CN107870217A - Detection method of the eltrombopag olamine intermediate about material and the impurity reference substance for this method - Google Patents
Detection method of the eltrombopag olamine intermediate about material and the impurity reference substance for this method Download PDFInfo
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Abstract
Entitled detection method of the eltrombopag olamine intermediate about material of the present invention and the impurity reference substance for this method, belong to Pharmaceutical Analysis technical field.The chromatographic column using phenyl and octadecylsilane chemically bonded silica as filler is respectively adopted in the present invention, using phosphate buffer and acetonitrile or phosphoric acid acetonitrile solution as mobile phase, using gradient elution mode, the 2 kinds of high efficient liquid phase analysis methods established can effectively determine the carboxylic acid of I 2' hydroxyl 3' amino phenylbenzene of eltrombopag olamine synthetic intermediate 3 and intermediate II 2 (3 respectively, 4 3,5-dimethylphenyls) 5 methyl 2, the content of the ketone of 4 pyrazoline 3 and above-mentioned intermediate related impurities, method specificity is good, high sensitivity, it is easy to operate, quality control for eltrombopag olamine finished product provides effective guarantee.The invention also discloses a kind of new impurity reference substance compound.
Description
Technical field
The invention belongs to Pharmaceutical Analysis technical field, and in particular to a kind of intermediate I determined for synthesizing eltrombopag olamine
2'- hydroxyls -3'- amino-phenylbenzene -3- carboxylic acids and intermediate II 2- (3,4- Dimethvl-phenyls) -5- methyl -2,4- dihydros-pyrrole
The Pharmaceutical Analysis method of the content of azoles -3- ketone related impuritieses.
The invention further relates to a kind of new impurity reference substance chemical combination in the analysis method for eltrombopag olamine intermediate II
Thing DH.
Background technology
Eltrombopag olamine, entitled 3'- { (2Z) -2- [1- (3,4- the 3,5-dimethylphenyl) -3- methyl -5- oxos -1,5- bis- of chemistry
Hydrogen -4H- pyrazoles -4- subunits] diazanyl } -2'- Hydroxybiphenyl -3- carboxylic acid diethanol amine, molecular formula C25H22N4O4·
2C2H7NO, molecular weight 564.63, its structural formula are as follows：
Eltrombopag olamine is oral platelet generation factor type medicine, the compound can with the TPO of transmembrane region by
Body phase interaction, produce signal cascade amplification, and then the propagation of inducing bone marrow megacaryocyte and differentiation.Eltrombopag olamine is by Pueraria lobota
Lan Su SmithKlines company is taken the lead in developing, and food and medicine Surveillance Authority of the U.S. (FDA) approval is obtained in November, 2008 in the U.S.
Listing, trade name Promacta, for treating, glucocorticoid medicine, immunoglobulin therapy be invalid or Postsplenectomy
The decrease of platelet of chronic idiopathic thrombocytopenic purpura patient, it is small molecule thrombopoietin receptor evident in efficacy
Activator.
The synthetic route of eltrombopag olamine is all in patent WO2010114943, WO2013072921, US20150087845A1 etc.
Had been reported that in more documents.Although technological parameter is different, synthetic route is basically identical, that is, passes through 2'- hydroxyl -3'- ammonia
Base-phenylbenzene -3- carboxylic acids (intermediate I) and 2- (3,4- Dimethvl-phenyls) -5- methyl -2,4- dihydro-pyrazol -3- ketone are (middle
Body II) it is prepared by the condensation coupling reaction between two intermediates.
Quality control of intermediates improves important role for reaction monitoring and yield, while also contributes to finished product
Quality.So far, the report of pertinent literature and patent without eltrombopag olamine analysis of medium method, and comprehensive system from
Beginning raw material is particularly to finished product and carries out quality control to the key intermediate during synthesis, is ground for the quality of eltrombopag olamine
Studying carefully the improvement with synthesis technique has important directive significance.
According to eltrombopag olamine intermediate I and intermediate II synthetic route, it can be caused in preparation and storage
Impurity includes the accessory substances such as raw material, intermediate product and position isomer, demethylation product, dimer.To improve middle mistake
Process control, drug quality is effectively analyzed, ensure drug safety, it is necessary to develop among a kind of convenient, effective detection eltrombopag olamine
Analysis method of the body about material.
The content of the invention
First aspect present invention provide a kind of high-efficient liquid phase chromatogram technique analysis measure eltrombopag olamine intermediate I be 2'- hydroxyls-
The method of 3'- amino-phenylbenzene -3- carboxylic acid related impurities contents.
By the structural analysis to intermediate I and its known impurities, for compound molecular weight it is universal it is smaller, be provided with benzene
Situations such as ring structure, existence position isomery, first selection, are with steric hindrance characteristic and for aromatic compound with uniqueness
The phenyl silane bonded silica gel of retention be filler.In addition, according to compound functional group dissociate feature, use pH value for
3.0~4.0 phosphate buffer is mobile phase A, and phosphoric acid acetonitrile solution is Mobile phase B, changes stream during graded
The pH value of dynamic phase system, the impurity for making to be provided simultaneously with amino and carboxyl can show good chromatogram retention behavior and separation is imitated
Fruit.
Specifically, analysis method of the present invention can be realized as follows：
(1) sample preparation
Need testing solution：Take this product appropriate, add methanol to dissolve and dilute and be made in every 1ml methanol solutions containing about in 0.2mg
Mesosome I solution, as need testing solution.
System suitability solution：Take intermediate I, impurity A L, impurity B L, impurity CL, impurity SQ, impurity YW, impurity B D, miscellaneous
Matter LS, impurity QA are each appropriate, accurately weighed, dissolves and diluted with methanol and are made in every 1ml methanol solutions containing about intermediate I and respectively
Impurity is respectively 0.02mg mixed solution.The structure of each impurity is as shown in table 1：
The structure of the intermediate I related impurities of table 1
(2) chromatographic condition
Instrument：High performance liquid chromatograph-UV-detector
Chromatographic column：Use chromatographic column of the phenyl silane bonded silica gel for filler；Chromatographic column particle diameter is 5 μm；Column length
Spend for 250mm；Chromatogram column internal diameter is 4.6mm.
Mobile phase A：PH value be 3.0~4.0 phosphate buffer (preferable ph 3.2~3.8, most preferably 3.5)；Phosphoric acid
Salt buffer is the sodium dihydrogen phosphate or phosphoric acid that concentration is 5~15mmol/L (preferably 8~12mmol/L, most preferably 10mmol/L)
The disodium hydrogen aqueous solution, adjusted using the phosphoric acid solution (>=85%) or potassium hydroxide aqueous solution (1mol/L) of this area normal concentration
PH value.
Mobile phase B：The volume fraction of phosphoric acid acetonitrile solution, wherein phosphoric acid in acetonitrile is 0.05%~0.15%；It is preferred that
Volume fraction is 0.08%~0.12%, and most preferably volume fraction is 0.1%.
The preferable gradient elution program according to the form below of the present invention one is carried out：
Flow velocity：1.0~1.5ml/min, preferably 1.1~1.3ml/min, most preferably 1.2ml/min；
Column temperature：45 DEG C~55 DEG C, preferably 48 DEG C~52 DEG C, most preferably 50 DEG C；
Detection wavelength：230nm；
Sampling volume：10μl.
Preferable gradient elution program
| Time (minute) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 87 | 13 |
| 3 | 87 | 13 |
| 26 | 64 | 36 |
| 38 | 40 | 60 |
| 39 | 87 | 13 |
| 45 | 87 | 13 |
(3) calculating of impurity
Impurity peaks in need testing solution chromatogram, each known impurities in intermediate I are calculated according to by area normalization method
With the content of unknown impuritie.
It is 2- (3,4- that second aspect of the present invention, which provides a kind of high-efficient liquid phase chromatogram technique analysis measure eltrombopag olamine intermediate II,
Dimethvl-phenyl) -5- methyl -2,4- dihydro-pyrazol -3- ketone related impurities contents method.Specifically, it is of the invention above-mentioned
Method can be realized as follows：
(1) sample preparation
Need testing solution：Take this product appropriate, add methanol ultrasonic dissolution and dilute be made in every 1ml methanol solutions containing about
The solution of 0.5mg intermediate IIs, as need testing solution.
System suitability solution：Take intermediate II, impurity SM1, impurity F J, impurity DH each appropriate, it is accurately weighed, use methanol
Dissolve and dilute that to be made in every 1ml methanol solutions containing about intermediate I and each impurity be respectively 0.05mg mixed solution.Each impurity
Structure it is as shown in table 2：
The structure of the intermediate II related impurities of table 2
(2) chromatographic condition
Instrument：High performance liquid chromatograph-UV-detector
Chromatographic column：Use chromatographic column of the octadecylsilane chemically bonded silica for filler；Chromatographic column particle diameter is 3.5 μm；Color
Spectrum column length is 150mm；Chromatogram column internal diameter is 4.6mm.
Mobile phase A：PH value be 2.5~3.5 (preferably 2.8~3.2, phosphate buffer most preferably 3.0)；Phosphate delays
Fliud flushing is the sodium dihydrogen phosphate or phosphoric acid hydrogen that concentration is 15~25mmol/L (preferably 18~22mmol/L, most preferably 20mmol/L)
Two sodium water solutions, use the phosphoric acid solution or potassium hydroxide solution (such as 1mol/L) of this area normal concentration (such as >=85%)
Adjust pH value.
Mobile phase B：Acetonitrile
The preferable gradient elution program according to the form below of the present invention one is carried out：
Flow velocity：0.8~1.2ml/min, preferably 0.9~1.1ml/min, most preferably 1.0ml/min；
Column temperature：30 DEG C~40 DEG C, preferably 32 DEG C~38 DEG C, most preferably 35 DEG C；
Sample room temperature：4 DEG C~8 DEG C, preferably 4 DEG C~6 DEG C, most preferably 4 DEG C；
Detection wavelength：243nm；
Sampling volume：10μl.
Preferable gradient elution program
| Time (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 85 | 15 |
| 5 | 60 | 40 |
| 22 | 35 | 65 |
| 23 | 20 | 80 |
| 32 | 20 | 80 |
| 33 | 85 | 15 |
| 40 | 85 | 15 |
(3) calculating of impurity
Impurity peaks in need testing solution chromatogram are each known in intermediate II according to being calculated by area normalization method
The content of impurity and unknown impuritie.
In summary, the claimed analysis method of the present invention has following good effect：Described analysis method energy
Ai Qu pool pa intermediates and its related impurities system are enough efficiently separated, there is good specificity, high sensitivity, easy to operate, analysis
Many advantages, such as time is moderate, the theory (QBD) that design is come from according to quality have carried out standard from synthetic route upstream to impurity spectrum
Really monitoring, make follow-up impurity transfer law and process parameter optimizing research more convenient, be the quality control of eltrombopag olamine finished product
System provides effective guarantee.
Third aspect present invention provides a kind of new impurity reference substance compound DH, and provides one kind to prepare this contaminated
Compound DH method, this method comprise the following steps：
(1) SM1, ethyl acetoacetate, sodium hydrogensulfite are dissolved in ethanol, purified water, stirring is warming up to backflow, instead
Answer 2~4 hours；
(2) 35-45 DEG C is cooled to, is stirred 0.5~1 hour；
(3) 20~30 DEG C are cooled to, insulation crystallization 4~5 hours；
(4) filter, filtrate is evaporated, and through column chromatography for separation, obtains compound DH；
Wherein, SM1, ethyl acetoacetate and sodium hydrogensulfite mass ratio are 1 in step (1):0.8:1.2；SM1 with
The mass volume ratio of ethanol is 1:4, SM1 and purified water mass volume ratio be 1:4.
Brief description of the drawings
The system suitability solution collection of illustrative plates of analysis method described in Fig. 1 embodiments 1；
The system suitability solution collection of illustrative plates of analysis method described in Fig. 2 embodiments 2.
Embodiment
The present invention is further illustrated below by specific embodiment.It should be understood to：Embodiments of the invention are only
For illustrating the present invention, rather than limitation of the present invention.The simple of the present invention is changed on the basis of technical solution of the present invention
Enter or protection scope of the present invention is belonged to using the technical scheme obtained by customary means or composition progress equivalent substitution.
Reagent or raw material used in the present invention can be prepared by existing literature or prior art, or purchase
Obtain.
The intermediate I method validation of embodiment 1
1st, chromatographic condition
Instrument：Waters2695-2489-2998 high performance liquid chromatographs-UV-detector-PDAD
Chromatographic column：It is filler with phenyl silane bonded silica gel [Xbridge Phenyl (4.6 × 250mm, 5 μm)]
Mobile phase A：10mmol/L sodium dihydrogen phosphates (with phosphorus acid for adjusting pH value to 3.5)
Mobile phase B：0.1% phosphoric acid acetonitrile solution
Gradient elution program
| Time (minute) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 87 | 13 |
| 3 | 87 | 13 |
| 26 | 64 | 36 |
| 38 | 40 | 60 |
| 39 | 87 | 13 |
| 45 | 87 | 13 |
Flow velocity：1.2ml/min；Column temperature：50℃；Detection wavelength：230nm；Sampling volume：10μl
2nd, method validation
(1) system suitability
Impurity positions solution：Take intermediate I, impurity A L, impurity B L, impurity CL, impurity SQ, impurity YW, impurity B D, impurity
LS, impurity QA are each appropriate, accurately weighed, put respectively in different measuring bottles, are dissolved and diluted with methanol every 1ml methanol solutions are made
In containing about intermediate I and each impurity be respectively 0.2mg solution.
System suitability solution：It is each appropriate that precision measures above-mentioned impurity positioning solution, puts in same measuring bottle, adds methanol dilution
It is respectively 0.02mg mixed solution to be made in every 1ml methanol solutions containing intermediate I and each impurity.
Above-mentioned solution detects according to chromatographic condition described in the embodiment of the present invention, impurity positioning solution and system suitability solution
In each impurity retention time and separating degree be shown in Table 3, system suitability collection of illustrative plates is as shown in Figure 1.As a result show, each impurity and main peak
Between and each impurity peaks between separating degree be all higher than 1.5, the purity angle of main peak and each impurity peaks is respectively less than purity threshold value.Therefore,
System suitability meets the requirements.
The intermediate I system suitability result of table 3
(2) quantitative limit
The measure of baseline noise：Precision measures the μ l of methanol 10, injects liquid chromatograph, the pin of continuous sample introduction 3, according to the present invention
Chromatographic condition described in embodiment detects.Blank baseline noise in the range of intermediate I and each impurity appearance time is recorded, and is calculated flat
Average.
The preparation of quantitative limit solution：Take intermediate I, impurity A L, impurity B L, impurity CL, impurity SQ, impurity YW, impurity B D,
Impurity LS, impurity QA are each appropriate, accurately weighed, put in same measuring bottle, are dissolved with methanol and progressively diluted, and implement according to the present invention
The example chromatographic condition detection, until being diluted to ratio (i.e. signal to noise ratio, the S/ of intermediate I and each impurity peak height and corresponding noise
N untill being about) 10, now sample concentration and the ratio of test sample measure concentration are quantitative limit, with the concentration continuous sample introduction 6
Pin, calculate peak height average value.
Result of the test shows, under analysis method of the present invention, the quantitative limit of each impurity supplies below 0.05%
Each known impurities and unknown impuritie of the content more than 0.05% can be detected simultaneously accurate quantitative analysis in test product.Therefore, method
Sensitivity meet the requirements.
The intermediate I of table 4 and each impurity quantitative limit result
| Title | Concentration (μ g/ml) | Average peak height (μ V) | Average noise (μ V) | Signal to noise ratio | Quantitative limit (%) |
| Intermediate I | 0.0209 | 131.0 | 6.4 | 10.6 | 0.01 |
| Impurity QA | 0.1004 | 451.3 | 4.1 | 14.1 | 0.05 |
| Impurity B D | 0.0407 | 127.3 | 3.5 | 12.3 | 0.02 |
| Impurity A L | 0.0205 | 231.7 | 2.0 | 11.8 | 0.01 |
| Impurity LS | 0.0208 | 294.3 | 0.6 | 11.9 | 0.01 |
| Impurity SQ | 0.0214 | 147.8 | 2.3 | 10.8 | 0.01 |
| Impurity YW | 0.0212 | 95.2 | 3.7 | 11.0 | 0.01 |
| Impurity B L | 0.0207 | 172.5 | 0.9 | 12.6 | 0.01 |
| Impurity CL | 0.0211 | 192.2 | 1.5 | 13.7 | 0.01 |
(3) stability of solution
Need testing solution：Take intermediate I appropriate, it is accurately weighed, dissolved and diluted with methanol and be made in every 1ml containing about centre
The 0.2mg of body I solution.
Above-mentioned solution is placed under room temperature condition, respectively at 0 hour, 1 hour, 2 hours, 5 hours, 8 hours, 13 hours,
Precision measures 10 μ l injection liquid chromatographs, is detected according to chromatographic condition described in the embodiment of the present invention.Calculate known impurities, unknown
The rate of change of impurity and total impurities, the results are shown in Table 5.
Result of the test shows that at ambient temperature, impurity YW and total impurities have faint increase to become to intermediate I measure solution
Gesture, total impurities rate of change is 0.02% in 5 hours, and general less than pharmacopoeia of each country standard and ICH acceptable ignores threshold value
0.05%, thus intermediate I solution is stable in 5 hours at ambient temperature, ie in solution should be surveyed after preparing in 5 hours
It is fixed.
The intermediate I stability of solution result of table 5
Note：ND：It is not detected by.
The intermediate II method validation of embodiment 2
1st, chromatographic condition
Instrument：Waters2695-2489-2998 high performance liquid chromatographs-UV-detector-PDAD
Chromatographic column：With octadecylsilane chemically bonded silica [Xbridge Shield RP-18 (4.6 × 150mm, 3.5 μm)]
For filler
Mobile phase A：20mmol/L sodium dihydrogen phosphates (with phosphorus acid for adjusting pH value to 3.0)
Mobile phase B：Acetonitrile
Gradient elution program
| Time (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 85 | 15 |
| 5 | 60 | 40 |
| 22 | 35 | 65 |
| 23 | 20 | 80 |
| 32 | 20 | 80 |
| 33 | 85 | 15 |
| 40 | 85 | 15 |
Flow velocity：1.0ml/min；Column temperature：35℃；Sample room temperature：4℃；Detection wavelength：243nm；Sampling volume：10μl
2nd, method validation
(1) system suitability
Impurity positions solution：Take intermediate II, impurity SM1, impurity F J, impurity DH each appropriate, it is accurately weighed, put respectively not
With measuring bottle in, dissolved with methanol and dilute that to be made in every 1ml methanol solutions containing about intermediate II and each impurity be respectively 0.5mg
Solution.
System suitability solution：It is each appropriate that precision measures above-mentioned impurity positioning solution, puts in same measuring bottle, adds methanol dilution
It is respectively 0.05mg mixed solution to be made in every 1ml methanol solutions containing intermediate II and each impurity.
Above-mentioned solution detects according to chromatographic condition described in the embodiment of the present invention, impurity positioning solution and system suitability solution
In each impurity retention time and separating degree be shown in Table 6, system suitability collection of illustrative plates is as shown in Figure 2.As a result show, each impurity and main peak
Between and each impurity peaks between separating degree be all higher than 1.5, the purity angle of main peak and each impurity peaks is respectively less than purity threshold value.Therefore,
System suitability meets the requirements.
The intermediate II system suitability result of table 6
(2) quantitative limit
The measure of baseline noise：Precision measures the μ l of methanol 10, injects liquid chromatograph, the pin of continuous sample introduction 3, according to the present invention
Chromatographic condition described in embodiment detects.Blank baseline noise in the range of intermediate II and each impurity appearance time is recorded, and is calculated
Average value.
The preparation of quantitative limit solution：Take intermediate II, impurity SM1, impurity F J, impurity DH each appropriate, it is accurately weighed, put same
In one measuring bottle, dissolved with methanol and progressively diluted, detected according to chromatographic condition described in the embodiment of the present invention, until being diluted to centre
Untill the ratio (i.e. signal to noise ratio, S/N) of body I and each impurity peak height and corresponding noise is about 10, now sample concentration and test sample
The ratio for determining concentration is quantitative limit, with the pin of concentration continuous sample introduction 6, calculates peak height average value.
Result of the test shows, under analysis method of the present invention, the quantitative limit of each impurity supplies below 0.02%
Each known impurities of content more than 0.02% and unknown impuritie can be detected simultaneously accurate quantitative analysis in test product.Therefore, method
Sensitivity meets the requirements.
The intermediate II of table 7 and each impurity quantitative limit result
| Title | Concentration (μ g/ml) | Average peak height (μ V) | Average noise (μ V) | Signal to noise ratio | Quantitative limit (%) |
| Intermediate II | 0.1048 | 148.0 | 14.7 | 10.1 | 0.02 |
| Impurity SM1 | 0.1019 | 76.8 | 6.3 | 12.1 | 0.02 |
| Impurity F J | 0.0514 | 188.3 | 15.0 | 12.6 | 0.01 |
| Impurity DH | 0.0511 | 249.3 | 21.0 | 11.9 | 0.01 |
(3) stability of solution
Need testing solution：Take intermediate II appropriate, it is accurately weighed, dissolved and diluted with methanol every 1ml methanol solutions are made
In solution containing about intermediate I 0.5mg.
Above-mentioned solution is under the conditions of 4 DEG C, respectively at 0 hour, 1 hour, 2 hours, 5 hours, 8 hours, 13 hours, precision amount
Take 10 μ l to inject liquid chromatograph, detected according to chromatographic condition described in the embodiment of the present invention.Calculate known impurities, unknown impuritie and
The rate of change of total impurities, the results are shown in Table 8.
Result of the test shows that intermediate II determines solution under the conditions of 4 DEG C, places 12 hours, total impurities rate of change is less than
0.02%, less than pharmacopoeia of each country standard and ICH it is general it is acceptable ignore threshold value 0.05%, thus intermediate II solution is 4
It is stable in 12 hours under the conditions of DEG C.
Table 8 intermediate II solution, 4 DEG C of stability test results
| Time/h | SM1% | FJ% | DH% | Other single impurity % | Total impurities % | Total impurities rate of change % |
| 0 | ND | ND | 0.17 | 0.02 | 0.30 | \ |
| 2 | ND | ND | 0.17 | 0.02 | 0.29 | -0.01 |
| 4 | ND | ND | 0.17 | 0.02 | 0.32 | 0.02 |
| 8 | ND | ND | 0.17 | 0.02 | 0.30 | 0.00 |
| 12 | ND | ND | 0.17 | 0.02 | 0.31 | 0.01 |
Note：ND：It is not detected by.
The impurity compound DH of embodiment 3 preparation
By 3.0g compound SM1,2.4g compound acetyls ethyl acetate, 3.6g sodium hydrogensulfites be dissolved in 12mL ethanol,
In 12ml purified waters, stirring is warming up to backflow, reacts 3 hours；Reaction finishes, and is cooled to 40 DEG C, stirs 1 hour；It is cooled to 25
DEG C, insulation crystallization 5 hours；Filter, filtrate is evaporated, and through column chromatography for separation, obtains impurity DH 1.1g.
ESI(+)：m/z 231.20；
[M+1]+；1H NMR：(400MHz, DMSO-d6) δ=7.40 (d, 1H), 7.39 (m, 1H), 7.13 (d, 1H), 5.63
(s, 1H), 4.10 (d, 2H), 2.22 (s, 3H), 2.19 (s, 3H), 2.12 (s, 3H), 1.31 (m, 3H).
Claims (10)
1. a kind of method of the related impurities content of high-efficient liquid phase chromatogram technique analysis measure eltrombopag olamine intermediate compound I, the centre
Body I is 2'- hydroxyls -3'- amino-phenylbenzene -3- carboxylic acids, it is characterised in that uses phenyl silane bonded silica gel as filler
Chromatographic column, using phosphate buffer as mobile phase A, phosphoric acid acetonitrile solution is Mobile phase B, using gradient elution mode.
2. according to the method for claim 1, it is characterised in that the particle diameter of the chromatographic column is 5 μm, length 250mm, interior
Footpath is 4.6mm.
3. according to the method for claim 1, it is characterised in that the pH value as the phosphate buffer of mobile phase A
For 3.0~4.0, preferably 3.2~3.8, most preferably 3.5；As in the phosphoric acid acetonitrile solution of Mobile phase B, phosphoric acid is in acetonitrile
In volume fraction be 0.05%~0.15%, preferably 0.08%~0.12%, most preferably 0.1%.
4. according to the method for claim 1, it is characterised in that gradient is：
5. according to the method for claim 1, it is characterised in that the flow velocity of mobile phase is 1.0~1.5ml/min, preferably 1.1
~1.3ml/min, most preferably 1.2ml/min；The column temperature of chromatographic column is 45 DEG C~55 DEG C, preferably 48 DEG C~52 DEG C, most preferably 50
℃；Detection wavelength is 230nm；Sampling volume is 10 μ l.
6. a kind of method of the related impurities content of high-efficient liquid phase chromatogram technique analysis measure eltrombopag olamine intermediate II, the centre
Body II is 2- (3,4- Dimethvl-phenyl) -5- methyl -2,4- dihydro-pyrazol -3- ketone, it is characterised in that using octadecyl silicon
Alkane bonded silica gel is the chromatographic column of filler, and using phosphate buffer as mobile phase A, acetonitrile is Mobile phase B, using gradient elution
Mode；The particle diameter of wherein described chromatographic column is 3.5 μm, length 150mm, internal diameter 4.6mm.
7. according to the method for claim 6, it is characterised in that the pH value as the phosphate buffer of mobile phase A
For 2.5~3.5, preferably 2.8~3.2, most preferably 3.0.
8. according to the method for claim 6, it is characterised in that gradient is：
9. according to the method for claim 6, it is characterised in that the flow velocity of mobile phase is 0.8~1.2ml/min, preferably 0.9
~1.1ml/min, most preferably 1.0ml/min；The column temperature of chromatographic column is 30 DEG C~40 DEG C, preferably 32 DEG C~38 DEG C, most preferably 35
℃；Sample room temperature is 4 DEG C~8 DEG C, preferably 4 DEG C~6 DEG C, most preferably 4 DEG C；Detection wavelength is 243nm；Sampling volume is 10 μ
l。
10. a kind of impurity reference substance DH for claim 6 methods described, it has following structure：
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| CN201611263699.6A Active CN107870217B (en) | 2016-09-26 | 2016-12-30 | Detection method of eltrombopag intermediate I related substances |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113358804A (en) * | 2020-03-04 | 2021-09-07 | 齐鲁制药有限公司 | Ion chromatography analysis method for determining genotoxic impurity nitrite in eltrombopag ethanolamine |
| CN115166125A (en) * | 2022-08-16 | 2022-10-11 | 郑州大学第一附属医院 | Method for rapidly determining concentration of eltrombopag in human plasma by adopting ultra-high performance liquid chromatography-tandem mass spectrometry |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100256212A1 (en) * | 2009-04-01 | 2010-10-07 | Pliva Hrvatska D.O.O. | Polymorphs of eltrombopag and eltrombopag salts and processes for preparation thereof |
| US20120164102A1 (en) * | 2009-06-11 | 2012-06-28 | Peng Cho Tang | Salts of bicyclo-substituted pyrazolon azo derivatives, preparation method and use thereof |
| WO2013049605A1 (en) * | 2011-09-28 | 2013-04-04 | Assia Chemical Industries Ltd. | Processes for the preparation of an intermediate in the synthesis of eltrombopag |
| CN103360317A (en) * | 2012-04-11 | 2013-10-23 | 齐鲁制药有限公司 | Dicyclo-substituted pyrazolone azo derivatives, as well as preparation method and use thereof |
| CN104592033A (en) * | 2014-12-30 | 2015-05-06 | 杭州和泽医药科技有限公司 | Synthetic method of promacta key intermediate |
-
2016
- 2016-12-30 CN CN201611263699.6A patent/CN107870217B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100256212A1 (en) * | 2009-04-01 | 2010-10-07 | Pliva Hrvatska D.O.O. | Polymorphs of eltrombopag and eltrombopag salts and processes for preparation thereof |
| US20120164102A1 (en) * | 2009-06-11 | 2012-06-28 | Peng Cho Tang | Salts of bicyclo-substituted pyrazolon azo derivatives, preparation method and use thereof |
| WO2013049605A1 (en) * | 2011-09-28 | 2013-04-04 | Assia Chemical Industries Ltd. | Processes for the preparation of an intermediate in the synthesis of eltrombopag |
| CN103360317A (en) * | 2012-04-11 | 2013-10-23 | 齐鲁制药有限公司 | Dicyclo-substituted pyrazolone azo derivatives, as well as preparation method and use thereof |
| CN104592033A (en) * | 2014-12-30 | 2015-05-06 | 杭州和泽医药科技有限公司 | Synthetic method of promacta key intermediate |
Non-Patent Citations (5)
| Title |
|---|
| D.MAHABOOB BASHA 等: "RP-UPLC-MS assay method for eltrombopag：application in pharmaceuticals，human plasma and urine samples", 《ASIAN JOURNAL OF CHEMISTRY》 * |
| DENG YANLI 等: "Metabolism and Disposition of Eltrombopag, an Oral, Nonpeptide Thrombopoietin Receptor Agonist, in Healthy Human Subjects", 《DRUG METABOLISM AND DISPOSITION》 * |
| RAMBABU MADDELA 等: "Liquid chromatography–tandem mass spectrometric assay for eltrombopag in 50 L of human plasma: A pharmacokinetic study", 《JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS》 * |
| 吕春华 等: "高效液相色谱法测定氧化型染发剂中11种染料", 《理化检验(化学分册)》 * |
| 孙宝佳 等: "艾曲波帕", 《中国药物化学杂志》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113358804A (en) * | 2020-03-04 | 2021-09-07 | 齐鲁制药有限公司 | Ion chromatography analysis method for determining genotoxic impurity nitrite in eltrombopag ethanolamine |
| CN115166125A (en) * | 2022-08-16 | 2022-10-11 | 郑州大学第一附属医院 | Method for rapidly determining concentration of eltrombopag in human plasma by adopting ultra-high performance liquid chromatography-tandem mass spectrometry |
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| CN107870217B (en) | 2022-03-04 |
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