Analysis method of enzaloutamine

Abstract

The invention provides an analysis method of Enzalocamide. Specifically, the analysis method of the enzaloutamine disclosed by the invention comprises the following step: analyzing a to-be-detected object containing the enzaloutamine by adopting a high performance liquid chromatography. According to the high performance liquid chromatography analysis method, all peaks can be completely separated, the accuracy is high, the stability is high, the sensitivity is good, a technical basis is provided for quality detection of an Enzaloutamide raw material medicine, based on the AQbD concept, specificity, quantitation limit, detection limit, linearity, precision, accuracy, solution stability and method durability verification is carried out on the method, and the method has good application prospects. The result shows that the method is high in specificity, good in reproducibility and high in analysis efficiency.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical analysis, and particularly relates to an analytical method for enzalutamide. Background Art

[0002] Enzalutamide belongs to an oral selective inhibitor of androgen receptor (AR), and is used for the treatment of metastatic hormone-sensitive prostate cancer and non-metastatic castration-resistant prostate cancer, with the trade name Xtandi. Its chemical name is 4-{3-[4-cyano-3-(trifluoromethyl)phenyl]-5,5-dimethyl-4-oxo-2-thioxoimidazol-1-yl}-2-fluoro-N-methylbenzamide, and its structural formula is shown in Formula I.

[0003]

[0004] The International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use (ICH) proposed Quality by Design (QbD) in ICH Q8 released in 2009, and applied this concept to the establishment, evaluation and optimization of analytical methods, which is called Analytical Quality by Design (AQbD). By applying scientific knowledge, experimental design and statistical analysis, the functional relationship between key quality attributes and key analytical parameters is studied to find the analytical design space. Compared with the traditional trial-and-error method with single-factor variables, the AQbD concept has the advantages of being more scientific, more comprehensive in investigating influencing factors, and more economical and efficient. The QbD concept is mainly used in the research and development and production of new drugs, but it is less applied in the research and development of drug analytical methods.

[0005] At present, the detection methods for enzalutamide are not included in the pharmacopoeias of various countries. Therefore, it is urgent to develop a detection method for enzalutamide, which is of great significance for the quality control in the production process of its drugs. Summary of the Invention

[0006] The technical problem to be solved by the present invention is to overcome the problem of few detection methods for enzalutamide in the prior art, so as to provide an analytical method for enzalutamide. This method can achieve complete separation between peaks, with high accuracy, strong stability and good sensitivity.

[0007] The present invention provides an analytical method for enzalutamide, which comprises the following steps: analyzing the analyte containing enzalutamide by high performance liquid chromatography.

[0008] Among them, the conditions of the high performance liquid chromatography are as follows:

[0009] The chromatographic column is a chromatographic column with octadecylsilyl bonded silica as the stationary phase or a chromatographic column with octylsilyl bonded silica as the stationary phase;

[0010] Mobile phase A is a phosphoric acid solution; mobile phase B is a nitrile solvent;

[0011] The elution gradient of the high performance liquid chromatography method includes:

[0012] (1) The volume fraction of the nitrile solvent increases from an initial volume fraction of 15% - 40% to a second volume fraction of 50% - 60%; the increasing rate of change is 0.5 - 1.5% / min.

[0013] In one embodiment, the chromatographic analysis further includes step (2) where the volume fraction of the nitrile solvent increases from the second volume fraction of 50% - 60% to a third volume fraction of 90 - 95%; the increasing rate of change is 5 - 10% / min.

[0014] In one embodiment, the chromatographic analysis further includes step (3), where the volume fraction of the nitrile solvent is maintained at 90% - 95% within 0 - 10 min.

[0015] In one embodiment, in step (1), the initial volume fraction can be 20% - 35%, such as 20%, 33% or 35%.

[0016] In one embodiment, in step (1) or (2), the second volume fraction can be 58%.

[0017] In one embodiment, in step (1), the increasing rate of change can be 0.5% / min, 1.0% / min or 1.5% / min.

[0018] In one embodiment, in step (2), the increasing rate of change can be 9.25% / min or 7.4% / min.

[0019] The analyte containing enzalutamide further contains one or more of the following substances:

[0020] Compound IM1:

[0021] Compound IM2:

[0022] Compound IM3:

[0023] Compound IM4:

[0024] Compound IM5:

[0025] Compound IM6:

[0026] Compound IM8:

[0027] In one embodiment, the mobile phase A is an aqueous phosphoric acid solution, and further preferably a 0.1% aqueous phosphoric acid solution.

[0028] In one embodiment, the nitrile solvent is acetonitrile.

[0029] In one embodiment, the packing particle size of the chromatographic column is 2.5 μm to 7.5 μm, and more preferably 5 μm.

[0030] In one embodiment, the length of the chromatographic column is 100 mm to 250 mm, more preferably 150 mm to 250 mm, and even more preferably 250 mm.

[0031] In one embodiment, the inner diameter of the chromatographic column is 2 mm to 5 mm, more preferably 2.1 mm to 4.6 mm, and even more preferably 4.6 mm.

[0032] In one embodiment, the stationary phase is octadecylsilane-bonded silica gel Kromasil 100-5C18; the Kromasil 100-5C18 has a specification of 4.6 mm × 250 mm, 5 μm.

[0033] In one embodiment, the chromatographic column with octylsilane-bonded silica gel as the stationary phase is Agilent ZORBAX Eclipse Plus C8; the Agilent ZORBAX Eclipse Plus C8 has a specification of 4.6 mm × 250 mm, 5 μm.

[0034] In one embodiment, the injection volume of the high performance liquid chromatography is 5 μL to 20 μL, more preferably 10 μL to 20 μL, and even more preferably 10 μL;

[0035] In one embodiment, the detector of the high performance liquid chromatography is an ultraviolet detector and / or a diode array detector, more preferably an ultraviolet detector or a diode array detector.

[0036] In one embodiment, the column temperature of the high performance liquid chromatography is the conventional column temperature of high performance liquid chromatography A in the art, preferably 20 to 40 °C, and more preferably 30 °C.

[0037] In one embodiment, the flow rate of the high performance liquid chromatography is the conventional flow rate of high performance liquid chromatography A in the art, preferably 0.8 mL / min - 2.0 mL / min, more preferably 1.0 mL / min - 1.1 mL / min, and even more preferably 1.0 mL / min.

[0038] In one embodiment, the detection wavelength of the high performance liquid chromatography A is 210 nm to 280 nm, more preferably 210 nm to 240 nm, and even more preferably 218 nm.

[0039] In one embodiment, the high performance liquid chromatography preferably has the following conditions:

[0040] The chromatographic column is Kromasil 100-5 C18;

[0041] The mobile phase: Mobile phase A is an aqueous phosphoric acid solution; Mobile phase B is acetonitrile;

[0042] The detection wavelength is 218 nm;

[0043] The column temperature is 30 °C;

[0044] The injection volume is 10 μL.

[0045] In one embodiment, the gradient of the high performance liquid chromatography preferably has the following options from Option 1 to Option 7:

[0046] Option 1,

[0047]

[0048] Option 2,

[0049]

[0050] Option 3,

[0051]

[0052]

[0053] Option 4

[0054]

[0055] Option 5

[0056]

[0057] Option 6

[0058]

[0059] Option 7

[0060]

[0061] Even more preferably

[0062]

[0063] The present invention provides an application of the above-mentioned analytical method for enzalutamide in calculating the contents of compound IM1, compound IM2, compound IM3, compound IM4, compound IM6 or compound IM8 in enzalutamide, which comprises the following steps: separating and detecting compound IM1, compound IM2, compound IM3, compound IM4, compound IM5, compound IM6 or compound IM8 by using the above-mentioned analytical method for enzalutamide, and calculating the contents of compound IM1, compound IM2, compound IM3, compound IM4, compound IM6 or compound IM8 in enzalutamide by using the self-control method of 1% main component with correction factor;

[0064] Compound IM1:

[0065] Compound IM2:

[0066] Compound IM3:

[0067] Compound IM4:

[0068] Compound IM5:

[0069] Compound IM6:

[0070] Compound IM8:

[0071] In a certain embodiment, in the self-control method of 1% main component with correction factor: the correction factor of compound IM1 is 1.56; and / or the correction factor of compound IM2 is 1.41; and / or the correction factor of compound IM3 is 1.06; and / or the correction factor of compound IM4 is 0.92; and / or the correction factor of compound IM5 is 0.89; and / or the correction factor of compound IM6 is 0.92; and / or the correction factor of compound IM8 is 0.93.

[0072] The present invention also provides a method for establishing an analytical model of enzalutamide, which comprises the following steps:

[0073] S1: Rough screening is carried out on each parameter of the enzalutamide analytical method;

[0074] S2: According to the QbD concept, on the basis of S1, key analytical parameters and key quality attributes are determined through risk assessment, and single-factor experiments on key analytical parameters are carried out;

[0075] S3: Parameter optimization is carried out to establish a functional relationship between key analytical parameters and key quality attributes, and a maximized willing analytical method is determined.

[0076] In a certain solution, in step S3, the parameter optimization is preferably carried out by using the response surface design of JMP software.

[0077] On the basis of not violating the common knowledge in this field, the above preferred conditions can be combined arbitrarily to obtain various preferred examples of the present invention.

[0078] The reagents and raw materials used in the present invention are all commercially available.

[0079] The positive and progressive effects of the present invention are as follows: The high-performance liquid chromatography analysis method provided by the present invention can achieve complete separation between peaks, with high accuracy, strong stability and good sensitivity, providing a technical basis for the quality detection of enzalutamide raw materials. Based on the AQbD concept, the specificity, quantification limit, detection limit, linearity, precision, accuracy, solution stability and method durability of this method were verified. The results show that this method has strong specificity, good reproducibility and high analysis efficiency. Description of the Drawings

[0080] Figure 1 It is the representative chromatogram of the method for enzalutamide raw materials - the results of the final method parameters.

[0081] Figure 2 It is the chromatogram of the method development for enzalutamide raw materials - the screenshot of QBD maximization willingness.

[0082] Figure 3 It is the ultraviolet absorption spectrum of enzalutamide raw materials.

[0083] Figure 4 It is the ultraviolet absorption spectrum of compound IM1.

[0084] Figure 5 It is the ultraviolet absorption spectrum of compound IM2.

[0085] Figure 6 It is the ultraviolet absorption spectrum of compound IM3.

[0086] Figure 7 It is the ultraviolet absorption spectrum of compound IM4.

[0087] Figure 8 It is the ultraviolet absorption spectrum of compound IM5.

[0088] Figure 9 It is the ultraviolet absorption spectrum of compound IM6. Detailed Embodiments

[0089] The present invention will be further described below by way of examples, but the present invention is not limited to the scope of the examples described herein. For the experimental methods without specific conditions indicated in the following examples, they are carried out according to conventional methods and conditions, or selected according to the product specifications.

[0090] Materials and Reagents

[0091] Name Level Batch Number Source Phosphoric Acid HPLC BCCF1913 Sigma-aldrich Acetonitrile HPLC WXBD4680V Sigma-aldrich Acetonitrile HPLC WXBD4680V Sigma-aldrich Acetonitrile HPLC WXBD4680V Sigma-aldrich Purified Water Ultra-pure Water PWG-01-20210911 Water Purifier Phosphoric Acid GR 20200708-00902 Sinopharm Acetonitrile HPLC V30A1H-10901 Honeywell Water N / A N / A Wahaha

[0092] Samples and Reference Substances

[0093]

[0094] Instruments and Equipment

[0095]

[0096]

[0097] Preparation of Solutions:

[0098] Preparation of Test Solution: Take the test enzalutamide sample, dilute the sample with the diluent to prepare a test solution with a concentration of 0.1 - 1.0 mg / mL;

[0099] Preparation of Reference Solution: Take the enzalutamide reference substance, dilute the sample with the diluent to prepare a reference solution with a concentration of 0.1 - 1.0 mg / mL.

[0100] Example 1 Selection of Chromatographic Column

[0101] Diluent: Acetonitrile: Purified Water = 90:10 (v / v)

[0102] Mixed Impurity Stock Solution: Weigh approximately 5 mg of each of the reference substances of Compound IM1, Compound IM2, Compound IM3, Compound IM4, Compound IM6, and Compound IM8, place them in the same 20 mL volumetric flask, dissolve with the diluent and make up to the mark, shake well, as the mixed impurity stock solution.

[0103] Impurity Stock Solution I: Pipette 2.5 mL of the mixed impurity stock solution into a 50 mL volumetric flask, make up to the mark with the diluent, shake well, as Impurity Stock Solution I.

[0104] Resolution Solution: Weigh approximately 25 mg of the enzalutamide bulk drug reference substance into a 25 mL volumetric flask, add 3 mL of Impurity Stock Solution I to this flask, then make up to the mark with the diluent, shake well, as the resolution solution.

[0105] Take the resolution solution, use the chromatographic columns in Table 1 respectively, and carry out the determination according to the chromatographic conditions in Table 2, record the chromatogram.

[0106] Analyze the structural characteristics and chemical properties of the enzalutamide bulk drug. Different characteristic functional groups are contained in its molecular structure, making the polarity of the compound and its binding ability with the stationary phase different. Combining with the performance characteristics of the chromatographic column, four different types of chromatographic columns (detailed information is shown in the following table) are selected for method development, and the specific parameters and results are as follows.

[0107] Table 1 Selection of Chromatographic Columns (Information on Four Different Types of Chromatographic Columns)

[0108]

[0109]

[0110] Table 2 Method Development Parameters for Enzalutamide API - Other Parameters Except Chromatographic Column

[0111]

[0112] According to the analysis results, it can be seen that:

[0113] When using Waters XBridge Phenyl column, the retention of each component is poor. The resolution between compound IM6 and the adjacent peak is 0.6, the resolution between compound IM2 and the adjacent peak is 0.8, and the resolution between compound IM3 and compound IM1 is 1.1. The resolutions are all less than 1.5, not meeting the detection requirements.

[0114] When using Agilent Zorbax Bonus - RP column, the main peak and each compound have good retention. The asymmetry factor of compound IM3 is 1.67, and the peak of compound IM8 is front - extended.

[0115] When using Agilent ZORBAX Eclipse Plus C8 column, the retention of each component is good, and the resolution between each known impurity and the adjacent peak is greater than 1.5.

[0116] When using Kromasil 100 - 5C18 column, the retention of each component is good, and the resolution between each known impurity and the adjacent peak is greater than 1.5.

[0117] In summary, both Agilent ZORBAX Eclipse Plus C8 column and Kromasil 100 - 5C18 column can meet the detection of enzalutamide API. Select Kromasil 100 - 5C18 column as a representative for method optimization and verification.

[0118] Example 2 Selection of Mobile Phase System

[0119] Take the resolution solution in Example 1 and inject samples for analysis according to the chromatographic conditions in Table 3, Table 4 and Table 5. To study the mobile phase system of the analysis method, compare the trifluoroacetic acid system and the phosphoric acid system. The specific parameters and analysis results are as follows:

[0120] Table 3 Selection of Mobile Phase

[0121]

[0122] Table 4 Method Development Parameters of Enzalutamide API - Trifluoroacetic Acid System

[0123]

[0124] Table 5 Method Development Parameters of Enzalutamide API - Phosphoric Acid System

[0125]

[0126]

[0127] According to the analysis results, under the trifluoroacetic acid system, the resolution between compound IM2 and compound IM4 is 1.6, and the resolution between the main peak and compound IM3 is 3.0. However, after 35 minutes, the baseline drifts significantly, and compound IM8 has strong retention and elutes at the end of the gradient peak. Under the phosphoric acid system, the resolution between compound IM2 and compound IM4 is 7.1, and the resolution between the main peak and compound IM3 is 8.4, and the baseline is relatively flat. Therefore, the phosphoric acid system is selected as the mobile phase system for the determination of enzalutamide API.

[0128] Example 3 Optimization of Mobile Phase Elution Gradient

[0129] Take the separated solution in Example 1 and inject it for analysis according to the chromatographic conditions in Tables 6 - 9.

[0130] In the initial stage of method development, a general gradient was used for analysis, and the specific parameters and results are as follows.

[0131] Table 6 Method Development Parameters of Enzalutamide API - General Gradient

[0132]

[0133] Under the initial gradient parameters, the resolution between compound IM2 and compound IM4 is 1.33, and the resolution is less than 1.5, which does not meet the detection requirements. And under this condition, the compounds are relatively concentrated, eluting between 21 - 27 minutes, and only compound IM5 is detected between 5 - 16 minutes. Therefore, the mobile phase elution gradient was optimized, and the specific parameters and results are as follows.

[0134] Table 7 Method Development Parameters of Enzalutamide API - Gradient Optimization

[0135]

[0136] After optimizing the gradient parameters, the distribution of the main peak and impurities is relatively reasonable, and the resolution between the main peak and each impurity is greater than 1.5, meeting the detection requirements. However, in this method system, the elution time of compound IM5 is 2.931 min, with weak retention, and basically no impurities are detected after 31 min. Therefore, the method needs to be optimized, and the specific parameters and results are as follows.

[0137] Table 8 Enzalutamide API Method Development Parameters - Gradient Optimization II

[0138]

[0139]

[0140] After optimizing the gradient parameters, the elution time of compound IM5 is 13.230 min, with good retention. The distribution of the main peak and other known impurities is also relatively reasonable, and the resolution between the main peak and each known impurity is greater than 1.5, meeting the detection requirements.

[0141] Based on the QbD concept, on the basis of determining the method gradient parameters (see Table 9), the analysis method objectives are converted into key method attributes that can be quantitatively evaluated. Data are obtained through experimental design, and a design space for the gradient change of the analysis method is established by combining experimental design and statistical analysis. Finally, a control strategy for the analysis method is established. Therefore, the elution program of the mobile phase is optimized, and the effects of the initial ratio and change rate of different mobile phases on the separation of each component and the chromatographic peak shape are investigated respectively. The specific parameters and results are as follows.

[0142] Table 9 Selection of Initial Proportion and Change Rate of Organic Phase

[0143]

[0144]

[0145] Table 10 Resolution Results under Different Elution Gradient Conditions

[0146]

[0147] Table 11 Asymmetry Factor Results under Different Elution Gradient Conditions

[0148]

[0149] Note: "n.a." in the table indicates that the peak is split and cannot be read.

[0150] Input the above results into the JMP software and fit by the least squares method to predict the maximum willingness. The results are as Figure 2 . Among them, the names represented by Y~Y9, X1, and X2 in the QBD maximum willingness diagram are shown in Table 12 below:

[0151] Schematic diagram of parameters in the 12QBD maximization willingness diagram

[0152]

[0153]

[0154] Comparison of the results of the minimum resolution between each impurity and the adjacent peak before screening and the 13QBD maximization willingness

[0155] Name Before Screening After QBD Screening Compound IM5 5.8 19.8 Compound IM6 4.8 8.2 Compound IM2 1.6 2.7 Compound IM4 1.6 2.7 Enzalutamide API 3.2 6.2 Compound IM3 3.2 6.2 Compound IM1 10.1 20.4 Compound IM8 20.6 26.5

[0156] As shown above, the maximization willingness was obtained according to software fitting, that is, when the initial proportion of the organic phase was 32.5% and the change rate was 1% per minute, the result was the best. Therefore, a gradient elution program method with an initial proportion of 33% (rounded) of the organic phase and a change rate of 1% per minute was used for analysis, and the minimum resolution between each impurity and the adjacent peak under the gradient elution program conditions before screening was compared. The results are shown in Table 13. Under the gradient elution program obtained by the QBD maximization willingness, the minimum resolution between each known impurity and the adjacent peak was better than that under the gradient elution program before screening. Therefore, the initial proportion of the gradient elution program for the determination of enzalutamide raw material drug was 33% and the change rate was 1%.

[0157] Example 4 Column temperature selection

[0158] During the method development process, the resolution solution in Example 1 was taken. When the method parameters in Table 8 were used, compound IM2 and compound IM4, and compound IM3 and compound IM1 were relatively close. The separation of the main peak and each impurity was investigated under different column temperatures (30 °C, 35 °C, 40 °C).

[0159] The resolution between compound IM2 and compound IM4, and compound IM1 and compound IM3 gradually decreased with the increase of the column temperature (the specific resolution results are shown in Table 14). Under the condition of a column temperature of 30 °C, the resolution was greater than 1.5, meeting the detection requirements. Under the conditions of a column temperature of 35 °C and 40 °C, the resolution between compound IM1 and compound IM3 was less than 1.5, not meeting the detection requirements. Therefore, 30 °C was finally selected as the column temperature for the determination of enzalutamide raw material drug.

[0160] Table 14 Impurity resolution under different column temperature conditions

[0161]

[0162] Example 5 Wavelength selection

[0163] According to the ultraviolet absorption spectra of enzalutamide raw material drug and compounds (see Figures 3 - 9) It has strong absorption at 218 nm, 235 nm, 254 nm and 270 nm. By comparing the correction factors of each compound at the four wavelengths (see Table 15), the correction factors of each known impurity are closer to 1.0 at 218 nm wavelength. Also, according to the results of the spiked solution impurities (see Table 16), at 218 nm wavelength, the number of detected impurities is 6, while at 235 nm wavelength it is 4, at 254 nm wavelength it is 1, and at 270 nm wavelength it is 1. Therefore, the impurity detection ability is stronger at 218 nm wavelength.

[0164] In summary, 218 nm was finally selected as the determination wavelength for enzalutamide raw material.

[0165] Table 15 Correction factors of each known impurity at different wavelengths

[0166] Impurity Name 218nm 235nm 254nm 270nm Compound IM1 1.51 1.42 6.63 2.30 Compound IM2 1.59 1.73 1.05 1.11 Compound IM3 1.12 1.13 1.14 1.13 Compound IM4 0.96 0.98 0.93 0.98 Compound IM5 0.89 24.3 2.87 0.52 Compound IM6 1.16 2.20 2.32 1.32

[0167] Table 16 Results of impurity contents in the spiked solution of enzalutamide raw material

[0168]

[0169]

[0170] Note: "N.D." in the table indicates that the compound was not detected, and "N / A" indicates not applicable here.

[0171] Summary of the finally determined method parameters and representative spectra in Example 6

[0172] The finally determined detection method for enzalutamide raw material is shown in Table 17, and the detection results are as Figure 1 , and the retention times of each peak are shown in Table 18.

[0173] Table 17 Final parameters of the method for enzalutamide raw material

[0174]

[0175]

[0176] Table 18 Elution times of each substance

[0177] Chromatographic Peak Number Name Retention Time (min) Peak 4 XT166-B 17.478 Peak 5 XT166-C-IM6 18.812 Peak 7 XT166-C-IM2 20.958 Peak 8 XT166-C-IM4 21.665 Peak 9 Main Component XT166-C 24.052 Peak 10 XT166-C-IM3 25.445 Peak 11 XT166-C-IM1 29.692 Peak 14 XT166-C-IM8 32.678

[0178] During the development of the enzalutamide API method, phenol was not studied. It was originally planned to control phenol as a genotoxic impurity, but after consulting the NTP data, it was found that phenol is a non-carcinogenic impurity, so it can be controlled as a common impurity. Therefore, the applicability of phenol was investigated in the final determination method of enzalutamide API. Phenol was well retained, with a separation degree of 15.8 between adjacent peaks, an asymmetry factor of 1.07, and a ratio of the response value of the main peak to phenol of 1.1. Therefore, phenol can be determined in the determination method of enzalutamide API and controlled as an unknown impurity.

[0179] According to the validation plan, HPLC-UV method was adopted, with Kromasil 100-5C18 (250mm*4.6mm, 5μm) as the chromatographic column, column temperature of 30°C, wavelength of 218nm, flow rate of 1.0mL / min, sample plate temperature of 4°C, injection volume of 10μL, and 0.1% phosphoric acid aqueous solution and acetonitrile as mobile phases for gradient elution. The determination method of enzalutamide raw materials was validated by the known methods of pharmaceutical validation. The specificity, detection limit, quantification limit, linearity / correction factor, accuracy, precision (repeatability, reproducibility of the transferor and recipient laboratories), range, and durability of the method all met the acceptable standards, proving that the method is applicable for the determination of enzalutamide raw materials in both the transferor and recipient laboratories.

Claims

1. An analytical method for enzalutamide, characterized in that It includes the following steps: Just analyze the analyte containing enzalutamide by high performance liquid chromatography; The chromatographic column is a chromatographic column with octadecylsilyl bonded silica gel as the stationary phase or a chromatographic column with octylsilyl bonded silica gel as the stationary phase; Mobile phase A is a phosphoric acid solution; Mobile phase B is a nitrile solvent; The elution gradient of the high performance liquid chromatography includes: (1) The volume fraction of the nitrile solvent increases from an initial volume fraction of 15% - 40% to a second volume fraction of 50% - 60%; The increasing change rate is 0.5 - 1.5% / min.

2. The analysis method according to claim 1, wherein The analysis method satisfies one or more of the following conditions: (1) The chromatographic analysis further includes step (2) where the volume fraction of the nitrile solvent increases from the second volume fraction of 50% - 60% to a third volume fraction of 90 - 95%; The increasing change rate is 5 - 10% / min; (2) The chromatographic analysis further includes step (3) where the volume fraction of the nitrile solvent is maintained at 90% - 95% within 0 - 10 min; (3) In the step (1), the initial volume fraction is 20% - 35%; (4) In the step (1), the increasing change rate is 0.5% / min, 1.0% / min or 1.5% / min; and (5) The analyte containing enzalutamide contains one or more of the following substances: Compound IM1: Compound IM2: Compound IM3: Compound IM4: Compound IM5: Compound IM6: Compound IM8:

3. The analysis method according to claim 1 or 2, characterized in that The analysis method satisfies one or more of the following conditions: (1) The mobile phase A is an aqueous phosphoric acid solution; (2) The nitrile solvent is acetonitrile; (3) The filler particle size of the chromatographic column is 2.5μm - 7.5μm; (4) The length of the chromatographic column is 100mm - 250mm; (5) The inner diameter of the chromatographic column is 2mm - 5mm; (6) The chromatographic column with octadecylsilyl bonded silica gel as the filler is Kromasil 100 - 5C18; (7) The chromatographic column with octylsilyl bonded silica gel as the filler is Agilent ZORBAX Eclipse Plus C8; (8) The injection volume of the high performance liquid chromatography is 5μL - 20μL; (9) The detector of the high performance liquid chromatography is an ultraviolet detector and / or a diode array detector; (10) The column temperature of the high performance liquid chromatography is 20 - 40°C; (11) The flow rate of the high performance liquid chromatography is 0.8mL / min - 2.0mL / min; (12) The detection wavelength of the high performance liquid chromatography A is 210nm - 280nm; (13) In the step (1), the initial volume fraction is 20%, 33% or 35%; (14) In the step (1) or (2), the second volume fraction is 58%; and (15) In the step (2), the increasing change rate is 9.25% / min or 7.4% / min.

4. The analysis method according to claim 3, characterized in that, The analysis method satisfies one or more of the following conditions: (1) The mobile phase A is 0.1% aqueous phosphoric acid solution; (2) The packing particle size of the chromatographic column is 5 μm; (3) The length of the chromatographic column is 150 mm to 250 mm; (4) The inner diameter of the chromatographic column is 2.1 mm to 4.6 mm; (5) The specification of Kromasil 100-5C18 is 4.6 mm × 250 mm, 5 μm; (6) The specification of Agilent ZORBAX Eclipse Plus C8 is 4.6 mm × 250 mm, 5 μm; (7) The injection volume of the high performance liquid chromatography is 10 μL to 20 μL; (8) The detector of the high performance liquid chromatography is an ultraviolet detector or a diode array detector; (9) The column temperature of the high performance liquid chromatography is 30 °C; (10) The flow rate of the high performance liquid chromatography is 1.0 mL / min - 1.1 mL / min; (11) The detection wavelength of high performance liquid chromatography A is 210 nm to 240 nm.

5. The analysis method according to claim 1, characterized in that The analysis method satisfies one or more of the following conditions: (1) The length of the chromatographic column is 50 mm; (2) The inner diameter of the chromatographic column is 4.6 mm; (3) The injection volume of the high performance liquid chromatography is 10 μL; (4) The flow rate of the high performance liquid chromatography is 1.0 mL / min; (5) The detection wavelength of high performance liquid chromatography A is 218 nm; (6) The gradient of the high performance liquid chromatography is Scheme 1 - Scheme 7: Scheme 1, Scheme 2, Scheme 3, Scheme 4 Scheme 5 Scheme 6 Scheme 7 6. The analysis method according to claim 1, characterized in that, The high performance liquid chromatography includes the following conditions: The chromatographic column is Kromasil 100-5C18; The mobile phase: Mobile phase A is an aqueous phosphoric acid solution; Mobile phase B is acetonitrile; The detection wavelength is 218 nm; The column temperature is 30 °C; The injection volume is 10 μL; The gradient of the high performance liquid chromatography is:

7. Use of the analysis method according to any one of 1-6 in calculating the content of compound IM1, compound IM2, compound IM3, compound IM4, compound IM6 or compound IM8 in enzalutamide, comprising the following steps: Using the analysis method described in any one of 1 - 6 to separate and detect Compound IM1, Compound IM2, Compound IM3, Compound IM4, Compound IM5, Compound IM6 or Compound IM8, and using the 1% main component self-control method with correction factor to calculate the contents of Compound IM1, Compound IM2, Compound IM3, Compound IM4, Compound IM6 or Compound IM8 in enzalutamide; Compound IM1: Compound IM2: Compound IM3: Compound IM4: Compound IM5: Compound IM6: Compound IM8:

8. The application according to claim 7, characterized in that In the 1% main component self-control method with correction factor: The correction factor of Compound IM1 is 1.56; and / or the correction factor of Compound IM2 is 1.41; and / or the correction factor of Compound IM3 is 1.06; and / or the correction factor of Compound IM4 is 0.92; and / or the correction factor of Compound IM5 is 0.89; and / or the correction factor of Compound IM6 is 0.92; and / or the correction factor of Compound IM8 is 0.

93.

9. A method for establishing an analytical model of enzalutamide, characterized in that, It includes the following steps: S1: Conduct a preliminary screening on each parameter of the enzalutamide analysis method; S2: Based on QbD concept, on the basis of S1, determine the critical analysis parameters and critical quality attributes through risk assessment, and conduct single factor experiments on the critical analysis parameters; S3: Parameter optimization, establishing the functional relationship between key analysis parameters and key quality attributes, and determining the analysis method for maximizing willingness.

10. The method according to claim 9, characterized in that, In step S3, the parameter optimization is carried out by using the response surface design of JMP software.

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