Method for detecting content of impurities in concentrated solution for edaravone dextroborneol injection

Abstract

The invention belongs to the technical field of drug analysis and detection, and mainly relates to a method for detecting nine impurities in edaravone in a concentrated solution for edaravone dextroborneol injection by high performance liquid chromatography. The method comprises the following steps: preparing a reference substance solution, a test solution and a system applicable solution containing nine impurities, respectively injecting the solutions into high performance liquid chromatography, and calculating the contents of the nine impurities in edaravone by adopting a main component external standard method, the column temperature is 40-45 DEG C; the mobile phase A is an ammonium dihydrogen phosphate solution, and the pH value is adjusted to 2.3-2.7; the mobile phase B is acetonitrile; and gradient elution. According to the present invention, through the cooperation of the specific chromatographic column, the mobile phase and the gradient elution program, the provided high performance liquid chromatography method for determining the contents of the nine edaravone impurities in the edaravone dextroborneol injection concentrated solution has advantages of high impurity separation degree, simpleness, rapidness, high sensitivity and good repeatability, and can effectively control the quality of edaravone.

Description

Technical Field

[0001] This application belongs to the technical field of pharmaceutical analysis and detection, and specifically relates to a method for detecting the content of impurities in edaravone and dextrorotatory borneol concentrated solution for injection. Background Art

[0002] Edaravone is a neuroprotective agent (free radical scavenger). Clinical studies have shown that N-acetylaspartic acid (NAA) is a specific marker for surviving nerve cells, and its content decreases sharply in the early stage of cerebral infarction. When edaravone is administered to patients in the acute phase of cerebral infarction, it can inhibit the reduction of local cerebral blood flow around the infarction, and the content of NAA in the brain on the 28th day after onset is significantly higher than that in the glycerol control group. Preclinical studies have shown that when edaravone is intravenously administered to rats after ischemia / ischemia-reperfusion, it can prevent the progression of cerebral edema and cerebral infarction, relieve the accompanying neurological symptoms, and inhibit delayed neuronal death. Mechanism studies have shown that edaravone can scavenge free radicals and inhibit lipid peroxidation, thereby inhibiting oxidative damage to brain cells, vascular endothelial cells, and nerve cells.

[0003] The concentrated solution of edaravone and dextrorotatory borneol for injection was first approved for marketing in China on July 29, 2020. It is a Class 1 innovative drug. The active ingredients of this product are edaravone and dextrorotatory borneol, and it can be used to improve the neurological symptoms, activities of daily living ability, and functional impairment caused by acute cerebral infarction.

[0004] At present, the quality standards of the concentrated solution of edaravone and dextrorotatory borneol for injection are not included in the pharmacopoeias of various countries. The related substances of edaravone are only included in the ChP2020 and JP18 standards for raw materials and single-component preparations (edaravone injection), and an isocratic elution program is used, and only one impurity I is controlled. There are nine impurities in edaravone in this injection, and the existing detection methods have disadvantages such as poor resolution and incomplete elution of impurities.

[0005] Therefore, it is of great significance to develop a method with good separation effect and capable of accurately detecting the content of nine impurities of edaravone in the concentrated solution of edaravone and dextrorotatory borneol for injection. Summary of the Invention

[0006] The technical problem to be solved by this application is to overcome the defects and deficiencies of the existing methods, and provide a method for detecting the content of nine impurities of edaravone in the concentrated solution of edaravone and dextrorotatory borneol for injection. Through the cooperation of a specific chromatographic column, mobile phase, and gradient elution program, this application provides a detection method for the content of nine impurities of edaravone with simple operation, strong specificity, and good separation effect; the detection method of this application has the detection effects of high sensitivity, good reproducibility, strong specificity, and good resolution, and can be used for the routine analysis and quality control of the related substances of edaravone in the concentrated solution of edaravone and dextrorotatory borneol for injection.

[0007] A method for detecting the content of impurities in edaravone dextrorotatory borneol concentrated solution for injection, the detection method comprising:

[0008] Step 1, preparing a reference solution: Take edaravone reference substance, weigh accurately, dissolve and dilute with a solvent to prepare a solution containing 2 μg / ml of edaravone;

[0009] Test solution: Take the sample of edaravone dextrorotatory borneol concentrated solution for injection, dilute with a solvent to prepare a test solution containing 1 mg / ml of edaravone;

[0010] System suitability solution: Take appropriate amounts of edaravone reference substance and nine impurity reference substances, weigh accurately, dissolve with a solvent and quantitatively dilute to prepare a mixed solution containing approximately 1 mg of edaravone, 1 μg of impurity I, 2 μg each of impurity III, impurity V and impurity VII, 10 μg of impurity VI, and 3 μg each of impurity VIII and impurity IX per 1 ml;

[0011] Step 2, detecting the solutions prepared in Step 1 by liquid chromatography. The liquid chromatography uses octadecylsilane chemically bonded silica gel as the filler, 0.05 mol / L ammonium dihydrogen phosphate solution as mobile phase A, adjust the pH value to 2.5 with phosphoric acid, use acetonitrile as mobile phase B, the flow rate of the mobile phase is 0.8 mL / min to 1.2 mL / min, the column temperature is 38 °C to 47 °C, the detection wavelength is 240 to 250 nm, and the injection volume is 5 to 20 μL;

[0012] Among them, the elution program is gradient elution, and the elution program is: at the 0th minute, mobile phase A is 90%, mobile phase B is 10%; at the 15th minute, mobile phase A is 80%, mobile phase B is 20%; at the 35th minute, mobile phase A is 40%, mobile phase B is 60%; at the 50th minute, mobile phase A is 40%, mobile phase B is 60%; at the 51st minute, mobile phase A is 90%, mobile phase B is 10%; at the 65th minute, mobile phase A is 90%, mobile phase B is 10%;

[0013] Step 3, calculating the content of nine impurities of edaravone by the external standard method of the main component;

[0014] Among the nine impurities, impurity I is 4,4'-bis-(3-methyl-1-phenyl-5-pyrazolone); impurity II is 1-(5-hydroxy-3-methyl-1-phenyl-1H-pyrazol-4-yl)ethane-1-sulfonic acid; impurity III is 3,3',3”-trimethyl-1,1',1”-triphenyl-tris-(2-pyrazolin-5-one); impurity IV is disodium 1-(1-phenyl-3-methyl-5-pyrazolone-4-yl)-1-carboxyethanesulfonate; impurity V is 2-(3-methyl-5-oxo-1-phenyl-4,5-dihydro-1H-pyrazol-4-yl)propane-2-sulfonic acid; impurity VI is 1-hydroxy-2-(3-methyl-5-oxo-1-phenyl-4,5-dihydro-1H-pyrazol-4-yl)propane-2-sulfonic acid; impurity VII is 1-(3-methyl-5-oxo-1-phenyl-4,5-dihydro-1H-pyrazol-4-yl)propan-2-one; impurity VIII is (1S,2R) / (1R,2S)-2-hydroxy-1-(3-methyl-5-oxo-1-phenyl-4,5-dihydro-1H-pyrazol-4-yl)propane-1-sulfonic acid; impurity IX is (1S,2S) / (1R,2R)-2-hydroxy-1-(3-methyl-5-oxo-1-phenyl-4,5-dihydro-1H-pyrazol-4-yl)propane-1-sulfonic acid.

[0015] The pH regulator is phosphoric acid, and the pH value of mobile phase A is 2.5.

[0016] In the chromatographic conditions, the chromatographic column is a C8 liquid chromatographic column.

[0017] In the chromatographic conditions, the chromatographic column is selected from Ultimate LP-C8, with a column length of 250 mm, an inner diameter of 4.6 mm, and a particle size of 5 μm; or Ultimate XB-C8, with a column length of 250 mm, an inner diameter of 4.6 mm, and a particle size of 5 μm; preferably, the chromatographic column is Ultimate LP-C8, with a column length of 250 mm, an inner diameter of 4.6 mm, and a particle size of 5 μm.

[0018] The solvent is mobile phase A: methanol, and their volume ratio is 75:25. The concentration of edaravone in the test solution is 1 mg / mL.

[0019] The beneficial effects of this application include but are not limited to:

[0020] This application provides a detection method for the contents of nine impurities of edaravone, which is simple to operate and has no mutual interference among impurities. This detection method is a liquid chromatography analysis method with relatively low cost.

[0021] Through the cooperation of a specific chromatographic column, mobile phase, and gradient elution program, this application ensures the accuracy of the detection results. This detection method has good resolution, good reproducibility, high specificity, and high detection efficiency.

[0022] The detection method provided by this application is suitable for detecting the contents of nine impurities of edaravone in a concentrated solution for injection of edaravone dextrancan, and can be used for the routine analysis and quality control of the concentrated solution for injection of edaravone dextrancan.

[0023] The detection method provided by this application adopts a specific gradient elution program. This detection method avoids the interference of other substances, and the target substance is well separated from the adjacent impurity peaks during detection, so that the contents of nine impurities of edaravone in the concentrated solution for injection of edaravone dextrancan can be accurately detected. Description of the Drawings

[0024] Figure 1 Liquid chromatography detection spectrum of the blank solvent in Example 1;

[0025] Figure 2 Liquid chromatography detection spectrum of the blank excipient in Example 1;

[0026] Figure 3 Liquid chromatography detection spectrum of the sample solution to be measured in Example 1;

[0027] Figure 4 Liquid chromatography detection spectrum of the system suitability solution in Example 1;

[0028] Figure 5 Liquid chromatography detection spectrum of the system suitability solution in Example 2;

[0029] Figure 6 Liquid chromatography detection spectrum of the system suitability solution in Comparative Example 1;

[0030] Figure 7 Liquid chromatography detection spectrum of the system suitability solution in Comparative Example 2. Detailed Description of the Invention

[0031] The following further illustrates the content of this application with specific examples, but should not be construed as a limitation to this application. All other examples obtained by those of ordinary skill in the art based on the examples in this application without creative efforts fall within the scope of protection of this application.

[0032] Example 1:

[0033] 1. Instruments and Equipment

[0034] Instrument: Liquid chromatograph (Agilent 1260).

[0035] Reagents: Ammonium dihydrogen phosphate (analytical grade, Macklin), phosphoric acid (analytical grade, Tianjin Hengxing), acetonitrile (chromatographic grade, Fisher), methanol (analytical grade, MREDA).

[0036] Principal component reference substance: Edaravone reference substance (National Institutes for Food and Drug Control, batch number 100620 - 202204)

[0037] Sample to be tested: Concentrated solution of edaravone and dextrancamphor for injection, specification: 5 ml∶edaravone 10 mg and dextrancamphor 2.5 mg (self - prepared, batch number: 24102301).

[0038] Blank excipient: Negative solution without edaravone (self - prepared: batch number 24091902).

[0039] Reference substances for impurities I - IX: Purchased externally

[0040] 2. Preparation of solutions

[0041] Preparation of the sample solution to be tested: Precisely measure 5 ml of this product, place it in a 10 - mL volumetric flask, dilute it to the mark with mobile phase A - methanol (75:25), shake well, and prepare a sample solution to be tested containing 1 mg / mL of edaravone;

[0042] Preparation of the reference solution: Take about 10 mg of edaravone reference substance, place it in a 50 - mL volumetric flask, dissolve and dilute it to the mark with mobile phase A - methanol (75:25), shake well; precisely measure 1 ml, place it in a 100 - ml volumetric flask, and dilute it to the mark with mobile phase A - methanol (75:25) to prepare a reference solution containing 2 μg / mL of edaravone;

[0043] Preparation of the blank excipient solution: Precisely measure 5 ml of the negative solution without edaravone, place it in a 10 - mL volumetric flask, add an appropriate amount of mobile phase A - methanol (75:25) and dilute it to the mark, shake well.

[0044] Preparation of the system suitability solution: Take appropriate amounts of edaravone reference substance and nine impurity reference substances, precisely weigh them, dissolve them with mobile phase A - methanol (75:25) and quantitatively dilute to prepare a mixed solution containing approximately 1 mg of edaravone, 1 μg of impurity I, 2 μg of impurity II, impurity III, impurity IV, impurity V, and impurity VII each, 10 μg of impurity VI, and 3 μg of impurity VIII and impurity IX in every 1 ml.

[0045] 3. Chromatographic conditions:

[0046] Chromatographic column: Ultimate LP - C8, 4.6 mm×250 mm, 5 μm, filled with octadecylsilyl silica gel;

[0047] Mobile phase A: 0.05 mol / L ammonium dihydrogen phosphate solution (adjust the pH value to 2.5 with phosphoric acid);

[0048] Mobile phase B: Acetonitrile;

[0049] The flow rate of the mobile phase is 1.0 mL per minute; the column temperature is 43 °C; the detection wavelength is 245 nm; the injection volume is 10 μL;

[0050] Gradient elution is carried out using mobile phase A and mobile phase B, and the gradient elution conditions are as follows:

[0051]

[0052] The external standard method with the main component is used to calculate the contents of nine impurities of edaravone.

[0053] Calculation formula:

[0054]

[0055] Note: W 对 : Sampling amount of the reference solution, mg;

[0056] V 供 : Dilution factor of the test solution;

[0057] V 对 : Dilution factor of the reference solution;

[0058] A 对 : Peak area of the main peak in the chromatogram of the reference solution;

[0059] A 已知 : Peak area of the known impurity in the chromatogram of the test solution;

[0060] A 总 : Total peak area in the chromatogram of the test solution;

[0061] A 主 : Peak area of the main peak in the chromatogram of the test solution;

[0062] A 单杂 : Peak area of other single impurities in the chromatogram of the test solution;

[0063] P 对 : Content of the reference substance, %;

[0064] C 标 : Labeled concentration of edaravone, 2 mg / ml;

[0065] f: Correction factor.

[0066] 4. Verification results and conclusions

[0067] 4.1 System suitability

[0068] Take the test sample solution, reference solution, mixed solution and blank excipient solution, and detect them according to the above chromatographic conditions. The chromatogram of the blank solvent is as Figure 1As shown, the solvent peak does not interfere with the detection of the present invention; the chromatogram of the blank excipient is as Figure 2 shown, the negative solution without edaravone does not interfere with the detection of the present invention; the chromatogram of the sample solution to be tested is as Figure 3 shown, the chromatogram of the system suitability solution of edaravone and nine impurities is as Figure 4 shown. The results are shown in Table 1. All impurities can be completely separated. The minimum resolution of the known impurities is 1.305, which is greater than 1.0, and the resolution between edaravone and the adjacent impurities is greater than 1.5. It shows that in the method of the present invention, the main peak and the adjacent impurity peaks, and between each impurity can achieve good separation, so as to more accurately evaluate the product quality.

[0069] Table 1 Chromatogram of the system suitability solution

[0070]

[0071] The detection specificity of this application is good. The blank solvent and the blank excipient do not interfere with the detection; the resolution between the main peak and the adjacent impurity peaks and each impurity meets the requirements.

[0072] 4.2 Sensitivity experiment

[0073] Take appropriate amounts of edaravone and nine impurities, weigh accurately, and prepare a solution with a signal-to-noise ratio of about 10 as the quantitative limit solution, and a solution with a signal-to-noise ratio of about 3 as the detection limit solution.

[0074] Table 2 Results of the sensitivity test

[0075]

[0076] The detection method of this application has good sensitivity. As can be seen from Table 2, the S / N values of each component in the quantitative limit solution are all between 10 and 30, and the S / N of each component in the detection limit solution is between 3 and 8.

[0077] 4.3 Linearity test

[0078] Take appropriate amounts of edaravone and nine impurities, weigh accurately, and prepare a solution within the range of the quantitative limit to 2.0% limit concentration for linear verification. The results are shown in the following table:

[0079] Table 3 Results of the linearity test

[0080] Name Concentration range (μg / mL) Linear correlation equation <![CDATA[R 2 > Edaravone 0.0399～19.9280 y = 43.9664x - 0.0465 1.0000 Impurity I 0.0195～19.4613 y = 39.1434x + 0.5824 1.0000 Impurity II 0.0194～19.3644 y = 21.6008x - 0.0345 1.0000 Impurity III 0.0188～18.8323 y = 40.7398x - 0.1251 1.0000 Impurity IV 0.0368～18.3878 y = 14.6503x - 0.1317 1.0000 Impurity V 0.0093～18.5667 y = 20.3100x - 0.0069 1.0000 Impurity VI 0.2361～18.8860 y = 20.3956x + 0.6169 1.0000 Impurity VII 0.0197～19.7184 y = 33.5662x - 0.1444 1.0000 Impurity VIII 0.0197～19.7150 y = 22.3494x + 0.1161 1.0000 Impurity IX 0.0192～19.2369 y = 23.6798x - 0.0242 1.0000

[0081] The detection method of this application has good linearity. As can be seen from Table 3, within the range of the quantitative limit to 2.0% limit concentration, the concentrations of edaravone and nine impurities have a good linear relationship with the corresponding peak areas, and R 2 is greater than 0.999.

[0082] 4.4 Accuracy test

[0083] The standard addition recovery test was carried out at low concentration (50% of the limit concentration), medium concentration (limit concentration), and high concentration (150% of the limit concentration).

[0084] Stock solution of impurities: Weigh accurately about 1 mg of impurity I, about 2 mg of impurity II, impurity IV, impurity V, III, and VII, and about 3 mg of VIII and IX respectively, place them in the same 100 ml volumetric flask, dissolve with mobile phase A - methanol (75:25) and dilute to the scale, shake well; prepare 2 portions in parallel.

[0085] Stock solution of impurity VI: Weigh accurately about 2 mg of impurity VI, place it in a 20 ml volumetric flask, dissolve with mobile phase A - mobile phase B (75:25) and dilute to the scale, shake well; prepare 2 portions in parallel.

[0086] Reference solution: Accurately measure 1 ml of the stock solution of impurities and 1 ml of the stock solution of impurity VI, place them in a 10 ml volumetric flask, dissolve with mobile phase A - mobile phase B (75:25) and dilute to the scale, shake well;

[0087] Test solution: Accurately measure 5 ml of this product, place it in a 10 mL volumetric flask, add an appropriate amount of mobile phase A - methanol (75:25) and dilute to the scale, shake well; prepare 2 portions in parallel.

[0088] Recovery at low concentration: Accurately measure 10 ml of this product, place it in a 20 mL volumetric flask, accurately measure 1 ml of the stock solution of impurities and 1 ml of the stock solution of impurity VI respectively, place them in the same volumetric flask, dissolve with mobile phase A - methanol (75:25) and dilute to the scale, shake well; prepare 3 portions in parallel.

[0089] Recovery at medium concentration: Accurately measure 5 ml of this product, place it in a 10 mL volumetric flask, accurately measure 1 ml of the stock solution of impurities and 1 ml of the stock solution of impurity VI respectively, place them in the same volumetric flask, dissolve with mobile phase A - methanol (75:25) and dilute to the scale, shake well; prepare 3 portions in parallel.

[0090] Recovery at high concentration: Accurately measure 5 ml of this product, place it in a 10 mL volumetric flask, accurately measure 1.5 ml of the stock solution of impurities and 1.5 ml of the stock solution of impurity VI respectively, place them in the same volumetric flask, dissolve with mobile phase A - methanol (75:25) and dilute to the scale, shake well; prepare 3 portions in parallel.

[0091] The measurement results are shown in the following table

[0092] Table 4 Results of the methodology recovery experiment

[0093] Name Average recovery rate / % RSD / % Impurity I 97.9 1.68 Impurity II 98.1 0.41 Impurity III 99.4 0.75 Impurity IV 99.0 1.04 Impurity V 100.9 0.54 Impurity VI 99.4 0.42 Impurity VII 94.2 1.72 Impurity VIII 97.6 3.38 Impurity IX 96.74 4.00

[0094] The accuracy of the detection method of this application is good. At the limit concentrations of 50%, 100%, and 150%, the average recovery rates of each impurity are between 92% and 105%, and the RSD values are all less than 10.0%, indicating good method accuracy.

[0095] 4.5 Durability Test

[0096] The detection method of this application has good durability. By examining the flow rate of 0.9 mL / min to 1.1 mL / min, the column temperature of 40 °C to 45 °C, and the pH of mobile phase A of 2.3 to 2.7, the resolution between each impurity and between the main peak and the adjacent impurity in the system suitability solution meets the requirements. There is no significant difference in the detection results of the contents of nine impurities in the spiked test solution at the limit concentration, and the RSD value is less than 10.0%, indicating good durability. The results are shown in the following table:

[0097] Table 5 Durability Test Results - Resolution of System Suitability Solution

[0098]

[0099]

[0100] Table 6 Durability Test Results - Detection Results of Spiked Test Solution / %

[0101]

[0102] Example 2:

[0103] The difference between this example and Example 1 is that the chromatographic column in this example is Ultimate XB-C8 (4.6 mm × 250 mm, 5 μm), and other chromatographic conditions are exactly the same. From the liquid chromatography detection results, the resolution between each impurity and between the main peak and the adjacent impurity in the system suitability solution meets the requirements and can meet the detection needs.

[0104] Comparative Example 1:

[0105] The difference between this comparative example and Example 1 is that the mobile phase B in this example is methanol, and other chromatographic conditions are exactly the same. From the liquid chromatography detection results, the retention time of the main peak in the system suitability solution is about 37 min, the peak elution is late, and there is a situation where one impurity cannot be eluted, indicating that this system is not suitable for the detection of the impurity content in this product.

[0106] Name Retention time RRT Theoretical plate number Tailing factor Resolution / 25.547 0.69 70128 1.011 / / 28.367 0.77 135231 0.976 8.141 / 29.896 0.81 139227 0.988 4.854 / 32.803 0.88 199997 1.001 9.469 / 35.007 0.94 181791 1.067 7.076 Edaravone 37.080 1.00 56035 1.065 4.342 / 40.113 1.08 248113 0.815 6.395 / 47.733 1.29 545187 1.055 26.245 / 53.666 1.45 972738 1.128 24.919

[0107] Comparative Example 2:

[0108] The difference between this comparative example and Example 1 is that the column temperature in this example is 30 °C, and other chromatographic conditions are exactly the same. From the liquid chromatography test results, it can be seen that impurity II and impurity VIII in the system suitability solution completely overlap and cannot meet the separation requirements, indicating that this system is not suitable for the detection of the impurity content in this product.

[0109]

Claims

1. A method for detecting the content of impurities in edaravone and dextrancamphor injection concentrated solution, characterized in that, The detection method includes the following: Step 1: Prepare the reference solution. Take edaravone reference substance, weigh accurately, dissolve and dilute with a solvent to prepare a solution containing 2 μg / ml of edaravone. Test solution: Take the concentrated solution of edaravone and camphor injection sample, dilute with a solvent to prepare a test solution containing 1 mg / ml of edaravone. System suitability solution: Take appropriate amounts of edaravone reference substance and nine impurity reference substances, weigh accurately, dissolve with a solvent and quantitatively dilute to prepare a mixed solution containing approximately 1 mg of edaravone, 1 μg of impurity I, 2 μg each of impurity III, impurity V and impurity VII, 10 μg of impurity VI, and 3 μg each of impurity VIII and impurity IX in every 1 ml. Step 2: Detect the solutions prepared in Step 1 by liquid chromatography. The liquid chromatography uses octadecylsilane chemically bonded silica gel as the filler, 0.05 mol / L ammonium dihydrogen phosphate solution as mobile phase A, adjust the pH value to 2.5 with phosphoric acid, use acetonitrile as mobile phase B, the flow rate of the mobile phase is 0.8 mL / min - 1.2 mL / min, the column temperature is 38°C - 47°C, the detection wavelength is 240 - 250 nm, and the injection volume is 5 - 20 μL. Among them, the elution program is gradient elution, and the elution program is as follows: at the 0th minute, mobile phase A is 90% and mobile phase B is 10%; at the 15th minute, mobile phase A is 80% and mobile phase B is 20%; at the 35th minute, mobile phase A is 40% and mobile phase B is 60%; at the 50th minute, mobile phase A is 40% and mobile phase B is 60%; at the 51st minute, mobile phase A is 90% and mobile phase B is 10%; at the 65th minute, mobile phase A is 90% and mobile phase B is 10%. Step 3: Calculate the contents of nine impurities of edaravone by the external standard method of the main component. The nine impurities are as follows: Impurity I is 4,4'-bis-(3-methyl-1-phenyl-5-pyrazolone); Impurity II is 1-(5-hydroxy-3-methyl-1-phenyl-1H-pyrazol-4-yl)ethane-1-sulfonic acid; Impurity III is 3,3',3''-trimethyl-1,1',1''-triphenyl-tri-(2-pyrazolin-5-one); Impurity IV is disodium 1-(1-phenyl-3-methyl-5-pyrazolone-4-yl)-1-carboxyethanesulfonate; Impurity V is 2-(3-methyl-5-oxo-1-phenyl-4,5-dihydro-1H-pyrazol-4-yl)propane-2-sulfonic acid; Impurity VI is 1-hydroxy-2-(3-methyl-5-oxo-1-phenyl-4,5-dihydro-1H-pyrazol-4-yl)propane-2-sulfonic acid; Impurity VII is 1-(3-methyl-5-oxo-1-phenyl-4,5-dihydro-1H-pyrazol-4-yl)propan-2-one; Impurity VIII is (1S,2R) / (1R,2S)-2-hydroxy-1-(3-methyl-5-oxo-1-phenyl-4,5-dihydro-1H-pyrazol-4-yl)propane-1-sulfonic acid; Impurity IX is (1S,2S) / (1R,2R)-2-hydroxy-1-(3-methyl-5-oxo-1-phenyl-4,5-dihydro-1H-pyrazol-4-yl)propane-1-sulfonic acid.

2. The content detection method for impurities in the concentrated solution of edaravone and dextrancamphorol for injection according to claim 1, characterized in that, The pH regulator is phosphoric acid, and the pH value of mobile phase A is 2.

5.

3. The content detection method for impurities in the concentrated solution for injection of edaravone and dextrorotatory borneol according to claim 1, wherein The chromatographic column in the chromatographic conditions is a C8 liquid chromatographic column.

4. The content detection method for impurities in the concentrated solution for injection of edaravone and dextrorotatory borneol according to claim 1, wherein, The chromatographic column in the chromatographic conditions is selected from Ultimate LP-C8, with a column length of 250 mm, an inner diameter of 4.6 mm, and a particle size of 5 μm; or Ultimate XB-C8, with a column length of 250 mm, an inner diameter of 4.6 mm, and a particle size of 5 μm; preferably, the chromatographic column is Ultimate LP-C8, with a column length of 250 mm, an inner diameter of 4.6 mm, and a particle size of 5 μm.

5. The content detection method for impurities in the concentrated solution of edaravone and dextrorotatory borneol for injection according to claim 1, wherein, The solvent is mobile phase A: methanol, and their volume ratio is 75:

25. The concentration of edaravone in the test solution is 1 mg / mL.

Images