CN114689761A - Method for detecting parecoxib sodium positional isomer through liquid chromatography - Google Patents
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Abstract
The invention provides a method for detecting parecoxib sodium positional isomer by liquid chromatography, wherein a liquid phase chiral chromatographic column used in the method is a polysaccharide derivative normal phase coating type chiral chromatographic column CHIRALPAK AD-H, and a mobile phase is a mixed solution of normal hexane, isopropanol and trifluoroacetic acid. The detection method provided by the invention has the advantages that the theoretical plate number of the peak of the liquid chromatogram is excellent, the separation degree is high, and the separation degree values under different chromatographic conditions are all more than 5.
Description
Technical Field
The invention belongs to the field of pharmaceutical analysis, and relates to a method for detecting parecoxib sodium positional isomer by liquid chromatography.
Background
Parecoxib sodium, with the chemical name of N- [ [4- (5-methyl-3-phenyl-4-isoxazolyl) phenyl ] sulfonyl ] propionamide sodium salt, is a specific cyclooxygenase-2 (COX-2) inhibitor. The primary medicine is approved to be marketed in China in 2008, is the first COX-2 inhibitor capable of being administered through intravenous injection and intramuscular injection, and is mainly used for short-term treatment of postoperative pain.
The parecoxib sodium positional isomer, with the chemical name of N- [ [3- (5-methyl-3-phenyl-4-isoxazolyl) phenyl ] sulfonyl ] propionamide sodium salt, is a meta isomer of a benzene ring generated in a sulfonation process, cannot be effectively separated from a main peak under a reverse phase chromatographic condition, has the same retention time as parecoxib sodium, but has different biological activities due to different functional group positions. Therefore, the purity of the product is strictly controlled, and the method has important clinical significance.
In the prior patent, CN104965041B discloses a high performance liquid chromatography detection method for parecoxib sodium isomer, a chromatographic column with silane bonded silica gel as a filler is adopted, n-hexane-isopropanol is used as a flow to carry out separation detection on parecoxib sodium, the separation degree of parecoxib sodium obtained by the detection method and the position isomer thereof is low, and a lifting space is still left.
Although the parecoxib sodium positional isomer is separated and detected by the existing test method, the separation degree of the parecoxib sodium positional isomer obtained by the detection method is low, and an improvement space still exists. Therefore, how to develop a position isomer detection method which is convenient to operate and high in separation degree has important significance for quality control of parecoxib sodium.
Disclosure of Invention
The invention aims to explore various conditions of parecoxib sodium positional isomer liquid chromatography analysis, and provides a method for detecting parecoxib sodium positional isomer through liquid chromatography, so as to solve the problems of low separation degree of parecoxib sodium and positional isomer, poor separation effect and low chromatographic column efficiency in the existing method.
The invention adopts the following technical scheme:
a method for detecting parecoxib sodium positional isomer by liquid chromatography, wherein the chromatographic column is a liquid phase chiral chromatographic column which is a polysaccharide derivative normal phase coating type chiral chromatographic column CHIRALPAK AD-H; the mobile phase is a mixed solution of n-hexane, isopropanol and trifluoroacetic acid; the detection wavelength is 215 nm-220 nm; the flow rate of the mobile phase is 0.8-1.2 mL/min; the temperature of the chromatographic column is 30-40 ℃.
The optional chromatographic column is a liquid chiral chromatographic column prepared from CHIRALPAK AD-H (4.6 mm × 250mm, 5 μm) polysaccharide derivative chiral chromatographic column.
The stationary phase of the optional chromatographic column is silica gel coated with amylose-tris (3, 5-xylylcarbamate).
The temperature of the chromatographic column is preferably 30 ℃, 35 ℃ and 40 ℃; the detection wavelength is preferably 215nm, 217nm and 220 nm; the flow rate of the mobile phase is preferably 0.8mL/min, 1.0mL/min, or 1.2 mL/min.
The mobile phase is the volume ratio of n-hexane to isopropanol (75-90) to (10-25), and the volume ratio of trifluoroacetic acid is 0.099 of the whole mobile phase.
The mobile phase is preferably that the volume ratio of normal hexane to isopropanol is (75-85) to (15-25), and the volume ratio of trifluoroacetic acid is 0.099 of the whole mobile phase.
The mobile phase further preferably has a volume ratio of n-hexane to isopropanol of (80-85) to (15-20).
Preparing a test solution:
test solution preparation: precisely weighing a to-be-measured product, placing the to-be-measured product in a volumetric flask, adding absolute ethyl alcohol to dissolve, shaking up, fixing the volume to a scale mark to serve as a stock solution, then placing a certain amount of the stock solution in the volumetric flask, adding a diluent n-hexane-isopropanol 90:10 to dilute, shaking up, and fixing the volume to the scale mark.
Compared with the prior art, the detection method provided by the invention has the advantages that the theoretical plate number of the peak of the liquid chromatogram is excellent, the separation degree is high, and the separation degree values under different chromatographic conditions are all more than 5.
Drawings
FIG. 1 is a liquid chromatogram of example 1.
FIG. 2 is a liquid chromatogram of example 2.
FIG. 3 is a liquid chromatogram of example 3.
FIG. 4 is a liquid chromatogram of example 4.
FIG. 5 is a liquid chromatogram of example 5.
FIG. 6 is a liquid chromatogram of example 6.
FIG. 7 is a liquid chromatogram of example 7.
FIG. 8 is a liquid chromatogram of example 8.
FIG. 9 is a liquid chromatogram of example 9.
Detailed Description
In order to better illustrate the present invention and facilitate the understanding of the technical solutions, the present invention is further described in detail below.
The specific information of the reagent instrument used in the embodiment of the invention is as follows:
liquid chromatograph: shimadzu 10AT HPLC is equipped with UV detector;
a chromatographic column: the polysaccharide derivative normal phase coating type chiral chromatographic column CHIRALPAK AD-H, 4.6mm × 250mm, 5 μm.
Examples 1-9 were prepared as follows:
control solution: and (3) precisely weighing 12.5mg of parecoxib sodium positional isomer reference substance, placing the parecoxib sodium positional isomer reference substance into a 25ml measuring flask, adding absolute ethyl alcohol to dissolve and dilute the parecoxib sodium positional isomer reference substance to a scale, and shaking up to prepare a parecoxib sodium positional isomer reference substance concentrated solution. Precisely measuring 0.5ml of parecoxib sodium positional isomer reference substance concentrated solution, putting the parecoxib sodium positional isomer reference substance concentrated solution into a 50ml measuring flask, adding a solvent n-hexane-isopropanol 90:10 to dilute the solution to a scale, and shaking up to obtain the parecoxib sodium positional isomer reference substance solution.
Test solution: taking about 25mg of parecoxib sodium, precisely weighing, placing in a 25ml measuring flask, adding absolute ethyl alcohol to dissolve and dilute to a scale, shaking up, and preparing into concentrated solution of parecoxib sodium test sample. 1ml of parecoxib sodium test sample concentrated solution is precisely measured, placed in a 20ml measuring flask, diluted to scale by adding solvent n-hexane-isopropanol (90: 10), and shaken up to be used as test sample solution.
The measurement method is as follows:
precisely measuring 100 mu l of system applicability test solution, injecting the solution into a liquid chromatograph, recording a chromatogram, wherein the appearance sequence of the peaks is a parecoxib sodium peak and a parecoxib sodium position isomer peak, the separation degree is more than 1.5, and the number of theoretical plates is not less than 2000 calculated according to the parecoxib sodium peak.
In the following examples, the volumes in the mobile phase are all calculated in mL.
Example 1
The test conditions are as follows: the model of the chromatographic column is AD-H, 150 multiplied by 4.6mm and 5 mu m, the mobile phase is a mixed solvent of normal hexane, isopropanol and trifluoroacetic acid with the volume ratio of 80:20:0.1, the detection wavelength is 215nm, the temperature of the chromatographic column is 35 ℃, the flow rate of the mobile phase is 1 mL/min, and 100 mu L of parecoxib sodium positional isomer reference solution, test solution and system applicability test solution are respectively injected into a liquid chromatograph. The specific results are shown in Table 1, and the liquid chromatogram is shown in FIG. 1.
TABLE 1
In the data of table 1, parecoxib sodium and parecoxib sodium positional isomers peak sequentially, and the separation degree is 7.870.
Example 2
The test conditions are as follows: the model of the chromatographic column is AD-H, 150 multiplied by 4.6mm and 5 mu m, the mobile phase is a mixed solvent of normal hexane, isopropanol and trifluoroacetic acid with the volume ratio of 80:20:0.1, the detection wavelength is 215nm, the temperature of the chromatographic column is 30 ℃, the flow rate of the mobile phase is 1 mL/min, and 100 mu L of parecoxib sodium positional isomer reference solution, test solution and system applicability test solution are respectively injected into a liquid chromatograph. The specific results are shown in Table 2, and the liquid chromatogram is shown in FIG. 2.
Example 3
The test conditions are as follows: the model of the chromatographic column is AD-H, 150 x 4.6mm, 5 mu m, the mobile phase is a mixed solvent of normal hexane, isopropanol and trifluoroacetic acid with the volume ratio of 80:20:0.1, the detection wavelength is 215nm, the temperature of the chromatographic column is 40 ℃, the flow rate of the mobile phase is 1 mL/min, and 100 mu L of each of parecoxib sodium positional isomer reference solution, test solution and system applicability test solution is injected into a liquid chromatograph. The specific results are shown in Table 2, and the liquid chromatogram is shown in FIG. 3.
Example 4
The test conditions are as follows: the model of the chromatographic column is AD-H, 150 multiplied by 4.6mm and 5 mu m, the mobile phase is a mixed solvent of normal hexane, isopropanol and trifluoroacetic acid with the volume ratio of 80:20:0.1, the detection wavelength is 220nm, the temperature of the chromatographic column is 35 ℃, the flow rate of the mobile phase is 1 mL/min, and 100 mu L of parecoxib sodium positional isomer reference solution, test solution and system applicability test solution are respectively injected into a liquid chromatograph. The specific results are shown in Table 2, and the liquid chromatogram is shown in FIG. 4.
Example 5
The test conditions are as follows: the model of the chromatographic column is AD-H, 150 multiplied by 4.6mm and 5 mu m, the mobile phase is a mixed solvent of normal hexane, isopropanol and trifluoroacetic acid with the volume ratio of 80:20:0.1, the detection wavelength is 217nm, the temperature of the chromatographic column is 35 ℃, the flow rate of the mobile phase is 1 mL/min, and 100 mu L of parecoxib sodium positional isomer reference solution, test solution and system applicability test solution are respectively injected into a liquid chromatograph. The specific results are shown in Table 2, and the liquid chromatogram is shown in FIG. 5.
Example 6
The test conditions are as follows: the model of the chromatographic column is AD-H, 150 multiplied by 4.6mm and 5 mu m, the mobile phase is a mixed solvent of normal hexane, isopropanol and trifluoroacetic acid with the volume ratio of 80:20:0.1, the detection wavelength is 215nm, the temperature of the chromatographic column is 35 ℃, the flow rate of the mobile phase is 0.8mL/min, and 100 mu L of parecoxib sodium positional isomer reference solution, test solution and system applicability test solution are respectively injected into a liquid chromatograph. The results are shown in Table 2 and the liquid chromatogram is shown in FIG. 6.
Example 7
The test conditions are as follows: the model of the chromatographic column is AD-H, 150 multiplied by 4.6mm and 5 mu m, the mobile phase is a mixed solvent of normal hexane, isopropanol and trifluoroacetic acid with the volume ratio of 80:20:0.1, the detection wavelength is 215nm, the temperature of the chromatographic column is 35 ℃, the flow rate of the mobile phase is 1.2 mL/min, and 100 mu L of parecoxib sodium positional isomer reference solution, test solution and system applicability test solution are respectively injected into a liquid chromatograph. The results are shown in Table 2 and the liquid chromatogram is shown in FIG. 7.
Example 8
The test conditions are as follows: the model of the chromatographic column is AD-H, 150 multiplied by 4.6mm and 5 mu m, the mobile phase is a mixed solvent of n-hexane, isopropanol and trifluoroacetic acid with the volume ratio of 85:15:0.1, the detection wavelength is 215nm, the temperature of the chromatographic column is 35 ℃, the flow rate of the mobile phase is 1 mL/min, and 100 mu L of parecoxib sodium positional isomer reference solution, test solution and system applicability test solution are respectively injected into a liquid chromatograph. The specific results are shown in Table 2, and the liquid chromatogram is shown in FIG. 8.
Example 9
The test conditions are as follows: the model of the chromatographic column is AD-H, 150 multiplied by 4.6mm and 5 mu m, the mobile phase is a mixed solvent of n-hexane, isopropanol and trifluoroacetic acid with the volume ratio of 75:25:0.1, the detection wavelength is 215nm, the temperature of the chromatographic column is 35 ℃, the flow rate of the mobile phase is 1 mL/min, and 100 mu L of parecoxib sodium positional isomer reference solution, test solution and system applicability test solution are respectively injected into a liquid chromatograph. The specific results are shown in Table 2, and the liquid chromatogram is shown in FIG. 9.
TABLE 2 (examples 2 to 9)
As can be seen from the data in the table 2, the scheme provided by the application has the advantages of good detection effect, excellent stability and accordance with actual requirements.
Example 10
Taking 12.5mg of parecoxib sodium positional isomer reference substance, precisely weighing, placing in a 25ml measuring flask, adding absolute ethyl alcohol to dissolve and dilute to a scale, and shaking up; precisely measuring 0.5ml of the solution, placing the solution into a 50ml measuring flask, adding a solvent n-hexane-isopropanol (90: 10) to dilute the solution to a scale, and shaking the solution uniformly to serve as a parecoxib sodium positional isomer reference substance stock solution; precisely measuring 0.5ml of the parecoxib sodium positional isomer reference substance stock solution, putting the parecoxib sodium positional isomer reference substance stock solution into a 50ml measuring flask, adding a solvent n-hexane-isopropanol (90: 10) to dilute the solution to a scale, and shaking the solution uniformly to obtain the parecoxib sodium positional isomer reference substance solution.
And precisely weighing about 12.5mg of parecoxib sodium, placing the parecoxib sodium into a 25ml measuring flask, adding absolute ethyl alcohol to dissolve and dilute the parecoxib sodium to a scale, and shaking up to prepare a parecoxib sodium reference solution.
The preparation method of the sample solution with 50 percent impurity limit sample addition comprises the following steps: accurately measuring 5ml of parecoxib sodium reference solution and 0.25ml of parecoxib sodium positional isomer reference solution, placing the parecoxib sodium reference solution and the parecoxib sodium positional isomer reference solution into a 50ml measuring flask, adding a solvent n-hexane-isopropanol (90: 10) to dilute to a scale, shaking uniformly, and adding the sample solution as 50% impurity limit.
The preparation method of the sample solution with 100 percent impurity limit sample addition comprises the following steps: accurately measuring 5ml of parecoxib sodium reference solution and 0.5ml of parecoxib sodium positional isomer reference solution, placing the parecoxib sodium reference solution and the parecoxib sodium positional isomer reference solution into a 50ml measuring flask, adding a solvent n-hexane-isopropanol (90: 10) to dilute to a scale, shaking uniformly, and adding the sample solution as 100% impurity limit.
The preparation method of the sample solution with 120% impurity limit sample addition comprises the following steps: accurately measuring 5ml of parecoxib sodium reference solution and 0.6ml of parecoxib sodium positional isomer reference solution, placing the parecoxib sodium reference solution and the parecoxib sodium positional isomer reference solution into a 50ml measuring flask, adding a solvent n-hexane-isopropanol (90: 10) to dilute to a scale, shaking uniformly, and adding the sample solution as a 120% impurity limit.
Each concentration was made in 3 parts in parallel.
The parecoxib sodium positional isomer reference solution, the test solution and the sample-adding test solution are measured accurately, each 100 mu L, and injected into a chromatograph to calculate the recovery rate, and the results are shown in the following table.
TABLE 3 accuracy test results
And (4) conclusion: the average recovery rate of the parecoxib sodium positional isomer is 99.6% within the limit range of 50% -120%, the RSD is 3.32%, the requirements that the recovery rate is 90% -108% and the RSD is less than 5% are met, and the accuracy of the method is good.
COMPARATIVE EXAMPLE 1 (CN 104965041B EXAMPLE 1)
A high performance liquid chromatography separation detection method for parecoxib sodium isomer is provided, wherein the volume ratio of mobile phase n-hexane-isopropanol is 70: 30. The column was an Agela technologessilica 4.6X 250mm, 5 μm. The flow rate of the mobile phase is 1.0ml/min, the temperature of the chromatographic column is 35 ℃, and the detection wavelength is 215 nm. Preparing a test solution: taking a proper amount of a test sample, precisely weighing, and preparing a solution containing 0.4mg of parecoxib sodium per 1ml by using a diluent as a test sample solution; preparing a reference substance solution: taking a proper amount of parecoxib sodium isomer reference substance and parecoxib sodium reference substance, precisely weighing and preparing a mixed solution containing 0.8 mu g of parecoxib sodium isomer and 0.4mg of parecoxib sodium per 1ml of diluent as a reference substance solution; and (3) detection: precisely measuring 10 μ l of reference solution, injecting into chromatograph, precisely measuring 10 μ l of test solution, and injecting into liquid chromatograph, wherein the diluent is n-hexane-isopropanol at volume ratio of 50: 50. The comparative results are shown in Table 4.
COMPARATIVE EXAMPLE 2 (CN 104965041B EXAMPLE 2)
A high performance liquid chromatography separation detection method for parecoxib sodium isomer is provided, wherein the volume ratio of n-hexane to isopropanol of a mobile phase is 80: 20. The column was an Agela technologessilica 4.6X 250mm, 5 μm. The flow rate of the mobile phase is 0.8ml/min, the temperature of the chromatographic column is 30 ℃, and the detection wavelength is 215 nm. Preparing a test solution: taking a proper amount of a test sample, precisely weighing, and preparing a solution containing 0.4mg of parecoxib sodium per 1ml by using a diluent as a test sample solution, wherein the diluent is n-hexane-isopropanol in a volume ratio of 50: 50; preparing a reference substance solution: taking a proper amount of parecoxib sodium isomer reference substance and parecoxib sodium reference substance, precisely weighing a mixed solution which is prepared by using a diluent and contains 0.8 mu g of parecoxib sodium isomer and 0.4mg of parecoxib sodium per 1ml as a reference substance solution, wherein the diluent is n-hexane-isopropanol, the volume ratio is 50:50, precisely measuring 10 mu l of the reference substance solution, injecting the reference substance solution into a chromatograph, precisely measuring 10 mu l of the sample solution, and injecting the sample solution into a liquid chromatograph. The comparative results are shown in Table 4.
COMPARATIVE EXAMPLE 3 (CN 104965041B example 3)
A high performance liquid chromatography separation detection method for parecoxib sodium isomer is provided, wherein the volume ratio of n-hexane to isopropanol of a mobile phase is 90: 10. The column was an Agela technologessilica 4.6X 250mm, 5 μm. The flow rate of the mobile phase is 0.7ml/min, the temperature of the chromatographic column is 40 ℃, and the detection wavelength is 215 nm. Preparing a test solution: taking a proper amount of a test sample, precisely weighing, and preparing a solution containing 0.4mg of parecoxib sodium per 1ml by using a diluent as a test sample solution; preparation of a reference solution: taking a proper amount of parecoxib sodium isomer reference substance and parecoxib sodium reference substance, precisely weighing and preparing a mixed solution containing 0.8 mu g of parecoxib sodium isomer and 0.4mg of parecoxib sodium per 1ml of diluent as a reference substance solution; precisely measuring 10 μ l of the reference solution, injecting into a chromatograph, precisely measuring 10 μ l of the sample solution, and injecting into a liquid chromatograph, wherein the diluent is n-hexane-isopropanol at a volume ratio of 50: 50. The comparative results are shown in Table 4.
COMPARATIVE EXAMPLE 4 (CN 104965041B EXAMPLE 4)
A high performance liquid chromatography separation detection method for parecoxib sodium isomer is disclosed, wherein the volume ratio of mobile phase n-hexane-isopropanol is 95: 5. The column was an Agela technologessilica 4.6X 250mm, 5 μm. The flow rate of the mobile phase is 0.6ml/min, the temperature of the chromatographic column is 30 ℃, and the detection wavelength is 215 nm. Preparing a test solution: taking a proper amount of a test sample, precisely weighing, and preparing a solution containing 0.4mg of parecoxib sodium per 1ml by using a diluent as a test sample solution; preparation of a reference solution: taking a proper amount of parecoxib sodium isomer reference substance and parecoxib sodium reference substance, precisely weighing and preparing a mixed solution containing 0.8 mu g of parecoxib sodium isomer and 0.4mg of parecoxib sodium per 1ml of diluent as a reference substance solution; precisely measuring 10 μ l of the reference solution, injecting into a chromatograph, precisely measuring 10 μ l of the sample solution, and injecting into a liquid chromatograph, wherein the diluent is n-hexane-isopropanol at a volume ratio of 50: 50. The comparative results are shown in Table 4.
TABLE 4
Taking example 1 as a comparison representation of the invention, the comparison shows that the detection method of the invention is superior to the existing detection method in both the theoretical plate number value of the main peak and the isomer peak and the separation degree value. Therefore, the method is more suitable for detecting the parecoxib sodium positional isomer.
The present invention is illustrated by the above examples and comparative examples to illustrate the operation and advantages of the method for detecting parecoxib sodium regioisomer by liquid chromatography, but the present invention is not limited to the above examples. It should be understood by those skilled in the art that any modifications, equivalent substitutions, improvements, etc. made herein are intended to fall within the scope and disclosure of the present invention.
Claims (10)
1. A method for detecting parecoxib sodium positional isomer by liquid chromatography is characterized by comprising the following steps:
liquid chromatography conditions: the chromatographic column is a liquid phase chiral chromatographic column which is a polysaccharide derivative normal phase coating type chiral chromatographic column CHIRALPAK AD-H; the mobile phase is a mixed solution of n-hexane, isopropanol and trifluoroacetic acid.
2. The method for detecting parecoxib sodium positional isomers through liquid chromatography according to claim 1, wherein the chromatographic column is a polysaccharide derivative normal phase coating type chiral chromatographic column CHIRALPAK AD-H, 4.6mm x 250mm, 5 μm; the volume ratio of the n-hexane to the isopropanol in the mobile phase is (75-90) to (10-25), and the volume ratio of the trifluoroacetic acid is 0.1% of the whole mobile phase.
3. The method for detecting parecoxib sodium positional isomers through liquid chromatography according to claim 2, wherein stationary phases of a chromatographic column are as follows: the silica gel surface was coated with amylose-tris (3, 5-xylylcarbamate).
4. The method for detecting parecoxib sodium positional isomers through liquid chromatography as claimed in claim 2, wherein the volume ratio of n-hexane to isopropanol in the mobile phase is (75-85): (15-25), and the volume ratio of trifluoroacetic acid is 0.1% of the whole mobile phase.
5. The method for detecting parecoxib sodium positional isomer according to claim 1 by liquid chromatography, wherein the detection wavelength is 215 nm-220 nm; the flow rate of the mobile phase is 0.8-1.2 mL/min; the temperature of the chromatographic column is 30-40 ℃.
6. The method for detecting parecoxib sodium positional isomers according to claim 5, wherein the detection wavelengths are 215nm, 217nm and 220 nm.
7. The method for detecting parecoxib sodium positional isomers according to claim 5, wherein the flow rate of the mobile phase is 0.8mL/min, 1.0mL/min, or 1.2 mL/min.
8. The method for detecting parecoxib sodium positional isomers according to claim 5, wherein the temperature of a chromatographic column is 30 ℃, 35 ℃ and 40 ℃.
9. The method for detecting parecoxib sodium positional isomers through liquid chromatography as claimed in claim 4, wherein the volume ratio of n-hexane to isopropanol in a mobile phase is (80-85) to (15-20).
10. The method for detecting parecoxib sodium positional isomers through liquid chromatography according to claim 1, comprising the following test solution preparation: precisely weighing a to-be-measured product, placing the to-be-measured product in a volumetric flask, adding absolute ethyl alcohol to dissolve, shaking up, fixing the volume to a scale mark to serve as a stock solution, then placing a certain amount of the stock solution in the volumetric flask, adding a diluent n-hexane-isopropanol 90:10 to dilute, shaking up, and fixing the volume to the scale mark.
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Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103604894A (en) * | 2013-11-07 | 2014-02-26 | 深圳万乐药业有限公司 | Method for separating and determining bortezomib chiral isomers through high-performance liquid chromatography |
| CN104965041A (en) * | 2015-06-11 | 2015-10-07 | 成都克莱蒙医药科技有限公司 | High performance liquid chromatography detection method for parecoxib sodium isomer |
| CN105301135A (en) * | 2015-11-20 | 2016-02-03 | 扬子江药业集团有限公司 | Method for detecting chiral isomer content of ticagrelor by high performance liquid chromatography |
| CN106362714A (en) * | 2016-10-14 | 2017-02-01 | 宁夏大学 | Chiral chromatography stationary phase with silver functionalized silicon base and preparation method thereof |
| CN107966511A (en) * | 2017-11-23 | 2018-04-27 | 中山奕安泰医药科技有限公司 | The method for detecting (R) -1- (1- naphthyls) ethamine chiral isomer |
| CN111060625A (en) * | 2019-12-31 | 2020-04-24 | 北京鑫开元医药科技有限公司 | Detection method of 2- (thiophene-2-yl) ethyl p-toluenesulfonate and isomer thereof |
| CN113075307A (en) * | 2021-03-08 | 2021-07-06 | 瑞阳制药股份有限公司 | Detection method of propane fumarate tenofovir isomer |
| CN113866282A (en) * | 2020-06-30 | 2021-12-31 | 天津药业研究院股份有限公司 | Separation and detection method for isomer impurities in 3-halogenated-7- (4-bromobenzoyl) -1-hydro-indole and application thereof |
| CN114062518A (en) * | 2020-07-29 | 2022-02-18 | 武汉武药制药有限公司 | Method for separating and measuring tartaric acid optical isomer content |
| CN114471496A (en) * | 2021-12-23 | 2022-05-13 | 济宁医学院 | Preparation method and application of multi-mode and multifunctional molecular cage high performance liquid chromatography stationary phase |
-
2022
- 2022-05-31 CN CN202210602882.3A patent/CN114689761B/en active Active
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103604894A (en) * | 2013-11-07 | 2014-02-26 | 深圳万乐药业有限公司 | Method for separating and determining bortezomib chiral isomers through high-performance liquid chromatography |
| CN104965041A (en) * | 2015-06-11 | 2015-10-07 | 成都克莱蒙医药科技有限公司 | High performance liquid chromatography detection method for parecoxib sodium isomer |
| CN105301135A (en) * | 2015-11-20 | 2016-02-03 | 扬子江药业集团有限公司 | Method for detecting chiral isomer content of ticagrelor by high performance liquid chromatography |
| CN106362714A (en) * | 2016-10-14 | 2017-02-01 | 宁夏大学 | Chiral chromatography stationary phase with silver functionalized silicon base and preparation method thereof |
| CN107966511A (en) * | 2017-11-23 | 2018-04-27 | 中山奕安泰医药科技有限公司 | The method for detecting (R) -1- (1- naphthyls) ethamine chiral isomer |
| CN111060625A (en) * | 2019-12-31 | 2020-04-24 | 北京鑫开元医药科技有限公司 | Detection method of 2- (thiophene-2-yl) ethyl p-toluenesulfonate and isomer thereof |
| CN113866282A (en) * | 2020-06-30 | 2021-12-31 | 天津药业研究院股份有限公司 | Separation and detection method for isomer impurities in 3-halogenated-7- (4-bromobenzoyl) -1-hydro-indole and application thereof |
| CN114062518A (en) * | 2020-07-29 | 2022-02-18 | 武汉武药制药有限公司 | Method for separating and measuring tartaric acid optical isomer content |
| CN113075307A (en) * | 2021-03-08 | 2021-07-06 | 瑞阳制药股份有限公司 | Detection method of propane fumarate tenofovir isomer |
| CN114471496A (en) * | 2021-12-23 | 2022-05-13 | 济宁医学院 | Preparation method and application of multi-mode and multifunctional molecular cage high performance liquid chromatography stationary phase |
Non-Patent Citations (3)
| Title |
|---|
| 倪静文等: "手性固定相HPLC法测定替格瑞洛片中替格瑞洛异构体含量", 《中国新药杂志》 * |
| 聂忠莉等: "基于高效液相色谱法的帕瑞昔布钠位置异构体测定", 《成都大学学报(自然科学版)》 * |
| 霍立茹等: "替格瑞洛中异构体杂质的控制", 《药物分析杂志》 * |
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