CN104678006B - Sunitinib malate related substance analysis method - Google Patents

Abstract

The invention relates to a sunitinib malate related substance analysis method which is carried out on high performance liquid chromatography and adopts Poroshell 120? Bonus? An RP chromatographic column, an ammonium acetate aqueous solution and an organic solvent acetonitrile as mobile phases, and a liquid Diode Array Detector (DAD) or an Ultraviolet (UV) detector for carrying out gradient elution detection, thereby effectively separating and detecting related substances B, C, E and other related substances. The analysis method has the characteristics of good specificity, high accuracy, simplicity, convenience and quickness.

Description

A method for analyzing related substances of sunitinib malate

technical field

The invention relates to the field of medicine and chemical industry, and more specifically relates to an optimized method for detecting related substances of sunitinib malate, a chemical raw material drug.

Background technique

Sunitinib malate, chemical name N-(2-diethylaminoethyl)-5[(Z)-(5-fluoro-2-oxo-1hydro-indole-3-ylidene)methyl ]-2,4-dimethyl-1 hydrogen-pyrrole-3-carboxamide malate, its chemical structure is:

It is a multi-target receptor tyrosine kinase (RTK) inhibitor, which is mainly used for the treatment of gastrointestinal stromal tumors and metastatic renal cell carcinoma that do not respond to or cannot tolerate standard therapy through oral administration. The drug has been approved for marketing by the US FDA and the European EMEA, and the product is Sutent. Chinese authorized patent CN1329390C discloses a preparation method of sunitinib malate.

Usually, sunitinib malate raw materials also contain sunitinib degradation-related substance B, related substance C and related substance E introduced during the process of preparation, and other related substances.

The structural formulas of related substances B, C and E are shown in Table 1

Table 1

In order to ensure the safe use of drugs, it is usually necessary to carry out quality control on drugs, and related substances are items that must be tested for most drugs. At present, the quality standards of sunitinib malate are not included in the current edition of "European Pharmacopoeia" and its pharmacopoeia forum, "US Pharmacopoeia" and its pharmacopoeia forum, and the 2010 edition of Chinese Pharmacopoeia. Therefore, it is necessary to develop effective detection methods for substances B, C, E and other unknown impurities.

In order to ensure the applicability and effectiveness of the detection method for the drug during the production and storage phases, it is usually necessary to verify the analytical method, including experiments on specificity, precision and accuracy, see Appendix XIXA of Part II of the Chinese Pharmacopoeia 2010 Edition Guidelines for Validation of Analytical Methods for Quality Standards of Drugs (National Pharmacopoeia Commission. Pharmacopoeia of the People's Republic of China [S]. Beijing: China Medical Science and Technology Press, 2010: 194～195). During the investigation of specificity, it is usually necessary to accelerate the destruction of drugs through acid (alkali) hydrolysis, high temperature, strong light irradiation or oxidation, etc., to study possible degradation products and degradation pathways of drugs, and to verify the specificity of analytical methods.

High-performance liquid chromatography usually has internal standard method, external standard method, principal component self-control method with correction factor, principal component self-control method without correction factor and area normalization method, see Chinese Pharmacopoeia 2010 edition second Appendix ⅤD High Performance Liquid Chromatography (National Pharmacopoeia Committee. Pharmacopoeia of the People's Republic of China [S]. Beijing: China Medical Science and Technology Press, 2010: 29-31). The accurate determination of related substances in the sample by the area normalization method assumes that all the measured components in the sample to be tested are detected and have the same response factor at the same UV detection wavelength. However, in fact, different compounds often have different response factors at the same UV detection wavelength, which is well known to those skilled in the art. Therefore, the measurement error of the peak area normalization method is large, and it is usually only suitable for roughly checking the impurity content in the test product.

Among them, the principal component self-control method without a correction factor means that when measuring the impurity content, if there is no impurity reference substance, the principal component self-control method without a correction factor can also be used. After configuring the control solution and adjusting the detection sensitivity with the main component self-control method with the addition of the correction factor, take appropriate amounts of the test solution and the control solution and inject them separately. The recording time of the former, unless otherwise specified, should be the peak of the main component 2 times the retention time, measure the peak area of each impurity on the chromatogram of the test solution and compare it with the peak area of the main component of the contrast solution to calculate the impurity content.

State Food and Drug Administration Imported Drug Registration Standard, Sunitinib Malate Capsules (Standard Number: JX20060174), discloses a detection method for related substances of sunitinib malate. The method uses octadecylsilane bonded silica gel as a filler (such as SupelcoDiscoveryC18, 150 × 4.6mm, 5 μm), the column temperature is 45°C, and the detection wavelength is 268nm, with 0.05mol/L ammonium acetate buffer solution (pH5 .0) is the mobile phase A, acetonitrile is the mobile phase B, and eluted according to the following gradient,

The related substances of sunitinib malate were detected in a high-performance liquid chromatography system, and the contents of all related substances were calculated by the area normalization method. Therefore, the method has the problems of poor specificity, low accuracy, long running time and low efficiency.

In order to solve the technical problems existing in the existing methods for detecting related substances of sunitinib malate, the present invention provides an accurate, fast and simple method for analyzing related substances of sunitinib malate bulk drug.

Contents of the invention

Summary of the invention

Through repeated experiments, the inventor screened different types of chromatographic columns and tried different mobile phase conditions, and finally obtained an analytical method suitable for detecting related substances in sunitinib malate bulk drug. The method is carried out on high-performance liquid chromatography, and the detector is a liquid diode array detector (DAD) or an ultraviolet (UV) detector, and is eluted as a mobile phase with ammonium acetate aqueous solution and organic solvent acetonitrile, and adopts Poroshell120BonusRP The chromatographic column is used for chromatographic separation and detection, and the method has the characteristics of superior specificity, high sensitivity, high accuracy and high efficiency, and solves the problems existing in the prior art.

Among them, the particle size of the filler particles of the Poroshell120BonusRP chromatographic column is about 2.7 μm, and the particles are composed of an inner solid core with a diameter of 1.7 μm and a porous outer layer with a thickness of 0.5 μm covering the inner solid core, and the stationary phase is the bond n-octadecane bound to the filler particles. Due to the special structure of the filler particles of the chromatographic column, it shows excellent specificity in the detection of related substances B, C and E of sunitinib malate raw material drug. The selection of Poroshell120BonusRP chromatographic column as the application chromatographic column for the analysis method of related substances of sunitinib malate API is the result of multiple screenings.

Definition of Terms

The term "packing particles" refers to the field of liquid chromatography columns. The liquid chromatography column is prepared by packing particles (which can be different materials, such as silica gel) with a stationary phase on the surface of the column. The structure of the particles, Attributes such as size and pressure resistance will affect the specific performance of the liquid chromatographic column when analyzing and detecting specific components.

The term "peak purity" refers to an investigation parameter used to judge whether a certain chromatographic peak is caused by only one substance in HPLC detection. It is generally believed that a certain chromatographic peak under investigation is pure when the peak purity is between 0.990 and 1.000. The chromatographic peak is that of a single substance.

Term " RT " refers to the retention time of the chromatographic peak in the HPLC detection, and the unit is minute (min); The relative retention time of the main peak is 1 with reference to the peak.

The term "v/v" means a volume ratio, and "w/w" means a mass ratio.

Detailed description of the invention

The related substance analysis method of sunitinib malate crude drug provided by the invention is characterized in that:

The method is carried out on a high performance liquid chromatograph;

The detector is a diode array detector (DAD) or an ultraviolet absorption detector (UV), and the detection wavelength is 268nm;

The liquid chromatographic column is Poroshell120BonusRP reversed-phase chromatographic column, and the temperature of the chromatographic column is 20℃～30℃;

The mobile phase is composed of ammonium acetate aqueous solution as a buffer and acetonitrile as an organic solvent, and is subjected to gradient elution, and the flow rate of the mobile phase is about 0.9mL/min～1.1mL/min;

Inject sunitinib malate test solution and sunitinib malate reference solution for detection, and calculate the content of related substances according to the principal component self-control method without correction factor.

In some embodiments, the Poroshell120BonusRP reversed-phase chromatographic column is filled with filler particles with a particle size of about 2.7 μm, wherein the filler particles are composed of an inner solid core with a diameter of 1.7 μm and a 0.5 μm thick porous layer coated on the inner solid core. The outer layer is formed, wherein the stationary phase is n-octadecane bonded to the filler particles.

In some embodiments, the ammonium acetate aqueous solution has a concentration of about 3.45 g/L-4.20 g/L and a pH of about 5.5-6.0.

In some embodiments, the method of the present invention uses a gradient elution method to separate and detect the detection sample, and the gradient elution method can be

Time (min) ammonium acetate aqueous solution (v/v, %) acetonitrile (v/v, %)

09010

123763

In some embodiments, the high performance liquid chromatograph can be Agilent 1260, Agilent 1290 or Waters UPLC.

In some embodiments, the preparation of the test solution is carried out under dark conditions. Take an appropriate amount of sunitinib malate for the test product, dissolve it in a mixed solution of water and acetonitrile, and dilute it to a final concentration of about 0.10 mg/mL to 1.0 mg/mL, more preferably the test product solution concentration is about 0.50 mg/mL; The reference substance solution is to take an appropriate amount of sunitinib malate for the test to be dissolved in a mixed solution of water and acetonitrile, and then diluted to a final concentration of about 0.2 μg/mL～2.0 μg/mL, more preferably the reference substance solution concentration is about 1.0 μg /mL. In the mixed solution of water and acetonitrile described therein, the volume ratio of water and acetonitrile can be 7:3.

In some embodiments, the related substance method provided by the present invention is characterized in that:

High performance liquid chromatography: Agilent1260;

The detector is a diode array detector (DAD), and the detection wavelength is 268nm;

The liquid chromatography column is a Poroshell120BonusRP reversed-phase column, and the column temperature is 25°C;

The flow rate is about 1.0 mL/min;

Gradient elution:

Time (min) ammonium acetate aqueous solution (v/v, %) acetonitrile (v/v, %)

09010

123763

Inject sunitinib malate test product solution and sunitinib malate reference substance solution for detection, and calculate the content of related substances by the principal component self-control method without correction factor;

Among them, the Poroshell120BonusRP reversed-phase chromatographic column is filled with filler particles with a particle size of about 2.7 μm, wherein the filler particles are composed of an inner solid core with a diameter of 1.7 μm and a porous outer layer with a thickness of 0.5 μm coated on the inner solid core. , wherein the stationary phase is n-octadecane bonded to the packing particles, the length of the column is 50 mm, and the inner diameter of the column is 4.6 mm;

Wherein the concentration of ammonium acetate aqueous solution is about 3.85g/L, and the pH is about 5.7;

Among them, the sunitinib malate test solution is a mixed solution of acetonitrile and water with a sunitinib malate concentration of about 0.50 mg/mL, and the sunitinib malate reference solution is a sunitinib malate concentration A mixed solution of acetonitrile and water at about 1.0 μg/mL, wherein in the mixed solution of water and acetonitrile, the volume ratio of water and acetonitrile can be 7:3.

The analysis method provided by the present invention is suitable for detecting related substances in sunitinib malate bulk drug, especially suitable for detecting related substance B, related substance C and related substance E therein.

Description of drawings

Fig. 1 shows the detection chromatogram of the blank solution in embodiment 1

Fig. 2 shows the detection chromatogram of the sunitinib malate reference substance solution in embodiment 1

Fig. 3 shows the detection chromatogram of sunitinib malate need testing solution in embodiment 1

Fig. 4 shows the detection chromatogram of the sensitizing solution in embodiment 1

Fig. 5 shows the detection chromatogram of the non-destructive solution in embodiment 2

Fig. 6 shows the detection chromatogram of the acid destruction solution in embodiment 2

Fig. 7 shows the detection chromatogram of the alkali destruction solution in embodiment 2

Fig. 8 shows the detection chromatogram of the oxidation destruction solution in embodiment 2

Fig. 9 shows the detection chromatogram of the light damage solution in embodiment 2

Fig. 10 shows the detection chromatogram of the high temperature destruction solution in embodiment 2

Fig. 11 shows the chromatogram that the oxidative destruction solution in embodiment 5 detects with comparative detection method

detailed description

In order to enable those skilled in the art to better understand the technical solutions of the present invention, some non-limiting examples are further disclosed below to further describe the present invention in detail.

The reagents used in the present invention can be purchased from the market or can be prepared by the methods described in the present invention.

Embodiment 1 system adaptability experiment

Solution preparation:

Blank solution (diluent): Prepare a mixed solution of water and acetonitrile with a volume ratio of 7:3.

Sunitinib malate test solution: under dark conditions, take an appropriate amount of sunitinib malate test product, accurately weighed, put in a volumetric flask, and prepare sunitinib malate with an appropriate amount of diluent A solution with a concentration of about 0.50 mg/mL.

Sunitinib malate reference substance solution: under dark conditions, accurately pipette an appropriate amount of sunitinib malate test solution, put it in a volumetric flask, and dilute with an appropriate amount of diluent until the concentration of sunitinib malate is About 1.0 μg/mL solution.

Sensitization solution: Take appropriate amount of sunitinib malate test product, related substance B, related substance C and related substance E, accurately weigh them, place in a volumetric flask and prepare the concentration of sunitinib malate with diluent It is about 0.50 mg/mL, containing a solution of related substance B at a concentration of about 1.6 μg/mL, related substance C at a concentration of about 0.6 μg/mL and related substance E at a concentration of about 0.6 μg/mL, respectively.

Injection detection:

The chromatographic conditions are as follows:

High performance liquid chromatography: Agilent1260;

The detector is a diode array detector (DAD), and the detection wavelength is 268nm;

The liquid chromatography column is a Poroshell120BonusRP reversed-phase column, and the column temperature is 25°C;

The flow rate is about 1.0 mL/min;

Gradient elution:

Time (min) ammonium acetate aqueous solution (v/v, %) acetonitrile (v/v, %)

09010

123763

Among them, the Poroshell120BonusRP reversed-phase chromatographic column is filled with filler particles with a particle size of about 2.7 μm, wherein the filler particles are composed of an inner solid core with a diameter of 1.7 μm and a porous outer layer with a thickness of 0.5 μm coated on the inner solid core. , wherein the stationary phase is n-octadecane bonded to the packing particles, the length of the column is 50 mm, and the inner diameter of the column is 4.6 mm;

Wherein the concentration of ammonium acetate aqueous solution is about 3.85g/L, and the pH is about 5.7;

Take the blank solution, sunitinib malate test solution, sunitinib malate reference solution and sensitization solution, inject 1 needle each, and record the chromatogram. Report the retention time (RT) of the main peak in sunitinib malate test solution, sunitinib malate reference solution and sensitization solution; the main peak in sunitinib malate test solution and sensitization solution The peak purity, the relative retention time (RRT) and peak area of related substances B, C and E, the separation of the main peak and related substances B, C and E from adjacent peaks. Fig. 1 shows that the blank solution has no interference to the detection of sunitinib malate test substance; Fig. 2 shows that in the detection chromatogram of sunitinib malate reference substance solution, the reference substance has a good response value and high sensitivity; Fig. 3 It shows that in the sunitinib malate test product, the main peak and each known single impurity (related substances B, C and E) and the resolution of adjacent peaks are all good, and Fig. 4 shows that each added known compound in the sensitizing solution is related. The peak area of the substances (related substances B, C and E) increased significantly, and the peak area and the amount of impurities had a linear relationship. The test results are shown in Table 1.

Table 1

According to the experimental results and maps,

Blank solution, need testing solution and sensitizing solution compare, blank solution has no interference at the retention time of main peak and each known single impurity (related substances B, C and E); need testing solution and sensitizing solution chromatogram The separation of the main peak and each known single impurity (related substances B, C and E) and adjacent peaks is not less than 1.5; the retention time of the main peak in the test solution is consistent with that in the sensitizing solution and the contrast solution; the test solution The peak purity factor of the main peak in the chromatogram of the sensitizing solution is all greater than 0.990; compared with the test solution, the peak area of the peak at the retention time of impurity B, impurity C and impurity E in the chromatogram of the sensitizing solution is significantly enhanced. Therefore, the method provided by the present invention has good specificity.

Example 2 Strong Degradation Destruction Experiment

Destruction sample solution preparation:

Diluent: Prepare a mixed solution of water and acetonitrile with a volume ratio of 7:3.

Stock solution of the test product: under dark conditions, take 135mg of the test product, accurately weigh it into a 50mL volumetric flask, dissolve it ultrasonically with the diluent and make it to volume, shake well, and obtain it.

Undestroyed test product solution: under light-proof conditions, accurately pipette 5 mL of the test product stock solution into a 25 mL volumetric flask, dilute to volume with diluent, shake well, and obtain.

Acid destruction solution: under the condition of avoiding light, accurately pipette 5mL of the stock solution of the test product into a 25mL volumetric flask, add 1mL of 6mol/L hydrochloric acid solution, shake well, seal it, and place it at room temperature for 24h, then add 6mol/L Neutralize with 1mL of sodium hydroxide solution, let it cool, dilute to volume with diluent, shake well, and obtain;

Alkali destruction solution: under the condition of avoiding light, accurately pipette 5mL of the stock solution of the test product into a 25mL volumetric flask, add 1mL of 6mol/L sodium hydroxide solution, shake well, seal it, and place it at room temperature for 24h, then add 6mol /L hydrochloric acid solution 1mL to neutralize, let cool, dilute to volume with diluent, shake well, that is;

Oxidative destruction solution: under light-proof conditions, accurately pipette 5mL of the stock solution of the test product into a 25mL volumetric flask, add 1mL of 15% hydrogen peroxide, leave it at room temperature for 24 hours, dilute to volume with diluent, shake well, and obtain;

Light damage solution: Take an appropriate amount of the test product, spread it into a weighing bottle, place it under the condition of visible light 4500Lux±500Lux, ultraviolet light 1.7W*h/m2 for 13 days, the total visible light intensity reaches 1200000Lux, and the ultraviolet light intensity reaches 200W /m2 or more, then take 27mg of the above sample, accurately weigh it, put it in a 100mL volumetric flask, and dilute to volume with diluent under dark conditions, shake well to get it;

High-temperature destruction solution: Take an appropriate amount of the test sample, spread it into a weighing bottle, place it in an oven at 105°C for 24 hours, take it out and let it cool, then take about 27mg of the above sample, weigh it accurately, put it in a 100mL measuring bottle, and keep it in the dark Under certain conditions, dilute to volume with diluent, shake well, and obtain;

Injection detection:

Chromatographic conditions are as in Example 1;

Inject 1 injection of the above-mentioned undamaged solution and each destroyed solution, and record the chromatogram. Report the separation of the main peak and adjacent peaks, the purity of the main peak and the percentage of the total miscellaneous peak area in each destruction solution.

Figure 5, Figure 6, Figure 7, Figure 8, Figure 9 and Figure 10 are the chromatograms of undamaged solution, acid damaged solution, alkali damaged solution, oxidative damaged solution, light damaged solution and high temperature damaged solution respectively, indicating the undamaged condition The results of the solution determination under different conditions and damage conditions.

The test results are shown in Table 2.

Table 2

Solution name Total Miscellaneous (%) main peak purity factor Separation of main peak and adjacent peaks non-destructive solution 0.37 1.0000 1.5 Acid Breaking Solution 1.06 1.000 1.5 Alkali Breaking Solution 1.45 1.000 2.5 Oxidative Destruction Solution 6.72 1.000 2.7 light damage solution 0.36 1.000 1.5 High temperature destruction solution 0.34 1.000 1.6

According to the results of collection of graphs 5-10 and table 4, it can be known that the method of the present invention detects the undamaged sunitinib malate solution and the destroyed sunitinib malate solution, the peak purity of the main peak is very high, It shows that the main peak does not contain other impurity peaks; the separation between the main peak and other impurity peaks is good, ≥1.5. Therefore the method of the present invention all has good specificity when measuring sunitinib malate, even the related substance of sunitinib malate degradation sample, can effectively detect and control the sunitinib malate in the preservation process. Ni's content of related substances.

Embodiment 3 Precision experiment

Solution preparation:

Diluent: Prepare a mixed solution of water and acetonitrile with a volume ratio of 7:3.

Sunitinib malate test solution: under dark conditions, take an appropriate amount of sunitinib malate test product, accurately weighed, put in a volumetric flask, and prepare sunitinib malate with an appropriate amount of diluent A solution with a concentration of about 0.50 mg/mL was prepared in parallel in 6 copies.

Sunitinib malate reference substance solution: under dark conditions, accurately pipette an appropriate amount of sunitinib malate test solution, put it in a volumetric flask, and dilute with an appropriate amount of diluent until the concentration of sunitinib malate is About 1.0 μg/mL solution.

Injection detection:

Chromatographic conditions are as in Example 1;

Take 2 needles of sunitinib malate reference substance solution, and take 6 parts of sunitinib malate test solution and inject 1 needle each. Calculate the content of each impurity in 6 parts of sunitinib malate test solution, the mean value of the content and the relative standard deviation value of the total impurity according to the principal component self-control method without correction factor, and record the retention time and retention time of each impurity The relative standard deviation of time.

The test results are shown in Table 3

table 3

According to the test results, it can be known that the method provided by the present invention detects the precision experiment of sunitinib malate related substances, the retention time of each impurity is basically stable, and the content of each impurity is calculated by the principal component self-control method without correction factor , the content of each impurity is basically the same, and the relative standard deviation of the total impurity is less than 5.0%, which has very good precision.

Embodiment 4 accuracy experiment

Solution preparation:

Diluent: Prepare a mixed solution of water and acetonitrile with a volume ratio of 7:3.

Sunitinib malate test product solution: under dark conditions, take about 27 mg of sunitinib malate test product, weigh it accurately, put it in a 50mL volumetric flask, dissolve it with appropriate amount of diluent and dilute to scale, prepared into a solution with a concentration of sunitinib malate of about 0.50 mg/mL.

Sunitinib malate reference solution: under dark conditions, accurately pipette 1mL of sunitinib malate test solution into a 50mL volumetric flask, dilute to volume with diluent, and shake well; then precisely pipette the above Put 5mL of the solution into a 50mL volumetric flask, dilute to volume with diluent, and prepare a solution with a concentration of sunitinib malate of about 1.0 μg/mL.

Related substances B, C and E stock solution: take about 16mg of related substance B, about 6mg of related substance C and about 6mg of related substance E, weigh them accurately, put them in a 1000mL volumetric flask, dissolve them with diluent and dilute to the mark, shake well .

Adding standard solution: under dark conditions, take about 27mg of sunitinib malate test product, weigh it accurately, and place it in a 50mL volumetric flask; then accurately pipette 5mL of the stock solutions of related substances B, C and E Put it into the above-mentioned 50mL volumetric flask, dilute to the mark with diluent, and prepare sunitinib malate with a concentration of about 0.50 mg/mL, a related substance B concentration of about 1.6 μg/mL, and a related substance C concentration of about 0.6 The solutions with μg/mL and related substance E concentration of about 0.6 μg/mL were prepared in parallel in 3 parts.

Injection detection:

Chromatographic conditions are as in Example 1;

Take the prepared sunitinib malate reference substance solution, sunitinib malate test solution and standard addition solution to detect by the chromatographic conditions of embodiment 1, each sample injection 1 needle, record the malate For the peak area of the control peak in the Nitinib reference substance solution, record the peak areas of related substance B, related substance C and related substance E in the sunitinib malate test solution and the standard addition solution, and calculate the respective peak areas according to the following formula Spike recovery of impurities:

Theoretical amount (mg) = W _x × P _x

In the formula: W _STD is the weighing amount of sunitinib malate in the sunitinib malate reference substance solution, mg;

Wx is the weighing of related substance B, related substance C and related substance E;

A _STD is the control peak area in the sunitinib malate reference substance solution;

A _S is the peak area of related substances (related substances B, C or E) in the sunitinib malate test solution;

A _X is the peak area of related substances (related substances B, C or E) in each spiked solution; D _Std is the dilution factor of sunitinib malate reference substance solution;

D _X is the dilution multiple of each spiked solution;

P _Std is the content of sunitinib malate test product, which is 99.4%, w/w;

P _X is the content of related substance B (98.7%, w/w), related substance C (99.7%, w/w) and related substance E (99.2%, w/w);

The experimental results are shown in Table 4

Table 4

Embodiment 5 Comparative Examples

Take the oxidative destruction solution in the strong degradation experiment of Example 2 as the test solution, according to the prior art State Food and Drug Administration import drug registration standard, Sunitinib Malate Capsules (standard number: JX20060174), the Shunimalate malate disclosed in The detection method for the related substances of Nitinib was detected, and 1 needle was injected, and the chromatogram was recorded. As shown in Figure 11, it shows that the impurities produced by oxidation damage cannot be separated from the main peak.

In summary,

The sunitinib malate related substance method of the present invention has good specificity, good precision, high accuracy and fast detection speed, and is applicable to the control of related substances during production and storage.

The method of the present invention has been described through preferred embodiments, and relevant persons can obviously make changes or appropriate changes and combinations to the methods and applications described herein within the content, spirit and scope of the present invention to realize and apply the technology of the present invention . Those skilled in the art can refer to the content of this article to appropriately improve the process parameters to achieve. In particular, it should be pointed out that all similar substitutions and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention.

Claims (6)

1. A sunitinib malate related substance analysis method is characterized in that: The method is carried out on a high performance liquid chromatograph; The detector is a diode array detector or an ultraviolet absorption detector, and the detection wavelength is 268nm; The liquid chromatographic column is Poroshell120BonusRP reversed-phase chromatographic column, and the temperature of the chromatographic column is 20℃～30℃; The mobile phase is composed of ammonium acetate aqueous solution as a buffer and acetonitrile as an organic solvent, and is subjected to gradient elution, and the flow rate of the mobile phase is 0.9mL/min～1.1mL/min; Inject sunitinib malate test product solution and sunitinib malate reference substance solution for detection, and calculate the content of related substances by the principal component self-control method without correction factor; The Poroshell120BonusRP reverse-phase chromatographic column described therein is filled with filler particles with a particle size of about 2.7 μm, wherein the filler particles are composed of an inner solid core with a diameter of 1.7 μm and a porous outer layer with a thickness of 0.5 μm coated on the inner solid core constituted, wherein the stationary phase is n-octadecane bonded to the filler particles; Wherein the concentration of the ammonium acetate aqueous solution is 3.45g/L～4.20g/L, pH5.5～6.0; The gradient program is as follows: The method described herein is used to detect related substance B, related substance C or related substance E in the sunitinib malate bulk drug. 2. analytical method according to claim 1, wherein said sunitinib malate need testing solution is to get sunitinib malate need testing water and the mixed solution dissolving of acetonitrile, dilution, final concentration is 0.10mg/mL～1.0mg/mL; The sunitinib malate reference substance solution is dissolved in a mixed solution of water and acetonitrile for the sunitinib malate test sample, diluted, and the final concentration is 0.2 μg/mL～ 2.0 μg/mL. 3. analytical method according to claim 2, the volume ratio of water and acetonitrile in the mixed solution of wherein said water and acetonitrile is 7:3. 4. An analytical method for related substances of sunitinib malate, characterized in that: The method is carried out on a high performance liquid chromatograph; The detector is a diode array detector, and the detection wavelength is 268nm; The liquid chromatography column is a Poroshell120BonusRP reversed-phase column, and the column temperature is 25°C; The flow rate is about 1.0 mL/min; Gradient elution: Inject sunitinib malate test product solution and sunitinib malate reference substance solution for detection, and calculate the content of related substances by the principal component self-control method without correction factor; Among them, the Poroshell120BonusRP reversed-phase chromatographic column is filled with filler particles with a particle size of about 2.7 μm, wherein the filler particles are composed of an inner solid core with a diameter of 1.7 μm and a porous outer layer with a thickness of 0.5 μm coated on the inner solid core. , wherein the stationary phase is n-octadecane bonded to the packing particles, the length of the column is 50 mm, and the inner diameter of the column is 4.6 mm; Wherein the concentration of ammonium acetate aqueous solution is about 3.85g/L, and the pH is about 5.7; Among them, the sunitinib malate test solution is a mixed solution of acetonitrile and water with a sunitinib malate concentration of about 0.50 mg/mL, and the sunitinib malate reference solution is a sunitinib malate concentration A mixed solution of acetonitrile and water of about 1.0 μg/mL, wherein in the mixed solution of water and acetonitrile, the volume ratio of water and acetonitrile is 7:3; The method described herein is used to detect related substance B, related substance C or related substance E in the sunitinib malate bulk drug. 5. The analysis method according to any one of claims 1-4, wherein said high performance liquid chromatograph is Agilent1260, Agilent1290 or WatersUPLC. 6. The analysis method according to claim 5, wherein the method is used to detect related substance B, related substance C and related substance E in the sunitinib malate bulk drug.