CN105699524A - Detection method for content of isomer impurities in Ticagrelor - Google Patents
Detection method for content of isomer impurities in Ticagrelor Download PDFInfo
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Abstract
The invention discloses a detection method for content of isomer impurities in Ticagrelor. The detection method comprises the following steps: a) preparing a reference substance solution; b) preparing a test sample solution; c) respectively detecting the reference substance solution and the test sample solution by adopting a high performance liquid chromatography method; d) calculating by peak area according to an external reference method or calculating according to a standard curve between the peak area and the concentration, thereby acquiring the content of isomer impurities in Ticagrelor. According to the new detection method for content of isomer impurities in Ticagrelor provided by the invention, the degree of separation between chromatographic peaks is high, the chromatographic peaks are not interfered to each other, the accurate detection for impurity A, impurity B and impurity C can be realized at the same time, an effective detection method is supplied for monitoring the content of isomer impurities in Ticagrelor, and the product quality of Ticagrelor and the medication safety of the patient can be further ensured.
Description
Technical field
The invention belongs to the analysis detection field in chemistry, be specifically related in a kind of ticagrelor the detection method of isomer impurities content。
Background technology
Ticagrelor (Ticagrelor trade name: BRILINTA), chemistry (1S by name, 2S, 3R, 5S)-3-[7-[[(1R, 2S)-2-(3,4-difluorophenyl) cyclopropyl] amino]-5-rosickyite base-3H-1,2,3-triazolyl [4,5-d] pyrimidin-3-yl]-5-(2-hydroxy ethoxy)-1,2-Pentamethylene. glycol (molecular formula: C23H28F2N6O4S), it is a kind of selective adenosine diphosphate (ADP) (ADP) receptor antagonist developed by AstraZeneca pharmaceutical Co. Ltd (AstraZenecaAB), by activating P2Y12-receptor, the platelet activation of reversible retardance ADP mediation and gathering。
Due to by production technology and the restriction controlling level, produce in the ticagrelor product of gained, often have plurality of impurities, such as, (1R, 2R, 3S, 5R)-3-[7-{ [(1R, 2S)-2-(3, 4-difluorophenyl) cyclopropyl] amino }-5-rosickyite base-3H-1, 2, 3-triazole [4, 5-d] pyrimidin-3-yl]-5-(2-hydroxy ethoxy) Pentamethylene .-1, 2-glycol, (1R, 2R, 3S, 5R)-3-[7-{ [(1S, 2R)-2-(3, 4-difluorophenyl) cyclopropyl] amino }-5-rosickyite base-3H-1, 2, 3-triazole [4, 5-d] pyrimidin-3-yl]-5-(2-hydroxy ethoxy) Pentamethylene .-1, 2-glycol, (1S, 2S, 3R, 5S)-3-[7-{ [(1S, 2R)-2-(3, 4-difluorophenyl) cyclopropyl] amino }-5-rosickyite base-3H-1, 2, 3-triazole [4, 5-d] pyrimidin-3-yl]-5-(2-hydroxy ethoxy) Pentamethylene .-1, 2-glycol etc. (referring to: Ticagrelor impurity list,Http:// wenku.***.com/view/69885bd8fd0a79563c1e72aa.html？Re=view, the time of disclosure is on October 17th, 2014), have a strong impact on the product quality of ticagrelor and the drug safety of patient。
But, for detection ticagrelor, only have one section of patent documentation CN104634887A at present and have detected a kind of impurity content in ticagrelor: (1S, 2S, 3R, 5S)-3-(7-((1S, 2R)-2-(3,4-difluorophenyl) cyclopropyl amino)-5-(rosickyite base)-3H-[1,2,3] triazole [4,5-d] pyrimidin-3-yl)-5-(2-hydroxyl-oxethyl) Pentamethylene .-1,2-glycol, is difficult to reflect all sidedly the product quality of ticagrelor。
In order to impurity more in ticagrelor is detected, meanwhile, also for the product quality more fully monitoring ticagrelor, it is further ensured that the drug safety of patient, it is necessary to invent a kind of new detection method。
Summary of the invention
It is an object of the invention to provide the detection method of isomer impurities content in a kind of ticagrelor。
The detection method of isomer impurities content in a kind of ticagrelor provided by the invention, it comprises the following steps:
A, take the reference substance of isomer impurities, prepare reference substance solution；
B, take ticagrelor sample to be checked, prepare need testing solution；
Reference substance solution, need testing solution are detected by c, employing efficient liquid-phase chromatography method respectively, and the testing conditions of described efficient liquid-phase chromatography method is:
Chromatographic column: fixing is amylose-three (3,5-xylyl carbamate) mutually；
The volume ratio of mobile phase: normal hexane-alcoholic solution, normal hexane and ethanol is 88%:12%～92%:8%, contains the diethylamine of 0.1%～0.5%ml/ml in described ethanol；
Detection wavelength: 200nm～300nm；
D, by external standard method with calculated by peak area, or calculate according to the standard curve between peak area and concentration, obtain the isomer impurities content in ticagrelor sample。
Further, described isomer impurities at least includes (1R, 2R, 3S, 5R)-3-[7-{ [(1R, 2S)-2-(3,4-difluorophenyl) cyclopropyl] amino }-5-rosickyite base-3H-1,2,3-triazole [4,5-d] pyrimidin-3-yl]-5-(2-hydroxy ethoxy) Pentamethylene .-1,2-glycol。
Further, described isomer impurities at least includes (1R, 2R, 3S, 5R)-3-[7-{ [(1R, 2S)-2-(3,4-difluorophenyl) cyclopropyl] amino }-5-rosickyite base-3H-1,2,3-triazole [4,5-d] pyrimidin-3-yl]-5-(2-hydroxy ethoxy) Pentamethylene .-1,2-glycol and (1R, 2R, 3S, 5R)-3-[7-{ [(1S, 2R)-2-(3,4-difluorophenyl) cyclopropyl] amino }-5-rosickyite base-3H-1,2,3-triazole [4,5-d] pyrimidin-3-yl]-5-(2-hydroxy ethoxy) Pentamethylene .-1,2-glycol；
Preferably, described isomer impurities includes (1R simultaneously, 2R, 3S, 5R)-3-[7-{ [(1R, 2S)-2-(3, 4-difluorophenyl) cyclopropyl] amino }-5-rosickyite base-3H-1, 2, 3-triazole [4, 5-d] pyrimidin-3-yl]-5-(2-hydroxy ethoxy) Pentamethylene .-1, 2-glycol, (1R, 2R, 3S, 5R)-3-[7-{ [(1S, 2R)-2-(3, 4-difluorophenyl) cyclopropyl] amino }-5-rosickyite base-3H-1, 2, 3-triazole [4, 5-d] pyrimidin-3-yl]-5-(2-hydroxy ethoxy) Pentamethylene .-1, 2-glycol and (1S, 2S, 3R, 5S)-3-[7-{ [(1S, 2R)-2-(3, 4-difluorophenyl) cyclopropyl] amino }-5-rosickyite base-3H-1, 2, 3-triazole [4, 5-d] pyrimidin-3-yl]-5-(2-hydroxy ethoxy) Pentamethylene .-1, 2-glycol。
Further, in step a, the solvent of preparation reference substance solution is the mobile phase in step c；In step b, the solvent preparing need testing solution is the mobile phase in step c。
Further, in step c, described chromatographic column isAD-H, length is 250mm, and internal diameter is 4.6mm, and the particle diameter of filler is 5 μm。
Further, in step c, the column temperature of described chromatographic column is 25 DEG C～35 DEG C。
Further, in step c, the volume ratio of normal hexane and ethanol is 90%:10%；Described ethanol contains the diethylamine of 0.2%ml/ml。
Further, in step c, the flow velocity of described mobile phase is 0.9ml/min～1.1ml/min；Preferably, the flow velocity of described mobile phase is 1.0ml/min。
Further, in step c, described detection wavelength is 230nm～250nm；Preferably, described detection wavelength is 242nm。
Further, in step c, sampling volume is 10 μ l～50 μ l；Preferably, sampling volume is 20 μ l。
The invention provides the new detection method of isomer impurities content in a kind of ticagrelor, separating degree between each chromatographic peak is high, noiseless each other, can realize impurity B and impurity A simultaneously, the accurate detection of impurity C, and, easy and simple to handle, it is easily controlled, testing cost is low, and there is good linear relationship, specificity, precision, stability, sensitivity and repeatability, average recovery is high, testing result is accurate, reliably, a kind of effective detection method is provided for the content of isomer impurities in monitoring ticagrelor medicine, further ensure the product quality of ticagrelor and the drug safety of patient。
Obviously, the foregoing according to the present invention, according to ordinary technical knowledge and the customary means of this area, without departing under the above-mentioned basic fundamental thought premise of the present invention, it is also possible to make the amendment of other various ways, replacement or change。
The detailed description of the invention of form by the following examples, is described in further detail the foregoing of the present invention again。But this should not being interpreted as, the scope of the above-mentioned theme of the present invention is only limitted to Examples below。All technology realized based on foregoing of the present invention belong to the scope of the present invention。
Accompanying drawing explanation
Fig. 1 is the HPLC figure of impurity A reference substance solution under testing conditions of the present invention。
Fig. 2 is the HPLC figure of impurity B reference substance solution under testing conditions of the present invention。
Fig. 3 is the HPLC figure of impurity C reference substance solution under testing conditions of the present invention。
Fig. 4 is the HPLC figure of impurity A under testing conditions of the present invention, impurity B, impurity C mixed solution。
Fig. 5 is the HPLC figure of ticagrelor need testing solution under testing conditions of the present invention。
Fig. 6 is the HPLC figure of ticagrelor and impurity A under testing conditions of the present invention, impurity B, impurity C mixed solution。
Fig. 7 is the HPLC figure of impurity A reference substance solution under contrast test 1 chromatographic condition。
Fig. 8 is the HPLC figure of impurity B reference substance solution under contrast test 1 chromatographic condition。
Fig. 9 is the HPLC figure of impurity C reference substance solution under contrast test 1 chromatographic condition。
Figure 10 is the HPLC figure of ticagrelor need testing solution under contrast test 1 chromatographic condition
Figure 11 is the HPLC figure of impurity A reference substance solution under contrast test 2 chromatographic condition。
Figure 12 is the HPLC figure of impurity B reference substance solution under contrast test 2 chromatographic condition。
Figure 13 is the HPLC figure of impurity C reference substance solution under contrast test 2 chromatographic condition。
Figure 14 is the HPLC figure of ticagrelor need testing solution under contrast test 2 chromatographic condition
Figure 15 is the HPLC figure of poly-doped impurity solution under contrast test 2 chromatographic condition
Figure 16 is the HPLC figure of impurity A reference substance solution under contrast test 3 chromatographic condition。
Figure 17 is the HPLC figure of impurity B reference substance solution under contrast test 3 chromatographic condition。
Figure 18 is the HPLC figure of impurity C reference substance solution under contrast test 3 chromatographic condition。
Figure 19 is the HPLC figure of ticagrelor need testing solution under contrast test 3 chromatographic condition。
Figure 20 is the HPLC figure of poly-doped impurity solution under contrast test 3 chromatographic condition。
Figure 21 is the mixed solution HPLC figure of ticagrelor and impurity under contrast test 3 chromatographic condition。
Figure 22 is the ultraviolet spectrogram of the 1st lower ticagrelor of test example 1。
Figure 23 is the canonical plotting of the 2nd lower impurity A reference substance of test example 1, and vertical coordinate is peak area, and abscissa is concentration。
Figure 24 is the canonical plotting of the 2nd lower impurity B reference substance of test example 1, and vertical coordinate is peak area, and abscissa is concentration。
Figure 25 is the canonical plotting of the lower impurity C reference substance of test example 1 the 2nd, and vertical coordinate is peak area, and abscissa is concentration。
Detailed description of the invention
The raw material, the equipment that use in the specific embodiment of the invention are known product, obtain by buying commercially available prod。
Embodiment 1,
The raw material, the equipment that use in the specific embodiment of the invention are known product, obtain by buying commercially available prod；
Such as, the lot number of ticagrelor is 20140801；Impurity A lot number is 20140213, content: 99.6%；Impurity B lot number is 20140227, content: 98.4%；Impurity C lot number is 20140307, content: 99.1%；Derive from Chengdu Baiyu Pharmaceutical Technology Co., Ltd.。
The name of impurity A is called (1R, 2R, 3S, 5R)-3-[7-{ [(1S, 2R)-2-(3,4-difluorophenyl) cyclopropyl] amino }-5-rosickyite base-3H-1,2,3-triazole [4,5-d] pyrimidin-3-yl]-5-(2-hydroxy ethoxy) Pentamethylene .-1,2-glycol。
The name of impurity B is called (1R, 2R, 3S, 5R)-3-[7-{ [(1R, 2S)-2-(3,4-difluorophenyl) cyclopropyl] amino }-5-rosickyite base-3H-1,2,3-triazole [4,5-d] pyrimidin-3-yl]-5-(2-hydroxy ethoxy) Pentamethylene .-1,2-glycol。
The name of impurity C is called (1S, 2S, 3R, 5S)-3-[7-{ [(1S, 2R)-2-(3,4-difluorophenyl) cyclopropyl] amino }-5-rosickyite base-3H-1,2,3-triazole [4,5-d] pyrimidin-3-yl]-5-(2-hydroxy ethoxy) Pentamethylene .-1,2-glycol。
AUW220D type precision electronic balance is purchased from Shimadzu Corporation；LC-20AT type efficient liquid-phase chromatographic pump is purchased from Shimadzu Corporation, LC-20AT type efficient liquid-phase chromatographic pump is purchased from Shimadzu Corporation, SPD-M20ADAD detector is purchased from Shimadzu Corporation, and SIL-20A automatic sampler is purchased from Shimadzu Corporation, and LcSolution work station is purchased from Shimadzu Corporation；Waters1525 type efficient liquid-phase chromatographic pump is purchased from water generation company,AD-H (250mm × 4.6mm, 5 μm) chromatographic column is purchased from Daicel company；AgilentZorbaxC18 (250mm × 4.6mm, 3.5 μm) is purchased from Agilent company limited。
Embodiment 1, the present invention detect the efficient liquid-phase chromatography method of chiral isomer impurity in ticagrelor
Chromatographic column:AD-H, 4.6mm × 250mm, 5 μm；
Mobile phase: normal hexane-ethanol (90:10), adds the diethylamine of 0.1%～0.5%ml/ml in ethanol
Solvent: mobile phase
Column temperature: 30 DEG C；Flow velocity: 1.0ml/min；UV detector (detection wavelength 242nm)。
Sampling volume: 20 μ L。
Detecting step:
Take impurity A, B and C reference substance in right amount, dissolve with solvent, be configured to the reference substance solution of the impure each 5 μ g of every 1mL。
Taking ticagrelor sample appropriate, after dissolving with ethanol, solubilizer dilutes, and is configured to every 1mL need testing solution containing about 0.5mg。
Take ticagrelor appropriate, after dissolving with ethanol, add impurity A, B and C reference substance solution, with solvent dilution, be configured to mixed solution。
Algoscopy: taking each 20 μ L of above-mentioned solution and inject chromatograph of liquid, record chromatogram, result is Fig. 1～Fig. 6 such as。
Fig. 1 is the HPLC figure of impurity A reference substance solution, and the retention time of impurity A is 65.553min。
Fig. 2 is the HPLC figure of impurity B reference substance solution, and the retention time of impurity B is 42.595min。
Fig. 3 is the HPLC figure of impurity C reference substance solution, and the retention time of impurity C is 53.713min。
Fig. 4 is the HPLC figure of poly-doped impurity reference substance solution, and the retention time of impurity A is 70.675min, and the retention time of impurity B is 45.761min, and the retention time of impurity C is 58.058min。
Fig. 5 is the HPLC figure of need testing solution, and the retention time of ticagrelor main peak is 40.045min。
Fig. 6 is the HPLC figure of test sample and impurity A, impurity B and impurity C mixed solution, the retention time of ticagrelor is 39.658min, the retention time of impurity B is 45.346min, the retention time of impurity C is 57.591min, the retention time of impurity A is 70.185min, ticagrelor and impurity B, and the separating degree between impurity respectively 2.688,5.153,4.472。
It is shown that under the chromatographic condition of the present invention, the separating degree between ticagrelor and impurity is high, it is possible to achieve ticagrelor detects while multiple chiral impurity content。
Contrast test 1:
Chromatographic column: AgilentZorbaxC184.6mm × 250mm, 3.5 μm；
Mobile phase A: 1. phosphate buffer (takes 1.0mol/L sodium dihydrogen phosphate (adjusting pH value to 3.0 with phosphoric acid) 10ml, add water to 900ml)-acetonitrile (90:10)；
Mobile phase B: 2. phosphate buffer (takes 1.0mol/L sodium dihydrogen phosphate (adjusting pH value to 3.0 with phosphoric acid) 10ml, add water to 300ml)-acetonitrile (30:70)；
Solvent: acetonitrile-water (50:50)
Column temperature: 35 DEG C；Flow velocity: 1.0ml/min；UV detector (detection wavelength 242nm)。
Condition of gradient elution is as follows:
Take impurity A reference substance appropriate, dissolve with solvent, be configured to every 1mL impurity A reference substance solution containing about 0.5mg。
Taking impurity B appropriate, solubilizer is configured to every 1mL impurity B reference substance solution containing about 0.5mg。
Taking impurity C appropriate, solubilizer is configured to every 1mL impurity C reference substance solution containing about 0.5mg。
Taking ticagrelor appropriate, solubilizer is configured to every 1mL need testing solution containing about ticagrelor 0.5mg。
Algoscopy: taking above-mentioned solution 20 μ L and inject chromatograph of liquid, record chromatogram, result is such as shown in Fig. 7～10。
Fig. 7 is the HPLC figure of impurity A reference substance solution under contrast test 1 chromatographic condition, and impurity A retention time is 29.493min。
Fig. 8 is HPLC figure, the retention time 29.450min of impurity B of impurity B reference substance solution under contrast test 1 chromatographic condition。
Fig. 9 is the HPLC figure of impurity C reference substance solution under contrast test 1 chromatographic condition, and the retention time of impurity C is 29.461min。
Figure 10 is HPLC figure, the retention time 29.457min of ticagrelor of need testing solution under contrast test 1 chromatographic condition。
It is shown that impurity A, impurity B, impurity C and ticagrelor retention time are almost without difference, the method can not carry out impurity A, impurity B and impurity C separation detection completely。
Contrast test 2:
Chromatographic column:OD-H, 4.6mm × 250mm, 5 μm；
Mobile phase: normal hexane-ethanol (70:30)
Solvent: mobile phase
Column temperature: 30 DEG C；Flow velocity: 1.0ml/min；UV detector (detection wavelength 242nm)。
Sampling volume: 20 μ L。
Take impurity A reference substance appropriate, dissolve with solvent, be configured to every 1mL impurity A reference substance solution containing about 0.5 μ g。
Take impurity B reference substance appropriate, dissolve with solvent, be configured to every 1mL impurity B reference substance solution containing about 10 μ g。
Take impurity C reference substance appropriate, dissolve with solvent, be configured to every 1mL impurity C reference substance solution containing about 20 μ g。
Take ticagrelor sample appropriate, dissolve with solvent, be configured to every 1mL need testing solution containing about 20 μ g。
Taking ticagrelor and impurity A, impurity B, impurity C reference substance solution appropriate, solubilizer is diluted to every 1ml mixed solution containing each composition 0.5 μ g。
Algoscopy: taking above-mentioned solution 20 μ L and inject chromatograph of liquid, record chromatogram, result is such as shown in Figure 11～17。
Figure 11 is the HPLC figure of impurity A reference substance solution under contrast test 2 chromatographic condition, and the retention time of impurity A is 12.500min。
Figure 12 is the HPLC figure of impurity B reference substance solution under contrast test 2 chromatographic condition, and the retention time of impurity B is 9.796min。
Figure 13 is the HPLC figure of impurity C reference substance solution under contrast test 2 chromatographic condition, and the retention time of impurity C is 11.189min。
Figure 14 is the HPLC figure of ticagrelor need testing solution under contrast test 2 chromatographic condition, and ticagrelor peak retention time is 9.415min。
Figure 15 is the HPLC figure of mixed solution under contrast test 2 chromatographic condition, impurity C retention time is 11.224min, and impurity A retention time is 12.519min, and separating degree is better, but the chromatographic peak of ticagrelor and impurity B is almost completely superposed, it is impossible to accurately checked for impurities B。
Contrast test 3:
Chromatographic column:AD-H, 4.6mm × 250mm, 5 μm；
Mobile phase: normal hexane-ethanol (85:15)
Solvent: mobile phase
Column temperature: 30 DEG C；Flow velocity: 1.0ml/min；UV detector (detection wavelength 242nm)。
Sampling volume: 20 μ L。
Take impurity A reference substance appropriate, dissolve with solvent, be configured to every 1mL impurity A reference substance solution containing about 25 μ g。
Take impurity B reference substance appropriate, dissolve with solvent, be configured to every 1mL impurity B reference substance solution containing about 50 μ g。
Take impurity C reference substance appropriate, dissolve with solvent, be configured to every 1mL impurity C reference substance solution containing about 5 μ g。
Taking impurity A, impurity B, impurity C reference substance solution in right amount, solubilizer is diluted to every 1ml poly-doped impurity solution containing each impurity 0.5 μ g。
Take ticagrelor sample appropriate, dissolve with solvent, be configured to every 1mL need testing solution containing about 0.5mg。
Taking ticagrelor and impurity A, impurity B, impurity C reference substance solution appropriate, solubilizer is diluted to every 1ml containing each impurity 0.5 μ g, the mixed solution of ticagrelor 0.5mg。
Algoscopy: taking above-mentioned solution 20 μ L and inject chromatograph of liquid, record chromatogram, result is such as shown in Figure 16～21。
Figure 16 is the HPLC figure of impurity A reference substance solution under contrast test 3 chromatographic condition, and the retention time of impurity A is 32.182min。
Figure 17 is the HPLC figure of impurity B reference substance solution under contrast test 3 chromatographic condition, and the retention time of impurity B is 21.799min。
Figure 18 is the HPLC figure of impurity C reference substance solution under contrast test 3 chromatographic condition, and the retention time of impurity C is 27.382min。
Figure 19 is the HPLC figure of ticagrelor need testing solution under contrast test 3 chromatographic condition, and ticagrelor peak retention time is 19.806min。
Figure 20 is the HPLC figure of poly-doped impurity solution under contrast test 3 chromatographic condition, and impurity B retention time is 22.099min, and impurity C retention time is 27.480min, and impurity A retention time is 32.733min。
Figure 21 is the HPLC figure of test sample and impurity mixed solution under contrast test 3 chromatographic condition, and impurity C retention time is 27.392min, and impurity A retention time is 32.541min, and separating degree is better, but the chromatographic peak of impurity B is masked by ticagrelor main peak hangover place。Therefore, the method for contrast test 3 can not accurate checked for impurities B。
In order to further illustrate beneficial effects of the present invention, the present invention provides tests below example。
Test example 1, detection method methodological study
In this test example, various tests all adopt following condition:
Chromatographic column:AD-H, 4.6mm × 250mm, 5 μm；
Mobile phase: normal hexane-ethanol (90:10), adds the diethylamine of 0.1%～0.5% in ethanol
Solvent: mobile phase
Column temperature: 30 DEG C；Flow velocity: 1.0ml/min；UV detector (detection wavelength 242nm)。
Sampling volume: 20 μ L。
1, detection wavelength
Taking ticagrelor, dissolve and dilute the solution making suitable concentration with solvent, carry out spectral scan according to ultraviolet visible spectrophotometry (Chinese Pharmacopoeia two annex IVA of version in 2010) within the scope of 200～400nm, ultraviolet spectrogram is as shown in figure 22。
Result shows, ticagrelor all has absorption at 200～300nm, impurity A, impurity B and impurity C are the chiral isomer of ticagrelor, its ultraviolet spectrogram indistinction, therefore selects to select to measure wavelength within the scope of 200～300nm, and ticagrelor detects at 242nm wavelength place, baseline noise is less, other impurity are noiseless to impurity A, impurity B and impurity C, therefore, and the detection wavelength that final selection 242nm detects as ticagrelor chiral isomer。
2, specificity test
Take ticagrelor appropriate, after dissolving with ethanol, add mobile phase dilution and make in every 1mL the solution containing about 0.5mg, as need testing solution。Separately take impurity A in ticagrelor, impurity B, impurity C in right amount, impurity reference substance solution, separately take the mixing of impurity reference substance solution, with mobile phase dilution preparation poly-doped impurity solution。Precision takes each 20 μ L of above-mentioned poly-doped impurity reference substance solution, need testing solution and solvent respectively, injects chromatograph of liquid, records chromatogram。Result is as shown in figs. 1 to 6。
It is shown that when detection method, in ticagrelor sample and ticagrelor all the other impurity to impurity A, impurity B, impurity C mensuration noiseless, it was demonstrated that the specificity of detection method is strong。
3, standard curve and the range of linearity
Precision measures impurity A, impurity B, impurity C reference substance solution in right amount, makes the reference substance solution of a series of concentration with mobile phase dilution。The accurate each 20 μ L of reference substance solution taking variable concentrations, inject chromatograph of liquid respectively, record chromatogram。Measuring peak area respectively, result is in Table 1。
Table 1, linear relationship
With the concentration of impurity reference substance solution for abscissa X, with its peak area for vertical coordinate Y, drawing standard curve, calculate impurity (1R, 2S) the equation of linear regression of-2-(3,4-difluorophenyl) cyclopropylamine and correlation coefficient r, standard curve is as shown in figure 25。
It is shown that the concentration of impurity A is good linear relationship with peak area within the scope of 0.275 μ g/mL～2.204 μ g/mL in detection method, linear equation: Y=22540.6837X-2199.8432, r=0.9994；The concentration of impurity B is good linear relationship with peak area within the scope of 0.268 μ g/mL～2.145 μ g/mL, linear equation: Y=25575.0896X-1514.7421, r=0.9996；The concentration of impurity C is good linear relationship with peak area within the scope of 0.254 μ g/mL～2.034 μ g/mL, linear equation: Y=26287.5744X-1548.7861, r=0.9995, it was demonstrated that the inventive method range of linearity is wide, and accuracy is high。
Additionally, from standard curve equation and figure it can be seen that slope is far longer than intercept, standard curve, close to initial point, illustrates that the assay of each impurity is suitable for the one point external standard method of the present invention。
4, precision test
Taking the poly-doped impurity reference substance solution 3# under test example 1 the 3rd, precision takes 20 μ L, injects high performance liquid chromatograph, continuous sample introduction 6 times, measures peak area respectively according to the detection method of the present invention, and result is in Table 2。
Table 2, Precision test result
Calculating the RSD obtaining impurity A peak area is: 1.45%, and the RSD of impurity B peak area is: 2.54%, and the RSD of impurity C peak area is: 2.26%, it was demonstrated that the detection method precision of the present invention is excellent。
5, quantitative limit
Measuring the 3rd lower poly-doped impurity reference substance solution 6# of test example appropriate, add mobile phase dilution, precision measures 20 μ l, injects chromatograph of liquid, measures peak area and baseline noise according to the detection method of the present invention, and result is in Table 3。
Table 3, quantitative limit result of the test
| Impurity title | Concentration (μ g/mL) | Quantitative limit (ng) |
| Impurity A | 0.138 | 2.76 |
| Impurity B | 0.134 | 2.68 |
| Impurity C | 0.127 | 2.54 |
The peak height of impurity A, impurity B and impurity C is each about 10 times of baseline noise, by signal to noise ratio S/N=10, what obtain impurity A is quantitatively limited to 2.76ng, impurity B be quantitatively limited to 2.68ng, impurity C is quantitatively limited to 2.54ng, prove that the detection sensitivity of the inventive method is high, it is possible to fully meet the requirement of impurity quantitative assay。
6, replica test
Precision weighs ticagrelor 6 parts, respectively about 10mg, puts in 20mL measuring bottle respectively, and solubilizer dissolves and is diluted to scale, obtains need testing solution。Precision measures each 20 μ L of above-mentioned 6 parts of need testing solutions, detects according to the detection method of the present invention, and by the external standard method content with calculated by peak area impurity A, impurity B and impurity C, result is in Table 4。
Table 4, replica test result
| Sample number into spectrum | 1 | 2 | 3 | 4 | 5 | 6 |
| Impurity A | Do not detect | Do not detect | Do not detect | Do not detect | Do not detect | Do not detect |
| Impurity B | Do not detect | Do not detect | Do not detect | Do not detect | Do not detect | Do not detect |
| Impurity C | 0.019% | 0.018% | 0.016% | 0.019% | 0.017% | 0.020% |
From the above results, the repeatability of detection method is good。
7, solution stability testing
Precision weighs ticagrelor 101mg, puts in 20mL measuring bottle, and solubilizer dissolves and is diluted to scale, obtains need testing solution。0h, 1h, 2h, 4h, 6h, 8h sample introduction 20 μ L after preparation, records chromatogram, investigates the steadiness of impurity A in its need testing solution, impurity B and impurity C, and result is in Table 5。
Table 5, need testing solution stability test result table
| Standing time | 0h | 2h | 4h | 6h | 8h |
| Impurity A | Do not detect | Do not detect | Do not detect | Do not detect | Do not detect |
| Impurity B | Do not detect | Do not detect | Do not detect | Do not detect | Do not detect |
| Impurity C | 0.019% | 0.020% | 0.018% | 0.019% | 0.017% |
From the above results, in latter 8 hours of preparation, in need testing solution, impurity A, impurity B all do not detect, and impurity C detects result without significant change, it was demonstrated that detection method need testing solution is stable。
8, recovery test
Precision weighs ticagrelor 9 parts, respectively about 10mg, puts in 20mL measuring bottle respectively, add each impurity concentration under test example the 3rd and be about each 3 parts of the reference substance solution 0.5mL of 20 μ g/mL, 1.0mL, 2.0mL, solubilizer dissolves and is diluted to scale, shakes up, respectively as response rate need testing solution。The accurate each 20 μ L sample introductions of the reference substance solution under 9 parts of response rate need testing solutions and test example the 4th that take measure respectively, record chromatogram, calculate impurity A, impurity B and the measured amount of impurity C, reference substance addition and the response rate, and result is in Table 6～8。
Computing formula:
In formula: a is the amount (μ g) of contained isomer impurities in test sample；
B is isomer impurities reference substance addition (μ g)；
C is the measured amount (μ g) of isomer impurities。
Table 6, impurity A recovery test result
It is shown that detection method measures the chiral isomer impurity A in ticagrelor, the response rate is between 100.6%～106.0%, and relative standard deviation is 1.75%, it was demonstrated that the detection method response rate of the present invention is good, and accuracy is high。
Table 7, impurity B recovery test result
It is shown that detection method measures the chiral isomer impurity B in ticagrelor, the response rate is between 98.9%～103.6%, and relative standard deviation is 1.90%, it was demonstrated that the detection method response rate of the present invention is good, and accuracy is high。
Table 8, impurity C recovery test result
It is shown that detection method measures the chiral isomer impurity C in ticagrelor, the response rate is between 98.4%～103.0%, and relative standard deviation is 1.74%, it was demonstrated that the detection method response rate of the present invention is good, and accuracy is high。
In sum, the invention provides the new detection method of isomer impurities content in a kind of ticagrelor, separating degree between each chromatographic peak is high, noiseless each other, can realize impurity B and impurity A simultaneously, the accurate detection of impurity C, and, easy and simple to handle, it is easily controlled, testing cost is low, and there is good linear relationship, specificity, precision, stability, sensitivity and repeatability, average recovery is high, testing result is accurate, reliably, a kind of effective detection method is provided for the content of isomer impurities in monitoring ticagrelor medicine, further ensure the product quality of ticagrelor and the drug safety of patient。
Claims (10)
1. the detection method of isomer impurities content in a ticagrelor, it is characterised in that: it comprises the following steps:
A, take the reference substance of isomer impurities, prepare reference substance solution；
B, take ticagrelor sample to be checked, prepare need testing solution；
Reference substance solution, need testing solution are detected by c, employing efficient liquid-phase chromatography method respectively, and the testing conditions of described efficient liquid-phase chromatography method is:
Chromatographic column: fixing is amylose-three (3,5-xylyl carbamate) mutually；
The volume ratio of mobile phase: normal hexane-ethanol, normal hexane and ethanol is 88%:12%～92%:8%, contains the diethylamine of 0.1%～0.5%ml/ml in described ethanol；
Detection wavelength: 200nm～300nm；
D, by external standard method with calculated by peak area, or calculate according to the standard curve between peak area and concentration, obtain the isomer impurities content in ticagrelor sample。
2. detection method according to claim 1, it is characterized in that: described isomer impurities at least includes (1R, 2R, 3S, 5R)-3-[7-{ [(1R, 2S)-2-(3,4-difluorophenyl) cyclopropyl] amino }-5-rosickyite base-3H-1,2,3-triazole [4,5-d] pyrimidin-3-yl]-5-(2-hydroxy ethoxy) Pentamethylene .-1,2-glycol。
3. detection method according to claim 2, it is characterized in that: described isomer impurities at least includes (1R, 2R, 3S, 5R)-3-[7-{ [(1R, 2S)-2-(3, 4-difluorophenyl) cyclopropyl] amino }-5-rosickyite base-3H-1, 2, 3-triazole [4, 5-d] pyrimidin-3-yl]-5-(2-hydroxy ethoxy) Pentamethylene .-1, 2-glycol and (1R, 2R, 3S, 5R)-3-[7-{ [(1S, 2R)-2-(3, 4-difluorophenyl) cyclopropyl] amino }-5-rosickyite base-3H-1, 2, 3-triazole [4, 5-d] pyrimidin-3-yl]-5-(2-hydroxy ethoxy) Pentamethylene .-1, 2-glycol；
Preferably, described isomer impurities includes (1R simultaneously, 2R, 3S, 5R)-3-[7-{ [(1R, 2S)-2-(3, 4-difluorophenyl) cyclopropyl] amino }-5-rosickyite base-3H-1, 2, 3-triazole [4, 5-d] pyrimidin-3-yl]-5-(2-hydroxy ethoxy) Pentamethylene .-1, 2-glycol, (1R, 2R, 3S, 5R)-3-[7-{ [(1S, 2R)-2-(3, 4-difluorophenyl) cyclopropyl] amino }-5-rosickyite base-3H-1, 2, 3-triazole [4, 5-d] pyrimidin-3-yl]-5-(2-hydroxy ethoxy) Pentamethylene .-1, 2-glycol and (1S, 2S, 3R, 5S)-3-[7-{ [(1S, 2R)-2-(3, 4-difluorophenyl) cyclopropyl] amino }-5-rosickyite base-3H-1, 2, 3-triazole [4, 5-d] pyrimidin-3-yl]-5-(2-hydroxy ethoxy) Pentamethylene .-1, 2-glycol。
4. detection method according to claim 1, it is characterised in that: in step a, the solvent of preparation reference substance solution is the mobile phase in step c；In step b, the solvent preparing need testing solution is the mobile phase in step c。
5. detection method according to claim 1, it is characterised in that: in step c, described chromatographic column isAD-H, length is 250mm, and internal diameter is 4.6mm, and the particle diameter of filler is 5 μm。
6. detection method according to claim 1, it is characterised in that: in step c, the column temperature of described chromatographic column is 25 DEG C～35 DEG C。
7. detection method according to claim 1, it is characterised in that: in step c, the volume ratio of normal hexane and ethanol is 90%:10%；Described ethanol contains the diethylamine of 0.2%ml/ml。
8. detection method according to claim 1, it is characterised in that: in step c, the flow velocity of described mobile phase is 0.9ml/min～1.1ml/min；Preferably, the flow velocity of described mobile phase is 1.0ml/min。
9. detection method according to claim 1, it is characterised in that: in step c, described detection wavelength is 230nm～250nm；Preferably, described detection wavelength is 242nm。
10. detection method according to claim 1, it is characterised in that: in step c, sampling volume is 10 μ l～50 μ l；Preferably, sampling volume is 20 μ l。
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107402267A (en) * | 2017-07-25 | 2017-11-28 | 江苏工程职业技术学院 | A kind of method of normal phase high performance liquid chromatography measure rope fluorine cloth Wei bulk drug diastereoisomer and impurity content |
| CN107449842A (en) * | 2017-07-25 | 2017-12-08 | 江苏工程职业技术学院 | A kind of method of normal phase high performance liquid chromatography measure rope fluorine cloth Wei bulk drug enantiomter content |
| CN107966489A (en) * | 2016-10-20 | 2018-04-27 | 中国科学院上海有机化学研究所 | A kind of method of isomer impurities content in detection sitafloxacin |
| CN109856255A (en) * | 2017-11-30 | 2019-06-07 | 四川海思科制药有限公司 | A kind of analysis method for the isomer impurities content controlling ticagrelor intermediate |
| CN113125587A (en) * | 2019-12-30 | 2021-07-16 | 成都百裕制药股份有限公司 | Tofacitinib intermediate and detection method of enantiomer thereof |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010136137A1 (en) * | 2009-05-27 | 2010-12-02 | Bayer Schering Pharma Aktiengesellschaft | Substituted piperidines |
| EP2589587A1 (en) * | 2011-11-04 | 2013-05-08 | Chemo Ibérica, S.A. | Synthesis of nitrogen substituted cyclopropanes |
| WO2013150495A2 (en) * | 2012-04-05 | 2013-10-10 | Dr. Reddy's Laboratories Limited | Preparation of ticagrelor |
| CN104634887A (en) * | 2013-11-11 | 2015-05-20 | 广东东阳光药业有限公司 | Method for separating and measuring ticagrelor and optical isomer of ticagrelor |
| CN105092721A (en) * | 2014-05-21 | 2015-11-25 | 天津市汉康医药生物技术有限公司 | High performance liquid chromatography detection analysis method for controlling ticagrelor isomer |
-
2016
- 2016-01-29 CN CN201610070436.7A patent/CN105699524B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010136137A1 (en) * | 2009-05-27 | 2010-12-02 | Bayer Schering Pharma Aktiengesellschaft | Substituted piperidines |
| EP2589587A1 (en) * | 2011-11-04 | 2013-05-08 | Chemo Ibérica, S.A. | Synthesis of nitrogen substituted cyclopropanes |
| WO2013150495A2 (en) * | 2012-04-05 | 2013-10-10 | Dr. Reddy's Laboratories Limited | Preparation of ticagrelor |
| CN104634887A (en) * | 2013-11-11 | 2015-05-20 | 广东东阳光药业有限公司 | Method for separating and measuring ticagrelor and optical isomer of ticagrelor |
| CN105092721A (en) * | 2014-05-21 | 2015-11-25 | 天津市汉康医药生物技术有限公司 | High performance liquid chromatography detection analysis method for controlling ticagrelor isomer |
Non-Patent Citations (2)
| Title |
|---|
| L. KALYANI. ET AL.: "A VALIDATED STABILITY-INDICATING HPLC METHOD FOR DETERMINATION OF TICAGRELOR IN BULK AND ITS FORMULATION", 《INT J PHARM》 * |
| NEERAJ KUMAR ET AL.: "Four process-related potential new impurities in ticagrelor:Identification, isolation, characterization using HPLC, LC/ESI–MSn,NMR and their synthesis", 《JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS》 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107966489A (en) * | 2016-10-20 | 2018-04-27 | 中国科学院上海有机化学研究所 | A kind of method of isomer impurities content in detection sitafloxacin |
| CN107402267A (en) * | 2017-07-25 | 2017-11-28 | 江苏工程职业技术学院 | A kind of method of normal phase high performance liquid chromatography measure rope fluorine cloth Wei bulk drug diastereoisomer and impurity content |
| CN107449842A (en) * | 2017-07-25 | 2017-12-08 | 江苏工程职业技术学院 | A kind of method of normal phase high performance liquid chromatography measure rope fluorine cloth Wei bulk drug enantiomter content |
| CN109856255A (en) * | 2017-11-30 | 2019-06-07 | 四川海思科制药有限公司 | A kind of analysis method for the isomer impurities content controlling ticagrelor intermediate |
| CN109856255B (en) * | 2017-11-30 | 2022-04-05 | 四川海思科制药有限公司 | Analysis method for controlling isomer impurity content of ticagrelor intermediate |
| CN113125587A (en) * | 2019-12-30 | 2021-07-16 | 成都百裕制药股份有限公司 | Tofacitinib intermediate and detection method of enantiomer thereof |
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