Separation method of S-type and R-type flurbiprofen

Abstract

The present invention provides a kind of S-type and R-type flurbiprofen splitting method, comprising the steps: the flurbiprofen to be split is dissolved in an organic solvent, and a test solution is prepared; the test solution is split using supercritical fluid chromatography; wherein, the conditions of supercritical fluid chromatography include: mobile phase A is a supercritical fluid, and mobile phase B is an alcohol including carbon atoms of 1 to 3; the stationary phase of the chromatographic column is a silica gel surface coated with amylose tris (3,5-dimethylphenylcarbamate) stationary phase, a silica gel surface coated with amylose tris [(S)-a-methylbenzylcarbamate] stationary phase, a silica gel surface coated with cellulose tris (4-chloro-3-methylphenylcarbamate) stationary phase or a silica gel surface coated with cellulose tris (4-methylbenzoate) stationary phase. The splitting method has a short separation time, a good separation effect, and is more environmentally friendly.

Description

Technical Field

[0001] The invention relates to the technical field of chromatographic separation and purification, in particular to a method for separating S-type and R-type flurbiprofen. Background Art

[0002] Flurbiprofen is a non-steroidal aromatic propionic acid chiral anti-inflammatory drug commonly used in clinical practice. The S-type flurbiprofen enantiomer has a good therapeutic effect on rheumatoid arthritis, osteoarthritis, etc., while the R-type flurbiprofen enantiomer has almost no anti-inflammatory activity, but has a good effect on Heilmer's disease and inhibiting tumor growth. Therefore, obtaining a high-purity single flurbiprofen enantiomer by chiral separation has important practical significance.

[0003] At present, the separation method for flurbiprofen enantiomer mainly includes chemical separation and preparation of S-type flurbiprofen, enzyme separation and preparation of R-type flurbiprofen, crystallization kinetics separation and preparation of R-type flurbiprofen, and high performance liquid chromatography separation and preparation of R and S-type flurbiprofen. Chemical method, enzyme method and crystallization kinetics separation can only obtain one of the configurations of flurbiprofen enantiomers, and it is impossible to obtain two configurations of enantiomers at the same time. Both S and R-type flurbiprofen have medicinal value, and only extracting the enantiomer of a single configuration undoubtedly causes a waste of resources. Although S and R-type flurbiprofen can be obtained simultaneously by adopting high performance liquid chromatography separation and preparation, its separation and preparation need to consume a large amount of organic reagents, and the post-processing procedure is complicated and can also cause environmental pollution.

[0004] Supercritical fluid chromatography (SFC) uses supercritical fluids to replace conventional organic reagents. With the unique physical and chemical properties of supercritical fluids, it is used in stereoisomer separation, analysis of petroleum products, separation of lipids and natural products, and pharmaceutical and food analysis. In large-scale separation and purification, it can achieve an exponential increase in preparation efficiency by using about 30% of the amount of organic reagents required by high-performance liquid phase method, which fully meets the requirements of product development at this stage, that is, while quickly processing a large number of samples, reducing the use of chemical reagents, achieving less cost investment and more benefit output. The critical temperature and pressure of supercritical fluids are compatible with chromatographs, non-destructive to the compounds to be analyzed and purified, low reactivity and low toxicity, non-flammable, and supercritical fluids themselves are abundant and inexpensive. However, when supercritical fluid chromatography is actually used to separate flurbiprofen enantiomers, insufficient separation often occurs. Summary of the invention

[0005] Based on this, the present invention provides a method for separating S-type and R-type flurbiprofen, which has short separation time, good separation effect and is more environmentally friendly.

[0006] The present invention is achieved through the following technical solutions.

[0007] A method for separating S-type and R-type flurbiprofen comprises the following steps:

[0008] The flurbiprofen to be separated is dissolved in an organic solvent to prepare a test solution;

[0009] The test solution is separated by supercritical fluid chromatography;

[0010] Among them, the conditions of supercritical fluid chromatography include: mobile phase A is a supercritical fluid, mobile phase B is an alcohol containing 1 to 3 carbon atoms; the stationary phase of the chromatographic column is a silica gel surface coated with amylose-tris (3,5-dimethylphenylcarbamate) stationary phase, a silica gel surface coated with amylose-tris [(S)-a-methylbenzylcarbamate] stationary phase, a silica gel surface coated with cellulose-tris (4-chloro-3-methylphenylcarbamate) stationary phase or a silica gel surface coated with cellulose-tris (4-methylbenzoate) stationary phase.

[0011] In one embodiment, the conditions of supercritical fluid chromatography also include: using isocratic elution, the volume ratio of the mobile phase A to the mobile phase B is (80-99):(1-20).

[0012] In one embodiment, the volume ratio of the mobile phase A to the mobile phase B is (90-99):(1-10).

[0013] In one embodiment, the supercritical fluid is supercritical carbon dioxide.

[0014] In one embodiment, the alcohol having 1 to 3 carbon atoms is selected from one or more of ethanol and propanol.

[0015] In one embodiment, the conditions of supercritical fluid chromatography also include: the temperature of the supercritical fluid when passing through the pump is -5°C to 5°C, the density of the supercritical fluid is 890kg / m 3 ~960kg / m 3 .

[0016] In one embodiment, the conditions of supercritical fluid chromatography also include: a back pressure of 10 MPa to 14 MPa.

[0017] In one embodiment, the conditions of supercritical fluid chromatography also include: column temperature of 30°C to 35°C; detection wavelength of 245nm to 265nm; flow rate of 2mL / min to 10mL / min;

[0018] The dimensions of the chromatographic column include: a column length of 100 mm to 500 mm, and an inner diameter of 4.6 mm or 10 mm.

[0019] In one embodiment, the organic solvent is selected from one or more of ethanol and methanol.

[0020] In one embodiment, in the test solution, the concentration of the flurbiprofen to be separated is 4 mg / mL to 200 mg / mL.

[0021] Compared with the prior art, the separation method of S-type and R-type flurbiprofen of the present invention has the following beneficial effects:

[0022] The present invention provides a method for separating S-type and R-type flurbiprofen using a supercritical fluid chromatography system for the first time. The present invention selects a mixed fluid of a supercritical fluid and an alcohol having 1 to 3 carbon atoms as a mobile phase, and simultaneously selects silica gel surface coated with amylose-tris(3,5-dimethylphenylcarbamate), silica gel surface coated with amylose-tris[(S)-a-methylbenzylcarbamate], silica gel surface coated with cellulose-tris(4-chloro-3-methylphenylcarbamate) or silica gel surface coated with cellulose-tris(4-methylbenzoate) as a stationary phase, so as to achieve separation and semi-preparation of S-type and R-type in flurbiprofen, and the separation time is short, the separation effect is good, the purity can reach more than 99%, and the present invention has broad application prospects in the field of separation of S-type and R-type in flurbiprofen.

[0023] In addition, the present invention uses supercritical fluid instead of organic reagents, which reduces the harm to the human body and the environment, and also greatly reduces the solvent residue in the subsequent drug production, which is more green and environmentally friendly. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 The separation spectrum of the flurbiprofen sample provided in Example 1 of the present invention in a supercritical fluid chromatograph system;

[0025] Figure 2 The separation spectrum of the flurbiprofen sample provided in Example 2 of the present invention in a supercritical fluid chromatograph system;

[0026] Figure 3 The separation spectrum of the flurbiprofen sample provided in Example 3 of the present invention in a supercritical fluid chromatograph system;

[0027] Figure 4 The separation spectrum of the flurbiprofen sample provided in Example 4 of the present invention in a supercritical fluid chromatograph system;

[0028] Figure 5 The separation spectrum of the flurbiprofen sample provided in Example 5 of the present invention in a supercritical fluid chromatograph system;

[0029] Figure 6The separation spectrum of the flurbiprofen sample provided in Example 6 of the present invention in a supercritical fluid chromatograph system;

[0030] Figure 7 The separation spectrum of the flurbiprofen sample provided in Example 7 of the present invention in a supercritical fluid chromatograph system;

[0031] Figure 8 This is a purity detection diagram of type S flurbiprofen provided in Example 8 of the present invention;

[0032] Fig. 9 This is a purity detection diagram of R-type flurbiprofen provided in Example 8 of the present invention;

[0033] Fig.10 This is the separation spectrum of the flurbiprofen sample provided in Comparative Example 1 of the present invention in a supercritical fluid chromatograph system. DETAILED DESCRIPTION

[0034] In order to facilitate the understanding of the present invention, the present invention will be described more fully below with reference to the relevant drawings. The preferred embodiments of the present invention are given in the drawings. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive.

[0035] In addition, the terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Therefore, the features defined as "first" and "second" may explicitly or implicitly include at least one of the features. In the description of the invention, the meaning of "multiple" is at least two, such as two, three, etc., unless otherwise clearly and specifically defined. In the description of the present invention, the meaning of "several" is at least one, such as one, two, etc., unless otherwise clearly and specifically defined.

[0036] The words "preferably", "more preferably", etc. in the present invention refer to embodiments of the present invention that may provide certain beneficial effects in certain circumstances. However, other embodiments may also be preferred under the same circumstances or other circumstances. In addition, the description of one or more preferred embodiments does not imply that other embodiments are not applicable, nor is it intended to exclude other embodiments from the scope of the present invention.

[0037] When a numerical range is disclosed herein, the above range is considered to be continuous and includes the minimum and maximum values ​​of the range, as well as every value between such minimum and maximum values. Further, when a range refers to an integer, every integer between the minimum and maximum values ​​of the range is included. In addition, when multiple ranges are provided to describe features or characteristics, the ranges can be combined. In other words, unless otherwise indicated, all ranges disclosed herein should be understood to include any and all subranges included therein.

[0038] Unless otherwise specified, all percentages, fractions and ratios are calculated by the total mass of the composition of the present invention. Unless otherwise specified, all masses of the listed ingredients are given to the content of active substances, so they do not include solvents or by-products that may be contained in commercially available materials. The term "mass percentage content" herein can be represented by the symbol "%". Unless otherwise specified, all molecular weights in this article are weight average molecular weights expressed in Daltons. Unless otherwise specified, all formulations and tests in this article occur in an environment of 25°C. "Including", "including", "containing", "containing", "having" or other variants herein are intended to cover non-closed inclusions, and these terms are not distinguished. The term "comprising" means that other steps and ingredients that do not affect the final result can be added. The composition and method / process of the present invention include, consist of and are essentially composed of the necessary elements and restrictions described herein and any additional or optional ingredients, components, steps or restrictions described herein. The terms "efficacy", "performance", "effect" and "efficacy" are not distinguished between herein.

[0039] The weight of the relevant components mentioned in the description of the embodiments of the present invention may not only refer to the specific content of each component, but also indicate the weight ratio between the components. Therefore, as long as the content of the relevant components is proportionally enlarged or reduced according to the description of the embodiments of the present invention, it is within the scope disclosed in the description of the embodiments of the present invention. Specifically, the weight described in the description of the embodiments of the present invention may be a mass unit known in the chemical industry such as ug, mg, g, kg, etc.

[0040] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used herein in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used herein includes any and all combinations of one or more related listed items.

[0041] The present invention provides a method for separating S-type and R-type flurbiprofen, comprising the following steps:

[0042] The flurbiprofen to be separated is dissolved in an organic solvent to prepare a test solution;

[0043] The test solution was separated by supercritical fluid chromatography;

[0044] Among them, the conditions of supercritical fluid chromatography include: mobile phase A is a supercritical fluid, mobile phase B is an alcohol containing 1 to 3 carbon atoms; the stationary phase of the chromatographic column is a silica gel surface coated with amylose-tris (3,5-dimethylphenylcarbamate) stationary phase, a silica gel surface coated with amylose-tris [(S)-a-methylbenzylcarbamate] stationary phase, a silica gel surface coated with cellulose-tris (4-chloro-3-methylphenylcarbamate) stationary phase or a silica gel surface coated with cellulose-tris (4-methylbenzoate) stationary phase.

[0045] Preferably, the stationary phase of the chromatographic column is a silica gel surface coated with amylose-tris (3,5-dimethylphenylcarbamate) stationary phase or a silica gel surface coated with amylose-tris [(S)-a-methylbenzylcarbamate] stationary phase. More preferably, the stationary phase of the chromatographic column is a silica gel surface coated with amylose-tris (3,5-dimethylphenylcarbamate) stationary phase.

[0046] The inventors have found that the best separation of flurbiprofen can be achieved by using a supercritical fluid and an alcohol having 1 to 3 carbon atoms as the mobile phase and selecting a silica gel surface coated with amylose-tris(3,5-dimethylphenylcarbamate) or a silica gel surface coated with amylose-tris[(S)-a-methylbenzylcarbamate] as the stationary phase of the chromatographic column.

[0047] In a specific example, the conditions of supercritical fluid chromatography also include: using isocratic elution, and the volume ratio of mobile phase A to mobile phase B is (80-99):(1-20).

[0048] In a specific example, the volume ratio of mobile phase A to mobile phase B is (90-99):(1-10).

[0049] Preferably, the volume ratio of mobile phase A to mobile phase B is (93-97):(3-7). More preferably, the volume ratio of mobile phase A to mobile phase B is 95:5.

[0050] In one specific example, the supercritical fluid is supercritical carbon dioxide.

[0051] In a specific example, the alcohol having 1 to 3 carbon atoms is selected from one or more of ethanol and propanol.

[0052] Preferably, the alcohol having 1 to 3 carbon atoms is ethanol or isopropanol.

[0053] In a specific example, the mobile phase B further includes trifluoroacetic acid. More specifically, the volume concentration of trifluoroacetic acid is 0.05% to 0.15%.

[0054] In a specific example, the conditions of supercritical fluid chromatography also include: the temperature of the supercritical fluid when passing through the pump is -5°C to 5°C, the density of the supercritical fluid is 890 kg / m 3 ~960kg / m 3 .

[0055] In a specific example, the conditions of supercritical fluid chromatography also include: a back pressure of 10 MPa to 14 MPa.

[0056] It can be understood that in the present invention, the back pressure includes but is not limited to 10 MPa, 11 MPa, 12 MPa, 13 MPa, and 14 MPa.

[0057] In a specific example, the conditions of supercritical fluid chromatography also include: a column temperature of 30° C. to 35° C. It can be understood that in the present invention, the column temperature includes but is not limited to 30° C., 31° C., 32° C., 33° C., 34° C., and 35° C. Preferably, the column temperature is 35° C.

[0058] In a specific example, the conditions of supercritical fluid chromatography also include: the detection wavelength is 245nm to 265nm. It can be understood that in the present invention, the detection wavelength includes but is not limited to 245nm, 246nm, 247nm, 248nm, 249nm, 250nm, 251nm, 252nm, 253nm, 254nm, 255nm, 256nm, 257nm, 258nm, 259nm, 260nm, 261nm, 262nm, 263nm, 264nm, 265nm.

[0059] In a specific example, the conditions of supercritical fluid chromatography also include: a flow rate of 2 mL / min to 10 mL / min. It can be understood that in the present invention, the flow rate includes but is not limited to 2 mL / min, 3 mL / min, 4 mL / min, 5 mL / min, 6 mL / min, 7 mL / min, 8 mL / min, 9 mL / min, and 10 mL / min.

[0060] In a specific example, the conditions of supercritical fluid chromatography also include: a column length of 100 mm to 500 mm, and an inner diameter of 4.6 mm or 10 mm.

[0061] More specifically, the dimensions of the chromatographic column include: a column length of 250 mm and an inner diameter of 4.6 mm or 10 mm.

[0062] In a specific example, the conditions of supercritical fluid chromatography also include: the injection volume is 1 μL to 500 μL.

[0063] In a specific example, the organic solvent is selected from one or more of ethanol and methanol.

[0064] In a specific example, in the test solution, the concentration of the flurbiprofen to be separated is 4 mg / mL to 200 mg / mL.

[0065] More specifically, the chromatographic column has a specification of 4.6×250 mm, a flow rate of 2 mL / min, an injection volume of 1 μL to 5 μL, and a concentration of flurbiprofen to be separated of 20 mg / mL to 50 mg / mL; or

[0066] The specification of the chromatographic column is 10×250 mm, the flow rate is 10 mL / min, the injection volume is 100 μL to 500 μL, and the concentration of flurbiprofen to be separated is 20 mg / mL to 50 mg / mL.

[0067] The separation method of S-type and R-type flurbiprofen of the present invention is further described in detail below in conjunction with specific examples. The raw materials used in the following examples, unless otherwise specified, are all commercially available products.

[0068] Example 1

[0069] This embodiment provides a method for separating and preparing S-type and R-type flurbiprofen using a supercritical fluid chromatography system, as follows:

[0070] The chromatograph selected was a supercritical fluid chromatograph produced by Jiangsu Hanbang Technology Co., Ltd. The chromatographic column specification was 4.6×250 mm, the mobile phase flow rate was 1 mL / min, the back pressure was set to 12 MPa, the flurbiprofen concentration was 4 mg / mL, the injection volume was 1 μL, and the detection wavelength was 254 nm.

[0071] Table 1 shows other relevant conditions selected in the method for separating and preparing S-type and R-type flurbiprofen using a supercritical fluid chromatography system in Example 1.

[0072] Table 1 SFC separation conditions in Example 1

[0073]

[0074] The specific steps corresponding to Example 1 are as follows: the flurbiprofen sample is dissolved in ethanol, ultrasonically assisted to prepare a 4 mg / mL solution, and Chiralcel AS-H5μm 4.6×250mm (silica gel surface coated with amylose-tris[(S)-a-methylbenzylcarbamate]) is selected as the chromatographic column on the supercritical fluid chromatograph of Jiangsu Hanbang Technology Co., Ltd. The volume ratio of supercritical carbon dioxide and mobile phase B in the mobile phase, the total flow rate, the back pressure and the column temperature of the chromatographic column are controlled according to Table 1, and the temperature of the carbon dioxide entering the pump is ensured to be -5°C to 5°C, and the density is 890kg / m 3 ~960kg / m 3 , flurbiprofen was separated, and the separation effect diagram is shown in Figure 1 shown.

[0075] Example 2

[0076] This embodiment provides a method for separating and preparing S-type and R-type flurbiprofen using a supercritical fluid chromatography system, as follows:

[0077] The chromatograph selected was a supercritical fluid chromatograph produced by Jiangsu Hanbang Technology Co., Ltd. The chromatographic column specification was 4.6×250 mm, the mobile phase flow rate was 1 mL / min, the back pressure was set to 12 MPa, the flurbiprofen concentration was 4 mg / mL, the injection volume was 2 μL, and the detection wavelength was 254 nm.

[0078] Table 2 shows other relevant conditions selected in the method for separating and preparing S-type and R-type flurbiprofen using a supercritical fluid chromatography system in Example 2.

[0079] Table 2 SFC separation conditions in Example 2

[0080]

[0081] The specific steps corresponding to Example 2 are as follows: the flurbiprofen sample is dissolved in ethanol, ultrasonically assisted to prepare a 4 mg / mL solution, and Optichiral AB-55 μm 4.6×250 mm is selected as the chromatographic column on the supercritical fluid chromatograph of Jiangsu Hanbang Technology Co., Ltd. The volume ratio of supercritical carbon dioxide and mobile phase B in the mobile phase, the total flow rate, the back pressure and the column temperature of the chromatographic column are controlled according to Table 2, and the temperature of the carbon dioxide entering the pump is ensured to be -5°C to 5°C, and the density is 890 kg / m 3 ~960kg / m 3 , flurbiprofen was separated, and the separation effect diagram is shown in Figure 2 shown.

[0082] Example 3

[0083] This embodiment provides a method for separating and preparing S-type and R-type flurbiprofen using a supercritical fluid chromatography system, as follows:

[0084] The chromatograph selected was a supercritical fluid chromatograph produced by Jiangsu Hanbang Technology Co., Ltd. The chromatographic column specification was 4.6×250 mm, the mobile phase flow rate was 1 mL / min, the back pressure was set to 12 MPa, the flurbiprofen concentration was 4 mg / mL, the injection volume was 2 μL, and the detection wavelength was 254 nm.

[0085] Table 3 shows other relevant conditions selected in the method for separating and preparing S-type and R-type flurbiprofen using a supercritical fluid chromatography system in Example 3.

[0086] Table 3 SFC separation conditions in Example 3

[0087]

[0088] The specific steps corresponding to Example 3 are as follows: the flurbiprofen sample is dissolved in ethanol, ultrasonically assisted to prepare a 4 mg / mL solution, and Optichiral AB-55 μm 4.6×250 mm is selected as the chromatographic column on the supercritical fluid chromatograph of Jiangsu Hanbang Technology Co., Ltd. The volume ratio of supercritical carbon dioxide and mobile phase B in the mobile phase, the total flow rate, the back pressure and the column temperature of the chromatographic column are controlled according to Table 3, and the temperature of the carbon dioxide entering the pump is ensured to be -5°C to 5°C, and the density is 890 kg / m 3 ~960kg / m 3 , flurbiprofen was separated, and the separation effect diagram is shown in Figure 3 shown.

[0089] Example 4

[0090] This embodiment provides a method for separating and preparing S-type and R-type flurbiprofen using a supercritical fluid chromatography system, as follows:

[0091] The chromatograph selected was a supercritical fluid chromatograph produced by Jiangsu Hanbang Technology Co., Ltd. The chromatographic column specification was 4.6×250 mm, the mobile phase flow rate was 1 mL / min, the back pressure was set to 12 MPa, the flurbiprofen concentration was 4 mg / mL, the injection volume was 2 μL, and the detection wavelength was 254 nm.

[0092] Table 4 shows other relevant conditions selected in the method for separating and preparing S-type and R-type flurbiprofen using a supercritical fluid chromatography system in Example 4.

[0093] Table 4 SFC separation conditions in Example 4

[0094]

[0095] The specific steps corresponding to Example 4 are as follows: the flurbiprofen sample is dissolved in ethanol, ultrasonically assisted to prepare a 4 mg / mL solution, and Optichiral AB-55 μm 4.6×250 mm is selected as the chromatographic column on the supercritical fluid chromatograph of Jiangsu Hanbang Technology Co., Ltd. The volume ratio of supercritical carbon dioxide and mobile phase B in the mobile phase, the total flow rate, the back pressure and the column temperature of the chromatographic column are controlled according to Table 4, and the temperature of the carbon dioxide entering the pump is ensured to be -5°C to 5°C, and the density is 890 kg / m 3 ~960kg / m 3 , flurbiprofen was separated, and the separation effect diagram is shown in Figure 4 shown.

[0096] Example 5

[0097] This embodiment provides a method for separating and preparing S-type and R-type flurbiprofen using a supercritical fluid chromatography system, as follows:

[0098] The chromatograph selected was a supercritical fluid chromatograph produced by Jiangsu Hanbang Technology Co., Ltd. The chromatographic column specification was 4.6×250 mm, the mobile phase flow rate was 1 mL / min, the back pressure was set to 12 MPa, the flurbiprofen concentration was 4 mg / mL, the injection volume was 5 μL, and the detection wavelength was 254 nm.

[0099] Table 5 shows other relevant conditions selected in the method for separating and preparing S-type and R-type flurbiprofen using a supercritical fluid chromatography system in Example 5.

[0100] Table 5 SFC analysis test conditions in Example 5

[0101]

[0102] The specific steps corresponding to Example 5 are as follows: the flurbiprofen sample is dissolved in ethanol, ultrasonically assisted to prepare a 4 mg / mL solution, and Optichiral AB-55 μm 4.6×250 mm is selected as the chromatographic column on the supercritical fluid chromatograph of Jiangsu Hanbang Technology Co., Ltd. The volume ratio of supercritical carbon dioxide and mobile phase B in the mobile phase, the total flow rate, the back pressure and the column temperature of the chromatographic column are controlled according to Table 5, and the temperature of the carbon dioxide entering the pump is ensured to be -5°C to 5°C, and the density is 890 kg / m 3 ~960kg / m 3 , flurbiprofen was separated, and the separation effect diagram is shown in Figure 5 As shown. A method for rapid detection of S-type and R-type flurbiprofen can be provided, and the detection can be completed within 7 minutes.

[0103] Example 6

[0104] This embodiment provides a method for separating and preparing S-type and R-type flurbiprofen using a supercritical fluid chromatography system, as follows:

[0105] The chromatograph selected was a supercritical fluid chromatograph produced by Jiangsu Hanbang Technology Co., Ltd. The chromatographic column specification was 10×250 mm, the mobile phase flow rate was 10 mL / min, the back pressure was set to 12 MPa, the flurbiprofen concentration was 25 mg / mL, the injection volume was 200 μL, and the detection wavelength was 254 nm.

[0106] Table 6 shows other relevant conditions selected in the method for separating and preparing S-type and R-type flurbiprofen using a supercritical fluid chromatography system in Example 6.

[0107] Table 6 SFC preparation and separation conditions in Example 6

[0108]

[0109] The specific steps corresponding to Example 6 are as follows: the flurbiprofen sample is dissolved in ethanol, ultrasonically assisted to prepare a 200 mg / mL solution, and ChiralGel AH7μm 10×250mm (silica gel surface coated with amylose-tris (3,5-dimethylphenylcarbamate)) is selected as the chromatographic column on the supercritical fluid chromatograph of Jiangsu Hanbang Technology Co., Ltd. The volume ratio of supercritical carbon dioxide and mobile phase B in the mobile phase, the total flow rate, the back pressure and the column temperature of the chromatographic column are controlled according to Table 6, and the temperature of the carbon dioxide entering the pump is ensured to be -5°C to 5°C, and the density is 890kg / m 3 ~960kg / m 3 , flurbiprofen was separated, and the separation effect is shown in the figure Figure 6 According to the linear amplification principle, the chromatographic column with a size of 4.6×250 mm can be enlarged to a chromatographic column with a size of 10×250 mm, and the milligram-level preparation of flurbiprofen samples can be achieved.

[0110] Example 7

[0111] This embodiment provides a method for separating and preparing S-type and R-type flurbiprofen using a supercritical fluid chromatography system, as follows:

[0112] The chromatograph selected was a high performance liquid chromatograph produced by Jiangsu Hanbang Technology Co., Ltd. The column specifications were 10×250 mm, the mobile phase flow rate was 10 mL / min, the flurbiprofen concentration was 25 mg / mL, the injection volume was 550 μL, and the detection wavelength was 254 nm.

[0113] Table 7 shows other relevant conditions selected in the method for separating and extracting high-purity flurbiprofen using a high performance liquid chromatography system in Example 7.

[0114] Table 7 SFC preparation conditions in Example 7

[0115]

[0116] The specific steps corresponding to Example 7 are as follows: the flurbiprofen sample is dissolved in ethanol, ultrasonically assisted to prepare a 25 mg / mL solution, and ChiralGel AH 7 μm 10 × 250 mm is selected as the chromatographic column on the high performance liquid chromatograph of Jiangsu Hanbang Technology Co., Ltd., and the volume ratio of A and B in the mobile phase, the total flow rate and the column temperature of the chromatographic column are controlled according to Table 7 to separate flurbiprofen. The production efficiency is improved by stacking needle injection. The separation effect diagram is shown in FIG. Figure 7 shown.

[0117] Example 8

[0118] This example is a detection spectrum of the S-type and R-type flurbiprofen prepared in Example 7. The detection method is as follows:

[0119] The chromatograph used was Waters Acquity UPC 2 The chromatographic column specifications were Optichiral AB-5 5μm4.6×250mm, the mobile phase flow rate was 1mL / min, the injection volume was 10μL, and the detection wavelength was 254nm.

[0120] Table 8 shows the use of Waters Acquity UPC in Example 8 2 Other relevant conditions selected in the detection method of the S-type and R-type flurbiprofen prepared in Detection Example 7.

[0121] Table 8 SFC detection conditions in Example 8

[0122]

[0123] The specific steps corresponding to Example 8 are as follows: the collected S-type and R-type flurbiprofen samples were purified by Waters AcquityUPC 2 The chromatographic column specification is Optichiral AB-5 5μm 4.6×250mm. The volume ratio of A and B in the mobile phase, the total flow rate and the chromatographic column temperature are controlled according to Table 8. The prepared and collected samples are tested to determine their purity. According to the test chart, the purity of S-type and R-type flurbiprofen is greater than 99%. Figure 8-Figure 9 shown.

[0124] Comparative Example 1

[0125] This comparative example provides a method for analyzing and detecting high-purity flurbiprofen using a high performance liquid chromatography system, as follows:

[0126] The chromatograph selected was a high performance liquid chromatograph produced by Jiangsu Hanbang Technology Co., Ltd. The chromatographic column specification was 4.6×250 mm, the mobile phase flow rate was 0.6 mL / min, the concentration of flurbiprofen was 4 mg / mL, the injection volume was 1 μL, and the detection wavelength was 254 nm.

[0127] Table 9 shows other relevant conditions selected in the method for separating and extracting high-purity flurbiprofen using a high performance liquid chromatography system in Comparative Example 1.

[0128] Table 9 HPLC analysis detection conditions in Comparative Example 1

[0129]

[0130] The specific steps corresponding to Comparative Example 1 are as follows: Flurbiprofen sample is dissolved in ethanol, ultrasonically assisted to prepare a 4 mg / mL solution, and Chiralpak AD 5 μm 4.6 × 250 mm is selected as the chromatographic column on the high performance liquid chromatograph of Jiangsu Hanbang Technology Co., Ltd., and the volume ratio, total flow rate and chromatographic column temperature of the mobile phase are configured according to Table 9. Flurbiprofen is separated, and the separation effect is shown in the figure below. Fig.10 As shown, the entire analytical detection method needs to be completed within 17 minutes with a low sample load.

[0131] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0132] The above-described embodiments only express several implementation methods of the present invention, which are convenient for understanding the technical solutions of the present invention in detail, but they cannot be understood as limiting the scope of protection of the invention patent. It should be pointed out that for ordinary technicians in this field, several modifications and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. It should be understood that the technical solutions obtained by those skilled in the art through logical analysis, reasoning or limited experiments on the basis of the technical solutions provided by the present invention are all within the protection scope of the claims attached to the present invention. Therefore, the protection scope of the patent of the present invention shall be based on the contents of the attached claims, and the description and drawings can be used to interpret the contents of the claims.

Claims

1. A method for separating S-type and R-type flurbiprofen, characterized in that: The steps include: The flurbiprofen to be separated is dissolved in an organic solvent to prepare a test solution; The test solution is separated by supercritical fluid chromatography; The conditions of supercritical fluid chromatography include: mobile phase A is a supercritical fluid, mobile phase B is an alcohol containing 1 to 3 carbon atoms; the stationary phase of the chromatographic column is a silica gel surface coated with amylose-tris (3, 5-dimethylphenylcarbamate) stationary phase, a silica gel surface coated with amylose-tris [(S)-a-methylbenzylcarbamate] stationary phase or OptichiralAB-5 5μm 4.6×250mm; The conditions of the supercritical fluid chromatography also include: using isocratic elution, the volume ratio of the mobile phase A to the mobile phase B is (80-99):(1-20); The supercritical fluid is supercritical carbon dioxide; The alcohol having 1 to 3 carbon atoms is selected from one or more of ethanol and propanol; The conditions of supercritical fluid chromatography also include: column temperature of 30°C to 35°C; detection wavelength of 245nm to 265nm; The organic solvent is selected from one or more of ethanol and methanol.

2. The method for separating S-type and R-type flurbiprofen according to claim 1, characterized in that: The volume ratio of the mobile phase A to the mobile phase B is (90-99):(1-10).

3. The method for separating S-type and R-type flurbiprofen according to claim 1, characterized in that: The conditions of supercritical fluid chromatography also include: the temperature of the supercritical fluid when passing through the pump is -5°C to 5°C, the density of the supercritical fluid is 890kg / m 3 ~960kg / m 3 .

4. The method for separating S-type and R-type flurbiprofen according to claim 1, characterized in that: The conditions of supercritical fluid chromatography also include: a back pressure of 10 MPa to 14 MPa.

5. The method for separating S-type and R-type flurbiprofen according to claim 1, characterized in that: The conditions of supercritical fluid chromatography also include: a flow rate of 2 mL / min to 10 mL / min; The dimensions of the chromatographic column include: a column length of 100 mm to 500 mm, and an inner diameter of 4.6 mm or 10 mm.

6. The method for separating S-type and R-type flurbiprofen according to any one of claims 1 to 5, characterized in that: In the test solution, the concentration of the flurbiprofen to be separated is 4 mg / mL to 200 mg / mL.

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