CN116500173A - Method for determining impurity content in rivaroxaban drug - Google Patents
Method for determining impurity content in rivaroxaban drug Download PDFInfo
- Publication number
- CN116500173A CN116500173A CN202310796354.0A CN202310796354A CN116500173A CN 116500173 A CN116500173 A CN 116500173A CN 202310796354 A CN202310796354 A CN 202310796354A CN 116500173 A CN116500173 A CN 116500173A
- Authority
- CN
- China
- Prior art keywords
- mobile phase
- rivaroxaban
- imp
- formula
- volume percentage
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- KGFYHTZWPPHNLQ-AWEZNQCLSA-N rivaroxaban Chemical compound S1C(Cl)=CC=C1C(=O)NC[C@@H]1OC(=O)N(C=2C=CC(=CC=2)N2C(COCC2)=O)C1 KGFYHTZWPPHNLQ-AWEZNQCLSA-N 0.000 title claims abstract description 82
- 229960001148 rivaroxaban Drugs 0.000 title claims abstract description 82
- 239000012535 impurity Substances 0.000 title claims abstract description 67
- 239000003814 drug Substances 0.000 title claims abstract description 50
- 229940079593 drug Drugs 0.000 title claims abstract description 42
- 238000000034 method Methods 0.000 title claims abstract description 38
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 111
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims abstract description 72
- 239000000243 solution Substances 0.000 claims abstract description 53
- 238000001514 detection method Methods 0.000 claims abstract description 43
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims abstract description 36
- 239000011259 mixed solution Substances 0.000 claims abstract description 27
- 238000010828 elution Methods 0.000 claims abstract description 24
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 24
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 15
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims abstract description 8
- 239000000377 silicon dioxide Substances 0.000 claims abstract description 7
- 239000003153 chemical reaction reagent Substances 0.000 claims description 23
- 239000012488 sample solution Substances 0.000 claims description 22
- 239000006228 supernatant Substances 0.000 claims description 10
- 238000012360 testing method Methods 0.000 claims description 9
- 238000009210 therapy by ultrasound Methods 0.000 claims description 8
- 239000000047 product Substances 0.000 claims description 6
- 238000005119 centrifugation Methods 0.000 claims description 5
- 238000004090 dissolution Methods 0.000 claims description 5
- 238000000691 measurement method Methods 0.000 claims description 3
- 238000003556 assay Methods 0.000 claims 2
- 238000003908 quality control method Methods 0.000 abstract description 6
- 238000000926 separation method Methods 0.000 abstract description 6
- 230000000052 comparative effect Effects 0.000 description 7
- 238000001816 cooling Methods 0.000 description 7
- 238000007865 diluting Methods 0.000 description 7
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 230000006378 damage Effects 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- FGDQGIKMWOAFIK-UHFFFAOYSA-N acetonitrile;phosphoric acid Chemical compound CC#N.OP(O)(O)=O FGDQGIKMWOAFIK-UHFFFAOYSA-N 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- 239000000523 sample Substances 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- XUKUURHRXDUEBC-SXOMAYOGSA-N (3s,5r)-7-[2-(4-fluorophenyl)-3-phenyl-4-(phenylcarbamoyl)-5-propan-2-ylpyrrol-1-yl]-3,5-dihydroxyheptanoic acid Chemical compound C=1C=CC=CC=1C1=C(C=2C=CC(F)=CC=2)N(CC[C@@H](O)C[C@H](O)CC(O)=O)C(C(C)C)=C1C(=O)NC1=CC=CC=C1 XUKUURHRXDUEBC-SXOMAYOGSA-N 0.000 description 3
- 239000003513 alkali Substances 0.000 description 3
- 238000004587 chromatography analysis Methods 0.000 description 3
- 238000012494 forced degradation Methods 0.000 description 3
- 238000005286 illumination Methods 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 230000003647 oxidation Effects 0.000 description 3
- 238000007254 oxidation reaction Methods 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 239000000945 filler Substances 0.000 description 2
- 230000003472 neutralizing effect Effects 0.000 description 2
- 238000002604 ultrasonography Methods 0.000 description 2
- 238000012795 verification Methods 0.000 description 2
- 230000005526 G1 to G0 transition Effects 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 230000008832 photodamage Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
The invention belongs to the technical field of pharmacy, and provides a method for measuring impurity content in rivaroxaban. The method comprises the steps of dissolving rivaroxaban drug to be detected, and then performing high performance liquid chromatography detection, wherein octadecylsilane chemically bonded silica chromatographic column is used as a chromatographic column in the high performance liquid chromatography detection, a mixed solution of acetonitrile and phosphoric acid solution is used as a mobile phase A, and acetonitrile is used as a mobile phase B; and combining gradient elution conditions to distinguish impurities in rivaroxaban medicaments to be detected one by one. The examples show that: the determination method has good specificity, the minimum separation degree of each impurity can reach 3.6, the accuracy is high, and the quantitative limit concentration of each impurity is about 0.2 mug/mL. Therefore, the determination method provided by the invention can simply, conveniently, accurately and efficiently evaluate the impurity content in the rivaroxaban drug, and has important significance for quality control and clinical application safety of the rivaroxaban drug.
Description
Technical Field
The invention relates to the technical field of pharmacy, in particular to a method for measuring impurity content in rivaroxaban.
Background
In the development process of the medicine, an impurity detection method is an important evaluation parameter for quality control and medicine safety, and the method must pass through method verification to meet the requirements of specificity, precision, accuracy and other method verification, so that the reliability of an impurity detection result can be ensured, and the method can be used for guiding the quality control of the medicine.
According to the guidance of BP2022 rivaroxaban tablets, a high performance liquid chromatography is adopted to measure related substances, wherein the chromatographic condition is that octadecylsilane chemically bonded silica gel is used as a filler (5.0 mm multiplied by 55mm,3.0 mu m) for a stationary phase; mobile phase a was acetonitrile-0.067% phosphoric acid solution (8:92); mobile phase B is acetonitrile; the detection wavelength is 250nm; the flow rate is 1.0mL per minute; and the sample injection volume is 5 mu L. The elution gradient is as shown in table 1:
TABLE 1 elution procedure
The total number of corresponding impurities is 7, and the impurities, CAS numbers, molecular structural formulas and limits are shown in Table 2.
TABLE 2 impurity, CAS number, molecular Structure and Limit
However, in the detection of substances related to actual drugs, the detection method of BP2022 rivaroxaban tablet cannot be controlled most accurately, and some impurities cannot be completely separated and cannot be quantified accurately when the detection is performed.
Disclosure of Invention
In view of the above, the invention aims to provide a method for measuring the impurity content in rivaroxaban, which increases the accuracy of impurity detection in rivaroxaban, strengthens the quality control of rivaroxaban-containing medicines and improves the safety of clinical application of rivaroxaban medicines.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a method for measuring impurity content in rivaroxaban drug, which comprises the following steps:
dissolving rivaroxaban drug to be tested to obtain a test sample solution;
performing high performance liquid chromatography detection on the sample solution to obtain chromatographic information of rivaroxaban drug to be detected;
calculating to obtain the impurity content of rivaroxaban drug to be detected based on the chromatographic information of rivaroxaban standard and the chromatographic information of rivaroxaban drug to be detected;
the parameters of the high performance liquid chromatography detection include:
the chromatographic column is octadecylsilane chemically bonded silica chromatographic column, 4.0mm×55mm,3 μm;
the mobile phase system consists of a mobile phase A and a mobile phase B;
the mobile phase A is a mixed solution of acetonitrile and a phosphoric acid solution, wherein the volume concentration of the phosphoric acid solution is 0.057-0.077%, and the volume ratio of the acetonitrile to the phosphoric acid solution in the mixed solution is 5-10: 95-90;
the mobile phase B is acetonitrile;
the elution mode is gradient elution;
the conditions of the gradient elution are as follows:
0-5 min: the volume percentage of the mobile phase A is 95%, and the volume percentage of the mobile phase B is 5%;
5 min-15 min: the volume percentage of the mobile phase A is changed from 95% of uniform velocity to 75%, and the volume percentage of the mobile phase B is changed from 5% of uniform velocity to 25%;
15-30 min: the volume percentage of the mobile phase A is changed from 75% of uniform velocity to 60%, and the volume percentage of the mobile phase B is changed from 25% of uniform velocity to 40%;
30-31 min: the volume percentage of the mobile phase A is changed from 60% of uniform velocity to 95%, and the volume percentage of the mobile phase B is changed from 40% of uniform velocity to 5%;
31-40 min: the volume percentage of the mobile phase A is 95%, and the volume percentage of the mobile phase B is 5%;
the detection wavelength is 245 nm-255 nm;
the impurities are Imp-B, imp-D, imp-E, imp-G, imp-H, imp-I and Imp-J;
the Imp-B has a structure shown in a formula 1;
formula 1;
the Imp-D has a structure shown in a formula 2;
formula 2;
the Imp-E has a structure shown in a formula 3;
formula 3;
the Imp-G has a structure shown in a formula 4;
formula 4;
the Imp-H has a structure shown in a formula 5;
formula 5;
the Imp-I has a structure shown in a formula 6;
formula 6;
the Imp-J has a structure shown in formula 7;
formula 7.
Preferably, the rivaroxaban drug to be tested comprises rivaroxaban tablets.
Preferably, the dissolved reagent is a mixed solution of acetonitrile and phosphoric acid solution, wherein the volume concentration of the phosphoric acid solution is 0.057-0.077%, and the volume ratio of acetonitrile to phosphoric acid solution in the mixed solution is 1:1.
preferably, the dosage ratio of rivaroxaban drug to be tested and dissolved reagent is 0.5g: 200-250 mL.
Preferably, the dissolution mode is ultrasonic, and the time of ultrasonic is 10min.
Preferably, after the dissolution, the method further comprises centrifuging the product obtained by the dissolution, and taking supernatant as a sample solution; the rotational speed of the centrifugation is 5000rpm, and the time is 10min.
Preferably, the parameters of the high performance liquid chromatography detection further include: the flow rate is 0.8mL/min to 1.2mL/min.
Preferably, the parameters of the high performance liquid chromatography detection further include: the column temperature is 40-50 ℃.
The invention provides a method for measuring impurity content in rivaroxaban drug, which comprises the following steps: dissolving rivaroxaban drug to be tested to obtain a test sample solution; performing high performance liquid chromatography detection on the sample solution to obtain chromatographic information of rivaroxaban drug to be detected; calculating to obtain the impurity content of rivaroxaban drug to be detected based on the chromatographic information of rivaroxaban standard and the chromatographic information of rivaroxaban drug to be detected; the parameters of the high performance liquid chromatography detection include: the chromatographic column is octadecylsilane chemically bonded silica chromatographic column, 4.0mm×55mm,3 μm; the mobile phase system consists of a mobile phase A and a mobile phase B; the mobile phase A is a mixed solution of acetonitrile and a phosphoric acid solution, wherein the volume concentration of the phosphoric acid solution is 0.057-0.077%, and the volume ratio of the acetonitrile to the phosphoric acid solution in the mixed solution is 5-10: 95-90; the mobile phase B is acetonitrile; the elution mode is gradient elution; the conditions of the gradient elution are as follows: 0-5 min: the volume percentage of the mobile phase A is 95%, and the volume percentage of the mobile phase B is 5%;5 min-15 min: the volume percentage of the mobile phase A is changed from 95% of uniform velocity to 75%, and the volume percentage of the mobile phase B is changed from 5% of uniform velocity to 25%; 15-30 min: the volume percentage of the mobile phase A is changed from 75% of uniform velocity to 60%, and the volume percentage of the mobile phase B is changed from 25% of uniform velocity to 40%; 30-31 min: the volume percentage of the mobile phase A is changed from 60% of uniform velocity to 95%, and the volume percentage of the mobile phase B is changed from 40% of uniform velocity to 5%; 31-40 min: the volume percentage of the mobile phase A is 95%, and the volume percentage of the mobile phase B is 5%; the detection wavelength is 245 nm-255 nm; the impurities are Imp-B, imp-D, imp-E, imp-G, imp-H, imp-I and Imp-J; the Imp-B has a structure shown in a formula 1;
formula 1;
the Imp-D has a structure shown in a formula 2;
formula 2;
the Imp-E has a structure shown in a formula 3;
formula 3;
the Imp-G has a structure shown in a formula 4;
formula 4;
the Imp-H has a structure shown in a formula 5;
formula 5;
the Imp-I has a structure shown in a formula 6;
formula 6;
the Imp-J has a structure shown in formula 7;
formula 7.
The determination method provided by the invention can distinguish impurities in rivaroxaban medicine to be detected one by one, so that the impurity content is accurately controlled in the medicine production process, and the medicine quality is ensured. The data of the examples show that: the determination method provided by the invention has good specificity, the minimum separation degree of each impurity can reach 3.6, the accuracy is high, and the quantitative limit concentration of each impurity is 0.2 mug/mL. Therefore, the determination method provided by the invention can simply, conveniently, accurately and efficiently evaluate the impurity content in the rivaroxaban drug, and has important significance for quality control and clinical application safety of the rivaroxaban drug.
Drawings
FIG. 1 shows the results of the chromatographic test device in example 1 of the present invention;
FIG. 2 is a graph showing the quantitative limit of impurities in example 2 of the present invention;
FIG. 3 shows the results of the detection of the specificity (alkaline destruction solution) of example 3 of the present invention;
FIG. 4 shows the results of the chromatographic test device in example 4 of the present invention;
FIG. 5 shows the results of the chromatographic test device of comparative example 1 of the present invention.
Detailed Description
The invention provides a method for measuring impurity content in rivaroxaban drug, which comprises the following steps:
dissolving rivaroxaban drug to be tested to obtain a test sample solution;
performing high performance liquid chromatography detection on the sample solution to obtain chromatographic information of rivaroxaban drug to be detected;
calculating to obtain the impurity content of rivaroxaban drug to be detected based on the chromatographic information of rivaroxaban standard and the chromatographic information of rivaroxaban drug to be detected;
the parameters of the high performance liquid chromatography detection include:
the chromatographic column is octadecylsilane chemically bonded silica chromatographic column, 4.0mm×55mm,3 μm;
the mobile phase system consists of a mobile phase A and a mobile phase B;
the mobile phase A is a mixed solution of acetonitrile and a phosphoric acid solution, wherein the volume concentration of the phosphoric acid solution is 0.057-0.077%, and the volume ratio of the acetonitrile to the phosphoric acid solution in the mixed solution is 5-10: 95-90; the mobile phase B is acetonitrile;
the elution mode is gradient elution;
the conditions of the gradient elution are as follows:
0-5 min: the volume percentage of the mobile phase A is 95%, and the volume percentage of the mobile phase B is 5%;
5 min-15 min: the volume percentage of the mobile phase A is changed from 95% of uniform velocity to 75%, and the volume percentage of the mobile phase B is changed from 5% of uniform velocity to 25%;
15-30 min: the volume percentage of the mobile phase A is changed from 75% of uniform velocity to 60%, and the volume percentage of the mobile phase B is changed from 25% of uniform velocity to 40%;
30-31 min: the volume percentage of the mobile phase A is changed from 60% of uniform velocity to 95%, and the volume percentage of the mobile phase B is changed from 40% of uniform velocity to 5%;
31-40 min: the volume percentage of the mobile phase A is 95%, and the volume percentage of the mobile phase B is 5%;
the detection wavelength is 245 nm-255 nm;
the impurities are Imp-B, imp-D, imp-E, imp-G, imp-H, imp-I and Imp-J;
the Imp-B has a structure shown in a formula 1;
formula 1;
the Imp-D has a structure shown in a formula 2;
formula 2;
the Imp-E has a structure shown in a formula 3;
formula 3;
the Imp-G has a structure shown in a formula 4;
formula 4;
the Imp-H has a structure shown in a formula 5;
formula 5;
the Imp-I has a structure shown in a formula 6;
formula 6;
the Imp-J has a structure shown in formula 7;
formula 7.
In the present invention, the raw materials used in the present invention are preferably commercially available products unless otherwise specified.
According to the invention, rivaroxaban medicine to be detected is dissolved to obtain a sample solution.
In the invention, the rivaroxaban drug to be tested preferably comprises rivaroxaban tablets. In the invention, the dissolved reagent is preferably a mixed solution of acetonitrile and phosphoric acid solution, wherein the volume concentration of the phosphoric acid solution is preferably 0.057-0.077%, and more preferably 0.067%; the volume ratio of acetonitrile to phosphoric acid solution in the mixed solution is preferably 1:1.
in the invention, the dosage ratio of rivaroxaban drug to be tested and dissolved reagent is preferably 0.5g: 200-250 mL. In the present invention, the dissolution means is preferably ultrasound, and the time of the ultrasound is preferably 10min. In the present invention, it is preferable that the method further comprises centrifuging the dissolved product to obtain a supernatant as a sample solution. In the present invention, the rotational speed of the centrifugation is preferably 5000rpm, and the time is preferably 10min.
After obtaining the sample solution, the invention carries out high performance liquid chromatography detection on the sample solution to obtain the chromatographic information of rivaroxaban drug to be detected.
In the present invention, the parameters of the high performance liquid chromatography detection include:
the chromatographic column is octadecylsilane chemically bonded silica chromatographic column, and in the embodiment of the invention, purosphere is particularly preferably Purosphere STAR RP-18 endsupported, 4.0mm multiplied by 55mm and 3 mu m;
the mobile phase system consists of a mobile phase A and a mobile phase B;
the mobile phase A is a mixed solution of acetonitrile and a phosphoric acid solution, wherein the volume concentration of the phosphoric acid solution is 0.057-0.077%, preferably 0.067%; the volume ratio of acetonitrile to phosphoric acid solution in the mixed solution is 5-10: 95-90, preferably 8:92;
the mobile phase B is acetonitrile;
the elution mode is gradient elution;
the conditions of the gradient elution are as follows:
0-5 min: the volume percentage of the mobile phase A is 95%, and the volume percentage of the mobile phase B is 5%;
5 min-15 min: the volume percentage of the mobile phase A is changed from 95% of uniform velocity to 75%, and the volume percentage of the mobile phase B is changed from 5% of uniform velocity to 25%;
15-30 min: the volume percentage of the mobile phase A is changed from 75% of uniform velocity to 60%, and the volume percentage of the mobile phase B is changed from 25% of uniform velocity to 40%;
30-31 min: the volume percentage of the mobile phase A is changed from 60% of uniform velocity to 95%, and the volume percentage of the mobile phase B is changed from 40% of uniform velocity to 5%;
31-40 min: the volume percentage of the mobile phase A is 95%, and the volume percentage of the mobile phase B is 5%;
the detection wavelength is 245nm to 255nm, preferably 250nm.
In the invention, "15 min-30 min" in the gradient elution: the volume percentage of the mobile phase A is changed from 75% to 60% at uniform speed, and the volume percentage of the mobile phase B is changed from 25% to 40% at uniform speed, which is a key gradient, so that the impurities I and J can be separated.
In the present invention, the parameters of the high performance liquid chromatography detection further include: the flow rate is preferably 0.8mL/min to 1.2mL/min, and more preferably 1.0mL/min.
In the present invention, the parameters of the high performance liquid chromatography detection further include: the column temperature is preferably 40-50 ℃. In the invention, the column temperature can enable the instrument to be balanced more quickly, and the running stability of the method is improved.
After obtaining the chromatographic information of rivaroxaban drug to be detected, the method calculates the impurity content in the rivaroxaban drug to be detected based on the chromatographic information of rivaroxaban standard and the chromatographic information of rivaroxaban drug to be detected.
In the present invention, after obtaining the chromatographic information of rivaroxaban drug to be tested, the impurity peak type is preferably determined according to the relative retention time of table 3.
TABLE 3 impurity peak determination information
In the invention, the chromatographic information of the rivaroxaban standard is taken as a standard, and the impurities in the rivaroxaban drug to be detected are quantitatively analyzed.
In the invention, the concentration of the rivaroxaban standard substance is preferably 0.2-0.8 mug/mL.
The method for determining the impurity content in rivaroxaban drug provided by the invention is described in detail below with reference to examples, but they should not be construed as limiting the scope of the invention.
Example 1
The method for detecting the impurities in rivaroxaban tablets comprises the following steps:
the preparation method of the sample solution comprises the following steps: taking 4 rivaroxaban tablets, placing the tablets into a 200mL measuring flask, adding a proper amount of dissolved reagent (mixed solution of acetonitrile and phosphoric acid solution with volume concentration of 0.067% and volume ratio of 1:1), carrying out ultrasonic treatment for 10min, cooling, diluting to a scale with the dissolved reagent, shaking uniformly, centrifuging at 5000rpm for 10min, and taking supernatant as a sample solution.
High performance liquid chromatography detection is carried out on the sample solution to obtain the chromatographic information of rivaroxaban tablets
The conditions of the high performance liquid chromatography detection method include:
chromatographic column: purosphere cube RP-18 endsupported, 4.0mm 55mm,3 μm;
a detector: UV;
detection wavelength: 250nm;
column temperature: 50 ℃;
mobile phase a: the volume ratio of acetonitrile-phosphoric acid solution with the volume concentration of 0.067% is 8:92;
mobile phase B: acetonitrile;
flow rate: 1.0mL/min;
the gradient elution conditions for the column are shown in table 4:
TABLE 4 gradient elution conditions
In this embodiment, the sample injection amount of the sample solution for each detection is 5 μl, and the obtained chromatogram is shown in fig. 1.
The impurity peak type was determined according to the information of table 3.
And calculating the content of impurities in the rivaroxaban tablet according to the chromatogram peak area of the rivaroxaban standard substance solution.
The impurity components corresponding to the specific chromatographic peaks in fig. 1 are shown in table 5.
TABLE 5 impurity components corresponding to specific chromatographic peaks in FIG. 1
As can be seen from FIG. 1 and Table 5, the chromatographic method adopted in the present example evaluates the impurity content in rivaroxaban tablet, the method has good specificity, the minimum separation degree between each impurity can reach 3.6, the sensitivity is high, and the signal to noise ratio of each impurity is greater than 50 at about 0.2 mug/mL.
Example 2
The quantitative limits of the respective impurities were examined according to the measurement method of example 1, and the results are shown in fig. 2.
Preparing mixed solution of various impurities: impurity B, D, E, G, H, I, J and rivaroxaban are weighed and diluted quantitatively with dissolved reagent (mixed solution of acetonitrile and phosphoric acid solution with volume concentration of 0.067% in volume ratio of 1:1) to each mL of solution containing 0.2 mug of each impurity.
Table 6 shows the quantitative limit results.
TABLE 6 quantitative limit results
As can be seen from Table 6 and FIG. 2, the chromatographic method used in this example evaluated rivaroxaban tablets for high impurity sensitivity, with each impurity having a signal to noise ratio greater than 50 at about 0.2 μg/mL.
Example 3
The method was examined specifically according to the measurement method of example 1, wherein the detection result of the alkali-damaged solution is shown in fig. 3.
Acid breaking solution: taking 1 rivaroxaban tablet, placing in a 50mL measuring flask, adding 5mL of water, adding 5mL of 0.1mol/L hydrochloric acid solution, standing at room temperature for 1h, neutralizing with 0.1mol/L sodium hydroxide solution, adding a proper amount of dissolved reagent (mixed solution of acetonitrile and 0.067% phosphoric acid solution with volume ratio of 1:1), carrying out ultrasonic treatment for 10min, cooling, diluting to a scale with the dissolved reagent, shaking uniformly, centrifuging (5000 rpm,10 min), and taking supernatant.
Base destruction solution: taking 1 rivaroxaban tablet, placing in a 50mL measuring flask, adding 5mL of water, adding 5mL of 0.1mol/L sodium hydroxide solution, standing at room temperature for 1h, neutralizing with 0.1mol/L hydrochloric acid solution, adding a proper amount of dissolved reagent (mixed solution of acetonitrile and 0.067% phosphoric acid solution with volume ratio of 1:1), carrying out ultrasonic treatment for 10min, cooling, diluting to a scale with the dissolved reagent, shaking uniformly, centrifuging (5000 rpm,10 min), and taking supernatant.
Oxidative destruction solution: taking 1 rivaroxaban tablet, placing into a 50mL measuring flask, adding 5mL of water, adding 5mL of 3% hydrogen peroxide, standing for 1h at room temperature, adding a proper amount of dissolved reagent (mixed solution of acetonitrile and phosphoric acid solution with volume concentration of 0.067% and volume ratio of 1:1), carrying out ultrasonic treatment for 10min, cooling, diluting to a scale with the dissolved reagent, shaking uniformly, centrifuging (5000 rpm,10 min), and taking supernatant.
High temperature destruction solution: taking 1 tablet of rivaroxaban tablet placed at a high temperature of 60 ℃ for 5 days, placing the tablet into a 50mL measuring flask, adding a proper amount of dissolved reagent (a mixed solution of acetonitrile and phosphoric acid with volume concentration of 0.067% in a volume ratio of 1:1), carrying out ultrasonic treatment for 10min, cooling, diluting to a scale with the dissolved reagent, shaking uniformly, taking a proper amount of centrifugation (10000 rpm,10 min), and taking supernatant.
Light damage solution: taking 1 piece of rivaroxaban tablet 5 days in an illumination box, placing in a 50mL measuring flask, adding a proper amount of dissolved reagent (mixed solution of acetonitrile and phosphoric acid solution with volume concentration of 0.067% and volume ratio of 1:1), carrying out ultrasonic treatment for 10min, cooling, diluting to a scale with the dissolved reagent, shaking uniformly, taking a proper amount of centrifugation (10000 rpm,10 min), and taking supernatant.
Table 7 shows the results of the forced degradation test.
TABLE 7 forced degradation test results
As can be seen from table 7: the product does not produce impurities under the conditions of illumination, high temperature, oxidation and acid, and produces 1 unknown impurity under the condition of alkali damage; under the conditions of acid, alkali and oxidation forced degradation, the separation degree between a main peak and a known impurity peak and adjacent peaks is more than 1.5, which meets the requirements, and shows that the product is relatively stable under the conditions of oxidation, acid, alkali, illumination and high temperature, and the method has a stable indication effect. The measuring method provided by the embodiment has good specificity and high sensitivity for various impurities. Therefore, the determination method provided by the invention can simply, conveniently, accurately and efficiently evaluate the impurity content in the rivaroxaban tablet, and has important significance for quality control and clinical application safety of the rivaroxaban tablet.
Example 4
The method for detecting the impurities in rivaroxaban tablets comprises the following steps:
the preparation method of the sample solution comprises the following steps: taking 4 rivaroxaban tablets, placing the tablets into a 200mL measuring flask, adding a proper amount of dissolved reagent (mixed solution of acetonitrile and phosphoric acid solution with volume concentration of 0.067% and volume ratio of 1:1), carrying out ultrasonic treatment for 10min, cooling, diluting to a scale with the dissolved reagent, shaking uniformly, centrifuging at 5000rpm for 10min, and taking supernatant as a sample solution.
High performance liquid chromatography detection is carried out on the sample solution to obtain the chromatographic information of rivaroxaban tablets
The conditions of the high performance liquid chromatography detection method include:
chromatographic column: purosphere cube RP-18 endsupported, 4.0mm 55mm,3 μm;
a detector: UV;
detection wavelength: 250nm;
column temperature: 40 ℃;
mobile phase a: the volume ratio of acetonitrile-phosphoric acid solution with the volume concentration of 0.067% is 8:92;
mobile phase B: acetonitrile;
flow rate: 1.0mL/min.
The gradient elution conditions of the column are shown in table 4.
The detection results are shown in FIG. 4.
The impurity components corresponding to the specific chromatographic peaks in fig. 4 are shown in table 8.
TABLE 8 impurity Components corresponding to specific chromatographic peaks in FIG. 4
As can be seen from table 8: each impurity is separated.
Comparative example 1
The preparation method of the solution of the comparative example comprises the following steps: the impurity B, D, E, G, H, I, J and rivaroxaban are weighed and diluted quantitatively with a dissolving reagent (the mixed solution of acetonitrile and phosphoric acid solution with the volume concentration of 0.067% and the volume ratio of 60:40) to obtain a solution containing 0.4 mug of each impurity and 0.2mg of rivaroxaban per mL.
The conditions of the high performance liquid chromatography detection method are as follows:
chromatographic column: octadecylsilane chemically bonded silica (Purosphere cube STAR RP-18endcapped,4.0 mm. Times.55 mm,3 μm) is used as filler;
a detector: UV;
detection wavelength: 250nm;
column temperature: 45 ℃;
mobile phase a: the volume ratio of acetonitrile-phosphoric acid solution with the volume concentration of 0.067% is 8:92;
mobile phase B: acetonitrile;
flow rate: 1.0mL/min.
The gradient elution conditions of the column are shown in table 9.
TABLE 9 gradient elution conditions for the chromatography column of comparative example 1
In the comparative example, the sample injection amount of each detection of the sample solution is 5 μl, the obtained chromatogram is shown in fig. 5, and as can be seen from fig. 5, when the concentration of each impurity in the comparative example is equivalent to 0.2% of the sample, the separation degree of the impurity I and the impurity J is 1.1, and the baseline separation is not achieved, so that the method specificity is poor, and therefore, the quality of the impurity in the rivaroxaban tablet cannot be accurately controlled. Meanwhile, the dissolved reagent of this comparative example was an acetonitrile-phosphoric acid aqueous solution having a volume concentration of 0.067% (wherein the volume ratio of acetonitrile to phosphoric acid aqueous solution is 60:40), resulting in poor impurity B peak type.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (8)
1. The method for determining the impurity content in rivaroxaban is characterized by comprising the following steps of:
dissolving rivaroxaban drug to be tested to obtain a test sample solution;
performing high performance liquid chromatography detection on the sample solution to obtain chromatographic information of rivaroxaban drug to be detected;
calculating to obtain the impurity content of rivaroxaban drug to be detected based on the chromatographic information of rivaroxaban standard and the chromatographic information of rivaroxaban drug to be detected;
the parameters of the high performance liquid chromatography detection include:
the chromatographic column is octadecylsilane chemically bonded silica chromatographic column, 4.0mm×55mm,3 μm;
the mobile phase system consists of a mobile phase A and a mobile phase B;
the mobile phase A is a mixed solution of acetonitrile and a phosphoric acid solution, wherein the volume concentration of the phosphoric acid solution is 0.057-0.077%, and the volume ratio of the acetonitrile to the phosphoric acid solution in the mixed solution is 5-10: 95-90;
the mobile phase B is acetonitrile;
the elution mode is gradient elution;
the conditions of the gradient elution are as follows:
0-5 min: the volume percentage of the mobile phase A is 95%, and the volume percentage of the mobile phase B is 5%;
5 min-15 min: the volume percentage of the mobile phase A is changed from 95% of uniform velocity to 75%, and the volume percentage of the mobile phase B is changed from 5% of uniform velocity to 25%;
15-30 min: the volume percentage of the mobile phase A is changed from 75% of uniform velocity to 60%, and the volume percentage of the mobile phase B is changed from 25% of uniform velocity to 40%;
30-31 min: the volume percentage of the mobile phase A is changed from 60% of uniform velocity to 95%, and the volume percentage of the mobile phase B is changed from 40% of uniform velocity to 5%;
31-40 min: the volume percentage of the mobile phase A is 95%, and the volume percentage of the mobile phase B is 5%;
the detection wavelength is 245 nm-255 nm;
the impurities are Imp-B, imp-D, imp-E, imp-G, imp-H, imp-I and Imp-J;
the Imp-B has a structure shown in a formula 1;
formula 1;
the Imp-D has a structure shown in a formula 2;
formula 2;
the Imp-E has a structure shown in a formula 3;
formula 3;
the Imp-G has a structure shown in a formula 4;
formula 4;
the Imp-H has a structure shown in a formula 5;
formula 5;
the Imp-I has a structure shown in a formula 6;
formula 6;
the Imp-J has a structure shown in formula 7;
formula 7.
2. The assay of claim 1, wherein the rivaroxaban drug to be tested comprises rivaroxaban tablet.
3. The measurement method according to claim 1, wherein the dissolved reagent is a mixed solution of acetonitrile and phosphoric acid solution, wherein the volume concentration of the phosphoric acid solution is 0.057-0.077%, and the volume ratio of acetonitrile to phosphoric acid solution in the mixed solution is 1:1.
4. the assay according to claim 1 or 3 wherein the ratio of rivaroxaban drug to dissolved reagent to be tested is 0.5g: 200-250 mL.
5. The method according to claim 1, wherein the dissolution is performed by ultrasonic treatment for 10min.
6. The method according to claim 1 or 5, wherein after the dissolving, further comprising centrifuging the dissolved product to obtain a supernatant as a sample solution; the rotational speed of the centrifugation is 5000rpm, and the time is 10min.
7. The method according to claim 1, wherein the parameters of the high performance liquid chromatography detection further comprise: the flow rate is 0.8mL/min to 1.2mL/min.
8. The method according to claim 1, wherein the parameters of the high performance liquid chromatography detection further comprise: the column temperature is 40-50 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310796354.0A CN116500173A (en) | 2023-07-03 | 2023-07-03 | Method for determining impurity content in rivaroxaban drug |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310796354.0A CN116500173A (en) | 2023-07-03 | 2023-07-03 | Method for determining impurity content in rivaroxaban drug |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116500173A true CN116500173A (en) | 2023-07-28 |
Family
ID=87318836
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310796354.0A Pending CN116500173A (en) | 2023-07-03 | 2023-07-03 | Method for determining impurity content in rivaroxaban drug |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116500173A (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107941936A (en) * | 2017-11-17 | 2018-04-20 | 重庆华邦制药有限公司 | The method and application of separation determination razaxaban and its impurity |
| CN110849994A (en) * | 2019-11-25 | 2020-02-28 | 湖南九典制药股份有限公司 | Method for separating related substances in rivaroxaban |
| CN112557543A (en) * | 2020-12-09 | 2021-03-26 | 江苏嘉逸医药有限公司 | Method for measuring rivaroxaban and related substances thereof |
| CN112903846A (en) * | 2021-01-20 | 2021-06-04 | 上海普康药业有限公司 | Analysis method for determining rivaroxaban and impurities thereof |
-
2023
- 2023-07-03 CN CN202310796354.0A patent/CN116500173A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107941936A (en) * | 2017-11-17 | 2018-04-20 | 重庆华邦制药有限公司 | The method and application of separation determination razaxaban and its impurity |
| CN110849994A (en) * | 2019-11-25 | 2020-02-28 | 湖南九典制药股份有限公司 | Method for separating related substances in rivaroxaban |
| CN112557543A (en) * | 2020-12-09 | 2021-03-26 | 江苏嘉逸医药有限公司 | Method for measuring rivaroxaban and related substances thereof |
| CN112903846A (en) * | 2021-01-20 | 2021-06-04 | 上海普康药业有限公司 | Analysis method for determining rivaroxaban and impurities thereof |
Non-Patent Citations (2)
| Title |
|---|
| 尹秀娥;胡小燕;侯德粉;张嘉月;董乔;: "利伐沙班有关物质的HPLC测定", 中国医药工业杂志, no. 12 * |
| 袁利杰;刘辉;杨本霞;郑子栋;: "HPLC法测定利伐沙班片的含量和有关物质", 药物分析杂志, no. 04 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107941936B (en) | Method for separating and determining rivaroxaban and impurities thereof and application | |
| CN103698424A (en) | Detecting method of detecting organic solvent in slightly-soluble aluminum salt drug | |
| CN109580825A (en) | The detection method of p-methyl benzenesulfonic acid Ester in racecadotril | |
| CN116500173A (en) | Method for determining impurity content in rivaroxaban drug | |
| CN105486776A (en) | Gas chromatography method of metoclopramide | |
| CN110687228A (en) | Method for detecting related substances in simethicone | |
| CN106525994A (en) | Method for determination of related substances of paracetamol and tramadol hydrochloride capsules | |
| CN116338022A (en) | Method for analyzing related substances in indobufen starting material 1 | |
| CN105675754B (en) | The method of high effective liquid chromatography for measuring Li Gelieting enantiomter contents | |
| CN108593646A (en) | Method for rapidly detecting mannitol content of intermediate of compound mannitol injection | |
| CN103901147A (en) | Method for measuring dissolution rate of andrographolide pills | |
| CN118566395B (en) | Method, application and system for determining impurity content in toluene sulfonic acid Lu Meipai protuberance orally disintegrating tablet | |
| CN108072709A (en) | The method for measuring enantiomter content in amber love song Ge Lieting bulk pharmaceutical chemicals | |
| CN116297908B (en) | Analysis method of indobufen isomer impurity | |
| CN116973487B (en) | Method for measuring impurity content of saxagliptin and application | |
| CN117233312A (en) | Detection method of related substances in hydroquinone cream and application thereof | |
| CN112684035B (en) | Method for detecting residual quantity of oxalic acid in spiramycin | |
| CN115236255B (en) | Method for detecting related substances of loxoprofen sodium | |
| CN112083086B (en) | Micro-determination method for migration and dissolution of pigment red 254 in medical light-resistant material | |
| CN107884496A (en) | The content assaying method of butanedioic acid in a kind of amber love song Ge Lieting | |
| CN107870210A (en) | The method for determining Li Gelieting contents in Li Gelieting bulk drugs | |
| Foppa et al. | Development, validation and stability study of pediatric atenolol syrup | |
| CN118169280A (en) | Method for measuring dissolution rate of phylloquinone drops | |
| CN118032953A (en) | Method for detecting toxic impurities of oseltamium phosphate Wei Jiyin | |
| CN117554528A (en) | Method for detecting memantine hydrochloride and donepezil in mefenacet hydrochloride Jin Gangduo donepezil drug |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |