CN105259282A - Method for separating and determining rivaroxaban related substances through liquid chromatography - Google Patents
Method for separating and determining rivaroxaban related substances through liquid chromatography Download PDFInfo
- Publication number
- CN105259282A CN105259282A CN201510603166.7A CN201510603166A CN105259282A CN 105259282 A CN105259282 A CN 105259282A CN 201510603166 A CN201510603166 A CN 201510603166A CN 105259282 A CN105259282 A CN 105259282A
- Authority
- CN
- China
- Prior art keywords
- separating
- buffer salt
- razaxaban
- mobile phase
- related substance
- 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
Landscapes
- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
Abstract
The invention belongs to the field of analytical chemistry, and discloses a method for separating and determining rivaroxaban related substances and the content thereof through liquid chromatography. The method is characterized in that the content of rivaroxaban and related substances thereof can be quantitatively determined through using a chromatographic column with octadecylsilane chemically bonded silica as a packing material and using a certain ratio of buffer salt solution-organic phase as a mobile phase, so the quality of rivaroxaban can be effectively controlled. The method has the advantages of strong specificity, high accuracy and simple operation.
Description
Technical field
The invention belongs to analytical chemistry field, be specifically related to the method for liquid chromatography for separating and determining razaxaban and related substance thereof.
Background technology
Razaxaban clinically for hip joint or the replacement knee in arthroplasty adult patients of selecting a time, to prevent venous thronbosis.This medical instrument have security high, can oral, without the advantage such as cross resistance, bad reaction be little.Razaxaban chemistry is by name: 5-Chloro-thiophene-2-carboxylicacid{2-oxo-3-[4-(3-oxo-morpholin-4-yl)-phenyl]-oxazolidin-5-yl-methyl}-amide, molecular formula is C
19h
18clN
3o
5s.Razaxaban structural formula is:
In the art production process of razaxaban, some important intermediate may be removed not exclusively, thus affects pharmaceutical purity and quality; And in the storage and transportation of medicine, medicine also may produce degradation impurity through specific condition degradeds such as illumination, high temperature, high humiditys; Above-mentioned related substance, all without therapeutic action, also may affect stability and the curative effect of medicine, even be detrimental to health.Related substance for razaxaban major control has 6, related substance 1:2-{2-Oxo-3-[4-(3-oxo-morpholin-4-yl)-phenyl]-oxazolidin-5-yl}-isoindole-1 respectively, 3-dione, related substance 2:2-{2-Hydroxy-3-[4-(3-oxo-morpholin-4-yl)-phenylamino]-propyl}-isoindole-1, 3-dione, related substance 3:Di-imidazol-1-yl-methanone, related substance 4:Dimethyl-pyridin-4-yl-amine, related substance 5:5-Chloro-thiophene-2-carboxylicacid, related substance 6:4-[4-(5-Methylamine-2-oxo-oxazolidin-3-yl)-phenyl]-morpholin-3-one.
related substance 1 related substance 2
Related substance 3 related substance 4 related substance 5 related substance 6
Impurity removal in razaxaban is incomplete, will introduce bulk drug finished product, thus affect pharmaceutical purity and quality.Therefore, realize separation and the quantitative measurement of razaxaban and related substance thereof, have important practical significance in the production and quality control thereof of razaxaban.
Summary of the invention
For the impurity introduced in razaxaban synthesis and storage, need to carry out quality control in bulk drug and preparation, therefore, realize the separation of razaxaban and related substance thereof, its synthesis and quality control aspect are had important practical significance.
The method of liquid chromatography analysis razaxaban related substance of the present invention adopts octadecylsilane chemically bonded silica to be the chromatographic column of filler, with a certain proportion of buffer salt solution-ion-pairing agent-organic phase for mobile phase.
Above-mentioned said chromatographic column is filler with octadecylsilane chemically bonded silica, is selected from the chromatographic column of the brands such as Kromasil, Apollo and Alltima.
Above-mentioned said organic phase is selected from following compound: methyl alcohol, acetonitrile, propyl alcohol, isopropyl alcohol, particular methanol.
Above-mentioned said method, its mobile phase buffer salt solution-ion-pairing agent-organic phase adopts gradient elution.
In above-mentioned said method, in buffer salt solution, buffer salt is selected from phosphate, formates, acetate, citrate, perchlorate, preferably phosphate.
The concentration of the buffer salt wherein comprised in buffer salt solution is 10 ~ 100mmol/L, and preferred concentration is 20mmol/L.
In above-mentioned said method, ion-pairing agent is selected from following reagent: perfluorooctane sulfonate, sodium heptanesulfonate, sodium dodecylsulphonate, lauryl sodium sulfate, preferred perfluorooctane sulfonate.
Wherein the concentration of ion-pairing agent is 10 ~ 100mmol/L, and preferred concentration is 20mmol/L.
Method of separating and assaying of the present invention, can realize in accordance with the following methods:
1) get razaxaban sample appropriate, by acetonitrile or mobile phase sample dissolution, be mixed with the sample solution that every 1mL contains razaxaban and related substance 0.1 ~ 1.5mg thereof;
2) arranging flow rate of mobile phase is 0.5 ~ 1.5mL/min, the preferred 1.0mL/min of flow rate of mobile phase, and determined wavelength is 205 ~ 280nm, and best detection wavelength is 210nm, and column oven temperature is 30 ~ 50 DEG C, and column oven temperature the best is 40 DEG C;
3) get 1) sample solution 10 ~ 50 μ L, injection liquid chromatography, completes the separation determination of razaxaban and related substance.
Wherein:
The model of high performance liquid chromatograph, has no special requirements, and the chromatograph that the present invention adopts is Shimadzu high performance liquid chromatograph: LC-20AT, CBM-20A, SIL-20AC, SPD-M20A, CTO-10ASvp
Chromatographic column: C
18(Kromasil, 250 × 4.6mm, 5 μm)
Mobile phase A: 20mmol/L diammonium hydrogen phosphate buffer solution-20mmol/L perfluorooctane sulfonate (pH3.0)
Mobile phase B: methyl alcohol
Flow velocity: 1.0mL/min
Determined wavelength: 210nm
Column temperature: 40 DEG C
Sampling volume: 10 μ L
Gradient condition:
| T | 0 | 10 | 25 | 60 | 70 | 71 | 80 |
| B% | 30 | 30 | 34 | 40 | 40 | 30 | 30 |
The present invention adopts C
18(Kromasil, 250 × 4.6mm, 5 μm), can effectively be separated razaxaban and related substance thereof.The invention solves the separation determination problem of razaxaban and related substance thereof, thus ensure that the quality of razaxaban, realize quality controllable (the results are shown in accompanying drawing 1 ~ 5) of razaxaban bulk drug and preparation thereof.
Accompanying drawing explanation
Razaxaban when Fig. 1 is embodiment 1 and related substance HPLC thereof scheme;
Razaxaban HPLC when Fig. 2 is embodiment 1 schemes;
Razaxaban when Fig. 3 is embodiment 2 and related substance HPLC thereof scheme;
The HPLC figure of razaxaban when Fig. 4 is embodiment 2;
Solvent HPLC when Fig. 5 is embodiment 2 schemes;
embodiment:
Following examples are used for understanding the present invention further, but are not limited to the scope of this enforcement.
embodiment 1
Instrument and condition
High performance liquid chromatograph: Shimadzu: LC-20AT, CBM-20A, SIL-20AC, SPD-M20A, CTO-10ASvp;
Chromatographic column: C
18(Kromasil, 250 × 4.6mm, 5 μm);
Mobile phase A: 20mmol/L diammonium hydrogen phosphate-20mmol/L perfluorooctane sulfonate buffer solution (pH3.0)
Mobile phase B: acetonitrile
Flow velocity: 1.0mL/min
Determined wavelength: 210nm
Column temperature: 40 DEG C
Sampling volume: 10 μ L
Gradient condition:
| T | 0 | 10 | 25 | 60 | 70 | 71 | 80 |
| B% | 8 | 8 | 18 | 30 | 30 | 8 | 8 |
Experimental procedure
Get razaxaban and related substance thereof appropriate, use acetonitrile sample dissolution respectively, be mixed with the sample solution being about 1.0mg/mL containing razaxaban and related substance thereof.Get above-mentioned razaxaban and related substance solution thereof appropriate, be mixed with system suitability solution; Efficient liquid phase chromatographic analysis is carried out, record chromatogram by above-mentioned condition.To the results are shown in retention time in accompanying drawing 1 ~ 2, Fig. 1 be the chromatographic peak of 57.332min is razaxaban, and all the other chromatographic peaks are the chromatographic peak of razaxaban related substance; In Fig. 2, retention time is the chromatographic peak of 57.805min is razaxaban.
embodiment 2
Instrument and condition
High performance liquid chromatograph: Shimadzu: LC-20AT, CBM-20A, SIL-20AC, SPD-M20A, CTO-10ASvp;
Chromatographic column: C
18(Kromasil, 250 × 4.6mm, 5 μm);
Mobile phase A: 20mmol/L diammonium hydrogen phosphate-20mmol/L perfluorooctane sulfonate buffer solution (pH3.0)
Mobile phase B: methyl alcohol
Flow velocity: 1.0mL/min
Determined wavelength: 210nm
Column temperature: 40 DEG C
Sampling volume: 10 μ L
Gradient condition:
| T | 0 | 10 | 25 | 60 | 70 | 71 | 80 |
| B% | 30 | 30 | 34 | 40 | 40 | 30 | 30 |
Experimental procedure
Get razaxaban and related substance thereof appropriate, use acetonitrile sample dissolution respectively, be mixed with the sample solution being about 1.0mg/mL containing razaxaban and related substance thereof.Get above-mentioned razaxaban and related substance solution thereof appropriate, be mixed with system suitability solution; Efficient liquid phase chromatographic analysis is carried out, record chromatogram by above-mentioned condition.To the results are shown in retention time in accompanying drawing 3 ~ 5, Fig. 3 be the chromatographic peak of 59.068min is razaxaban, and all the other chromatographic peaks are the chromatographic peak of razaxaban related substance; In Fig. 4, retention time is the chromatographic peak of 59.343min is razaxaban; Fig. 5 is blank solvent chromatogram.
stability of solution
Get the test liquid of razaxaban and related substance thereof, with the method in embodiment 2, respectively at 0,2,4,6,8,10,12,24 hour sample introduction, investigate the stability of solution when sample size measures, from result, this solution is stable in 24 hours.
durability
Because the above-mentioned chromatographic condition determined is gradient elution, and determine corresponding flow velocity, column temperature, mobile phase pH and chromatographic column model, therefore these conditions are done corresponding fine setting, investigate the durability of chromatographic condition.
Change in flow is within the scope of ± 0.1mL/min, and the peak shape of its related substance of razaxaban does not change, retention time have corresponding reach and after move; Column temperature change is within the scope of ± 5 DEG C, and the peak shape of each material and retention time are all without larger change; The pH change of mobile phase is in ± 0.2 scope, and the peak shape of each material and retention time are also without larger change; During the durability of chromatographic column is investigated, Apollo, Kromasil each material retention time and degree of separation all without larger markization, chromatographic column good tolerance.
Claims (13)
1. a method for liquid chromatography for separating and determining razaxaban related substance, is characterized in that: octadecylsilane chemically bonded silica is the chromatographic column of filler, with a certain proportion of buffer salt solution-ion-pairing agent-organic phase for mobile phase.
2. method of separating and assaying according to claim 1, chromatographic column is selected from the chromatographic column that brand is Kromasil, Apollo and Alltima.
3. method of separating and assaying according to claim 1, said organic phase is selected from the one in following compound: methyl alcohol, acetonitrile, propyl alcohol, isopropyl alcohol.
4. method of separating and assaying according to claim 3, said organic phase is methyl alcohol.
5. method of separating and assaying according to claim 1, said buffer salt solution is selected from following buffer salt: phosphate, formates, acetate, citrate, perchlorate etc.
6. method of separating and assaying according to claim 5, said buffer salt preferably phosphate, the preferred 20mmol/L of the concentration of buffer salt in buffer salt solution.
7. method of separating and assaying according to claim 1, said ion-pairing agent is selected from following ion-pairing agent: perfluorooctane sulfonate, sodium heptanesulfonate, sodium dodecylsulphonate, lauryl sodium sulfate etc.
8. method of separating and assaying according to claim 7, said ion-pairing agent is perfluorooctane sulfonate, the preferred 20mmol/L of concentration.
9. method of separating and assaying according to claim 1, is characterized in that, comprises following step:
1). get razaxaban sample appropriate, use acetonitrile sample dissolution respectively, be mixed with the sample solution that every 1mL contains razaxaban and related substance 0.1 ~ 1.5mg thereof;
2). arranging flow rate of mobile phase is 0.5 ~ 1.5mL/min, and determined wavelength is 205 ~ 280nm, and chromatographic column column oven temperature is 30 ~ 50 DEG C.
3). mobile phase A is 10 ~ 100mmol/L ammonium dihydrogen phosphate, 10 ~ 100mmol/L perfluorooctane sulfonate, adjusts pH to 2.5 ~ 4.0 with phosphoric acid; Mobile phase B is acetonitrile or methyl alcohol; The gradient of mobile phase arranges as follows:
10. method for separating and analyzing according to claim 5, buffer salt preferably phosphoric acid ammonium dihydrogen.
11. method for separating and analyzing according to claim 6, the pH value of buffer salt solution preferably 3.0.
12. method for separating and analyzing according to claim 9, step 2) the preferred 1.0mL/min of said flow rate of mobile phase.
13. method for separating and analyzing according to claim 9, step 2) said determined wavelength is 210nm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510603166.7A CN105259282A (en) | 2015-09-20 | 2015-09-20 | Method for separating and determining rivaroxaban related substances through liquid chromatography |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510603166.7A CN105259282A (en) | 2015-09-20 | 2015-09-20 | Method for separating and determining rivaroxaban related substances through liquid chromatography |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105259282A true CN105259282A (en) | 2016-01-20 |
Family
ID=55099049
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510603166.7A Pending CN105259282A (en) | 2015-09-20 | 2015-09-20 | Method for separating and determining rivaroxaban related substances through liquid chromatography |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105259282A (en) |
Cited By (6)
| 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 |
| CN108152412A (en) * | 2017-12-20 | 2018-06-12 | 乐普药业股份有限公司 | A kind of method with liquid chromatography for separating and determining razaxaban and its in relation to substance |
| CN110057942A (en) * | 2019-05-20 | 2019-07-26 | 海南皇隆制药股份有限公司 | A kind of detection method of the related substance of razaxaban and its preparation |
| CN110118836A (en) * | 2019-05-29 | 2019-08-13 | 北京悦康科创医药科技股份有限公司 | The method of genetoxic impurity in high effective liquid chromatography for measuring razaxaban |
| CN110954603A (en) * | 2018-09-27 | 2020-04-03 | 辽宁远大诺康生物制药有限公司 | Method for determining rivaroxaban and related substances thereof by using high performance liquid chromatography |
| CN112114057A (en) * | 2019-06-21 | 2020-12-22 | 北京万全德众医药生物技术有限公司 | Method for separating and measuring rivaroxaban and related substances thereof by using liquid chromatography |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012035057A2 (en) * | 2010-09-14 | 2012-03-22 | Medichem S.A. | Process for determining the suitability for distribution of a batch of a thiophene-2-carboxamide derivative |
| CN102822167A (en) * | 2010-01-04 | 2012-12-12 | 埃南蒂亚有限公司 | Process for the preparation of rivaroxaban and intermediates thereof |
| CN103822997A (en) * | 2014-03-20 | 2014-05-28 | 山东新时代药业有限公司 | Analysis and detection method for rivaroxaban intermediate |
| CN104892593A (en) * | 2015-06-19 | 2015-09-09 | 汕头经济特区鮀滨制药厂 | Synthetic methods of related substances F and G of rivaroxaban |
| CN105004802A (en) * | 2015-06-19 | 2015-10-28 | 重庆华邦制药有限公司 | Method for separating and determining rivaroxaban and impurities thereof, and application thereof |
-
2015
- 2015-09-20 CN CN201510603166.7A patent/CN105259282A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102822167A (en) * | 2010-01-04 | 2012-12-12 | 埃南蒂亚有限公司 | Process for the preparation of rivaroxaban and intermediates thereof |
| WO2012035057A2 (en) * | 2010-09-14 | 2012-03-22 | Medichem S.A. | Process for determining the suitability for distribution of a batch of a thiophene-2-carboxamide derivative |
| CN103822997A (en) * | 2014-03-20 | 2014-05-28 | 山东新时代药业有限公司 | Analysis and detection method for rivaroxaban intermediate |
| CN104892593A (en) * | 2015-06-19 | 2015-09-09 | 汕头经济特区鮀滨制药厂 | Synthetic methods of related substances F and G of rivaroxaban |
| CN105004802A (en) * | 2015-06-19 | 2015-10-28 | 重庆华邦制药有限公司 | Method for separating and determining rivaroxaban and impurities thereof, and application thereof |
Cited By (8)
| 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 |
| CN108152412A (en) * | 2017-12-20 | 2018-06-12 | 乐普药业股份有限公司 | A kind of method with liquid chromatography for separating and determining razaxaban and its in relation to substance |
| CN110954603A (en) * | 2018-09-27 | 2020-04-03 | 辽宁远大诺康生物制药有限公司 | Method for determining rivaroxaban and related substances thereof by using high performance liquid chromatography |
| CN110954603B (en) * | 2018-09-27 | 2022-04-15 | 远大生命科学(辽宁)有限公司 | Method for determining rivaroxaban and related substances thereof by using high performance liquid chromatography |
| CN110057942A (en) * | 2019-05-20 | 2019-07-26 | 海南皇隆制药股份有限公司 | A kind of detection method of the related substance of razaxaban and its preparation |
| CN110118836A (en) * | 2019-05-29 | 2019-08-13 | 北京悦康科创医药科技股份有限公司 | The method of genetoxic impurity in high effective liquid chromatography for measuring razaxaban |
| CN110118836B (en) * | 2019-05-29 | 2021-10-29 | 北京悦康科创医药科技股份有限公司 | Method for determining genotoxic impurities in rivaroxaban by high performance liquid chromatography |
| CN112114057A (en) * | 2019-06-21 | 2020-12-22 | 北京万全德众医药生物技术有限公司 | Method for separating and measuring rivaroxaban and related substances thereof by using liquid chromatography |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105259282A (en) | Method for separating and determining rivaroxaban related substances through liquid chromatography | |
| CN103353491B (en) | A method of with liquid chromatography for separating and determining alogliptin benzoate raw material and its preparation | |
| CN106596798B (en) | Analysis method of related substances in vortioxetine hydrobromide | |
| CN104316606A (en) | Method for separation and determination of vildagliptin related substances by liquid chromatography method | |
| CN104965041A (en) | High performance liquid chromatography detection method for parecoxib sodium isomer | |
| CN108152412A (en) | A kind of method with liquid chromatography for separating and determining razaxaban and its in relation to substance | |
| CN103698436B (en) | Method for detecting enantiomer in pramipexole dihydrochloride and method for separating enantiomer from pramipexole dihydrochloride | |
| CN104569269A (en) | Method for testing related substances of levocetirizine hydrochloride intermediate | |
| CN101701942A (en) | Method for separating and measuring entecavir and optical isomer thereof by liquid chromatography | |
| CN103760280A (en) | Method for separating and measuring asenapine intermediate related substances by liquid chromatography | |
| CN103353492A (en) | Method of separating and measuring solifenacin succinate raw material and preparation thereof by using liquid chromatography | |
| CN103364500B (en) | A method of with liquid chromatography for separating and determining bilastine raw material and its preparation | |
| CN105301156A (en) | Related substance analysis method for terlipressin for injection | |
| CN104569202A (en) | Method for separating and testing bazedoxifene and related substances of bazedoxifene | |
| CN106680386A (en) | Method for separating and testing related substance of carbocisteine raw material medicine and preparation by using liquid chromatography | |
| CN104297366A (en) | Liquid phase analysis method of maleic acid asenapine and impurities thereof | |
| CN104133008A (en) | Method using liquid chromatographic method for analysis of paliperidone intermediate and related substances | |
| CN104614468A (en) | Method for separating imidafenacin and related substances thereof by using high performance liquid chromatography | |
| CN109521102A (en) | Method of separating and assaying of the hydrobromic acid Vortioxetine finished product in relation to substance | |
| CN103760258A (en) | Method for separating and measuring asenapine maleate related substances by liquid chromatography | |
| CN108872405B (en) | HPLC analysis detection method for relative substances of lodoxylamine tromethamine | |
| CN108254463B (en) | Analysis method for detecting ornithine aspartate impurities | |
| CN103487532B (en) | A method of with liquid chromatography for separating and determining Vilazodone Hydrochloride raw material and its preparation | |
| CN102375044B (en) | Method for analyzing related substance from hydrochloric acid bendamustine intermediate Z6 | |
| CN104614467A (en) | Method for separating etoricoxib and related substances thereof by using high performance liquid chromatography |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20160120 |
|
| RJ01 | Rejection of invention patent application after publication |