CN113866318A - Method for detecting (6-aminopyridine-2-yl) (1-methylpiperidine-4-yl) methanone dihydrochloride - Google Patents
Method for detecting (6-aminopyridine-2-yl) (1-methylpiperidine-4-yl) methanone dihydrochloride Download PDFInfo
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Abstract
The invention belongs to the technical field of organic matter detection and analysis, and discloses a method for detecting (6-aminopyridine-2-yl) (1-methylpiperidine-4-yl) methanone dihydrochloride. The detection method comprises the following steps: detecting (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride by adopting a liquid chromatography; the conditions for liquid chromatography include: the adopted mobile phase comprises a mobile phase A and a mobile phase B; the mobile phase A is phosphate buffer solution; the mobile phase B is acetonitrile; the mobile phase is subjected to gradient elution according to certain conditions. The detection method can accurately detect the purity of (6-aminopyridine-2-yl) (1-methylpiperidine-4-yl) methanone dihydrochloride in a test sample.
Description
Technical Field
The invention belongs to the technical field of organic matter detection and analysis, and particularly relates to a method for detecting (6-aminopyridine-2-yl) (1-methylpiperidine-4-yl) methanone dihydrochloride.
Background
(6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride, CAS number: 613678-10-7, which is an important intermediate for producing the drug rasemiptan, and the purity and the impurity thereof directly influence the purity and the impurity size of the drug, thereby directly influencing the curative effect of the drug.
At present, no relevant documents and reports of a purity detection method of (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride are found, and in order to strengthen quality control of (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride and further control quality of rasemiptan, the method for detecting purity and impurity content of (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride is particularly important.
Disclosure of Invention
The present invention is directed to solving at least one of the problems of the prior art described above. Therefore, the invention provides a detection method of (6-aminopyridine-2-yl) (1-methylpiperidine-4-yl) methanone dihydrochloride, which has the advantages of simplicity, accuracy, good repeatability, rapidness, reliability and the like.
The invention provides a method for detecting (6-aminopyridin-2-yl) (1-methylpiperidine-4-yl) methanone dihydrochloride, which comprises the following steps:
detecting (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride by adopting a liquid chromatography;
the conditions of the liquid chromatography include: the adopted mobile phase comprises a mobile phase A and a mobile phase B; the mobile phase A is phosphate buffer solution; the mobile phase B is acetonitrile;
the mobile phase was eluted with a gradient as follows:
0 minute, 65-75% of mobile phase A and 25-35% of mobile phase B;
5 minutes, 65-75% of mobile phase A and 25-35% of mobile phase B;
for 25 minutes, the content of the mobile phase A is 15-25%, and the content of the mobile phase B is 75-85%;
for 30 minutes, the content of the mobile phase A is 15-25%, and the content of the mobile phase B is 75-85%;
35 minutes, 65-75% of mobile phase A and 25-35% of mobile phase B;
and the content of the mobile phase A is 65-75% and the content of the mobile phase B is 25-35% in 40 minutes.
Preferably, the detection method comprises the following steps:
dissolving (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride in a diluent to obtain a reference solution;
dissolving a test sample containing (6-aminopyridine-2-yl) (1-methylpiperidine-4-yl) methanone dihydrochloride in a diluent to obtain a test sample solution;
taking the reference solution, detecting by liquid chromatography, and recording a chromatogram map A;
taking the test solution, detecting by liquid chromatography, and recording a chromatogram map B;
calculating the purity of (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride according to the chromatogram A and the chromatogram B;
the conditions of the liquid chromatography include: the adopted mobile phase comprises a mobile phase A and a mobile phase B; the mobile phase A is phosphate buffer solution; the mobile phase B is acetonitrile;
the mobile phase was eluted with a gradient as follows:
0 minute, 65-75% of mobile phase A, 25-35% of mobile phase B, preferably 70% of mobile phase A and 20% of mobile phase B;
5 minutes, 65-75% of mobile phase A, 25-35% of mobile phase B, 70% of mobile phase A and 20% of mobile phase B are preferred;
for 25 minutes, the content of the mobile phase A is 15-25%, the content of the mobile phase B is 75-85%, preferably the content of the mobile phase A is 20%, and the content of the mobile phase B is 70%;
for 30 minutes, the content of the mobile phase A is 15-25%, the content of the mobile phase B is 75-85%, preferably the content of the mobile phase A is 20%, and the content of the mobile phase B is 70%;
35 minutes, 65-75% of mobile phase A, 25-35% of mobile phase B, 70% of mobile phase A and 20% of mobile phase B are preferred;
for 40 minutes, the content of the mobile phase A is 65-75%, the content of the mobile phase B is 25-35%, and the content of the mobile phase A is 70% and the content of the mobile phase B is 20% are preferred.
Preferably, the diluent is a mixture of water and acetonitrile, and the volume ratio of the water to the acetonitrile in the diluent is (1-2) to (1-2), more preferably 1: 1; the concentration of the (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride in the control solution is 0.25 to 1mg/mL, and more preferably 0.5 mg/mL; in the test solution, the concentration of the test is 0.25-1mg/mL, more preferably 0.5 mg/mL.
Preferably, the detection method further comprises the steps of a system suitability test, the system suitability test being: dissolving (6-aminopyridine-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride, (6-bromo-2-pyridyl) (1-methyl-4-piperidyl) methanone and (1-methylpiperidin-4-yl) (pyrrolidine-1-yl) methanone by using a diluent to obtain a system applicability solution; and detecting the system applicability solution by adopting liquid chromatography, and recording a chromatogram map C. The purpose of the system suitability test is primarily to determine that the chromatographic system used for the analysis is valid and suitable.
Preferably, the concentration of the (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride in the system-suitable solution is 0.25 to 1mg/mL, more preferably 0.5 mg/mL; the concentration of the (6-bromo-2-pyridyl) (1-methyl-4-piperidyl) methanone is 0.15-0.5mg/mL, and more preferably 0.25 mg/mL; the concentration of the (1-methylpiperidin-4-yl) (pyrrolidin-1-yl) methanone is 0.15 to 0.5mg/mL, more preferably 0.25 mg/mL.
Preferably, the result of said detection is: the number of theoretical plates should not be less than 2000 calculated as (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride, and the degree of separation of the main peak from adjacent impurities should be greater than 1.5.
Preferably, the detection method further comprises the step of performing a blank test, wherein the blank test is: and detecting the diluent by liquid chromatography, recording a chromatogram D, deducting a chromatogram peak in the chromatogram D from the chromatogram B, and calculating according to an area normalization method. The blank test aims to eliminate the influence caused by the diluent and can further improve the detection accuracy.
Preferably, the phosphate buffer is a potassium dihydrogen phosphate buffer; the concentration of the potassium dihydrogen phosphate buffer solution is 0.01-0.03mol/L, and more preferably 0.01 mol/L; the pH of the potassium dihydrogen phosphate buffer is 5.0 to 8.0, and more preferably 7.0. The pH of the phosphate buffer was chosen to help improve the peak profile, which is optimal especially when the pH is 7.0.
Preferably, the conditions of the liquid chromatography further comprise:
a chromatographic column: WondaSil C18 Superb (5um,4.6 x 250mm) (W) and its equivalent performance C18 chromatography column;
sample introduction amount: 5-30. mu.L, more preferably 10. mu.L;
flow rate of mobile phase: 0.5-0.7mL/min, more preferably 0.6 mL/min;
column temperature: 20-30 ℃, more preferably 26 ℃;
detection wavelength: 210-220nm, more preferably 215 nm;
a detector: an ultraviolet detector.
The invention also provides application of the detection method in preparing the lamidotril.
In particular to application of the detection method of (6-aminopyridin-2-yl) (1-methylpiperidine-4-yl) methanone dihydrochloride in preparation of rasemiptan.
Compared with the prior art, the invention has the following beneficial effects:
1. according to the invention, a phosphate buffer solution is used as a mobile phase A and acetonitrile is used as a mobile phase B, and gradient elution is carried out according to a certain proportion, so that (6-aminopyridin-2-yl) (1-methylpiperidine-4-yl) methanone dihydrochloride, known impurities and unknown impurities thereof can be effectively separated, the separation effect is improved, and the detection accuracy, the detection speed and the detection sensitivity are improved.
2. The detection method has the advantages of simplicity, convenience, accuracy, good repeatability, rapidness, reliability and the like, and is favorable for enhancing the quality control of (6-aminopyridin-2-yl) (1-methylpiperidine-4-yl) methanone dihydrochloride and further controlling the quality of the rasemiptan.
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FIG. 1 is a liquid chromatogram of a solution suitable for use in the system of example 1 of the present invention;
FIG. 2 is a first liquid chromatogram of a control solution of example 1 of the present invention;
FIG. 3 is a second liquid chromatogram of a control solution of example 1 of the present invention;
FIG. 4 is a liquid chromatogram of a test solution of example 1 of the present invention;
FIG. 5 is a liquid chromatogram of a test solution in example 2 of the present invention;
FIG. 6 is a liquid chromatogram of a test solution in example 3 of the present invention;
FIG. 7 is a liquid chromatogram of a test solution of example 4 of the present invention;
FIG. 8 is a liquid chromatogram of a solution suitable for use in the system of example 5 of the present invention;
FIG. 9 is a liquid chromatogram of a system suitability solution of example 6 of the present invention;
FIG. 10 is a liquid chromatogram of a solution suitable for use in the comparative example 1 system of the present invention;
FIG. 11 is a liquid chromatogram of a solution suitable for use in the comparative example 2 system of the present invention;
FIG. 12 is a liquid chromatogram of a solution suitable for use in the comparative example 3 system of the present invention.
Detailed Description
In order to make the technical solutions of the present invention more apparent to those skilled in the art, the following examples are given for illustration. It should be noted that the following examples are not intended to limit the scope of the claimed invention.
The starting materials, reagents or apparatuses used in the following examples are conventionally commercially available or can be obtained by conventionally known methods, unless otherwise specified.
Taking four batches of (6-aminopyridin-2-yl) (1-methylpiperidine-4-yl) methanone dihydrochloride products produced by the same production specification instruction as test products 1-4, respectively detecting according to the detection methods of examples 1-4, and calculating the purity and impurities by adopting an area normalization method. Known impurities in the (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride product are: (6-bromo-2-pyridyl) (1-methyl-4-piperidinyl) methanone and (1-methylpiperidin-4-yl) (pyrrolidin-1-yl) methanone.
Example 1
A method for detecting (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride comprises the following steps:
1) preparing a reference substance solution: taking a proper amount of (6-aminopyridine-2-yl) (1-methylpiperidine-4-yl) methanone dihydrochloride reference substance, and dissolving the reference substance by using acetonitrile aqueous solution with the volume percentage concentration of 50% to obtain reference substance solution with the concentration of 0.5 mg/mL;
2) preparing a test solution: taking a proper amount of (6-aminopyridine-2-yl) (1-methylpiperidine-4-yl) methanone dihydrochloride test sample, and dissolving the test sample by using acetonitrile aqueous solution with the volume percentage concentration of 50% to obtain a reference substance solution with the concentration of 0.5 mg/mL;
3) preparing a system applicability solution: appropriate amounts of (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride as a control, (6-bromo-2-pyridinyl) (1-methyl-4-piperidinyl) methanone and (1-methylpiperidin-4-yl) (pyrrolidin-1-yl) methanone were placed in the same volumetric flask and dissolved in 50% by volume acetonitrile in water to yield a solution containing 0.5mg of (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride as a control, 0.25mg/mL of (6-bromo-2-pyridinyl) (1-methyl-4-piperidinyl) methanone and 0.25mg/mL of (1-methylpiperidin-4-yl) (pyrrolidin-1-yl) methanone per mL A solution of a ketone to obtain a system suitability solution;
4) blank test, precisely measuring 10 μ L acetonitrile aqueous solution with volume percentage concentration of 50%, injecting into a liquid chromatograph, and recording chromatogram; the blank has no interference;
5) and (3) testing the applicability of the system: precisely measuring 10 mu L of system applicability solution, injecting into a liquid chromatograph, and recording a chromatogram, wherein the number of theoretical plates is not less than 2000 calculated according to (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride, and the separation degree of a main peak and adjacent impurities is more than 1.5;
6) testing a test solution: precisely measuring 10 μ L of the test solution, injecting into a liquid chromatograph, recording chromatogram, deducting chromatographic peak of blank test from chromatogram of the test solution, and calculating the concentration of the test solution according to area normalization method.
The test conditions of the liquid chromatograph of the present embodiment were set as follows: the chromatographic column is an Shimadzu Wondasil C18 Superb chromatographic column with the specification of 4.6 x 250mm and the diameter of 5 mu m, and the column temperature is 26 ℃; gradient elution is carried out by adopting double mobile phases, wherein the mobile phase A is potassium dihydrogen phosphate buffer solution (the pH value is adjusted to 7.0 by sodium hydroxide solution) with the concentration of 0.01mol/L, the mobile phase B is acetonitrile, and the flow rate of the mobile phase is 0.6 mL/min; the detector is an ultraviolet detector, and the detection wavelength is 215 nm; the conditions of gradient elution are shown in table 1, and the volume ratio of mobile phase a and mobile phase B is constant and gradual.
TABLE 1 gradient elution conditions
| Time (%) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 70 | 30 |
| 5 | 70 | 30 |
| 25 | 20 | 80 |
| 30 | 20 | 80 |
| 35 | 70 | 30 |
| 40 | 70 | 30 |
The chromatogram of the solution suitable for use in the system of this example is shown in FIG. 1, where "1" in FIG. 1 represents a peak corresponding to (1-methylpiperidin-4-yl) (pyrrolidin-1-yl) methanone, "2" represents a peak corresponding to (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride, "3" represents a peak corresponding to (6-bromo-2-pyridyl) (1-methyl-4-piperidyl) methanone, and "4, 5, 6, 7" represent peaks corresponding to other impurities (the ordinate "Absorbance" in mAU in FIG. 1). It can be seen from FIG. 1 that the degree of separation between the 3 major peaks is greater than 1.5 and the number of trays is high. The integration results and the detection results are shown in tables 2 and 3.
TABLE 2 integration of the chromatograms
TABLE 3 test results of System suitability solutions
In this example, the control solution was subjected to two detections, two chromatograms of the control solution are shown in fig. 2 and 3, respectively, "2" in fig. 2 and 3 represents a peak corresponding to (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride, and "1, 3, and 4" represent peaks corresponding to other impurities (the ordinate "Absorbance" in fig. 2 and 3 represents Absorbance, and the unit is mAU). The results of integration of the two and the results of detection are shown in tables 4, 5, 6 and 7.
TABLE 4 integration results of the first chromatogram
TABLE 5 first test results for control solutions
TABLE 6 integration of the second chromatogram
TABLE 7 second test results for control solutions
The peak areas of the two determinations were analyzed and the Relative Standard Deviation (RSD) was calculated as: 0.02 percent and good repeatability. The purity (relative peak area) of the two determinations was analyzed and the Relative Standard Deviation (RSD) was calculated as: 0.01 percent and good repeatability.
The chromatogram of the sample solution of this example is shown in FIG. 4, where "2" in FIG. 4 represents a peak corresponding to (1-methylpiperidin-4-yl) (pyrrolidin-1-yl) methanone, "3" represents a peak corresponding to (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride, "4" represents a peak corresponding to the maximum single impurity, and "1, 5, 6" represents peaks corresponding to other impurities (the ordinate "Absorbance" in FIG. 4 represents Absorbance, in mAU). The integration results and the detection results are shown in tables 8 and 9.
TABLE 8 integration results of chromatograms
TABLE 9 test results of the test article solutions
Example 2
A method for detecting (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride comprises the following steps:
1) preparing a reference substance solution: taking a proper amount of (6-aminopyridine-2-yl) (1-methylpiperidine-4-yl) methanone dihydrochloride reference substance, and dissolving the reference substance by using acetonitrile aqueous solution with the volume percentage concentration of 50% to obtain reference substance solution with the concentration of 0.5 mg/mL;
2) preparing a test solution: taking a proper amount of (6-aminopyridine-2-yl) (1-methylpiperidine-4-yl) methanone dihydrochloride test sample, and dissolving the test sample by using acetonitrile aqueous solution with the volume percentage concentration of 50% to obtain a reference substance solution with the concentration of 0.5 mg/mL;
3) preparing a system applicability solution: appropriate amounts of (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride as a control, (6-bromo-2-pyridinyl) (1-methyl-4-piperidinyl) methanone and (1-methylpiperidin-4-yl) (pyrrolidin-1-yl) methanone were placed in the same volumetric flask and dissolved in 50% by volume acetonitrile in water to yield a solution containing 0.5mg of (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride as a control, 0.25mg/mL of (6-bromo-2-pyridinyl) (1-methyl-4-piperidinyl) methanone and 0.25mg/mL of (1-methylpiperidin-4-yl) (pyrrolidin-1-yl) methanone per mL A solution of a ketone to obtain a system suitability solution;
4) blank test, precisely measuring 10 μ L acetonitrile aqueous solution with volume percentage concentration of 50%, injecting into a liquid chromatograph, and recording chromatogram; the blank has no interference;
5) and (3) testing the applicability of the system: precisely measuring 10 mu L of system applicability solution, injecting into a liquid chromatograph, and recording a chromatogram, wherein the number of theoretical plates is not less than 2000 calculated according to (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride, and the separation degree of a main peak and adjacent impurities is more than 1.5;
6) testing a test solution: precisely measuring 10 μ L of the test solution, injecting into a liquid chromatograph, recording chromatogram, deducting chromatographic peak of blank test from chromatogram of the test solution, and calculating the concentration of the test solution according to area normalization method.
The test conditions of the liquid chromatograph of the present embodiment were set as follows: the chromatographic column is an Shimadzu Wondasil C18 Superb chromatographic column with the specification of 4.6 x 250mm and the diameter of 5 mu m, and the column temperature is 26 ℃; gradient elution is carried out by adopting double mobile phases, wherein the mobile phase A is potassium dihydrogen phosphate buffer solution (the pH value is adjusted to 7.0 by sodium hydroxide solution) with the concentration of 0.01mol/L, the mobile phase B is acetonitrile, and the flow rate of the mobile phase is 0.6 mL/min; the detector is an ultraviolet detector, and the detection wavelength is 215 nm; the conditions for gradient elution are shown in Table 10.
TABLE 10 gradient elution conditions
| Time (%) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 70 | 30 |
| 5 | 70 | 30 |
| 25 | 20 | 80 |
| 30 | 20 | 80 |
| 35 | 70 | 30 |
| 40 | 70 | 30 |
The chromatogram and integration results of the sample solution of this example are shown in FIG. 5, where "2" in FIG. 5 represents a peak corresponding to (1-methylpiperidin-4-yl) (pyrrolidin-1-yl) methanone, "3" represents a peak corresponding to (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride, "4" represents a peak corresponding to the maximum single impurity, and "1, 5" represent peaks corresponding to other impurities (the ordinate "Absorbance" in FIG. 5 represents Absorbance, in mAU). The integration results and the detection results are shown in tables 11 and 12.
TABLE 11 integration results of chromatograms
TABLE 12 test results of the test article solutions
Example 3
A method for detecting (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride comprises the following steps:
1) preparing a reference substance solution: taking a proper amount of (6-aminopyridine-2-yl) (1-methylpiperidine-4-yl) methanone dihydrochloride reference substance, and dissolving the reference substance by using acetonitrile aqueous solution with the volume percentage concentration of 50% to obtain reference substance solution with the concentration of 0.5 mg/mL;
2) preparing a test solution: taking a proper amount of (6-aminopyridine-2-yl) (1-methylpiperidine-4-yl) methanone dihydrochloride test sample, and dissolving the test sample by using acetonitrile aqueous solution with the volume percentage concentration of 50% to obtain a reference substance solution with the concentration of 0.5 mg/mL;
3) preparing a system applicability solution: appropriate amounts of (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride as a control, (6-bromo-2-pyridinyl) (1-methyl-4-piperidinyl) methanone and (1-methylpiperidin-4-yl) (pyrrolidin-1-yl) methanone were placed in the same volumetric flask and dissolved in 50% by volume acetonitrile in water to yield a solution containing 0.5mg of (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride as a control, 0.25mg/mL of (6-bromo-2-pyridinyl) (1-methyl-4-piperidinyl) methanone and 0.25mg/mL of (1-methylpiperidin-4-yl) (pyrrolidin-1-yl) methanone per mL A solution of a ketone to obtain a system suitability solution;
4) blank test, precisely measuring 10 μ L acetonitrile aqueous solution with volume percentage concentration of 50%, injecting into a liquid chromatograph, and recording chromatogram; the blank has no interference;
5) and (3) testing the applicability of the system: precisely measuring 10 mu L of system applicability solution, injecting into a liquid chromatograph, and recording a chromatogram, wherein the number of theoretical plates is not less than 2000 calculated according to (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride, and the separation degree of a main peak and adjacent impurities is more than 1.5;
6) testing a test solution: precisely measuring 10 μ L of the test solution, injecting into a liquid chromatograph, recording chromatogram, deducting chromatographic peak of blank test from chromatogram of the test solution, and calculating the concentration of the test solution according to area normalization method.
The test conditions of the liquid chromatograph of the present embodiment were set as follows: the chromatographic column is an Shimadzu Wondasil C18 Superb chromatographic column with the specification of 4.6 x 250mm and the diameter of 5 mu m, and the column temperature is 26 ℃; gradient elution is carried out by adopting double mobile phases, wherein the mobile phase A is potassium dihydrogen phosphate buffer solution (the pH value is adjusted to 7.0 by sodium hydroxide solution) with the concentration of 0.01mol/L, the mobile phase B is acetonitrile, and the flow rate of the mobile phase is 0.6 mL/min; the detector is an ultraviolet detector, and the detection wavelength is 215 nm; the conditions for gradient elution are shown in Table 13.
TABLE 13 gradient elution conditions
| Time (%) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 70 | 30 |
| 5 | 70 | 30 |
| 25 | 20 | 80 |
| 30 | 20 | 80 |
| 35 | 70 | 30 |
| 40 | 70 | 30 |
The chromatogram and integration results of the sample solution of this example are shown in fig. 6, where "2" in fig. 6 represents a peak corresponding to (1-methylpiperidin-4-yl) (pyrrolidin-1-yl) methanone, "3" represents a peak corresponding to (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride, "4" represents a peak corresponding to the maximum single impurity, and "1, 5, 6, 7" represents peaks corresponding to other impurities (the ordinate "Absorbance" in fig. 6 represents mAU). The integration results and the detection results are shown in tables 14 and 15.
TABLE 14 integration results of chromatograms
TABLE 15 test results of the test article solutions
Example 4
A method for detecting (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride comprises the following steps:
1) preparing a reference substance solution: taking a proper amount of (6-aminopyridine-2-yl) (1-methylpiperidine-4-yl) methanone dihydrochloride reference substance, and dissolving the reference substance by using acetonitrile aqueous solution with the volume percentage concentration of 50% to obtain reference substance solution with the concentration of 0.5 mg/mL;
2) preparing a test solution: taking a proper amount of (6-aminopyridine-2-yl) (1-methylpiperidine-4-yl) methanone dihydrochloride test sample, and dissolving the test sample by using acetonitrile aqueous solution with the volume percentage concentration of 50% to obtain a reference substance solution with the concentration of 0.5 mg/mL;
3) preparing a system applicability solution: appropriate amounts of (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride as a control, (6-bromo-2-pyridinyl) (1-methyl-4-piperidinyl) methanone and (1-methylpiperidin-4-yl) (pyrrolidin-1-yl) methanone were placed in the same volumetric flask and dissolved in 50% by volume acetonitrile in water to yield a solution containing 0.5mg of (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride as a control, 0.25mg/mL of (6-bromo-2-pyridinyl) (1-methyl-4-piperidinyl) methanone and 0.25mg/mL of (1-methylpiperidin-4-yl) (pyrrolidin-1-yl) methanone per mL A solution of a ketone to obtain a system suitability solution;
4) blank test, precisely measuring 10 μ L acetonitrile aqueous solution with volume percentage concentration of 50%, injecting into a liquid chromatograph, and recording chromatogram; the blank has no interference;
5) and (3) testing the applicability of the system: precisely measuring 10 mu L of system applicability solution, injecting into a liquid chromatograph, and recording a chromatogram, wherein the number of theoretical plates is not less than 2000 calculated according to (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride, and the separation degree of a main peak and adjacent impurities is more than 1.5;
6) testing a test solution: precisely measuring 10 μ L of the test solution, injecting into a liquid chromatograph, recording chromatogram, deducting chromatographic peak of blank test from chromatogram of the test solution, and calculating the concentration of the test solution according to area normalization method.
The test conditions of the liquid chromatograph of the present embodiment were set as follows: the chromatographic column is an Shimadzu Wondasil C18 Superb chromatographic column with the specification of 4.6 x 250mm and the diameter of 5 mu m, and the column temperature is 26 ℃; gradient elution is carried out by adopting double mobile phases, wherein the mobile phase A is potassium dihydrogen phosphate buffer solution (the pH value is adjusted to 7.0 by sodium hydroxide solution) with the concentration of 0.01mol/L, the mobile phase B is acetonitrile, and the flow rate of the mobile phase is 0.6 mL/min; the detector is an ultraviolet detector, and the detection wavelength is 215 nm; the conditions for gradient elution are shown in Table 16.
TABLE 16 gradient elution conditions
The chromatogram and integration result of the sample solution of this example are shown in FIG. 7, where "1" in FIG. 7 represents the peak corresponding to the maximum single impurity, "2" represents the peak corresponding to (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride, and "3, 4" represent the peaks corresponding to other impurities (the ordinate "Absorbance" in FIG. 7 represents Absorbance, in mAU). The integration results and the detection results are shown in tables 17 and 18.
TABLE 17 integration of chromatograms
TABLE 18 test results of the test article solutions
As can be seen from the detection results of the embodiments 1 to 4, different purities, known impurities and unknown impurities thereof can be detected for different batches of samples.
Example 5
Example 5 differs from example 1 in that the pH of mobile phase A potassium dihydrogen phosphate buffer was 8.5, and the other conditions were the same as in example 1. The system suitability test was performed on the system suitability solution of example 5, and the chromatogram thereof is shown in fig. 8. As can be seen from FIG. 8, when the pH of the potassium dihydrogen phosphate buffer is not within the range defined by the present invention, the peak of the known impurity, 1-methylpiperidin-4-yl (pyrrolidin-1-yl) methanone, is branched, the peak pattern is not good, and the judgment of the unknown impurity and the known impurity is affected. The integration results are shown in Table 19.
TABLE 19 integration results of chromatograms
Example 6
Example 6 differs from example 1 in that the pH of mobile phase A potassium dihydrogen phosphate buffer was 4.5, and the other conditions were the same as in example 1. The system suitability test was performed on the system suitability solution of example 6, and the chromatogram thereof is shown in fig. 9. As can be seen from FIG. 9, when the pH of the potassium dihydrogen phosphate buffer was outside the range defined in the present invention, the substances were not separated, the degree of separation was very poor, and the peak pattern was also poor. The integration results are shown in Table 20.
TABLE 20 integration of chromatograms
Comparative example 1
Comparative example 1 differs from example 1 in that comparative example 1 employs isocratic elution, the elution conditions of which are shown in table 21.
TABLE 21 isocratic elution conditions
| Time (%) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 50 | 50 |
| 30 | 50 | 50 |
The system suitability solution of the present comparative example was subjected to a system suitability test, and its chromatogram was shown in fig. 10. As can be seen from fig. 10, comparative example 1 is not suitable for use in the chromatographic system due to the difference in the degree of separation between substances and the difference in the peak pattern caused by the use of isocratic elution. The integration results are shown in Table 22.
TABLE 22 integration of chromatograms
Comparative example 2
Comparative example 2 is different from example 1 in that the gradient elution of comparative example 2 is not within the range defined by the present invention, and the gradient elution conditions are shown in Table 23.
TABLE 23 gradient elution conditions
| Time (%) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 60 | 40 |
| 5 | 60 | 40 |
| 25 | 30 | 70 |
| 30 | 30 | 70 |
| 35 | 60 | 40 |
| 40 | 60 | 40 |
The system suitability solution of this comparative example was subjected to a system suitability test, and its chromatogram was shown in fig. 11. As can be seen from fig. 11, comparative example 2 resulted in a difference in the degree of separation between substances, a difference in the peak pattern, and the chromatographic system was not suitable because the gradient elution conditions were not within the range defined by the present invention. The integration results are shown in Table 24.
TABLE 24 integration of chromatograms
Comparative example 3
Comparative example 3 is different from example 1 in that the mobile phase B of comparative example 3 uses methanol instead of acetonitrile, and other conditions and detection methods are the same as those of example 1. The system suitability solution of the present comparative example was subjected to a system suitability test, and its chromatogram was shown in fig. 12. As can be seen from fig. 12, in comparative example 3, the separation degree was good but the peak pattern and the baseline were poor due to the replacement of acetonitrile with methanol. The integration results are shown in Table 25.
TABLE 25 integration results of chromatograms
While the preferred embodiments of the present invention have been illustrated and described, it will be understood by those skilled in the art that the present invention is not limited to the details of the embodiments shown and described, but is capable of numerous equivalents and substitutions without departing from the spirit of the invention as set forth in the claims appended hereto.
Claims (10)
1. A method for detecting (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride is characterized by comprising the following steps of:
detecting (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride by adopting a liquid chromatography;
the conditions of the liquid chromatography include: the adopted mobile phase comprises a mobile phase A and a mobile phase B; the mobile phase A is phosphate buffer solution; the mobile phase B is acetonitrile;
the mobile phase was eluted with a gradient as follows:
0 minute, 65-75% of mobile phase A and 25-35% of mobile phase B;
5 minutes, 65-75% of mobile phase A and 25-35% of mobile phase B;
for 25 minutes, the content of the mobile phase A is 15-25%, and the content of the mobile phase B is 75-85%;
for 30 minutes, the content of the mobile phase A is 15-25%, and the content of the mobile phase B is 75-85%;
35 minutes, 65-75% of mobile phase A and 25-35% of mobile phase B;
and the content of the mobile phase A is 65-75% and the content of the mobile phase B is 25-35% in 40 minutes.
2. The detection method according to claim 1, characterized by comprising the steps of:
dissolving (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride in a diluent to obtain a reference solution;
dissolving a test sample containing (6-aminopyridine-2-yl) (1-methylpiperidine-4-yl) methanone dihydrochloride in a diluent to obtain a test sample solution;
detecting the reference substance solution by liquid chromatography, and recording chromatogram A;
taking the test solution, detecting by liquid chromatography, and recording chromatogram map B;
calculating the purity of (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride according to the chromatogram A and the chromatogram B;
the conditions of the liquid chromatography include: the adopted mobile phase comprises a mobile phase A and a mobile phase B; the mobile phase A is phosphate buffer solution; the mobile phase B is acetonitrile; the mobile phase was eluted with a gradient as follows:
0 minute, 65-75% of mobile phase A and 25-35% of mobile phase B;
5 minutes, 65-75% of mobile phase A and 25-35% of mobile phase B;
for 25 minutes, the content of the mobile phase A is 15-25%, and the content of the mobile phase B is 75-85%;
for 30 minutes, the content of the mobile phase A is 15-25%, and the content of the mobile phase B is 75-85%;
35 minutes, 65-75% of mobile phase A and 25-35% of mobile phase B;
and the content of the mobile phase A is 65-75% and the content of the mobile phase B is 25-35% in 40 minutes.
3. The detection method according to claim 2, wherein the diluent is a mixture of water and acetonitrile, and the volume ratio of water to acetonitrile in the diluent is (1-2): (1-2); in the reference solution, the concentration of the (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride is 0.25-1 mg/mL; in the test solution, the concentration of the test article is 0.25-1 mg/mL.
4. The inspection method of claim 2, further comprising the step of performing a system suitability test, the system suitability test being: dissolving (6-aminopyridine-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride, (6-bromo-2-pyridyl) (1-methyl-4-piperidyl) methanone and (1-methylpiperidin-4-yl) (pyrrolidine-1-yl) methanone by using a diluent to obtain a system applicability solution; and detecting the system applicability solution by adopting liquid chromatography, and recording a chromatogram map C.
5. The detection method according to claim 4, wherein the concentration of the (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride in the system-compatible solution is 0.25 to 1mg/mL, the concentration of the (6-bromo-2-pyridyl) (1-methyl-4-piperidyl) methanone is 0.15 to 0.5mg/mL, and the concentration of the (1-methylpiperidin-4-yl) (pyrrolidin-1-yl) methanone is 0.15 to 0.5 mg/mL.
6. The detection method according to claim 4, characterized in that the result of said detection is: the number of theoretical plates should not be less than 2000 calculated as (6-aminopyridin-2-yl) (1-methylpiperidin-4-yl) methanone dihydrochloride, and the degree of separation of the main peak from adjacent impurities should be greater than 1.5.
7. The method of claim 2, further comprising the step of performing a blank test, wherein the blank test is: detecting the diluent by liquid chromatography, and recording chromatogram map D.
8. The detection method according to claim 2, wherein the phosphate buffer is a potassium dihydrogen phosphate buffer; the concentration of the potassium dihydrogen phosphate buffer solution is 0.01-0.03 mol/L; the pH value of the potassium dihydrogen phosphate buffer solution is 5.0-8.0.
9. The detection method according to claim 1, wherein the conditions of the liquid chromatography further comprise:
sample introduction amount: 5-30 μ L;
flow rate of mobile phase: 0.5-0.7 mL/min;
column temperature: 20-30 ℃;
detection wavelength: 210-220 nm.
10. Use of the assay of any one of claims 1-9 in the preparation of rasemiptan.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115356411A (en) * | 2022-08-09 | 2022-11-18 | 河北一品生物医药有限公司 | Method for detecting related substances of L-piperidinecarboxylic acid hydrochloride by high performance liquid chromatography |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1642939A (en) * | 2002-03-29 | 2005-07-20 | 伊莱利利公司 | Pyridinoylpiperidines as 5-HTLF agonists |
| WO2020095171A1 (en) * | 2018-11-05 | 2020-05-14 | Glenmark Life Sciences Limited, Glenmark Pharmaceuticals Limited | Process for preparation of lasmiditan |
| WO2021001350A1 (en) * | 2019-07-01 | 2021-01-07 | Moehs Iberica, S.L. | 2,2-dimethyl-n-[6-(1-methyl-piperidin-4-carbonyl)-pyridin-2-yl]-propionamide, method for preparing (6-amino-pyridin-2-yl)-(1-methyl-piperidin-4-yl)-methanone using said compound, and use of said compound in the preparation of lasmiditan |
| WO2021007155A1 (en) * | 2019-07-09 | 2021-01-14 | Eli Lilly And Company | Processes and intermediate for the large-scale preparation of 2,4,6-trifluoro-n-[6-(1-methyl-piperidine-4-carbonyl)-pyridin-2-yl]-benzamide hemisuccinate, and preparation of 2,4,6-trifluoro-n-[6-(1-methyl-piperidine-4-carbonyl)-pyridin-2-yl]-benzamide acetate |
| WO2021116979A1 (en) * | 2019-12-13 | 2021-06-17 | Olon S.P.A. | Process for the preparation of lasmiditan and of a synthesis intermediate |
| CN113045540A (en) * | 2019-12-27 | 2021-06-29 | 上海天慈国际药业有限公司 | Preparation method of rasemiptan |
-
2021
- 2021-10-21 CN CN202111230346.7A patent/CN113866318B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1642939A (en) * | 2002-03-29 | 2005-07-20 | 伊莱利利公司 | Pyridinoylpiperidines as 5-HTLF agonists |
| WO2020095171A1 (en) * | 2018-11-05 | 2020-05-14 | Glenmark Life Sciences Limited, Glenmark Pharmaceuticals Limited | Process for preparation of lasmiditan |
| WO2021001350A1 (en) * | 2019-07-01 | 2021-01-07 | Moehs Iberica, S.L. | 2,2-dimethyl-n-[6-(1-methyl-piperidin-4-carbonyl)-pyridin-2-yl]-propionamide, method for preparing (6-amino-pyridin-2-yl)-(1-methyl-piperidin-4-yl)-methanone using said compound, and use of said compound in the preparation of lasmiditan |
| WO2021007155A1 (en) * | 2019-07-09 | 2021-01-14 | Eli Lilly And Company | Processes and intermediate for the large-scale preparation of 2,4,6-trifluoro-n-[6-(1-methyl-piperidine-4-carbonyl)-pyridin-2-yl]-benzamide hemisuccinate, and preparation of 2,4,6-trifluoro-n-[6-(1-methyl-piperidine-4-carbonyl)-pyridin-2-yl]-benzamide acetate |
| WO2021116979A1 (en) * | 2019-12-13 | 2021-06-17 | Olon S.P.A. | Process for the preparation of lasmiditan and of a synthesis intermediate |
| CN113045540A (en) * | 2019-12-27 | 2021-06-29 | 上海天慈国际药业有限公司 | Preparation method of rasemiptan |
Non-Patent Citations (2)
| Title |
|---|
| RAJU M DAVID: "Development and Validation of HPLC Method for the Determination of Lasmiditan Drug in Bulk and Tablet Dosage Form", 《JOURNAL OF PHARMACEUTICAL SCIENCES AND RESEARCH》 * |
| 成佳威 等: "2- 氨基-6-(1- 甲基哌啶-4- 基羰基) 吡啶的新合成工艺", 《中国医药工业杂志》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115356411A (en) * | 2022-08-09 | 2022-11-18 | 河北一品生物医药有限公司 | Method for detecting related substances of L-piperidinecarboxylic acid hydrochloride by high performance liquid chromatography |
| CN115356411B (en) * | 2022-08-09 | 2023-07-28 | 河北一品生物医药有限公司 | Method for detecting related substances of L-piperidine hydrochloride by high performance liquid chromatography |
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