) to detect the effluent, and according to the time gradient, use an automatic partial collector (Shanghai Huxi Analytical Instrument Factory Co., Ltd., instrument model SBS-100) respectively collect the eluate at intervals of 2 minutes between 90min and 160min, and collect the corresponding time period of the front peak according to the time period when the enantiomer double peaks appear in the ultraviolet detection spectrum, that is, within 90-125min The eluents of the eluents were combined to obtain the pre-peak eluate containing the target component D-α-cyclopentylmandelic acid monomer, and the eluents collected within the corresponding time period of the rear peak, that is, 125-160min, were combined to obtain the eluent containing The eluent of the back peak of the target component L-α-cyclopentylmandelic acid monomer. According to the ultraviolet detection spectrum (shown in Figure 12), the calculated resolution is 0.99. The pre-peak eluate containing the target component D-α-cyclopentylmandelic acid monomer and the post-peak eluate containing the target component L-α-cyclopentylmandelic acid monomer were detected by high performance liquid phase. The purity of each monomer eluate was greater than 98%. The detection condition of high performance liquid chromatography is the same as embodiment 1.

Recover the sample from the eluent: adjust the pH of the eluent from the front peak and the eluent from the back peak to 1-2 with hydrochloric acid, then extract 2-3 times with methyl tert-butyl ether, and combine the methyl tert-butyl The ether layer was washed with water to neutrality, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to remove the solvent to obtain D-α-cyclopentyl mandelic acid monomer and L-α-cyclopentyl monomer respectively. D-α-cyclopentylmandelic acid monomer has a purity of 99.6% and a recovery rate of 92.9%, and a L-α-cyclopentylmandelic acid monomer has a purity of 99.5% and a recovery rate of 93.4%.

Example 13

(1) Mix n-hexane: methyl tert-butyl ether: phosphate buffer containing hydroxypropyl-β-cyclodextrin (prepared with 0.1mol/L phosphate buffer solution, pH=2.7, containing 0.1mol/L Hydroxypropyl-β-cyclodextrin, with a substitution degree of 7.5) was configured in a separatory funnel at a volume ratio of 8.5:1.5:10, shaken up and allowed to stand to separate layers. After equilibrating for a period of time, the upper and lower phases were separated, the organic phase was used as the stationary phase, and the aqueous phase was used as the mobile phase.

(2) Weigh 200mg of racemic α-cyclopentylmandelic acid and dissolve it in 10mL of organic phase, and make a sample solution after dissolution, ready for use.

(3) Use semi-preparative high-speed countercurrent chromatography (Shanghai Tongtian Biotechnology Co., Ltd., instrument model TBE-200V) to separate racemic α-cyclopentylmandelic acid: the volume of the separation column is 200mL. The countercurrent chromatographic separation column is filled with stationary phase, the column temperature is 10°C, the speed controller is turned on, the rotation speed is 800rpm, and the mobile phase is pumped into the column at a flow rate of 2.0mL/min. After the two-phase solvent system reaches hydrodynamic equilibrium (that is, when the mobile phase flows out from the outlet of the chromatographic column), the injection valve starts to inject the sample. Then use a UV detector with a wavelength of 254nm (Shanghai Jinda Biochemical Instrument Co., Ltd., instrument model UVD-200UV) to detect the effluent, and according to the time gradient, use an automatic partial collector (Shanghai Huxi Analytical Instrument Factory Co., Ltd., instrument model SBS-100) respectively collect the eluate at intervals of 2 minutes between 90min and 180min, and collect the corresponding time period of the front peak according to the time period when the enantiomer double peaks appear in the ultraviolet detection spectrum, that is, 90-132min The eluents of the eluents were combined to obtain the pre-peak eluate containing the target component D-α-cyclopentylmandelic acid monomer, and the eluents collected within the corresponding time period of the rear peak, that is, 132-180min, were combined to obtain the eluent containing The eluent of the back peak of the target component L-α-cyclopentylmandelic acid monomer. According to the ultraviolet detection spectrum (shown in Figure 13), the calculated resolution is 0.98. The pre-peak eluate containing the target component D-α-cyclopentylmandelic acid monomer and the post-peak eluate containing the target component L-α-cyclopentylmandelic acid monomer were subjected to high performance liquid phase detection (high efficiency The liquid chromatograms are shown in Figure 14 and Figure 15 respectively), and the purity of the two monomer eluents are both greater than 98%. The detection condition of high performance liquid chromatography is the same as embodiment 1.

Recover the sample from the eluent: adjust the pH of the eluent from the front peak and the eluent from the back peak to 1-2 with hydrochloric acid, then extract 2-3 times with methyl tert-butyl ether, and combine the methyl tert-butyl The ether layer was washed with water to neutrality, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to remove the solvent to obtain D-α-cyclopentyl mandelic acid monomer and L-α-cyclopentyl monomer respectively. The purity of D-α-cyclopentylmandelic acid monomer is 99.4%, and the recovery rate is 93.6%; the purity of L-α-cyclopentylmandelic acid monomer is 99.7%, and the recovery rate is 91.4%.

Claims (10)

1. a chiral separation method for racemic ' alpha '-cyclopentyl amygdalic acid, is characterized in that described method carries out as follows:

(1) organic solvent A, organic solvent B and the phosphate buffered saline buffer that contains hydroxypropyl-beta-cyclodextrin are formed to solvent system according to volume ratio 1～9:1～9:10, mix, stratification, separatory obtains organic phase and water; The substitution value of described hydroxypropyl-beta-cyclodextrin is 4.5～8.0; The pH value of the described phosphate buffered saline buffer that contains hydroxypropyl-beta-cyclodextrin is 2.0～8.0; In the described phosphate buffered saline buffer that contains hydroxypropyl-beta-cyclodextrin, the concentration of hydroxypropyl-beta-cyclodextrin is 0.02～0.40mol/L; Described organic solvent A is the alkane of C1～C8; Described organic solvent B is the ester class of ether or the C3～C8 of C2～C8;

(2) organic phase of racemic ' alpha '-cyclopentyl step (1) gained for amygdalic acid is dissolved, make sample;

(3) adopt high-speed countercurrent chromatography resolution of racemic α-cyclopentyl amygdalic acid: the organic phase that the step (1) of take obtains is stationary phase, water is moving phase, countercurrent chromatography separation column is filled up to stationary phase, column temperature is 2～35 ℃, opening speed controller, rotating speed is 200～2000rpm, flow velocity with 0.1～3.0mL/min pumps into moving phase in post, after two phase solvent system reaches fluid dynamic equilibrium, when moving phase flows out from chromatographic column exit, the sample that step (2) is made is by sampling valve sample introduction, UV-detector detects effluent liquid under wavelength 190～400nm, according to time gradient, with auto partial sampler, collect respectively the elutriant of 30min to the gradient timed interval between 200min, and according to the time period of the bimodal appearance of enantiomorph in ultraviolet detection spectrogram, the elutriant that leading peak was collected in the corresponding time period merges and obtains the leading peak elutriant containing target components dextrorotation α-cyclopentyl amygdalic acid monomer, the elutriant that postpeak was collected in the corresponding time period merges and obtains the postpeak elutriant containing the left-handed α-cyclopentyl of target components amygdalic acid monomer, described leading peak refers to the bimodal middle elder generation of enantiomorph main peak out, described postpeak refer to enantiomorph bimodal in after main peak out,

(4) from step (3), collect in the leading peak elutriant obtaining and reclaim, obtain dextrorotation α-cyclopentyl amygdalic acid monomer; From step (3), collect in the postpeak elutriant obtaining and reclaim, obtain left-handed α-cyclopentyl amygdalic acid monomer.

2. the chiral separation method of racemic ' alpha '-cyclopentyl amygdalic acid as claimed in claim 1, is characterized in that, in step (1), described organic solvent A is normal hexane, normal heptane, Skellysolve A, hexanaphthene or sherwood oil.

3. the chiral separation method of racemic ' alpha '-cyclopentyl amygdalic acid as claimed in claim 1, is characterized in that, in step (1), described organic solvent B is ether, methyl tertiary butyl ether, ethyl acetate, methyl acetate or ethyl formate.

4. the chiral separation method of racemic ' alpha '-cyclopentyl amygdalic acid as claimed in claim 1, is characterized in that, in step (1), the pH value of the described phosphate buffered saline buffer that contains hydroxypropyl-beta-cyclodextrin is 2.0～4.0.

5. the chiral separation method of racemic ' alpha '-cyclopentyl amygdalic acid as claimed in claim 1, it is characterized in that, in step (1), in the described phosphate buffered saline buffer that contains hydroxypropyl-beta-cyclodextrin, the concentration of hydroxypropyl-beta-cyclodextrin is 0.05～0.40mol/L.

6. the chiral separation method of racemic ' alpha '-cyclopentyl amygdalic acid as claimed in claim 1, is characterized in that, in step (1), the substitution value of described hydroxypropyl-beta-cyclodextrin is 5.5～8.0.

7. the chiral separation method of racemic ' alpha '-cyclopentyl amygdalic acid as claimed in claim 1, it is characterized in that in step (1), described solvent system is that the phosphate buffered saline buffer containing hydroxypropyl-beta-cyclodextrin that normal hexane, methyl tertiary butyl ether and pH value are 2.7 is mixed with volume ratio 9:1:10, and the described concentration containing hydroxypropyl-beta-cyclodextrin in the phosphate buffered saline buffer of hydroxypropyl-beta-cyclodextrin is 0.1mol/L.

8. the chiral separation method of racemic ' alpha '-cyclopentyl amygdalic acid as claimed in claim 1, is characterized in that, in step (2), in described sample, the concentration of racemic ' alpha '-cyclopentyl amygdalic acid is 1～20mg/mL.

9. the chiral separation method of racemic ' alpha '-cyclopentyl amygdalic acid as claimed in claim 1, is characterized in that in step (3), and the described gradient timed interval is that every 0.5～5min collects once.

10. the chiral separation method of racemic ' alpha '-cyclopentyl amygdalic acid as claimed in claim 1, it is characterized in that in step (4), described recovery is carried out as follows: leading peak elutriant and postpeak elutriant are used respectively to salt acid for adjusting pH value to 1～2, again with methyl tertiary butyl ether extraction 2～3 times, merge methyl tert-butyl ether layers, wash with water to neutrality, anhydrous sodium sulfate drying, filtration, filtrate steaming removal solvent, obtains respectively dextrorotation α-cyclopentyl amygdalic acid monomer and left-handed α-cyclopentyl amygdalic acid monomer.

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