CN117825579A - High performance liquid chromatography analysis and detection method for 6 genotoxic impurities in telmisartan - Google Patents
High performance liquid chromatography analysis and detection method for 6 genotoxic impurities in telmisartan Download PDFInfo
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- 239000012535 impurity Substances 0.000 title claims abstract description 85
- RMMXLENWKUUMAY-UHFFFAOYSA-N telmisartan Chemical compound CCCC1=NC2=C(C)C=C(C=3N(C4=CC=CC=C4N=3)C)C=C2N1CC(C=C1)=CC=C1C1=CC=CC=C1C(O)=O RMMXLENWKUUMAY-UHFFFAOYSA-N 0.000 title claims abstract description 64
- 239000005537 C09CA07 - Telmisartan Substances 0.000 title claims abstract description 32
- 229960005187 telmisartan Drugs 0.000 title claims abstract description 32
- 238000001514 detection method Methods 0.000 title claims abstract description 29
- 231100000024 genotoxic Toxicity 0.000 title claims abstract description 20
- 230000001738 genotoxic effect Effects 0.000 title claims abstract description 20
- 238000004128 high performance liquid chromatography Methods 0.000 title claims abstract description 13
- 238000004458 analytical method Methods 0.000 title abstract description 13
- 238000000034 method Methods 0.000 claims abstract description 26
- 239000012488 sample solution Substances 0.000 claims abstract description 13
- 239000003814 drug Substances 0.000 claims abstract description 8
- 229940079593 drug Drugs 0.000 claims abstract description 7
- 239000000243 solution Substances 0.000 claims description 41
- 239000000523 sample Substances 0.000 claims description 28
- 239000013558 reference substance Substances 0.000 claims description 21
- 239000002904 solvent Substances 0.000 claims description 21
- 238000012360 testing method Methods 0.000 claims description 18
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 11
- 238000007865 diluting Methods 0.000 claims description 10
- 238000002347 injection Methods 0.000 claims description 9
- 239000007924 injection Substances 0.000 claims description 9
- 239000011259 mixed solution Substances 0.000 claims description 9
- 239000007864 aqueous solution Substances 0.000 claims description 8
- 239000000706 filtrate Substances 0.000 claims description 8
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 claims description 6
- AUALQMFGWLZREY-UHFFFAOYSA-N acetonitrile;methanol Chemical group OC.CC#N AUALQMFGWLZREY-UHFFFAOYSA-N 0.000 claims description 5
- 238000010828 elution Methods 0.000 claims description 5
- 238000001914 filtration Methods 0.000 claims description 5
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims description 5
- 239000000377 silicon dioxide Substances 0.000 claims description 5
- 239000012085 test solution Substances 0.000 claims description 5
- 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 claims description 4
- 238000010812 external standard method Methods 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 4
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 4
- 239000002245 particle Substances 0.000 claims description 4
- ROBLTDOHDSGGDT-UHFFFAOYSA-M sodium;pentane-1-sulfonate Chemical compound [Na+].CCCCCS([O-])(=O)=O ROBLTDOHDSGGDT-UHFFFAOYSA-M 0.000 claims description 4
- 239000000725 suspension Substances 0.000 claims description 4
- AAEQXEDPVFIFDK-UHFFFAOYSA-N 3-(4-fluorobenzoyl)-2-(2-methylpropanoyl)-n,3-diphenyloxirane-2-carboxamide Chemical compound C=1C=CC=CC=1NC(=O)C1(C(=O)C(C)C)OC1(C=1C=CC=CC=1)C(=O)C1=CC=C(F)C=C1 AAEQXEDPVFIFDK-UHFFFAOYSA-N 0.000 claims description 3
- 239000000945 filler Substances 0.000 claims description 3
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 3
- 238000012856 packing Methods 0.000 claims description 2
- 238000000926 separation method Methods 0.000 abstract description 12
- 230000035945 sensitivity Effects 0.000 abstract description 5
- 238000011002 quantification Methods 0.000 abstract description 3
- 239000012071 phase Substances 0.000 description 13
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 4
- 125000003118 aryl group Chemical group 0.000 description 4
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 230000000711 cancerogenic effect Effects 0.000 description 3
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- 238000005303 weighing Methods 0.000 description 3
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 2
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- 238000011084 recovery Methods 0.000 description 2
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- 206010064571 Gene mutation Diseases 0.000 description 1
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- 230000015572 biosynthetic process Effects 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
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- 238000012986 modification Methods 0.000 description 1
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- 231100000350 mutagenesis Toxicity 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 229940124531 pharmaceutical excipient Drugs 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000012088 reference solution Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 238000013112 stability test Methods 0.000 description 1
- 125000004079 stearyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Abstract
The invention provides a high performance liquid chromatography analysis and detection method for 6 genotoxic impurities in telmisartan, and aims to separate and detect the impurities A, B, C, D, E, F and other 6 genotoxic impurities in a telmisartan bulk drug by using high performance liquid chromatography. The limit of genotoxic impurities is low, and in detection, a high-concentration sample solution is usually required to be prepared, so that the set detection limit is reached, the problems of poor separation degree and the like caused by chromatographic column blockage and unknown peak interference are inevitably caused, and further, the accuracy and the quantification are not ensured. The method has the advantages of low cost, high sensitivity, good separation degree, simple operation, good stability, good reproducibility, high precision and the like, reduces the cost, effectively detects telmisartan and improves the safety of the product.
Description
Technical Field
The invention belongs to the technical field of medicine analysis, and relates to an analysis and detection method for 6 genotoxic impurities in telmisartan.
Background
The telmisartan possibly introduces impurities containing amino or N-oxidized aromatic structures in the structure in the synthesis process, and the structural formula shows that the impurities have amino or N-oxidized aromatic structures and are warning structures of genotoxic cancerogenic substances, the mutation potential and the cancerogenic potential of the impurities are classified into 3 types according to the amino or N-oxidized aromatic structures, if the impurities remain in the process, the impurities can be introduced into telmisartan medicines, and patients can directly or indirectly damage human DNA after taking the impurities, so that gene mutation or in-vivo mutagenesis is caused, the possibility of cancer is increased, and the administration safety is ensured and the impurities need to be strictly controlled according to the limits of genotoxic impurities.
The structural formula of the impurity is shown below.
At present, a chromatography-mass spectrometry combined method is generally selected for detecting genotoxic impurities, and has high sensitivity, but the method is complex to operate and has higher cost. At present, no related report that high performance liquid chromatography is adopted in the analysis and detection method of genotoxic impurities in telmisartan exists in China. The limit of genotoxic impurities is low, and in detection, a high-concentration sample solution is usually required to be prepared, so that the set detection limit is reached, the problems of poor separation degree and the like caused by chromatographic column blockage and unknown peak interference are inevitably caused, and further, the accuracy and the quantification are not ensured. The invention provides a technical reference which has the advantages of low cost, high sensitivity, simple operation, good separation degree, high accuracy and the like and can effectively control 6 genotoxic impurities in telmisartan.
Disclosure of Invention
The invention aims to provide a method for separating and measuring 6 genotoxic impurities in a telmisartan crude drug by using high performance liquid chromatography. The method has the advantages of low cost, high sensitivity, simple operation, good stability, good reproducibility and high precision.
The technical scheme adopted by the invention for solving the technical problems is as follows:
a method for measuring 6 genotoxic impurities in a telmisartan crude drug by high performance liquid chromatography separation, wherein the impurities are as follows:
the structural formula of the impurity A is as follows:
the structural formula of the impurity B is as follows:
the structural formula of the impurity C is as follows:
the structural formula of the impurity D is as follows:
the structural formula of the impurity E is as follows:
the structural formula of the impurity F is as follows:
the detection method adopts high performance liquid chromatography and comprises the following steps:
(1) Control solution: mixing the impurity A, B, C, D, E and the F reference substance in proper amounts, dissolving in solvent and diluting to obtain a mixed solution containing 0.15-10 μg per 1 ml.
(2) Preparing a test solution: taking a telmisartan sample to be detected, dissolving and diluting the telmisartan sample to be detected into a suspension solution containing 10 mg-50 mg of telmisartan in each 1ml by using a solvent, filtering, and taking a subsequent filtrate as a sample solution;
the solvent in the steps (1) to (2) is a methanol-water solution;
(3) Chromatographic conditions:
chromatographic column: octadecylsilane chemically bonded silica is used as a filler, the particle size of the octadecylsilane chemically bonded silica is 1.8-6 mu m, the inner diameter of the column is 2-8 mm, and the length of the column is 150-350 mm;
detection wavelength: 210 nm-300 nm;
gradient elution is carried out by adopting a mobile phase A and a mobile phase B;
the mobile phase A is aqueous solution with the pH value ranging from 3.0 to 5.0;
the mobile phase B is acetonitrile-methanol mixed solution;
the elution gradient is:
flow rate of mobile phase: 0.5-1.5 ml/mi;
column temperature: 25-50 ℃;
sample injection volume: 5-100 μl; preferably, the sample injection volume is 10 μl;
(4) And (3) measuring: and (3) injecting the reference substance solution and the test sample solution obtained in the steps (1) and (2) into a high performance liquid chromatograph, recording a chromatogram, and carrying out qualitative and/or quantitative detection on 6 genotoxic impurities in the test sample according to an external standard method.
Further, the solvent in the steps (1) and (2) is a methanol-water solution with the volume ratio of 50:50.
Further, the concentration of the sample solution in the step (2) is 10mg/ml to 50mg/ml of a sample containing telmisartan to be detected;
preferably, the concentration of the sample solution in the step (2) is 20mg/ml of the sample containing telmisartan to be detected.
Further, in the step (3), the column length was 250mm, the column inner diameter was 4.6mm, and the column packing particle size was 5. Mu.m.
Further, the aqueous solution of the mobile phase A in the step (3) is a mixed aqueous solution containing 2.0g of potassium dihydrogen phosphate and 3.8g of sodium pentanesulfonate per 1000 ml.
Further, the aqueous solution of mobile phase A in step (3) has a pH of 4.0.
Further, in the step (3), the mobile phase B is acetonitrile-methanol mixed solution with a volume ratio of 4:1.
Further, the detection wavelength in the step (3) is 230nm.
Further, the flow rate of the mobile phase in the step (3) is 1.0ml/min.
Further, the column temperature in step (3) is 40 ℃.
The beneficial effects of the invention are as follows:
according to the invention, the high performance liquid chromatography is utilized to detect 6 basic toxin impurities in the telmisartan bulk drug, the separation effect is good, the peak type is good, the detection sensitivity to impurity detection is excellent, the cost is reduced, meanwhile, telmisartan is effectively detected, and the safety of the product is improved.
Drawings
FIG. 1 is a blank solvent map;
FIG. 2 is a graph of a control solution;
FIG. 3 is a sample solution graph.
Detailed Description
The present invention is further illustrated by the following examples, which are not intended to limit the invention in any way.
Instrument: the liquid chromatograph comprises a liquid phase pump, an ultraviolet detector and a sample injector;
reagent: potassium dihydrogen phosphate (analytically pure), sodium pentanesulfonate (pharmaceutical excipients), phosphoric acid (analytically pure), acetonitrile;
chromatographic column: octadecyl bonded silica gel column.
Examples:
the measurement is carried out by high performance liquid chromatography (China pharmacopoeia 2020 edition, four-part rule 0512).
Solvent: methanol-water (volume ratio 50:50).
(1) The system applicability solution is prepared by taking a proper amount of telmisartan test sample, adding a proper amount of solvent, carrying out ultrasonic treatment for 5 minutes to uniformly disperse, respectively adding a proper amount of impurity A, B, C, D, E and F reference substance, diluting with the solvent to prepare a suspension solution containing 20mg of telmisartan, 0.375 mug of impurity A, 0.375 mug of impurity B, 0.375 mug of impurity C, 0.375 mug of impurity D, 0.375 mug of impurity E and 0.375 mug of impurity F in each 1ml, filtering (discarding at least 10ml of primary filtrate), and taking a subsequent filtrate.
(2) The reference substance solution is prepared by dissolving impurity A, B, C, D, E and F reference substance in appropriate amount, and diluting with solvent to obtain mixed solution containing 0.375 μg per 1 ml.
(3) Taking a proper amount of telmisartan sample to be detected from a test solution, adding a proper amount of solvent, carrying out ultrasonic treatment for 5 minutes to uniformly disperse, diluting with the solvent to prepare a suspension solution containing 20mg of telmisartan per 1ml, filtering (discarding at least 10ml of primary filtrate), and taking the subsequent filtrate.
(4) Chromatographic conditions
Chromatographic column: octadecylsilane chemically bonded silica (Kromasil 100-5-C18, 250 mm. Times.4.6 mm,5 μm) was used as a filler;
detection wavelength: 230nm;
mobile phase a: phosphate buffer (2.0 g of monopotassium phosphate, 3.8g of sodium pentanesulfonate, 1000ml of water are added to dissolve, the pH value is regulated to 4.0 by phosphoric acid) is taken as a mobile phase A,
mobile phase B: acetonitrile-methanol (volume ratio of 4:1) is taken as a mobile phase B,
gradient elution was performed according to the following table;
flow rate: 1.0ml per minute;
column temperature: 40 ℃;
sample cell temperature: 5 ℃;
sample injection volume: 10 μl.
In the system applicability requirement system applicability solution chromatograms, the peak outlet sequence is the impurity B, D, C, A, F and the E peak in sequence, and the separation degree between every two adjacent chromatographic peaks meets the requirement.
(5) The method comprises precisely measuring the solution of the sample and the solution of the reference substance, respectively injecting into a liquid chromatograph, recording the chromatograms, and carrying out qualitative and/or quantitative detection on 6 genotoxic impurities in the sample according to an external standard method.
Limit: the sum of the contents of the impurities A, B, C, D, E and F calculated by the external standard method in terms of peak area should not be more than 18.75ppm.
Results: in the system applicability solution chromatogram (fig. 2), the retention time and the separation degree of each component are shown in the following table:
retention time and degree of separation of the components
The patterns of the blank solvent, the reference substance and the sample to be tested are shown in figures 1 to 3, and according to the results, the detection method can accurately and sensitively detect the impurities A, B, C, D, E and F, the impurities and the separation degree of the impurities and telmisartan samples can meet the requirements of qualitative and quantitative detection accuracy. At present, no related report exists that the genotoxic impurities in telmisartan adopt a high performance liquid chromatography analysis detection method, the impurities have amino or N-oxidation aromatic structures, the residue of the impurities must be strictly controlled for the warning structure of genotoxic cancerogenic substances, and the method can effectively detect the impurities and provides important guarantee for medication safety.
In this example, the content of the detected impurity E in the sample solution was 4.1ppm, and the other impurities were not detected (FIG. 3).
The present example performs the following methodological verification:
(1) Specialization of
Methanol-water (volume ratio 50:50) was used as solvent. And respectively weighing a proper amount of each impurity reference substance, adding a solvent for dissolving and gradually diluting to a solution containing 0.375 mu g/ml of the impurity, and taking the solution as an impurity positioning solution. Taking a proper amount of a sample and each impurity reference substance, so that the concentration level of each impurity is 100% of the limit concentration of each impurity, filtering, and taking the subsequent filtrate as a system applicability solution. Analysis is performed by chromatographic conditions in specific embodiments. The result shows that the solvent has no interference to the detection of impurities; in the system applicability solution chromatogram, the separation degree between the known impurities and the adjacent peaks is more than 1.5. The specificity of the method is good.
(2) Linearity test
Weighing a proper amount of each impurity reference substance, adding a solvent for dissolution and gradually diluting to prepare a series of reference substance mixed solutions with the concentration of 30%, 50%, 80%, 100% and 200% of the limit concentration of each impurity, measuring according to chromatographic conditions in a specific embodiment, drawing a linear graph by taking the concentration as an abscissa and the peak area as an ordinate, calculating a regression equation and a correlation coefficient r, and calculating the ratio of the y-axis intercept of the impurity to a 100% response value. The results are shown in Table 1.
TABLE 1 Linear test results
(3) Limit of detection and limit of quantification
And (3) taking the mixed solution of the series of reference substances under the item (2), gradually diluting with a solvent, carrying out sample analysis, and calculating a detection limit and a quantitative limit according to the signal to noise ratio (S/N) of more than 3 and 10 respectively. The results showed that the detection limits of the impurities A, B, C, D, E and F were 1.7ppm and the quantitative limits were 5.6ppm. Under the quantitative limit of each impurity, 6 needles are continuously injected, and the result shows that the RSD (n=6) of the peak area 5 of the impurity A, B, C, D, E and the F is 6%, 5%, 3%, 2% and 3%, respectively, which indicates that the minimum quantitative value of the method can meet the use requirement.
(4) Accuracy test
Taking the solution with the linear impurity concentration of 100% under the item (2) as a reference solution.
Respectively precisely weighing 1g of telmisartan, placing in a 50ml measuring flask (containing 20mg/m 1), adding 12 parts in parallel, adding a proper amount of each impurity reference substance into each part, so that the concentration level of each impurity is respectively 30%, 50%, 100% and 200% of the limit concentration of each impurity, and each concentration level is respectively 3 parts, and preparing a part of test solution without adding the impurity reference substance as an un-added standard test solution; all the solutions are filtered, and the subsequent filtrate is taken as an accuracy solution. And (5) carrying out sample injection analysis according to chromatographic conditions in the specific embodiment, and calculating the recovery rate. The results showed average recovery of impurities A, B, C, D, E and F (n=12) of 97%, 101%, 104%, 102% and 106%, respectively; RSD is 3%, 5%, 2%, 1%, 5% and 2%, respectively. The accuracy of the method is good.
(5) Repeatability test
Preparing 6 parts of test sample solution in parallel according to the preparation method of the concentration level (100 percent of each impurity limit) solution under the item of the accuracy test (4); and taking the reference substance solution under the accuracy test item of (4) as the reference substance solution. And (3) carrying out sample injection analysis according to chromatographic conditions in a specific embodiment, and calculating the RSD of each impurity content. The results showed that the RSD (n=6) of impurity A, B, C, D, E and F contents were 2%, 3%, 1%, 2%, 3% and 2%, respectively. The reproducibility of the method is good.
(6) Intermediate precision test
Taking the same batch of test products, carrying out experiments according to the requirements of "(5) repeatability test" by different analyzers and different instruments at different times, and calculating the RSD of each impurity content in 12 solutions prepared by two researchers, wherein the result shows that the RSD (n=6) of the impurity A, B, C, D, E and the F content in 12 solutions prepared by two analyzers is 3%, 6% and 5% respectively. The intermediate precision of the method is good.
(7) Durability test
Taking (5) a reference substance solution and a repeatability solution for a repeatability test, carrying out sample injection analysis, respectively examining the tolerance condition of the method when the flow speed is changed by +/-0.2 ml/min, the column temperature is changed by +/-5 ℃ and the wavelength is changed by +/-2 nm, and calculating the RSD of the impurity A, B, C, D, E and the F content under each condition. The result shows that in the repetitive solution chromatograms under various conditions, the separation degree between each known impurity peak and the adjacent peak is more than 1.5, and the requirements are met; the RSD (n=7) of the impurity A, B, C, D, E and F contents are 4%, 2%, 9% and 4%, respectively. The method has good durability in the range of the flow rate of 0.8-1.2 ml/min, the column temperature of 35-45 ℃ and the wavelength of 228-232 nm.
(8) Solution stability test
Taking the reference substance solution and the repeatability solution for the repeatability test (5), standing at 5 ℃ for a period of time, and respectively carrying out sample injection analysis at 0h, 5h, 12h, 20h and 30 h. The results show that the peak areas of the impurities A, B, C, D, E and F in the control solution and the repetitive solution chromatograms are not changed obviously.
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 (10)
1. A method for separating and measuring 6 genotoxic impurities in telmisartan bulk drug by high performance liquid chromatography is characterized in that,
the impurities are as follows:
the structural formula of the impurity A is as follows:
the structural formula of the impurity B is as follows:
the structural formula of the impurity C is as follows:
the structural formula of the impurity D is as follows:
the structural formula of the impurity E is as follows:
the structural formula of the impurity F is as follows:
the detection method adopts high performance liquid chromatography and comprises the following steps:
(1) Control solution: dissolving and diluting impurity A, B, C, D, E and F reference substance with solvent to obtain mixed solution containing 0.15-10 μg per 1ml as reference substance solution;
(2) Preparing a test solution: taking a telmisartan sample to be detected, dissolving and diluting the telmisartan sample to be detected into a suspension solution containing 10 mg-50 mg of telmisartan in each 1ml by using a solvent, filtering, and taking a subsequent filtrate as a sample solution;
the solvent in the steps (1) to (2) is a methanol-water solution;
(3) Chromatographic conditions:
chromatographic column: octadecylsilane chemically bonded silica is used as a filler, the particle size of the octadecylsilane chemically bonded silica is 1.8-6 mu m,
the inner diameter of the column is 2-8 mm, and the length of the column is 150-350 mm;
detection wavelength: 210 nm-300 nm;
gradient elution is carried out by adopting a mobile phase A and a mobile phase B;
the mobile phase A is aqueous solution with the pH value ranging from 3.0 to 5.0;
the mobile phase B is acetonitrile-methanol mixed solution;
the elution gradient is:
flow rate of mobile phase: 0.5-1.5 ml/mi;
column temperature: 25-50 ℃;
sample injection volume: 5-100 μl; preferably, the sample injection volume is 10 μl;
(4) And (3) measuring: and (3) injecting the reference substance solution and the test sample solution obtained in the steps (1) and (2) into a high performance liquid chromatograph, recording a chromatogram, and carrying out qualitative and/or quantitative detection on 6 genotoxic impurities in the test sample according to an external standard method.
2. The method according to claim 1, wherein the solvent in the steps (1) and (2) is a methanol-water solution having a volume ratio of 50:50.
3. The detection method according to claim 1, wherein the concentration of the sample solution in the step (2) is 10mg/ml to 50mg/ml of the sample containing telmisartan to be detected;
preferably, the concentration of the sample solution in the step (2) is 20mg/ml of the sample containing telmisartan to be detected.
4. The method according to claim 1, wherein in the step (3), the column length is 250mm, the column inner diameter is 4.6mm, and the column packing has a particle diameter of 5. Mu.m.
5. The method according to claim 1, wherein the aqueous solution of mobile phase A in step (3) is a mixed aqueous solution containing 2.0g of potassium dihydrogen phosphate and 3.8g of sodium pentanesulfonate per 1000 ml.
6. The method according to claim 1, wherein the aqueous solution of mobile phase a in step (3) has a pH of 4.0.
7. The method according to claim 1, wherein the mobile phase B in the step (3) is an acetonitrile-methanol mixed solution with a volume ratio of 4:1.
8. The method according to claim 1, wherein the detection wavelength in the step (3) is 230nm.
9. The method according to claim 1, wherein the flow rate of the mobile phase in step (3) is 1.0ml/min.
10. The method according to claim 1, wherein the column temperature in step (3) is 40 ℃.
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