CN103018346A - High-performance liquid chromatography analysis method for impurities in vorinostat and drug composition thereof - Google Patents
High-performance liquid chromatography analysis method for impurities in vorinostat and drug composition thereof Download PDFInfo
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Abstract
The present invention relates to a drug analysis method, especially to a method for detecting impurities in vorinostat by using high-performance liquid chromatography. As we all know, impurities in a drug have individual characteristics, and the same drug with different production processes can have different impurity spectrums. The present invention mainly provides an impurity detection method for vorinostat prepared through a special synthesis process. In addition, the method is further applicable for detection analysis of impurities in a vorinostat drug composition prepared through the synthesis process, wherein the impurities are aniline and/or 7-phenylcarbamoylheptanoic acid. The method has characteristics of rapidness, simpleness, effectiveness and reliability.
Description
Technical field
The present invention relates to a kind of analytical approach of medicine, the method that particularly with high performance liquid chromatography the impurity in Vorinostat and the pharmaceutical composition thereof is detected.
Background technology
Because a kind of medicine is from the synthesis material medicine to the relevant preparation of preparation, again through storage, transportation, use, experience one section comparatively complicated and very long process, each process all may produce relevant impurity during this period, may bring initiation material, reagent, intermediate, accessory substance and isomeride etc. in producing; In storage and transportation, may produce the special impurities such as catabolite, polymkeric substance or crystal transfer.The process contaminants that impurity in medicine general reference produces in the production of medicine and storage and transport process or catabolite etc.The bad reaction that medicine produces in clinical use also has much relations except outside the Pass the pharmacologically active with major component has with the impurity that exists in the medicine, and the control of impurity is an importance of drug research and development in the medicine, also is the guarantee of clinical safety in utilization.Therefore, in order to guarantee the safe and effective of medicine, also to consider the production actual conditions simultaneously, in the research process of medicine, all impurity be detected the important indicator as the control drug quality both at home and abroad.It is one of thin spot in present China drug research and development that medicine impurity detects research.Want the level of General Promotion China drug research and development, conscientiously guarantee the security of public's medication, must pay attention to and strengthen the research of relative substance in the medicine.
Vorinostat (Vorinostat) molecular formula is as shown below; chemical name is " SAHA " or " Vorinostat (SAHA) "; in the world first inhibition of histone deacetylase (histone deacetylase of Merck company exploitation; HDAC) new type anticancer medicine, treatment increase the weight of, continue and recur or treat rear invalid CTCLs (CTCL) with two kinds of general medications.
Vorinostat
Patent W02011/061545 discloses a kind of method that adopts efficient liquid phase chromatographic analysis Vorinostat content and impurity thereof, and this method especially is fit to analyze Vorinostat content and the impurity thereof that adopts the preparation of patent WO2010/043904 synthesis technique.As described in patent WO2011/061545 patent, the major impurity that the Vorinostat that adopts patent WO2010/043904 synthesis technique to prepare contains is:
(1) N, N '-diphenyl-suberamide:
(2) N '-phenyl-suberamide:
(3) 7-phenyl amino formoxyl enanthic acid:
(4) N-hydroxy-n, N-two (N '-phenyl suberoyl amido) amine:
As everyone knows, the impurity of medicine has the Extraordinary feature, and different medicines has different impurity spectrums, and same medicine also may have different impurity spectrums with different production technologies.The synthetic route of Vorinostat is more, and related initiation material and intermediate all are not quite similar, so the detection of the impurity of the Vorinostat that the method that provides of patent WO2011/061545 can not applicable all process routes preparations.In other words, adopt the Vorinostat of different process route preparation that the method that is fit to its impurity detection all need be arranged.
The inventor is also to synthetic many-sided research, the new synthesis technique of establishment of having carried out of Vorinostat.Find by analysis and research, the impurity that the Vorinostat that the synthesis technique that adopts the inventor to determine prepares may contain is: aniline, diphenylamine, 7-phenyl amino formoxyl enanthic acid, N-hydroxy-n, N-two (N '-phenyl suberoyl amido) amine.Therefore, need to explore condition determination, for this synthesis technique set up fast, simple and effectively, the reliable impurity determination method in Vorinostat and the composition thereof.
Summary of the invention
The objective of the invention is to explore the various conditions of the efficient liquid phase chromatographic analysis of impurity in Vorinostat and the pharmaceutical composition thereof, provide a kind of fast, simple, reliable, the effective HPLC analytical method of impurity in Vorinostat and the pharmaceutical composition thereof.
The inventor in the process of exploitation Vorinostat, primary study the HPLC analytical method of Vorinostat and composition impurity thereof.In research process, find; if with aniline, diphenylamine, 7-phenyl amino formoxyl enanthic acid, N-hydroxy-n; N-two (N '-phenyl suberoyl amido) four kinds of impurity of amine detect under same chromatographic condition; need the larger gradient condition of span; easily produce complicated eluting peak, disturb the judgement to impurity.According to researching and analysing the result; the inventor detects impurity aniline, 7-phenyl amino formoxyl enanthic acid and Vorinostat by technical solution of the present invention; and with impurity diphenylamine and N-hydroxy-n; N-two (N '-phenyl suberoyl amido) amine detects by another kind of technical scheme, and this technical scheme will be described in detail in another piece invention.
Technical scheme of the present invention is as follows:
Adopt the impurity in high-efficient liquid phase chromatogram technique analysis Vorinostat and the composition thereof, its chromatographic condition is:
(1) chromatographic column: octadecylsilane chemically bonded silica is filling agent;
(2) mobile phase: acetonitrile: phosphate buffered solution 10～50: 50～90 (v/v), pH value 3～8;
(3) detect wavelength: 200～250nm;
Wherein, described " impurity " is aniline and/or 7-phenyl amino formoxyl enanthic acid.
The technical program detects the preferred 210nm of wavelength or 240nm.
Preferred 0.01～the 0.05mol/L of phosphate buffered solution, more preferably 0.02mol/L; Acetonitrile: preferred 25: 75 of phosphate buffered solution volume ratio (v/v); PH value preferred 6; Phosphate can be sodium dihydrogen phosphate, potassium dihydrogen phosphate, sodium hydrogen phosphate, dipotassium hydrogen phosphate, sodium phosphate, potassium phosphate, ammonium phosphate etc., preferably phosphoric acid sodium dihydrogen.
In addition, the technical program adopts phosphoric acid or triethylamine to regulate mobile phase pH value.
(1) selection of testing conditions
1. detect the initial option of wavelength
The inventor has investigated aniline, 7-phenyl amino formoxyl enanthic acid and the uv absorption of Vorinostat reference substance in methyl alcohol; concrete grammar is for getting each impurity and the Vorinostat reference substance is an amount of; add respectively the methyl alcohol dissolving and make the solution that every ml contains each reference substance 10 μ g; measure according to UV-VIS spectrophotometry (two appendix IV of Chinese Pharmacopoeia version in 2005 A); the results are shown in Table 1, see accompanying drawing 1, Fig. 2, Fig. 3.
Table 1 reference substance uv absorption wavelength
| The sample title | Uv absorption |
| Aniline | Maximal ultraviolet absorption 204nm, 233nm |
| 7-phenyl amino formoxyl enanthic acid | Maximal ultraviolet absorption 203,242nm |
| Vorinostat | Maximal ultraviolet absorption 241nm; The 210nm absorbance is close to the 240nm absorbance |
Conclusion: by Fig. 1-3 and table 1 as can be known, aniline, 7-phenyl amino formoxyl enanthic acid and Vorinostat have larger absorption near 210nm and 240nm.
2. the selection and optimization of mobile phase
The inventor, gropes the liquid-phase condition of Vorinostat as detecting wavelength with 210nm and 240nm, and experimental result sees Table 2, table 3, table 4, table 5, table 6.
Table 2 mobile phase is selected
As shown in Table 2 take 0.2% (ml/ml) phosphoric acid solution-acetonitrile-methyl alcohol (70: 25: 5) as mobile phase, the Vorinostat retention time is moderate, better with the impurity degree of separation, but impurity aniline overlaps with solvent peak under this chromatographic condition, is unfavorable for the quantitative test to impurity aniline.
Therefore the inventor continues on this basis flow and is optimized mutually.Prioritization scheme sees table 3 for details:
Table 3 Optimization of mobile phase
As shown in Table 3, under the chromatographic condition on basis, the retention time of aniline is bad take acetonitrile-0.1% (ml/ml) phosphoric acid solution.Although by changing the pH value of mobile phase, can change the retention time of aniline in the 3rd scheme, can not improve the peak type of aniline.So need to continue to optimize chromatogram flow phase.The scheme that continues to optimize sees Table 4
Table 4 continues to optimize mobile phase
As shown in Table 4, under the chromatographic condition on basis, the retention time of aniline is moderate, the optimization but the peak type is still needed take acetonitrile-0.02mol/L phosphate buffered solution.The inventor makes further research mutually by the mode flow of the pH value of conversion buffer solution, research finds acetonitrile-0.02mol/L sodium dihydrogen phosphate buffer in pH value to 3～8 scopes, and aniline all can obtain more suitable retention time and peak type preferably.
The inventor adopts acetonitrile-0.02mol/L sodium dihydrogen phosphate buffer (with triethylamine adjust pH to 3～8) to be mobile phase; impurity 7-phenyl amino formoxyl enanthic acid is studied, and discovery 7-phenyl amino formoxyl enanthic acid also can be realized better keeping under this condition and satisfied peak type.Result of study sees Table 5:
The selection of table 5 impurity 7-phenyl amino formoxyl enanthic acid mobile phase
Therefore, can reach a conclusion: impurity aniline and 7-phenyl amino formoxyl enanthic acid all can be in the peak type of acetonitrile-0.02mol/L sodium dihydrogen phosphate buffer (with triethylamine adjust pH to 3～8) for realizing under the condition of mobile phase keeping and being satisfied with preferably.
On this basis, the inventor adopts acetonitrile-0.02mol/L sodium dihydrogen phosphate buffer (with triethylamine adjust pH to 3～8) for mobile phase, the Vorinostat crude product to be studied.Result of study sees Table 6:
The selection of table 6 Vorinostat crude product mobile phase
As shown in Table 7, acetonitrile-0.02mol/L sodium dihydrogen phosphate buffer (with triethylamine adjust pH to 3～8) is under the spectral condition of mobile phase, is fit to the detection analysis to impurity aniline among the Fu Linuo and 7-phenyl amino formoxyl enanthic acid.
In addition, the inventor adopts potassium dihydrogen phosphate, sodium hydrogen phosphate, dipotassium hydrogen phosphate, sodium phosphate, potassium phosphate, ammonium phosphate that sodium dihydrogen phosphate is replaced, and all can realize the effective analysis to impurity aniline among the Fu Linuo and 7-phenyl amino formoxyl enanthic acid.
The inventor go back flow mutually in phosphatic content in the phosphate buffered solution analyze; when finding that phosphatic content in the phosphate buffered solution is in the scope of 0.01～0.05mol/L, all can realize the effective analysis to impurity aniline among the Fu Linuo and 7-phenyl amino formoxyl enanthic acid.
Through comparative analysis, the particularly preferred mobile phase composition of the present invention is: acetonitrile-0.02mol/L sodium dihydrogen phosphate (regulating pH value to 6.0 with triethylamine) (25: 75).
3. auxiliary material interference experiment
The inventor also adopts technical solution of the present invention that the pharmaceutic adjuvant of preparation Vorinostat composition commonly used on the market is studied, and finds that after deliberation pharmaceutic adjuvant commonly used does not disturb the technical program.
(2) detection side's science of law research
1, aniline methodological study
Chromatographic condition is: octadecylsilane chemically bonded silica is the chromatographic column of filling agent; Acetonitrile: 0.02mol/L sodium dihydrogen phosphate (regulating pH value to 6.0 with triethylamine) (25: 75) is mobile phase; Detect wavelength 210nm; 30 ℃ of column temperatures; Theoretical cam curve is not less than 2000.
By above-mentioned chromatographic condition the aniline reference substance solution is measured, with the peak area of each aniline sample size is carried out linear regression, the gained equation of linear regression is: y=76.659x-38.384; Related coefficient is 1.Under this chromatographic system, aniline is good in 5.94ng/ml～1188.0ng/ml concentration range internal linear relation, aniline quantitatively be limited to 0.59ng, detect and be limited to 0.12ng.
2,7-phenyl amino formoxyl enanthic acid methodological study
Chromatographic condition is: octadecylsilane chemically bonded silica is the chromatographic column of filling agent; Acetonitrile: 0.02mol/L sodium dihydrogen phosphate (regulating pH value to 6.0 with triethylamine) (25: 75) is mobile phase; Detect wavelength 210nm; 30 ℃ of column temperatures; Theoretical cam curve is not less than 2000.
By above-mentioned chromatographic condition 7-phenyl amino formoxyl enanthic acid reference substance solution is measured, with the peak area of each 7-phenyl amino formoxyl enanthic acid sample size is carried out linear regression, the gained equation of linear regression is: y=49.183x+450.28; Related coefficient is 1.Under this chromatographic system, 7-phenyl amino formoxyl enanthic acid is good in 63.3ng/ml～1265.6ng/ml concentration range internal linear relation, 7-phenyl amino formoxyl enanthic acid quantitatively be limited to 0.63ng, detect and be limited to 0.13ng.
Beneficial effect: the inventive method has good detection effect for the impurity aniline in the Vorinostat under the specific synthesis technique and 7-phenyl amino formoxyl enanthic acid.Good, highly sensitive in extremely low impurity level scope linear relationship, detection line can reach 0.12ng and 0.13ng respectively.Under this testing conditions, impurity aniline and 7-phenyl amino formoxyl enanthic acid and Vorinostat retention time are all shorter in addition, and what wherein retention time was the longest is Vorinostat, and approximately about 10 minutes, required analysis time is shorter.
Description of drawings
Fig. 1 is the uv absorption figure of reference substance aniline;
Fig. 2 is reference substance 7-phenyl amino formoxyl enanthic acid uv absorption figure;
Fig. 3 is reference substance Vorinostat uv absorption figure;
Fig. 4 is Vorinostat capsule stability test of long duration determination of related substances in March;
Fig. 5 is Vorinostat capsule stability test of long duration determination of related substances in June;
Fig. 6 is the former Vorinostat capsule determination of related substances of grinding;
Fig. 7 is aniline at acetonitrile: the chromatogram under 10: 90 (pH 4) conditions of phosphate buffer solution;
Fig. 8 is aniline at acetonitrile: the chromatogram under 20: 80 (pH6) conditions of sodium dihydrogen phosphate buffer;
Fig. 9 is aniline at acetonitrile: the chromatogram under 25: 75 (pH6) conditions of potassium dihydrogen phosphate buffer solution;
Figure 10 is 7-phenyl amino formoxyl enanthic acid at acetonitrile: the chromatogram under 25: 75 (pH6) conditions of disodium hydrogen phosphate buffer solution.
Embodiment
The present invention will be further described below by embodiment, but embodiment does not limit protection scope of the present invention.
Take self-control Vorinostat crude product as raw material
Chromatographic condition: octadecylsilane chemically bonded silica is the chromatographic column of filling agent; Acetonitrile: 0.05mol/L sodium dihydrogen phosphate (10: 90) is mobile phase (with phosphorus acid for adjusting pH value to 3.0); Detect wavelength 200nm; 30 ℃ of column temperatures; Theoretical cam curve is not less than 2000.
The preparation of impurity reference substance solution: get respectively 7-phenyl amino formoxyl enanthic acid, aniline reference substance in measuring bottle, add mobile phase and be diluted to scale, shake up as the impurity contrast solution.
The preparation of need testing solution: precision takes by weighing the Vorinostat crude product and puts in right amount in the volumetric flask, adds methyl alcohol dissolving, and adds mobile phase and be diluted to scale, shakes up, as need testing solution.
The preparation of contrast solution: precision measures need testing solution 1ml, puts in the 100ml measuring bottle, and mobile phase is diluted to scale, shakes up, in contrast solution.
Determination method: get respectively impurity reference substance solution, need testing solution, contrast solution injection liquid chromatography, and the record chromatogram.
Testing result shows that the Vorinostat retention time is moderate, and the peak type is better, and impurity aniline peak and 7-phenyl amino formoxyl enanthic acid peak and Vorinostat degree of separation are better.
Embodiment 2
Take self-control Vorinostat crude product as raw material
Chromatographic condition: octadecylsilane chemically bonded silica is the chromatographic column of filling agent; Acetonitrile: 0.03mol/L potassium dihydrogen phosphate (10: 90) is mobile phase (with phosphorus acid for adjusting pH value to 4.0); Detect wavelength 210nm; 30 ℃ of column temperatures; Theoretical cam curve is not less than 2000.
The preparation of impurity reference substance solution: get respectively 7-phenyl amino formoxyl enanthic acid, aniline reference substance in measuring bottle, add mobile phase and be diluted to scale, shake up as the impurity contrast solution.
The preparation of need testing solution: precision takes by weighing the Vorinostat crude product and puts in right amount in the volumetric flask, adds methyl alcohol dissolving, and adds mobile phase and be diluted to scale, shakes up, as need testing solution.
The preparation of contrast solution: precision measures need testing solution 1ml, puts in the 100ml measuring bottle, and mobile phase is diluted to scale, shakes up, in contrast solution.
Determination method: get respectively impurity reference substance solution, need testing solution, contrast solution injection liquid chromatography, and the record chromatogram.
Testing result shows that the Vorinostat retention time is moderate, and the peak type is better, and impurity aniline peak and 7-phenyl amino formoxyl enanthic acid peak and Vorinostat degree of separation are better.
Embodiment 3
Take self-control Vorinostat crude product as raw material
Chromatographic condition: octadecylsilane chemically bonded silica is the chromatographic column of filling agent; Acetonitrile: 0.02mol/L sodium dihydrogen phosphate (10: 90) is mobile phase (regulating pH value to 6.0 with triethylamine); Detect wavelength 240nm; 30 ℃ of column temperatures; Theoretical cam curve is not less than 2000.
The preparation of impurity reference substance solution: get respectively 7-phenyl amino formoxyl enanthic acid, aniline reference substance in measuring bottle, add mobile phase and be diluted to scale, shake up as the impurity contrast solution.
The preparation of need testing solution: precision takes by weighing the Vorinostat crude product and puts in right amount in the volumetric flask, adds methyl alcohol dissolving, and adds mobile phase and be diluted to scale, shakes up, as need testing solution.
The preparation of contrast solution: precision measures need testing solution 1ml, puts in the 100ml measuring bottle, and mobile phase is diluted to scale, shakes up, in contrast solution.
Determination method: get respectively impurity reference substance solution, need testing solution, contrast solution injection liquid chromatography, and the record chromatogram.
Testing result shows that the Vorinostat retention time is moderate, and the peak type is better, and impurity aniline peak and 7-phenyl amino formoxyl enanthic acid peak and Vorinostat degree of separation are better.
Embodiment 4
Take self-control Vorinostat crude product as raw material
Chromatographic condition: octadecylsilane chemically bonded silica is the chromatographic column of filling agent; Acetonitrile: 0.01mol/L dipotassium hydrogen phosphate solution (10: 90) is mobile phase (with phosphorus acid for adjusting pH value to 8.0); Detect wavelength 250nm; 30 ℃ of column temperatures; Theoretical cam curve is not less than 2000.
The preparation of impurity reference substance solution: get respectively 7-phenyl amino formoxyl enanthic acid, aniline reference substance in measuring bottle, add mobile phase and be diluted to scale, shake up as the impurity contrast solution.
The preparation of need testing solution: precision takes by weighing the Vorinostat crude product and puts in right amount in the volumetric flask, adds methyl alcohol dissolving, and adds mobile phase and be diluted to scale, shakes up, as need testing solution.
The preparation of contrast solution: precision measures need testing solution 1ml, puts in the 100ml measuring bottle, and mobile phase is diluted to scale, shakes up, in contrast solution.
Determination method: get respectively impurity reference substance solution, need testing solution, contrast solution injection liquid chromatography, and the record chromatogram.
Testing result shows that the Vorinostat retention time is moderate, and the peak type is better, and impurity aniline peak and 7-phenyl amino formoxyl enanthic acid peak and Vorinostat degree of separation are better.
Embodiment 5
Take self-control Vorinostat crude product as raw material
Chromatographic condition: octadecylsilane chemically bonded silica is the chromatographic column of filling agent; Acetonitrile: 0.05mol/L disodium phosphate soln (20: 80) is mobile phase (with phosphorus acid for adjusting pH value to 3.0); Detect wavelength 200nm; 30 ℃ of column temperatures; Theoretical cam curve is not less than 2000.
The preparation of impurity reference substance solution: get respectively 7-phenyl amino formoxyl enanthic acid, aniline reference substance in measuring bottle, add mobile phase and be diluted to scale, shake up as the impurity contrast solution.
The preparation of need testing solution: precision takes by weighing the Vorinostat crude product and puts in right amount in the volumetric flask, adds methyl alcohol dissolving, and adds mobile phase and be diluted to scale, shakes up, as need testing solution.
The preparation of contrast solution: precision measures need testing solution 1ml, puts in the 100ml measuring bottle, and mobile phase is diluted to scale, shakes up, in contrast solution.
Determination method: get respectively impurity reference substance solution, need testing solution, contrast solution injection liquid chromatography, and the record chromatogram.
Testing result shows that the Vorinostat retention time is moderate, and the peak type is better, and impurity aniline peak and 7-phenyl amino formoxyl enanthic acid peak and Vorinostat degree of separation are better.
Embodiment 6
Take self-control Vorinostat crude product as raw material
Chromatographic condition: octadecylsilane chemically bonded silica is the chromatographic column of filling agent; Acetonitrile: 0.03mol/L dipotassium hydrogen phosphate solution (20: 80) is mobile phase (with phosphorus acid for adjusting pH value to 4.0); Detect wavelength 210nm; 30 ℃ of column temperatures; Theoretical cam curve is not less than 2000.
The preparation of impurity reference substance solution: get respectively 7-phenyl amino formoxyl enanthic acid, aniline reference substance in measuring bottle, add mobile phase and be diluted to scale, shake up as the impurity contrast solution.
The preparation of need testing solution: precision takes by weighing the Vorinostat crude product and puts in right amount in the volumetric flask, adds methyl alcohol dissolving, and adds mobile phase and be diluted to scale, shakes up, as need testing solution.
The preparation of contrast solution: precision measures need testing solution 1ml, puts in the 100ml measuring bottle, and mobile phase is diluted to scale, shakes up, in contrast solution.
Determination method: get respectively impurity reference substance solution, need testing solution, contrast solution injection liquid chromatography, and the record chromatogram.
Testing result shows that the Vorinostat retention time is moderate, and the peak type is better, and impurity aniline peak and 7-phenyl amino formoxyl enanthic acid peak and Vorinostat degree of separation are better.
Embodiment 7
Take self-control Vorinostat crude product as raw material
Chromatographic condition: octadecylsilane chemically bonded silica is the chromatographic column of filling agent; Acetonitrile: 0.02mol/L disodium phosphate soln (20: 80) is mobile phase (with phosphorus acid for adjusting pH value to 6.0); Detect wavelength 240nm; 30 ℃ of column temperatures; Theoretical cam curve is not less than 2000.
The preparation of impurity reference substance solution: get respectively 7-phenyl amino formoxyl enanthic acid, aniline reference substance in measuring bottle, add mobile phase and be diluted to scale, shake up as the impurity contrast solution.
The preparation of need testing solution: precision takes by weighing the Vorinostat crude product and puts in right amount in the volumetric flask, adds methyl alcohol dissolving, and adds mobile phase and be diluted to scale, shakes up, as need testing solution.
The preparation of contrast solution: precision measures need testing solution 1ml, puts in the 100ml measuring bottle, and mobile phase is diluted to scale, shakes up, in contrast solution.
Determination method: get respectively impurity reference substance solution, need testing solution, contrast solution injection liquid chromatography, and the record chromatogram.
Testing result shows that the Vorinostat retention time is moderate, and the peak type is better, and impurity aniline peak and 7-phenyl amino formoxyl enanthic acid peak and Vorinostat degree of separation are better.
Embodiment 8
Take self-control Vorinostat crude product as raw material
Chromatographic condition: octadecylsilane chemically bonded silica is the chromatographic column of filling agent; Acetonitrile: 0.01mol/L potassium phosphate solution (20: 80) is mobile phase (with phosphorus acid for adjusting pH value to 8.0); Detect wavelength 250nm; 30 ℃ of column temperatures; Theoretical cam curve is not less than 2000.
The preparation of impurity reference substance solution: get respectively 7-phenyl amino formoxyl enanthic acid, aniline reference substance in measuring bottle, add mobile phase and be diluted to scale, shake up as the impurity contrast solution.
The preparation of need testing solution: precision takes by weighing the Vorinostat crude product and puts in right amount in the volumetric flask, adds methyl alcohol dissolving, and adds mobile phase and be diluted to scale, shakes up, as need testing solution.
The preparation of contrast solution: precision measures need testing solution 1ml, puts in the 100ml measuring bottle, and mobile phase is diluted to scale, shakes up, in contrast solution.
Determination method: get respectively impurity reference substance solution, need testing solution, contrast solution injection liquid chromatography, and the record chromatogram.
Testing result shows that the Vorinostat retention time is moderate, and the peak type is better, and impurity aniline peak and 7-phenyl amino formoxyl enanthic acid peak and Vorinostat degree of separation are better.
Embodiment 9 Vorinostat stability study test of long duration
, put under 25 ± 2 ℃ of temperature, relative humidity RH60% ± 10% condition by commercially available back and placed 6 months to make the Vorinostat capsule by oneself as testing sample with the inventor, and in 3rd month, 6 samplings at the end of month, stable with 0 month contrast investigation Vorinostat.
Chromatographic condition is: octadecylsilane chemically bonded silica is the chromatographic column of filling agent; Acetonitrile: 0.02mol/L potassium dihydrogen phosphate (regulating pH value to 6.0 with triethylamine) (25: 75) is mobile phase; Detect wavelength 210nm; 30 ℃ of column temperatures; Theoretical cam curve is not less than 2000.
The preparation of impurity reference substance solution: get respectively 7-phenyl amino formoxyl enanthic acid, aniline reference substance in measuring bottle, add mobile phase and be diluted to scale, shake up as the impurity contrast solution.
The preparation of need testing solution: get the content under the Vorinostat capsule content uniformity item, mix, take by weighing in right amount and put in the capacity measuring bottle, add methyl alcohol dissolving, and add mobile phase and be diluted to scale, shake up, as need testing solution.
The preparation of contrast solution: precision measures need testing solution 1ml, puts in the 100ml measuring bottle, and mobile phase is diluted to scale, shakes up, in contrast solution.
Determination method: get respectively impurity reference substance solution, need testing solution, contrast solution injection liquid chromatography, and the record chromatogram.
Conclusion: the study on the stability result of this product shows, this product under the accelerated test condition, place 6 months basicly stable.The results are shown in following table:
Table 7 long-term test results
(25 ± 2 ℃/RH60% of temperature ± 10%)
| Investigate the date | 7-phenyl amino formoxyl enanthic | Aniline | Remarks | |
| 0 month | 0.04 | 0.00 | ||
| March | 0.04 | 0.00 | The test sample chromatogram is seen accompanying drawing Fig. 4 | |
| June | 0.04 | 0.00 | The test sample chromatogram is seen accompanying drawing Fig. 5 |
Take the former Vorinostat capsule that grinds as testing sample.
Chromatographic condition is: octadecylsilane chemically bonded silica is the chromatographic column of filling agent; Acetonitrile: 0.02mol/L sodium dihydrogen phosphate (regulating pH value to 6.0 with triethylamine) (25: 75) is mobile phase; Detect wavelength 240nm; 30 ℃ of column temperatures; Theoretical cam curve is not less than 2000.
The preparation of impurity reference substance solution: get respectively 7-phenyl amino formoxyl enanthic acid, aniline reference substance in measuring bottle, add mobile phase and be diluted to scale, shake up as the impurity contrast solution.
The preparation of need testing solution: get the content under the Vorinostat capsule content uniformity item, mix, take by weighing in right amount and put in the capacity measuring bottle, add methyl alcohol dissolving, and add mobile phase and be diluted to scale, shake up, as need testing solution.
The preparation of contrast solution: precision measures need testing solution 1ml, puts in the 100ml measuring bottle, and mobile phase is diluted to scale, shakes up, in contrast solution.
Determination method: get respectively impurity reference substance solution, need testing solution, contrast solution injection liquid chromatography, and record chromatogram (the test sample chromatogram is seen accompanying drawing Fig. 6).
Embodiment 11
With the inventor take self-control Vorinostat capsule as testing sample.
Chromatographic condition: octadecylsilane chemically bonded silica is the chromatographic column of filling agent; Acetonitrile: 0.03mol/L sodium radio-phosphate,P-32 solution (25: 75) is mobile phase (with phosphorus acid for adjusting pH value to 4.0); Detect wavelength 200nm; 30 ℃ of column temperatures; Theoretical cam curve is not less than 2000.
The preparation of impurity reference substance solution: get respectively 7-phenyl amino formoxyl enanthic acid, aniline reference substance in measuring bottle, add mobile phase and be diluted to scale, shake up as the impurity contrast solution.
The preparation of need testing solution: get the content under the Vorinostat capsule content uniformity item, mix, take by weighing in right amount and put in the capacity measuring bottle, add methyl alcohol dissolving, and add mobile phase and be diluted to scale, shake up, as need testing solution.
The preparation of contrast solution: precision measures need testing solution 1ml, puts in the 100ml measuring bottle, and mobile phase is diluted to scale, shakes up, in contrast solution.
Determination method: get respectively impurity reference substance solution, need testing solution, contrast solution injection liquid chromatography, and the record chromatogram.
Testing result shows that the Vorinostat retention time is moderate, and the peak type is better, and impurity aniline peak and 7-phenyl amino formoxyl enanthic acid peak and Vorinostat degree of separation are better.
Embodiment 12
With the inventor take self-control Vorinostat capsule as testing sample.
Chromatographic condition: octadecylsilane chemically bonded silica is the chromatographic column of filling agent; Acetonitrile: 0.05mol/L potassium phosphate solution (25: 75) is mobile phase (with phosphorus acid for adjusting pH value to 3.0); Detect wavelength 210nm; 30 ℃ of column temperatures; Theoretical cam curve is not less than 2000.
The preparation of impurity reference substance solution: get respectively 7-phenyl amino formoxyl enanthic acid, aniline reference substance in measuring bottle, add mobile phase and be diluted to scale, shake up as the impurity contrast solution.
The preparation of need testing solution: get the content under the Vorinostat capsule content uniformity item, mix, take by weighing in right amount and put in the capacity measuring bottle, add methyl alcohol dissolving, and add mobile phase and be diluted to scale, shake up, as need testing solution.
The preparation of contrast solution: precision measures need testing solution 1ml, puts in the 100ml measuring bottle, and mobile phase is diluted to scale, shakes up, in contrast solution.
Determination method: get respectively impurity reference substance solution, need testing solution, contrast solution injection liquid chromatography, and the record chromatogram.
Testing result shows that the Vorinostat retention time is moderate, and the peak type is better, and impurity aniline peak and 7-phenyl amino formoxyl enanthic acid peak and Vorinostat degree of separation are better.
Embodiment 13
With the inventor take self-control Vorinostat capsule as testing sample.
Chromatographic condition: octadecylsilane chemically bonded silica is the chromatographic column of filling agent; Acetonitrile: 0.01mol/L potassium phosphate solution (25: 75) is mobile phase (with phosphorus acid for adjusting pH value to 8.0); Detect wavelength 210nm; 30 ℃ of column temperatures; Theoretical cam curve is not less than 2000.
The preparation of impurity reference substance solution: get respectively 7-phenyl amino formoxyl enanthic acid, aniline reference substance in measuring bottle, add mobile phase and be diluted to scale, shake up as the impurity contrast solution.
The preparation of need testing solution: get the content under the Vorinostat capsule content uniformity item, mix, take by weighing in right amount and put in the capacity measuring bottle, add methyl alcohol dissolving, and add mobile phase and be diluted to scale, shake up, as need testing solution.
The preparation of contrast solution: precision measures need testing solution 1ml, puts in the 100ml measuring bottle, and mobile phase is diluted to scale, shakes up, in contrast solution.
Determination method: get respectively impurity reference substance solution, need testing solution, contrast solution injection liquid chromatography, and the record chromatogram.
Testing result shows that the Vorinostat retention time is moderate, and the peak type is better, and impurity aniline peak and 7-phenyl amino formoxyl enanthic acid peak and Vorinostat degree of separation are better.
Embodiment 14
With the inventor take self-control Vorinostat capsule as testing sample.
Chromatographic condition: octadecylsilane chemically bonded silica is the chromatographic column of filling agent; Acetonitrile: 0.03mol/L sodium dihydrogen phosphate (50: 50) is mobile phase (with phosphorus acid for adjusting pH value to 3.0); Detect wavelength 210nm; 30 ℃ of column temperatures; Theoretical cam curve is not less than 2000.
The preparation of impurity reference substance solution: get respectively 7-phenyl amino formoxyl enanthic acid, aniline reference substance in measuring bottle, add mobile phase and be diluted to scale, shake up as the impurity contrast solution.
The preparation of need testing solution: get the content under the Vorinostat capsule content uniformity item, mix, take by weighing in right amount and put in the capacity measuring bottle, add methyl alcohol dissolving, and add mobile phase and be diluted to scale, shake up, as need testing solution.
The preparation of contrast solution: precision measures need testing solution 1ml, puts in the 100ml measuring bottle, and mobile phase is diluted to scale, shakes up, in contrast solution.
Determination method: get respectively impurity reference substance solution, need testing solution, contrast solution injection liquid chromatography, and the record chromatogram.
Testing result shows that the Vorinostat retention time is moderate, and the peak type is better, and impurity aniline peak and 7-phenyl amino formoxyl enanthic acid peak and Vorinostat degree of separation are better.
With the inventor take self-control Vorinostat capsule as testing sample.
Chromatographic condition: octadecylsilane chemically bonded silica is the chromatographic column of filling agent; Acetonitrile: 0.02mol/L potassium dihydrogen phosphate (50: 50) is mobile phase (regulating pH value to 4.0 with triethylamine); Detect wavelength 200nm; 30 ℃ of column temperatures; Theoretical cam curve is not less than 2000.
The preparation of impurity reference substance solution: get respectively 7-phenyl amino formoxyl enanthic acid, aniline reference substance in measuring bottle, add mobile phase and be diluted to scale, shake up as the impurity contrast solution.
The preparation of need testing solution: get the content under the Vorinostat capsule content uniformity item, mix, take by weighing in right amount and put in the capacity measuring bottle, add methyl alcohol dissolving, and add mobile phase and be diluted to scale, shake up, as need testing solution.
The preparation of contrast solution: precision measures need testing solution 1ml, puts in the 100ml measuring bottle, and mobile phase is diluted to scale, shakes up, in contrast solution.
Determination method: get respectively impurity reference substance solution, need testing solution, contrast solution injection liquid chromatography, and the record chromatogram.
Testing result shows that the Vorinostat retention time is moderate, and the peak type is better, and impurity aniline peak and 7-phenyl amino formoxyl enanthic acid peak and Vorinostat degree of separation are better.
Embodiment 16
With the inventor take self-control Vorinostat capsule as testing sample.
Chromatographic condition: octadecylsilane chemically bonded silica is the chromatographic column of filling agent; Acetonitrile: 0.01mol/L disodium phosphate soln (50: 50) is mobile phase (with phosphorus acid for adjusting pH value to 6.0); Detect wavelength 210nm; 30 ℃ of column temperatures; Theoretical cam curve is not less than 2000.
The preparation of impurity reference substance solution: get respectively 7-phenyl amino formoxyl enanthic acid, aniline reference substance in measuring bottle, add mobile phase and be diluted to scale, shake up as the impurity contrast solution.
The preparation of need testing solution: get the content under the Vorinostat capsule content uniformity item, mix, take by weighing in right amount and put in the capacity measuring bottle, add methyl alcohol dissolving, and add mobile phase and be diluted to scale, shake up, as need testing solution.
The preparation of contrast solution: precision measures need testing solution 1ml, puts in the 100ml measuring bottle, and mobile phase is diluted to scale, shakes up, in contrast solution.
Determination method: get respectively impurity reference substance solution, need testing solution, contrast solution injection liquid chromatography, and the record chromatogram.
Testing result shows that the Vorinostat retention time is moderate, and the peak type is better, and impurity aniline peak and 7-phenyl amino formoxyl enanthic acid peak and Vorinostat degree of separation are better.
Embodiment 17
With the inventor take self-control Vorinostat capsule as testing sample.
Chromatographic condition: octadecylsilane chemically bonded silica is the chromatographic column of filling agent; Acetonitrile: 0.05mol/L potassium phosphate solution (50: 50) is mobile phase (with phosphorus acid for adjusting pH value to 8.0); Detect wavelength 240nm; 30 ℃ of column temperatures; Theoretical cam curve is not less than 2000.
The preparation of impurity reference substance solution: get respectively 7-phenyl amino formoxyl enanthic acid, aniline reference substance in measuring bottle, add mobile phase and be diluted to scale, shake up as the impurity contrast solution.
The preparation of need testing solution: get the content under the Vorinostat capsule content uniformity item, mix, take by weighing in right amount and put in the capacity measuring bottle, add methyl alcohol dissolving, and add mobile phase and be diluted to scale, shake up, as need testing solution.
The preparation of contrast solution: precision measures need testing solution 1ml, puts in the 100ml measuring bottle, and mobile phase is diluted to scale, shakes up, in contrast solution.
Determination method: get respectively impurity reference substance solution, need testing solution, contrast solution injection liquid chromatography, and the record chromatogram.
Testing result shows that the Vorinostat retention time is moderate, and the peak type is better, and impurity aniline peak and 7-phenyl amino formoxyl enanthic acid peak and Vorinostat degree of separation are better.
Embodiment 18
Take commercial aniline as product to be tested.
Chromatographic condition is: octadecylsilane chemically bonded silica is the chromatographic column of filling agent; Acetonitrile: 0.1% (v/v) phosphoric acid solution (10: 90) is mobile phase (regulating pH value to 4.0 with triethylamine); Detect wavelength 210nm; 30 ℃ of column temperatures; Theoretical cam curve is not less than 2000.
Precision takes by weighing aniline and puts in the volumetric flask, adds methyl alcohol and makes dissolving and be diluted to scale, shakes up, the accurate test sample injection liquid chromatography of drawing, record chromatogram (seeing accompanying drawing Fig. 7).
Conclusion: by spectrogram as can be known, retention time 10.8 minutes, the peak type is relatively poor.
Embodiment 19
Take commercial aniline as product to be tested.
Chromatographic condition is: octadecylsilane chemically bonded silica is the chromatographic column of filling agent; Acetonitrile: 0.02% (mol/L) sodium dihydrogen phosphate (20: 80) is mobile phase (regulating pH value to 6.0 with triethylamine); Detect wavelength 210nm; 30 ℃ of column temperatures; Theoretical cam curve is not less than 2000.
Precision takes by weighing aniline and puts in the volumetric flask, adds methyl alcohol and makes dissolving and be diluted to scale, shakes up, the accurate test sample injection liquid chromatography of drawing, record chromatogram (seeing accompanying drawing Fig. 8).
Conclusion: by spectrogram as can be known, retention time 13.4 minutes, the peak type is better.
Take commercial aniline as product to be tested.
Chromatographic condition is: octadecylsilane chemically bonded silica is the chromatographic column of filling agent; Acetonitrile: 0.02% (mol/L) phosphate potassium dihydrogen solution (25: 75) is mobile phase (with phosphorus acid for adjusting pH value to 6.0); Detect wavelength 210nm; 30 ℃ of column temperatures; Theoretical cam curve is not less than 2000.
Precision takes by weighing aniline and puts in the volumetric flask, adds methyl alcohol and makes dissolving and be diluted to scale, shakes up, the accurate test sample injection liquid chromatography of drawing, record chromatogram (seeing accompanying drawing Fig. 9).
Conclusion: by spectrogram as can be known, retention time 9.2 minutes, the peak type is better.
Embodiment 21
Take homemade 7-phenyl amino formoxyl enanthic acid as product to be tested
Chromatographic condition is: octadecylsilane chemically bonded silica is the chromatographic column of filling agent; Acetonitrile: 0.02% (mol/L) sodium hydrogen phosphate (25: 75) is mobile phase (with phosphorus acid for adjusting pH value to 4.0); Detect wavelength 210nm; 30 ℃ of column temperatures; Theoretical cam curve is not less than 2000.
Get 7-phenyl amino formoxyl enanthic acid and put in the volumetric flask, add methyl alcohol and make dissolving and be diluted to scale, shake up, the accurate test sample injection liquid chromatography of drawing, record chromatogram (seeing accompanying drawing Figure 10).
Conclusion: by spectrogram as can be known, retention time 8.0 minutes, the peak type is better.
Claims (8)
1. the analytical approach of impurity in Vorinostat and the pharmaceutical composition thereof is characterized in that adopting high performance liquid chromatography, and chromatographic condition is:
(1) chromatographic column: octadecylsilane chemically bonded silica is filling agent;
(2) mobile phase: acetonitrile: phosphate buffered solution 10～50: 50～90 (v/v), pH value 3～8;
(3) detect wavelength: 200～250nm.
Wherein said impurity is aniline and/or 7-phenyl amino formoxyl enanthic acid.
2. the method for claim 1 is characterized in that described wavelength is 210nm or 240nm.
3. the method for claim 1, the phosphate content that it is characterized in that described phosphate buffered solution is 0.01～0.05mol/L.
4. method as claimed in claim 3, the phosphate content that it is characterized in that phosphate buffered solution is 0.02mol/L.
5. such as claim 1,2,3,4 described methods, it is characterized in that acetonitrile: phosphate buffered solution is 25: 75 (v/v).
6. method as claimed in claim 5 is characterized in that described phosphate buffered solution pH value is 6.
7. method as claimed in claim 6 is characterized in that described phosphate is sodium dihydrogen phosphate.
8. such as method as described in the claim 1,2,3,4,6 or 7, it is characterized in that adopting phosphoric acid or triethylamine to regulate the pH of mobile phase.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105259261A (en) * | 2015-10-09 | 2016-01-20 | 扬子江药业集团江苏海慈生物药业有限公司 | Measuring method for aniline content in drug |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040122101A1 (en) * | 2002-03-04 | 2004-06-24 | Miller Thomas A. | Polymorphs of suberoylanilide hydroxamic acid |
| WO2010043904A2 (en) * | 2008-10-15 | 2010-04-22 | Generics [Uk] Limited | Improved process |
| CN101874793A (en) * | 2009-04-29 | 2010-11-03 | 北京本草天源药物研究院 | Vorinostat solid preparation |
| CN101939289A (en) * | 2008-02-07 | 2011-01-05 | 基因里克斯(英国)有限公司 | Novel process for the preparation of vorinostat |
| WO2011061545A1 (en) * | 2009-11-23 | 2011-05-26 | Generics [Uk] Limited | Hplc method for analyzing vorinostat |
-
2011
- 2011-09-20 CN CN2011102788364A patent/CN103018346A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040122101A1 (en) * | 2002-03-04 | 2004-06-24 | Miller Thomas A. | Polymorphs of suberoylanilide hydroxamic acid |
| CN101939289A (en) * | 2008-02-07 | 2011-01-05 | 基因里克斯(英国)有限公司 | Novel process for the preparation of vorinostat |
| WO2010043904A2 (en) * | 2008-10-15 | 2010-04-22 | Generics [Uk] Limited | Improved process |
| CN101874793A (en) * | 2009-04-29 | 2010-11-03 | 北京本草天源药物研究院 | Vorinostat solid preparation |
| WO2011061545A1 (en) * | 2009-11-23 | 2011-05-26 | Generics [Uk] Limited | Hplc method for analyzing vorinostat |
Non-Patent Citations (4)
| Title |
|---|
| KATAN PATEL ET AL.: "Simultaneous determination of decitabine and vorinostat (Suberoylanalide hydroxamic acid, SAHA) by liquid chromatography tandem mass spectrometry for clinical studies", 《JOURNAL OF CHROMATOGRAPHY B》 * |
| LIHONG DU ET AL.: "High turbulence liquid chromatography online extraction and tandem mass spectrometry for the simultaneous determination of suberoylanilide hydroxamic acid and its two metabolites in human serum", 《RAPID COMMUNICATIONS IN MASS SPECTROMETRY》 * |
| 胡杨 等: "伏立诺他的合成", 《中国医药工业杂志》 * |
| 苏国琛 等: "辛二酰苯胺异差劲肟酸合成的实验设计", 《实验室科学》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105259261A (en) * | 2015-10-09 | 2016-01-20 | 扬子江药业集团江苏海慈生物药业有限公司 | Measuring method for aniline content in drug |
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