Method for detecting olaparil related substances

Abstract

The invention belongs to the technical field of pharmaceutical analytical chemistry, and discloses a method for detecting olaparil related substances. Olaparil is subjected to related substance detection by adopting a high performance liquid chromatography, an octadecylsilane chemically bonded silica chromatographic column is used, and a mobile phase A is a water phase which is a phosphate buffer solution; a mobile phase B is an organic phase: acetonitrile; and carrying out gradient elution. According to the novel liquid phase detection method for the olaparil related substances, potential olaparil processes and degraded impurities can be effectively detected, the separation degree between olaparil and adjacent impurities and the separation degree between impurities meet the requirements, and methodology verification shows that the sensitivity is high, and specificity and repeatability are good. And the method has important significance for realizing quality control of olaparil.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical analytical chemistry, and more specifically to a method for detecting Olaparib and related substances. Background Art

[0002] Ovarian cancer (OC) is the most malignant tumor among gynecological tumors. Most patients are diagnosed at an advanced stage. Ovarian cancer is highly malignant, has a high recurrence rate, and is difficult to treat. Currently, the main clinical treatment for advanced ovarian cancer patients is surgery combined with chemotherapy, but the recurrence rate within 1 to 2 years is as high as 70% to 80%. In addition, the mortality rate of ovarian cancer ranks first among female reproductive system tumors. After multiple recurrences, the resistance to chemotherapy drugs increases, and the 5-year survival rate is only 47.4%.

[0003] As the world's first approved PARP inhibitor, Olaparib uses defects in the tumor DNA damage response pathway to preferentially kill cancer cells. The drug was approved for marketing by the U.S. Food and Drug Administration in December 2014 for patients with advanced ovarian cancer who have received more than two anti-tumor treatments and carry germline mutations in the breast cancer susceptibility gene (BRCA). It was subsequently approved in the European Union, Japan and other places, and its indications have been continuously expanded, currently covering breast cancer, ovarian cancer, fallopian tube cancer, peritoneal cancer, prostate cancer and pancreatic cancer.

[0004] Olaparib, English name: Olaparib, CAS number is 763113-22-0, molecular formula is C 24 H 23 FN4O3, molecular weight 434.46; white to slightly yellow powder; almost insoluble in water, non-hygroscopic; logP = 1.49; structural formula is as follows:

[0005]

[0006] Patent CN116930347A discloses an impurity detection method for Olaparib API, which can simultaneously determine three specific impurities in Olaparib API. However, this method uses citrate as the mobile phase, which may cause damage to the instrument and the chromatographic column. Although this method has good separation, the tailing factor of the chromatographic peak and the partial overlap with adjacent impurities still need to be improved.

[0007] Patent CN117470978A discloses a method for detecting related substances of Olaparib API, which can simultaneously determine seven impurities. However, the methodological validation limit of this method is relatively loose, and the chromatogram of the system suitability solution has the problem of partial peak overlap, which needs further optimization.

[0008] Based on the existing reported detection methods for Olaparib-related substances, the present invention selects acetonitrile and phosphoric acid solution as mobile phases, and establishes a method with high separation, good accuracy and simple operation. In order to ensure the safety, effectiveness and quality control of Olaparib drugs, a scientific and reasonable analytical method that can effectively control the intrinsic quality of drugs is established. Summary of the invention

[0009] In view of this, the present invention provides a detection method for Olaparib and related substances, which adopts specific liquid chromatography analysis conditions to separate Olaparib and four related substances, and can effectively achieve the separation and quantitative determination of Olaparib and the four related substances, so as to reduce the impact of related substances on the quality of the final Olaparib product.

[0010] In order to achieve the above object, the present invention provides a method for detecting related substances of Olaparib, characterized in that high performance liquid chromatography is used, and the conditions include:

[0011] Chromatographic column: C18 column;

[0012] Filler: octadecylsilane bonded silica gel;

[0013] Mobile phase A: 0.01-1.0 wt% phosphoric acid solution;

[0014] Mobile phase B: 70-100 wt% acetonitrile solution;

[0015] Flow rate: 0.8~1.2ml / min;

[0016] Detection wavelength: 270~280nm;

[0017] Injection volume: 1-20 μL;

[0018] Gradient elution was used.

[0019] Preferably, the inner diameter of the C18 chromatographic column is 1 to 5 mm, the length is 100 to 300 mm, and the filler particle size is 1 to 20 μm.

[0020] Preferably, the gradient elution, in terms of volume percentage, has a gradient elution procedure as follows:

[0021] Time (minutes) Mobile phase A (%) Mobile phase B (%) 0～16 65～75 35～25 17～25 60～70 40～30 26～28 15～5 85～95 29～32 85～95 15～5 33～36 65～75 35～25 .

[0022] Further preferably, the gradient elution, in terms of volume percentage, has a gradient elution procedure as follows:

[0023]

[0024] Furthermore, the olaparib-related substances include olaparib and known impurities, and the known impurities include one or more of olaparib acid, defluoroolaparib, n-propylolaparib, and olaparib pip dimer, and the structural formula and chemical name are as follows:

[0025]

[0026]

[0027] Further, a method for detecting olaparib-related substances comprises the following steps:

[0028] (1) Sample preparation:

[0029] Preparation of reference substance stock solution: Accurately weigh Olaparib into a 10 mL volumetric flask, add diluent to dissolve and dilute to the scale, shake well to obtain the reference substance stock solution;

[0030] Preparation of related substance stock solutions: accurately weigh equal amounts of olaparib acid, defluoroolaparib, and n-propylolaparib into a 10 mL volumetric flask, add diluent to dissolve and dilute to the mark, and shake well; accurately weigh the same amount of olaparib pip dimer into a 10 mL volumetric flask, add appropriate amount of DMSO to dissolve it, then add pure acetonitrile to dilute to the mark, and shake well; the olaparib related substance stock solution is obtained;

[0031] Preparation of system suitability solution: Accurately measure the olaparib reference substance stock solution and olaparib related substance solution and place them in the same 10mL volumetric flask, dilute to the mark with diluent to serve as the system suitability solution;

[0032] (2) High Performance Liquid Chromatography Determination: Accurately measure the system suitability solution and inject it into the high performance liquid chromatograph. Perform the test according to the high performance liquid chromatography conditions described above, record the chromatogram and perform data analysis.

[0033] Preferably, in step (1), the concentration of Olaparib in the reference substance stock solution is 0.1-0.2 mg / mL, and the concentration of Olaparib in the related substance stock solution is 1-40 μg / mL.

[0034] Preferably, in step (1), the diluent is acetonitrile and water prepared in a volume ratio of (1-10):(1-10).

[0035] The beneficial effect of the above technical solution is that there is no significant difference in the influence of the composition ratio of the above diluent on the peak time and separation degree.

[0036] Preferably, in step (1), the solid-liquid ratio of the olaparib pip dimer to the DMSO is 50 mg:1 mL.

[0037] Furthermore, the impurity limit requirements are formulated according to the "9102 Guidelines for Drug Impurity Analysis" for the results of HPLC determination of the related substances of Olaparib, and the impurity peak areas in the results of HPLC determination of the related substances of Olaparib are calculated according to the external standard method of impurity reference substances.

[0038] Preferably, in the results of the high performance liquid chromatography determination, a single impurity does not exceed 0.15%, and a total impurity does not exceed 0.5%.

[0039] Preferably, the impurity reference substance external standard method comprises the following steps: (1) preparing linear solutions of different concentrations of each known impurity reference substance, and performing high performance liquid chromatography determination, and obtaining regression equations of each impurity and the main peak by using the numerical linear regression equation of the concentration of the linear solution and the numerical value of the corresponding peak area; (2) adding a known amount of the impurity reference substance to the sample to be tested to prepare a spiked test solution and its reference substance solution, and performing high performance liquid chromatography determination, and calculating the content of each impurity in the sample to be tested from the peak area (or peak height) of each impurity in the test solution; the correlation coefficient in each regression equation is ≥0.995.

[0040] Preferably, in step (2), in the results of the high performance liquid chromatography determination, the separation between the impurity peaks in the olaparib-related substances and between the impurity peaks and the main peak is >2.0, and the tailing factor of the olaparib and related substance peaks is not greater than 1.5.

[0041] The method of the present invention establishes a liquid phase analysis method for the four known related substances in the olaparib preparation, the method has strong specificity, high sensitivity, and accurate quantitative determination results; the analysis method is simple and fast, and all related substances can be separated well in a short time, and the separation between each chromatographic peak is greater than 1.5, and the related substances in olaparib and its preparation can be fully detected, the safety of the drug is improved, and the basis for formulating the quality standard of the olaparib preparation is improved. At the same time, the method of the present invention is easy to implement, the instruments and reagents used are all conventional products, there are no harsh conditions, the cost is low, and the conditions of most laboratories can be met.

[0042] It can be seen from the above technical solution that, compared with the prior art, the beneficial effects achieved by the present invention are:

[0043] 1. The present invention provides a method for detecting related substances of Olaparib, which is analyzed and detected by high performance liquid chromatography. It is a simple detection method with good separation effect, high precision and accuracy. The result is accurate and reliable, with a wide detection range, high sensitivity and strong accuracy. It is a commonly used detection method with extremely high recognition.

[0044] 2. The present invention provides a method for detecting related substances of Olaparib, which can accurately reflect the content of related substances in Olaparib and is suitable for the detection and quality control of related substances of Olaparib, thereby providing a basis for the formulation of reasonable quality standards, so as to better control and master product quality and ensure the safety of food and medicine.

[0045] 3. The present invention provides a method for detecting related substances of Olaparib, which optimizes the chromatographic conditions. Compared with the existing method for detecting related substances of Olaparib, the obtained results, whether the peak shape of impurities or the separation degree, have been significantly improved, thereby improving the detection accuracy. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying creative work.

[0047] Figure 1 Solution chromatography for system suitability Figure 1 ;

[0048] Figure 2 This is the chromatogram of Olaparib and related substances;

[0049] Figure 3 Solution chromatography for system suitability Figure 2 ;

[0050] Figure 4 is the linear relationship diagram of olaparib acid;

[0051] Figure 5 This is the linear relationship diagram of desfluoro-olaparib;

[0052] Figure 6 This is the linear relationship diagram of Olaparib pip dimer;

[0053] Figure 7 This is the linear relationship diagram of n-propyl olaparib. DETAILED DESCRIPTION

[0054] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0055] Reagent selection: Olaparib reference substance (Guangzhou Jiatu Technology Co., Ltd., catalog number: C4X-1135, purity: 99.7%), Olaparib acid standard substance (Guangzhou Jiatu Technology Co., Ltd., catalog number: C4X-113518, purity: 98.6%); Desfluoro Olaparib standard substance (Guangzhou Jiatu Technology Co., Ltd., catalog number: C4X-11356, purity: 99.95%); Olaparib pip dimer standard substance (Guangzhou Jiatu Technology Co., Ltd., catalog number: C4X-113511, Purity: 96.18%); n-propyl olaparib standard (Guangzhou Jiatu Technology Co., Ltd., product number: C4X-113518, purity: 99.17%); dimethyl sulfoxide (DMSO, BIOTOPPED, purity: 99.99%), phosphoric acid (Tianjin Fengchuan Chemical Reagent Technology Co., Ltd., phosphoric acid content not less than 85%), acetonitrile (Thermo Fisher Scientific (China) Co., Ltd., purity: 99.95%); distilled water; Wahaha purified water; acetonitrile is chromatographic grade, and other chemical reagents are analytical grade.

[0056] Embodiment 1:

[0057] Instruments and conditions:

[0058] Chromatograph: LC-2030C 3D Plus, Shimadzu Instrument (Suzhou) Co., Ltd.;

[0059] Detector: UV detector;

[0060] Chromatographic column: octadecylsilane bonded silica gel as filler (Dimosail, Inspire-C18 250 mm × 4.6 mm, 5 μm);

[0061] Column temperature: 25°C

[0062] Flow rate: 0.8ml / min

[0063] Injection volume: 5 μL

[0064] Detection wavelength: 276nm

[0065] Chromatographic conditions:

[0066]

[0067]

[0068] Sample preparation:

[0069] Preparation of reference substance stock solution: Accurately weigh 10 mg of Olaparib into a 10 mL volumetric flask, add acetonitrile: water (1:1) to dissolve and dilute to the scale, shake well to obtain the reference substance stock solution.

[0070] Preparation of related substance reference stock solution: Accurately weigh 10.0 mg of olaparib acid, defluoroolaparib, n-propylolaparib and olaparib pip dimer into a 10 mL volumetric flask, add acetonitrile: water (1:1) to dissolve and dilute to the scale; the olaparib related substance stock solution is obtained.

[0071] Preparation of system suitability solution: Accurately measure 0.3 mL of Olaparib reference stock solution and 0.03 mL of Olaparib related substance solution and place them in the same 10 mL volumetric flask. Dilute with acetonitrile: water (1:1) to make a solution containing approximately 30 μg of Olaparib and 3 μg of Olaparib acid and other related substances per 1 mL as the system suitability solution.

[0072] Experimental steps:

[0073] Take the system suitability solution and inject it into the liquid chromatograph according to the above chromatographic analysis conditions, and record the chromatogram. The results are shown in the attached Figure 1 The relative retention time of olaparib acid was 10.579min, the relative retention time of defluoroolaparib was 9.853min, the olaparib pip dimer did not elute, and the elute time of n-propylolaparib was 15.863min; the separation degree between each related substance peak and the main component peak was ≥1.5, and the separation effect was good, but this sample processing method did not allow the four related substances to be detected at the same time, and further optimization was needed.

[0074] Embodiment 2:

[0075] Instruments and conditions:

[0076] Chromatograph: LC-2030C 3D Plus, Shimadzu Instrument (Suzhou) Co., Ltd.;

[0077] Detector: UV detector;

[0078] Chromatographic column: octadecylsilane bonded silica gel as filler (Dimosail, Inspire-C18 250 mm × 4.6 mm, 5 μm);

[0079] Column temperature: 25°C

[0080] Injection volume: 5 μL

[0081] Flow rate: 0.8 mL / min

[0082] Detection wavelength: 276nm

[0083] Chromatographic conditions:

[0084]

[0085] Experimental steps:

[0086] Preparation of reference substance stock solution: Accurately weigh 10 mg of Olaparib into a 10 mL volumetric flask, add acetonitrile: water (1:1) to dissolve and dilute to the scale, shake well to obtain the reference substance stock solution.

[0087] Preparation of related substance reference stock solution: accurately weigh 10.0 mg of olaparib acid, defluoroolaparib, and n-propylolaparib into a 10 mL volumetric flask, add acetonitrile: water (1:1) to dissolve and dilute to the scale; accurately weigh 10.0 mg of olaparib pip dimer into a 10 mL volumetric flask, add 0.20 mL of DMSO solution to dissolve it, and then add pure acetonitrile to dilute to the scale; the olaparib related substance stock solution is obtained.

[0088] Inject the reference solution of each relevant substance separately, analyze according to the above chromatographic conditions, and record the chromatogram. The results are shown in the attached Figure 2 , Figure 2 The chromatograms of Olaparib and its related substances and blank solution. Figure 2 It shows that the method of the present invention can effectively detect Olaparib and its related substances, and can accurately perform quantitative detection to calculate the limits of the related substances.

[0089] Embodiment 3:

[0090] Instruments and conditions:

[0091] Chromatograph: LC-2030C 3D Plus, Shimadzu Instrument (Suzhou) Co., Ltd.;

[0092] Detector: UV detector;

[0093] Chromatographic column: octadecylsilane bonded silica gel as filler (Dimosail, Inspire-C18 250 mm × 4.6 mm, 5 μm);

[0094] Column temperature: 25°C

[0095] Injection volume: 5 μL

[0096] Flow rate: 0.8 mL / min

[0097] Detection wavelength: 276nm

[0098] Chromatographic conditions:

[0099]

[0100] Sample preparation:

[0101] Preparation of reference substance stock solution: Accurately weigh 10 mg of Olaparib into a 10 mL volumetric flask, add acetonitrile: water (1:1) to dissolve and dilute to the scale, shake well to obtain the reference substance stock solution.

[0102] Preparation of related substance reference stock solution: accurately weigh 10.0 mg of olaparib acid, defluoroolaparib, and n-propylolaparib into a 10 mL volumetric flask, add acetonitrile: water (1:1) to dissolve and dilute to the scale; accurately weigh 10.0 mg of olaparib pip dimer into a 10 mL volumetric flask, add 0.20 ml of DMSO solution to dissolve it, and then add pure acetonitrile to dilute to the scale; the olaparib related substance stock solution is obtained.

[0103] Preparation of system suitability solution: Accurately measure 0.3 mL of Olaparib reference stock solution and 0.03 mL of Olaparib related substance solution and place them in the same 10 mL volumetric flask. Dilute with acetonitrile: water (1:1) to make a solution containing approximately 30 μg of Olaparib and 3 μg of Olaparib acid and other related substances per 1 mL as the system suitability solution.

[0104] Experimental steps:

[0105] Take the system suitability solution in Example 3 and inject it into the liquid chromatograph according to the above chromatographic analysis conditions, and record the chromatogram. The results are shown in the attached Figure 3 The relative retention time of olaparib acid was 10.579min, the relative retention time of defluoroolaparib was 9.853min, the relative retention time of olaparib pip dimer was 27.353min, and the elution time of n-propylolaparib was 15.863min; the separation degrees of each related substance peak and the main component peak were ≥1.5, and the separation effect was good.

[0106] Example 4: The present invention verifies the following items of the analysis method of Olaparib and its related substances.

[0107] Instruments and conditions:

[0108] Chromatograph: LC-2030C 3D Plus, Shimadzu Instrument (Suzhou) Co., Ltd.;

[0109] Detector: UV detector;

[0110] Chromatographic column: octadecylsilane bonded silica gel as filler (Dimosail, Inspire-C18 250 mm × 4.6 mm, 5 μm);

[0111] Column temperature: 25°C

[0112] Injection volume: 5 μL

[0113] 1. Linearity and range

[0114] (1) Sample preparation

[0115] Pipette 0.02, 0.04, 0.08, 0.16, 0.32, and 0.64 mL of the reference substance stock solution of Olaparib related substances under "Example 2" into a 10 mL volumetric flask, and dilute to the mark with acetonitrile: water (1:1).

[0116] (2) Test results

[0117] The results showed that olaparib acid in the range of 2 to 64 μg mL -1 The linearity is good within the range, and the linear regression equation is y=10992x-12098, R 2 =0.9987;

[0118] Desfluoroolaparib is 2-64 μg mL -1 The linearity is good within the range, and the linear regression equation is y=14072x-3843.5, R 2 =1;

[0119] Olaparib pip dimer is 2~64μg·mL -1 The linearity is good within the range, and the linear regression equation is y=6989.6x+4789.5, R 2 =0.9996;

[0120] Olaparib propyl is 2~64μg·mL -1 The linearity is good within the range, and the linear regression equation is y=6935.5x-3644.7, R 2 =1. (Test results are shown in the attached Figure 4-7 )

[0121] 2. Accuracy

[0122] (1) Sample preparation

[0123] Preparation of blank excipient solution for Olaparib tablets: According to the prescription ratio of Olaparib tablets, mix various excipients according to the actual usage amount; add acetonitrile: water (1:1) to dissolve and dilute to the scale, shake well, and obtain.

[0124] Pipette 0.10ml, 0.13ml, and 0.15ml of the stock solution of each relevant substance reference substance under "Example 2" into a 10mL volumetric flask, add the blank excipient solution of Olaparib tablets to dilute to the scale, and prepare 3 copies in parallel.

[0125] (2) Test results

[0126] Table 1 Results of accuracy test of related substances of Olaparib

[0127]

[0128]

[0129] As shown in Table 1, the recoveries of all relevant substances meet the requirements, and the RSD values ​​are all less than 10%, which is in line with the regulations. The accuracy of this method is good.

[0130] 3. Stability

[0131] (1) Solution preparation

[0132] Pipette 0.15 mL of each Olaparib-related substance reference solution under "Example 2" into a 10 mL volumetric flask, add acetonitrile: water (1:1) to dilute to scale. After standing at room temperature for 0, 2, 4, 6, 8, 10 and 12 hours, accurately measure 5 μL and inject into the liquid chromatograph.

[0133] (2) Test results

[0134] Table 2 Solution stability of Olaparib-related substances

[0135]

[0136]

[0137] As shown in Table 2, the four related substances of Olaparib are stable in acetonitrile: water (1:1) within 12 hours.

[0138] 4. Limit of detection and limit of quantification

[0139] (1) Solution preparation

[0140] The reference solutions of the related substances of Olaparib under "Example 2" were taken, continuously diluted with acetonitrile: water (1:1) and injected into the chromatograph. When the detection signal / baseline noise (S / N) was ≥10, the solution could be used as the quantitative limit solution. When the detection signal / baseline noise (S / N) was ≥3, the solution could be used as the detection limit solution.

[0141] (2) Test results

[0142] Table 3 Detection limit and quantification limit of olaparib related substances

[0143]

[0144] 5. Durability

[0145] Investigate the tolerance of the detection method when there are slight changes in the chromatographic conditions. Investigate the durability of the method for detecting related substances under conditions such as changing the flow rate, column temperature, and detector temperature.

[0146] (1) Solution preparation and parameter setting

[0147] Solution: Take 0.9 mL of each Olaparib-related substance reference solution under "Example 2" in a 10 mL volumetric flask, dilute to the mark with acetonitrile: water (1:1), and inject into the chromatograph.

[0148] Parameter settings:

[0149] Table 4 Validation of relevant substance analysis methods - Durability parameter range

[0150]

[0151] (2) Test results

[0152] Table 5 Durability-Flow Rate

[0153]

[0154] It can be seen from Table 5 that changing the flow rate has a significant impact on the determination results of the contents of related substances of Olaparib.

[0155] Table 6 Durability-column temperature

[0156]

[0157] It can be seen from Table 6 that changing the column temperature has little effect on the determination results of the content of related substances of Olaparib.

[0158] Table 7 Durability-Wavelength

[0159]

[0160] It can be seen from Table 7 that changing the wavelength has little effect on the determination results of the content of related substances of Olaparib.

[0161] Investigation of the column temperature, flow rate and wavelength in the chromatographic conditions showed that changing the flow rate had a significant impact on the determination results of the content of related substances in Olaparib. After comprehensive consideration, the following choices were made: column temperature 25°C, flow rate 0.8mL / min, wavelength 276nm.

[0162] The above description of the disclosed embodiments enables one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for detecting olaparib-related substances, characterized in that: High performance liquid chromatography was used under the following conditions: Chromatographic column: C18 column; Filler: octadecylsilane bonded silica gel; Mobile phase A: 0.01-1.0 wt% phosphoric acid solution; Mobile phase B: 70-100 wt% acetonitrile solution; Flow rate: 0.8~1.2ml / min; Detection wavelength: 270~280nm; Injection volume: 1-20 μL; Gradient elution was used.

2. The method for detecting olaparib-related substances according to claim 1, characterized in that: The inner diameter of the C18 chromatographic column is 1-5 mm, the length is 100-300 mm, and the filler particle size is 1-20 μm.

3. The method for detecting related substances of Olaparib according to claim 1, characterized in that: The gradient elution, in terms of volume percentage, has the following procedure:

4. The method for detecting olaparib-related substances according to claim 1, characterized in that: The following steps are involved: (1) Sample preparation: Preparation of reference substance stock solution: Accurately weigh Olaparib into a 10 mL volumetric flask, add diluent to dissolve and dilute to the scale, shake well to obtain the reference substance stock solution; Preparation of related substance stock solutions: accurately weigh equal amounts of olaparib acid, defluoroolaparib, and n-propylolaparib into a 10 mL volumetric flask, add diluent to dissolve and dilute to the mark, and shake well; accurately weigh the same amount of olaparib pip dimer into a 10 mL volumetric flask, add appropriate amount of DMSO to dissolve it, then add pure acetonitrile to dilute to the mark, and shake well; the olaparib related substance stock solution is obtained; Preparation of system suitability solution: Accurately measure the reference substance stock solution and related substance stock solution and place them in the same 10mL volumetric flask, dilute to the mark with diluent, and use this as the system suitability solution; (2) High performance liquid chromatography determination: accurately measure the system suitability solution, inject it into a high performance liquid chromatograph, perform the detection according to any one of the detection methods described in claims 1 to 3, record the chromatogram and perform data analysis.

5. The method for detecting olaparib-related substances according to claim 4, characterized in that: In step (1), the reference substance stock solution contains 0.1-2 mg of Olaparib per 1 mL, and the related substance stock solution contains 1-40 μg of Olaparib per 1 mL.

6. The method for detecting olaparib-related substances according to claim 4, characterized in that: In step (1), the diluent is prepared by acetonitrile and water in a volume ratio of (1-10):(1-10).

7. The method for detecting olaparib-related substances according to claim 4, characterized in that: In step (1), the solid-liquid ratio of the olaparib pip dimer to the DMSO is 50 mg:1 mL.

8. The method for detecting olaparib-related substances according to claim 4, characterized in that: In step (1), in the preparation of the system suitability solution, the volume ratio of the reference substance stock solution to the related substance stock solution is 10:1, and the concentration of Olaparib in the system suitability solution is 30 μg / mL.

9. The method for detecting olaparib-related substances according to claim 4, characterized in that: In step (2), the results of the high performance liquid chromatography determination show that the single impurity does not exceed 0.15%, and the total impurity does not exceed 0.5%.

10. The method for detecting related substances of Olaparib according to claim 4, characterized in that: In step (2), in the results of the high performance liquid chromatography determination, the separation between the impurity peaks in the Olaparib related substances and between the impurity peaks and the main peak is >2.0, and the tailing factor of the Olaparib and related substance peaks is not greater than 1.5.

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