Method for detecting polymer impurities in product containing cefuroxime sodium and application of method

Abstract

The invention relates to a method for detecting polymer impurities in a product containing cefuroxime sodium and application of the method. The detection method comprises the following steps: detecting a product containing the cefuroxime sodium by adopting a reversed-phase liquid chromatography-mass spectrometry method to obtain the variety and content of polymer impurities in the product containing the cefuroxime sodium, mobile phases of the reversed-phase liquid chromatography comprise a mobile phase A and a mobile phase B, the mobile phase A is an ammonium acetate buffer solution, and the mobile phase B is acetonitrile. According to the detection method, the polymer impurities can be comprehensively and accurately controlled, 15 polymer impurities can be directly detected, all the detected polymer impurity types belong to one another, and the structure speculation of 12 polymer peaks is completed. The detection method is strong in specificity, high in accuracy and good in reproducibility, can effectively control the quality of the cefuroxime sodium for injection, and has positive significance on storage of the cefuroxime sodium for injection and polymer control during clinical use.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical analysis, and particularly relates to a method for detecting polymer impurities in a product containing cefuroxime sodium and an application thereof. Background Art

[0002] Cefuroxime sodium for injection is a freeze-dried powder injection of cefuroxime sodium, the English name is Cefuroxime, the molecular formula is C 16 H 15 N4NaO8S, with a relative molecular mass of 446.4, is chemically named (6R,7R)-7-[2-furyl(methoxyimino)acetylamino]3-carbamoyloxymethyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt, and its structural formula is shown in formula (1):

[0003]

[0004] Cefuroxime sodium belongs to the second-generation, broad-spectrum cephalosporin class, and is widely used to treat infections caused by sensitive strains of specific microorganisms, such as respiratory tract infections, ear, nose, and throat infections, and urinary tract infections. From a safety perspective, its sensitization strength is related to the polymer impurity content.

[0005] Cephalosporin antibiotics are easily degraded to produce polymer impurities during production and storage, especially under high temperature and light conditions. They are also very likely to produce polymer impurities when used in combination with other solvents in clinical practice. Their structure is complex and difficult to control.

[0006] CN114252516A discloses a method for detecting polymers in cefuroxime sodium. This method uses a common octadecylsilane-bonded silica gel chromatographic column to separate three mixed impurity peaks, which are then subjected to mass spectrometry analysis to identify five possible polymer impurities. However, this method does not separate individual polymer impurities, nor does it define their specific structure and determination method, resulting in poor specificity.

[0007] CN114166963A discloses a two-dimensional detection method for impurities in cefuroxime sodium. Similar to CN114252516A, a test solution is prepared using the cefuroxime sodium raw material, and then subjected to HPLC-UV-MS detection. Three impurities in cefuroxime sodium are separated by one-dimensional chromatography, and a polymer in cefuroxime sodium is subjected to a secondary separation under two-dimensional chromatography conditions. However, the method also suffers from the problem of poor specificity.

[0008] CN117589902A discloses a method for simultaneously detecting related substances and polymers of cefuroxime sodium. However, the method only performs a qualitative analysis of four polymer impurities concentrated within the retention time range of 45.5 minutes to 49.5 minutes at the end of the elution gradient. A comprehensive analysis and control of the polymer impurities is not performed, and thus the polymer impurities cannot be fully controlled.

[0009] CN118671211A discloses a method for simultaneously determining cefuroxime sodium-related substances and polymer impurities. The main chromatographic conditions used are: mobile phase A, 0.68 g of sodium acetate, 5.8 g of glacial acetic acid, diluted with water to 1000 mL, and the pH value adjusted to 3.4 with glacial acetic acid; mobile phase B, acetonitrile. This method detects cefuroxime sodium-related substances and polymer impurities, but does not provide a comprehensive analysis of polymer impurities.

[0010] Therefore, in order to comprehensively control polymer impurities in products containing cefuroxime sodium and reduce the risks of clinical use, it is urgent to develop a method with strong specificity, high accuracy and good reproducibility that can comprehensively detect polymer impurities in products containing cefuroxime sodium. Summary of the Invention

[0011] In response to the shortcomings of the prior art, the present invention aims to provide a method for detecting polymer impurities in products containing cefuroxime sodium and its application. This method can comprehensively and accurately control polymer impurities with strong specificity, high accuracy, and good reproducibility.

[0012] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0013] In a first aspect, the present invention provides a method for detecting polymer impurities in a product containing cefuroxime sodium, the detection method comprising: detecting the product containing cefuroxime sodium using a reversed-phase liquid chromatography-mass spectrometry method to obtain the type and content of polymer impurities in the product containing cefuroxime sodium.

[0014] The mobile phase of the reversed-phase liquid chromatography comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is an ammonium acetate buffer and the mobile phase B is acetonitrile.

[0015] The present invention uses ammonium acetate buffer as mobile phase A and acetonitrile as mobile phase B, thereby improving the separation of various polymer impurities and enhancing specificity. Furthermore, ammonium acetate is volatile, and the separated impurities can be directly qualitatively analyzed using HPLC-MS without adjusting the mobile phase. This method is simple to operate and highly durable.

[0016] Preferably, the pH value of the ammonium acetate buffer is 3-4.

[0017] Among them, the specific point values ​​in 3-4 can be selected as 3, 3.2, 3.4, 3.6, 3.8, 4, etc.

[0018] In the present invention, when the pH value of the ammonium acetate buffer is adjusted to 3-4, the separation degree of various polymer impurities can be better improved, and the specificity is stronger.

[0019] Preferably, the chromatographic column for reverse phase liquid chromatography is an octylsilane bonded silica gel chromatographic column.

[0020] Preferably, the model of the chromatographic column is Welch Ultimate XB-C8 chromatographic column.

[0021] The present invention creatively discovered that a Welch Ultimate XB-C8 chromatographic column can better improve the sensitivity and repeatability of polymer impurity detection.

[0022] Preferably, the column temperature of the reverse phase liquid chromatography column is 25-35°C, for example, 25°C, 27°C, 29°C, 31°C, 33°C, 35°C, etc.

[0023] Preferably, the detection wavelength of the reverse phase liquid chromatography is 270-275 nm, for example, it can be 271 nm, 272 nm, 273 nm, 274 nm, 275 nm, etc.

[0024] Preferably, the injection flow rate of the reversed-phase liquid chromatography is 1-2 mL / min, for example, 1 mL / min, 1.2 mL / min, 1.4 mL / min, 1.6 mL / min, 1.8 mL / min, 2 mL / min, etc.

[0025] Preferably, the injection volume of the reverse phase liquid chromatography is 15-25 μL, for example, it can be 15 μL, 17 μL, 19 μL, 21 μL, 23 μL, 25 μL, etc.

[0026] Preferably, the reverse phase liquid chromatography adopts a gradient elution method, and the procedure of the gradient elution is:

[0027] From 0 to 5 minutes, the volume fraction of mobile phase A is 75-85%, and the volume fraction of mobile phase B is 15-25%.

[0028] From 5 to 40 minutes, the volume fraction of mobile phase A was changed from 75-85% to 65-75% at a constant speed, and the volume fraction of mobile phase B was changed from 15-25% to 25-35% at a constant speed.

[0029] At 40-45 minutes, the volume fraction of mobile phase A was 65-75%, and the volume fraction of mobile phase B was 25-35%.

[0030] At 45-45.1 min, the volume fraction of mobile phase A was changed from 65-75% to 75-85% at a constant speed, and the volume fraction of mobile phase B was changed from 25-35% to 15-25% at a constant speed.

[0031] At 45.1-50 min, the volume fraction of mobile phase A was 75-85%, and the volume fraction of mobile phase B was 15-25%.

[0032] Among them, the specific point values ​​in 75-85% can be selected from 75%, 77%, 79%, 81%, 83%, 85%, etc., the specific point values ​​in 15-25% can be selected from 15%, 17%, 19%, 21%, 23%, 25%, etc., the specific point values ​​in 65-75% can be selected from 65%, 67%, 69%, 71%, 73%, 75%, etc., and the specific point values ​​in 25-35% can be selected from 25%, 27%, 29%, 31%, 33%, 35%, etc.

[0033] In the present invention, the above-mentioned specific elution procedure can better improve the sensitivity and repeatability of polymer impurity detection.

[0034] Preferably, the ion source of the mass spectrometer comprises an electrospray ion source.

[0035] Preferably, the mass spectrometry modes include positive ion mode and negative ion mode.

[0036] Preferably, the retention time of the polymer impurities is 15-35 min, for example, 15 min, 20 min, 25 min, 30 min, 35 min, etc.

[0037] Preferably, the detection method adopts an external standard method and uses cefuroxime as a reference substance to perform quantitative analysis on polymer impurities.

[0038] Preferably, the cefuroxime sodium polymer impurities include 31 H 29 N7O 14 S2, C 25 H 24 N6O 12 S.C. 30 H 28 N6O 13 S2, C 30 H 28 N6O 14 S2, C 31 H 29 N7O 14 S2, C 30 H 29 N7O 12 S2, C 25 H24 N6O 12 S or C 31 H 32 N8O 14 Any one or a combination of at least two of S2.

[0039] In a second aspect, the present invention provides a use of the method for detecting polymer impurities in a product containing cefuroxime sodium as described in the first aspect in detecting a product containing cefuroxime sodium.

[0040] Preferably, the product comprises an injection containing cefuroxime sodium (cefuroxime sodium for injection).

[0041] The numerical range described in the present invention includes not only the point values ​​listed above, but also any point values ​​between the above numerical ranges that are not listed. Due to space limitations and for the sake of simplicity, the present invention no longer exhaustively lists the specific point values ​​included in the range.

[0042] Compared with the prior art, the present invention has the following beneficial effects:

[0043] The present invention provides a method for detecting polymer impurities in a product containing cefuroxime sodium. The method can comprehensively and accurately control polymer impurities, directly detect 15 polymer impurities, classify the types of all detected polymer impurities, and complete structural inference of 12 polymer peaks.

[0044] The detection method provided by the present invention has strong specificity, high accuracy and good reproducibility, can effectively control the quality of cefuroxime sodium for injection, and has positive significance for the storage of cefuroxime sodium for injection and the control of polymers during clinical use.

[0045] The detection method provided by the present invention is simple to operate and has good durability. The mobile phase does not need to be adjusted and the separated impurities can be directly qualitatively analyzed by HPLC-MS.

[0046] Furthermore, the present invention uses a Welch Ultimate XB-C8 chromatographic column, which can better improve the sensitivity and repeatability of polymer impurity detection.

[0047] Furthermore, the specific elution procedure of the present invention can better separate various impurities and improve the detection capability of polymer impurities. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 This is the liquid chromatogram of Comparative Example 3, wherein A is the liquid chromatogram and B is an enlarged view of the elution time segment from 30 min to 52 min.

[0049] Figure 2It is the liquid chromatogram of the test solution 1-1 in Example 1.

[0050] Figure 3 It is the liquid chromatogram of the test solution 1-2 in Example 1.

[0051] Figure 4 It is the liquid chromatogram of the test solution 2-1 in Example 1.

[0052] Figure 5 It is the liquid chromatogram of the test solution 2-2 in Example 1.

[0053] Figure 6 It is the liquid chromatogram of the test solution 2-3 in Example 1.

[0054] Figure 7 It is the liquid chromatogram of the test solution 2-4 in Example 1.

[0055] Figure 8 This is the liquid chromatogram of the test solution in Test Example 4 with a column temperature of 25°C.

[0056] Figure 9 This is the liquid chromatogram of the test solution in Test Example 4 with a column temperature of 35°C.

[0057] Figure 10 This is the liquid chromatogram of the test solution at a wavelength of 271 nm in Test Example 4.

[0058] Figure 11 This is the liquid chromatogram of the test solution at a wavelength of 275 nm in Test Example 4. DETAILED DESCRIPTION

[0059] The technical solution of the present invention is further described below by way of specific embodiments. It should be understood by those skilled in the art that the embodiments are merely to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0060] The test samples selected in the following examples are configured as follows:

[0061] Test solution 1-1: Take an appropriate amount of cefuroxime sodium that has been stored at high temperature (60°C) for 20 days, dissolve it in water and dilute it to make a solution containing approximately 10 mg of cefuroxime per 1 ml.

[0062] Test solution 1-2: Take an appropriate amount of cefuroxime sodium that has been placed under light conditions (4500±500 Lux) for 20 days, dissolve it in water and dilute it to make a solution containing approximately 10 mg of cefuroxime per 1 ml.

[0063] Test solution 2-1: Take one vial of cefuroxime sodium for injection, add 6 ml of diluent (water) to dissolve it, let it stand at room temperature for 24 hours, accurately measure an appropriate amount, and use the diluent to make a solution containing approximately 20 mg of cefuroxime per 1 ml.

[0064] Test solution 2-2: Take one vial of cefuroxime sodium for injection, add 6 ml of diluent (0.9% sodium chloride solution) to dissolve it, let it stand at room temperature for 24 hours, accurately measure an appropriate amount, and use the diluent to make a solution containing approximately 20 mg of cefuroxime per 1 ml.

[0065] Test solution 2-3: Take one vial of cefuroxime sodium for injection, add 6 ml of diluent (10% glucose solution) to dissolve it, and let it stand at room temperature for 24 hours. Accurately measure an appropriate amount and use the diluent to make a solution containing approximately 20 mg of cefuroxime per 1 ml.

[0066] Test solution 2-4: Dissolve 1 vial of cefuroxime sodium for injection in 6 ml of diluent (sodium lactate Ringer's solution). Allow to stand at room temperature for 24 hours. Accurately measure an appropriate amount and use the diluent to prepare a solution containing approximately 20 mg of cefuroxime per ml.

[0067] Example 1

[0068] This embodiment provides a method for detecting polymer impurities in a product containing cefuroxime sodium. The test conditions are as follows:

[0069] (1) Liquid chromatography conditions:

[0070] Column: Welch Ultimate XB-C8, 4.6 mm × 250 mm, 5.0 μm;

[0071] Mobile phase: Mobile phase A is 5 mmol / L ammonium acetate solution (pH adjusted to 3.4 with acetic acid); mobile phase B is acetonitrile;

[0072] Detection wavelength: 273nm;

[0073] Column temperature: 30°C;

[0074] Flow rate: 1.5 mL / min;

[0075] Injection volume: 20 μL;

[0076] The elution gradient program is shown in Table 1:

[0077] Table 1

[0078] Time (min) Mobile phase A (%) Mobile phase B (%) 0.0 80 20 5.0 80 20 40.0 70 30 45.0 70 30 45.1 80 20 50.0 80 20

[0079] (2) Mass spectrometry conditions are shown in Table 2:

[0080] Table 2

[0081] Ion source Electrospray ionization (ESI) model Positive ions and negative ions Scan range 50-1200 m / z Curtain Air (CUR) 35psi Spray voltage (IS) 5500V Ionization temperature (TEM) 550℃ Atomizing gas (GS1) 55psi Atomizing gas (GS2) 55psi Declustering voltage (DP) 80V Entry voltage (EP) 10V Collision energy (CE) 46V

[0082] (3) Test samples:

[0083] Accurately weigh 20 μL of test solution 1-1, test solution 1-2, test solution 2-1, test solution 2-2, test solution 2-3, and test solution 2-4, and inject them into the liquid chromatograph in sequence for detection and analysis.

[0084] Example 2

[0085] This embodiment provides a method for detecting polymer impurities in a product containing cefuroxime sodium. The test conditions are as follows:

[0086] (1) Liquid chromatography conditions:

[0087] Column: Welch Ultimate XB-C8, 4.6 mm × 250 mm, 5.0 μm;

[0088] Mobile phase: Mobile phase A is 5 mmol / L ammonium acetate solution (pH adjusted to 3.0 with acetic acid); mobile phase B is acetonitrile;

[0089] Detection wavelength: 270nm;

[0090] Column temperature: 35°C;

[0091] Flow rate: 1 mL / min;

[0092] Injection volume: 15 μL;

[0093] The elution gradient program is shown in Table 3:

[0094] Table 3

[0095] Time (min) Mobile phase A (%) Mobile phase B (%) 0.0 75 25 5.0 75 25 40.0 65 35 45.0 65 35 45.1 75 25 50.0 75 25

[0096] (2) Mass spectrometry conditions are shown in Table 4:

[0097] Table 4

[0098] Ion source Electrospray ionization (ESI) model Positive ions and negative ions Scan range 50-1200 m / z Curtain Air (CUR) 35psi Spray voltage (IS) 5500V Ionization temperature (TEM) 550℃ Atomizing gas (GS1) 55psi Atomizing gas (GS2) 55psi Declustering voltage (DP) 80V Entry voltage (EP) 10V Collision energy (CE) 46V

[0099] (3) Test samples:

[0100] Accurately weigh 20 μL of the test solution 1-1 and inject it into the liquid chromatograph for detection and analysis.

[0101] Example 3

[0102] This embodiment provides a method for detecting polymer impurities in a product containing cefuroxime sodium. The test conditions are as follows:

[0103] (1) Liquid chromatography conditions:

[0104] Column: Welch Ultimate XB-C8, 4.6 mm × 250 mm, 5.0 μm;

[0105] Mobile phase: Mobile phase A is 5 mmol / L ammonium acetate solution (pH adjusted to 4.0 with acetic acid); mobile phase B is acetonitrile; detection wavelength: 275 nm;

[0106] Column temperature: 25°C;

[0107] Flow rate: 2 mL / min;

[0108] Injection volume: 25 μL;

[0109] The elution gradient program is shown in Table 5:

[0110] Table 5

[0111]

[0112]

[0113] (2) Mass spectrometry conditions are shown in Table 6:

[0114] Table 6

[0115] Ion source Electrospray ionization (ESI) model Positive ions and negative ions Scan range 50~1200m / z Curtain Air (CUR) 35psi Spray voltage (IS) 5500V Ionization temperature (TEM) 550℃ Atomizing gas (GS1) 55psi Atomizing gas (GS2) 55psi Declustering voltage (DP) 80V Entry voltage (EP) 10V Collision energy (CE) 46V

[0116] (3) Test samples:

[0117] Accurately weigh 20 μL of the test solution 1-1 and inject it into the liquid chromatograph for detection and analysis.

[0118] Example 4

[0119] This example provides a method for detecting polymer impurities in a product containing cefuroxime sodium. The test sample is test solution 1-1. The method differs from Example 1 only in that the chromatographic column is replaced with an Agilent ZORBAX Eclipse XDB-C8, 4.6 mm × 250 mm, 5 μm column. The remaining steps remain the same as in Example 1.

[0120] Example 5

[0121] This example provides a method for detecting polymer impurities in a product containing cefuroxime sodium. The test sample is the test solution 1-1. The only difference from Example 1 is that the chromatographic column is replaced with a Shimpack GISS C18, 2.1 mm × 100 mm, 1.9 μm. The remaining steps are consistent with Example 1.

[0122] Example 6

[0123] This example provides a method for detecting polymer impurities in a product containing cefuroxime sodium. The test sample is test solution 1-1. The only difference from Example 1 is the elution gradient program shown in Table 7. The remaining steps remain the same as Example 1.

[0124] Table 7

[0125]

[0126]

[0127] Example 7

[0128] This example provides a method for detecting polymer impurities in a product containing cefuroxime sodium. The test sample is test solution 1-1. The only difference from Example 1 is the elution gradient program shown in Table 8. The remaining steps remain the same as Example 1.

[0129] Table 8

[0130] Time (min) Mobile phase A (%) Mobile phase B (%) 0.0 70 30 5.0 70 30 40.0 60 40 45.0 60 40 45.1 70 30 50.0 70 30

[0131] Comparative Example 1

[0132] This comparative example provides a method for detecting polymer impurities in a product containing cefuroxime sodium. The difference between the method and Example 1 is that the mobile phase A is replaced with an equal volume of sodium acetate solution (the pH value is adjusted to 3.4 with acetic acid), and the remaining steps are consistent with Example 1.

[0133] Comparative Example 2

[0134] This comparative example provides a method for detecting polymer impurities in a product containing cefuroxime sodium. The difference between the comparative example and Example 1 is that the mobile phase B is replaced with an equal volume of methanol, and the remaining steps are consistent with Example 1.

[0135] Comparative Example 3

[0136] This comparative example provides a method for detecting polymer impurities in a product containing cefuroxime sodium. The method disclosed in the prior art CN117589902A is used to detect polymer impurities. The test conditions are as follows:

[0137] (1) Liquid chromatography conditions:

[0138] Chromatographic column: Agilent ZORBAX Eclipse XDB-C8, 4.6 mm × 250 mm, 5.0 μm;

[0139] Mobile phase: Mobile phase A (0.68 g of sodium acetate and 5.8 g of glacial acetic acid were dissolved in water and diluted to 1000 mL. The pH was adjusted to 3.4 with glacial acetic acid and ultrasonicated for 30 min as mobile phase A); mobile phase B was acetonitrile;

[0140] Detection wavelength: 273nm;

[0141] Column temperature: 30°C;

[0142] Flow rate: 1.5 mL / min;

[0143] Injection volume: 20 μL;

[0144] The elution gradient program is shown in Table 9:

[0145] Table 9

[0146] Time (min) Mobile phase A (%) Mobile phase B (%) 0 95 5 5 95 5 30 80 20 50 65 35

[0147] (2) Mass spectrometry conditions: consistent with those in implementation 1;

[0148] (3) Test samples:

[0149] Accurately weigh 20 μL of the test solution 1-1 and inject it into the liquid chromatograph for detection and analysis.

[0150] Test Example 1

[0151] This test example analyzes the test results of Example 1 and Comparative Example 3:

[0152] (1) First, the liquid chromatogram in Comparative Example 3 is as follows Figure 1 As shown in Figure 10 (where A is a liquid chromatogram and B is an enlarged view of the elution time section from 30 min to 52 min), the impurity information and elution order in the chromatogram are shown in Table 10:

[0153] Table 10

[0154]

[0155] Combined with the structural analysis of impurity H and polymer impurity I, based on the separation principle of reverse HPLC that the smaller the polarity, the later the peak, the impurity eluting after impurity H is initially classified as a polymer impurity. Figure 1 As shown in the figure, the peak numbered 1 is the polymer impurity 1 which overlaps with another polymer impurity with a molecular weight of 920.35 (positive ion 920.35, negative ion 901.2) after HPLC-MS scanning, and peak 2 and peak 3 are not completely separated.

[0156] (2) The liquid chromatogram of the test solution 1-1 (placed under high temperature conditions for 20 days) in Example 1 is as follows Figure 2As shown, the liquid chromatogram of the test solution 1-2 (placed under light conditions for 20 days) is as follows Figure 3 As shown, the liquid chromatogram of the test solution 2-1 (cefuroxime sodium for injection diluted with water) is as follows Figure 4 As shown, the liquid chromatogram of the test solution 2-2 (cefuroxime sodium for injection diluted with 0.9% sodium chloride solution) is as follows Figure 5 As shown, the liquid chromatogram of the test solution 2-3 (cefuroxime sodium for injection diluted with 10% glucose solution) is as follows Figure 6 As shown, the liquid chromatogram of the test solution 2-4 (cefuroxime sodium for injection diluted with sodium lactate Ringer's solution) is as follows Figure 7 shown.

[0157] The impurity information and elution order in the chromatogram are shown in Table 11 and Table 12:

[0158] Table 11

[0159]

[0160] Table 12

[0161]

[0162]

[0163] The molecular ion peak attribution and structural speculation of each impurity detected by LC-MS analysis are shown in Table 13 and Table 14:

[0164] Table 13

[0165]

[0166]

[0167] Table 14

[0168]

[0169]

[0170]

[0171] As can be seen from the data in Tables 13 and 14, the retention time of the test solution is in the range of 13-35 min, and 15 polymer impurities such as polymers, dimers and trimers with molecular weights between monomers and dimers are detected. The types of all polymer impurities are classified, and the structures of 12 polymer impurities are speculated. Among them, the peaks numbered 7 (polymer impurity 1) and 8 are two peaks separated from the peak numbered 1 in Comparative Example 3. It can be seen that the liquid chromatography conditions involved in the present invention have good separation of the polymer impurities in the product containing cefuroxime sodium, and 15 polymer impurities are successfully separated, which is much higher than the separation degree of each impurity in Comparative Example 3.

[0172] Test Example 2

[0173] This test example tests the sensitivity of the detection methods of various embodiments and comparative examples.

[0174] (1) Test sample:

[0175] Take an appropriate amount of cefuroxime reference substance, weigh accurately, dissolve in water and dilute to a signal-to-noise ratio (S / N) of 10:1 and 3:1.

[0176] (2) Test conditions:

[0177] The test conditions of the detection methods of the embodiments and comparative examples were respectively adopted to carry out detection and analysis.

[0178] (3) Test results:

[0179] The quantitative limit and detection limit results of cefuroxime in the detection methods of various embodiments and comparative examples are shown in Table 15:

[0180] Table 15

[0181]

[0182] Cefuroxime has a similar structure to cefuroxime sodium and cefuroxime sodium impurities (such as polymers such as dimers and trimers). Therefore, its response value can be used to indirectly infer the detection capability of polymer impurities.

[0183] From the data in Examples 1-3 in Table 15, it can be seen that the sensitivity of each polymer impurity is calculated as cefuroxime, the limit of quantification is 0.04 μg / mL-0.05 μg / mL, which is equivalent to 0.0004%-0.0005% of the concentration of the test solution (calculated as 10 mg / mL of cefuroxime), and the limit of detection is 0.02 μg / mL-0.025 μg / mL, which is equivalent to 0.0002%-0.00025% of the concentration of the test solution (calculated as 10 mg / mL of cefuroxime), which is very sensitive.

[0184] Comparison of the data in Example 1 with Comparative Examples 1-2 shows that the detection method has higher sensitivity when using ammonium acetate buffer as mobile phase A and acetonitrile as mobile phase B. Comparison of the data in Example 1 with Comparative Example 3 shows that the detection method of the present invention has lower limits of quantitation and detection than the detection methods disclosed in the prior art.

[0185] Comparison of the data in Example 1 with those in Examples 4-5 shows that the Welch Ultimate XB-C8 column has a higher detection sensitivity. Comparison of the data in Example 1 with those in Examples 6-7 shows that the specific gradient elution procedure can further reduce the limit of quantitation and the limit of detection, thereby improving the detection capability.

[0186] Test Example 3

[0187] This test example tests the repeatability of the detection methods of various embodiments and comparative examples.

[0188] (1) Test sample:

[0189] Reference substance solution: Take an appropriate amount of cefuroxime reference substance, accurately weigh it, dissolve it in water and dilute it to make a solution containing 20 μg / mL of cefuroxime, and prepare 2 parallel portions.

[0190] Test solution: Take an appropriate amount of cefuroxime sodium for injection, accurately weigh it, dissolve it in water and dilute it to make a solution containing 10 mg / mL of cefuroxime, and prepare 6 parallel portions.

[0191] (2) Test method:

[0192] The test conditions of the detection methods of the embodiments and comparative examples were used in turn to perform detection and analysis, and the content of polymer impurity 1 and the total polymer impurity content in 6 parallel samples of the test solution measured by different test methods were calculated, and the relative standard deviation (RSD) was calculated;

[0193] Relative standard deviation (RSD) = standard deviation (SD) / arithmetic mean (X) of impurity content in 6 parallel samples × 100%.

[0194] (3) Test results:

[0195] The RSD results of polymer impurities detected by the detection methods of various examples and comparative examples are shown in Table 16:

[0196] Table 16

[0197]

[0198] From the data of Examples 1-3 in Table 16, it can be seen that the RSDs of the polymer impurity 1 and the total polymer impurity contents are all within 10%, indicating that the detection method of the present invention has good repeatability.

[0199] Comparison of the data in Example 1 with Comparative Examples 1-2 shows that the detection method has better repeatability when using ammonium acetate buffer as mobile phase A and acetonitrile as mobile phase B. Comparison of the data in Example 1 with Comparative Example 3 shows that the detection method of the present invention can more stably detect polymer impurities compared to detection methods disclosed in the prior art.

[0200] Comparison of the data in Example 1 with those in Examples 4-5 shows that the Welch Ultimate XB-C8 column has higher repeatability for detecting polymer impurities in the cefuroxime sodium product. Comparison of the data in Example 1 with those in Examples 6-7 shows that the specific gradient elution procedure can more stably detect the content of polymer impurities in the cefuroxime sodium product.

[0201] Test Example 4

[0202] This test example tests the durability of the column temperature and wavelength of the detection method of Example 1.

[0203] (1) Test sample:

[0204] Reference solution: Accurately weigh an appropriate amount of cefuroxime reference solution, dissolve in water, and dilute to a 10 μg / mL cefuroxime solution. Prepare two replicates.

[0205] Test solution: Take an appropriate amount of cefuroxime sodium for injection, accurately weigh it, dissolve it in water and dilute it to make a solution containing 10 mg / mL of cefuroxime, and prepare 2 parallel aliquots.

[0206] (2) Test method:

[0207] (2.1) Column temperature durability test:

[0208] Referring to the test method of Example 1, two parallel samples of the test solution were tested at 25°C, 30°C and 35°C column temperatures, and the liquid chromatograms at 25°C and 35°C were as follows: Figure 8 and Figure 9 The average content of polymer impurity 1 and the average content of total polymer impurities at each temperature are shown in Table 17. It can be seen that the content of polymer impurity 1 and total polymer impurities are consistent. This shows that when parameters such as the chromatographic column, mobile phase, and elution program are consistent, column temperature has little effect on the test results. This test method is universal and has a wide range of applications.

[0209] Table 17

[0210] Column temperature Polymer impurity 1 content (%) Total polymer impurity content (%) 25℃ 0.003 0.03 30℃ 0.003 0.03 35℃ 0.003 0.03 average value 0.003 0.03

[0211] (2.2) Wavelength durability test:

[0212] Referring to the test method of Example 1, the sample was tested at wavelengths of 271 nm, 273 nm and 275 nm, respectively. The liquid chromatography spectra at wavelengths of 271 nm and 275 nm were as follows: Figure 10 and Figure 11 The average content of polymer impurity 1 and the average content of total polymer impurities at each wavelength are shown in Table 18. It can be seen that the content of polymer impurity 1 and total polymer impurities are consistent. This shows that wavelength has little effect on the test results when parameters such as the chromatographic column, mobile phase, and elution program are consistent. The test method is universal and has a wider range of applications.

[0213] Table 18

[0214] wavelength Polymer impurity 1 content (%) Total polymer impurity content (%) 271nm 0.003 0.03 273nm 0.003 0.03 275nm 0.003 0.03 average value(%) 0.003 0.03

[0215] The applicant declares that the present invention is illustrated by the above-described embodiments, but the present invention is not limited to the above-described embodiments. This does not mean that the present invention must rely on the above-described embodiments in order to be implemented. Those skilled in the art should understand that any improvements to the present invention, equivalent replacements for raw materials in the present invention, additions of auxiliary ingredients, and selection of specific methods, etc., fall within the scope of protection and disclosure of the present invention.

[0216] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the scope of protection of the present invention.

[0217] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any appropriate manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. A method for detecting polymer impurities in a product containing cefuroxime sodium, characterized in that: The detection method comprises: using a reverse phase liquid chromatography-mass spectrometry method to detect a product containing cefuroxime sodium, and obtaining the type and content of polymer impurities in the product containing cefuroxime sodium; The mobile phase of the reversed-phase liquid chromatography comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is an ammonium acetate buffer and the mobile phase B is acetonitrile.

2. The detection method according to claim 1, wherein The pH value of the ammonium acetate buffer is 3-4.

3. The detection method according to claim 1 or 2, characterized in that The chromatographic column of the reversed-phase liquid chromatography is an octylsilane bonded silica gel chromatographic column; Preferably, the model of the chromatographic column is Welch Ultimate XB-C8 chromatographic column.

4. The detection method according to any one of claims 1 to 3, characterized in that The column temperature of the reversed-phase liquid chromatography is 25-35°C; Preferably, the detection wavelength of the reversed-phase liquid chromatography is 270-275 nm; Preferably, the injection flow rate of the reversed-phase liquid chromatography is 1-2 mL / min; Preferably, the injection volume of the reverse phase liquid chromatography is 15-25 μL.

5. The detection method according to any one of claims 1 to 4, characterized in that The reverse phase liquid chromatography adopts the gradient elution mode, and the procedure of the gradient elution is: From 0 to 5 minutes, the volume fraction of mobile phase A was 75-85%, and the volume fraction of mobile phase B was 15-25%; From 5 to 40 minutes, the volume fraction of mobile phase A was changed from 75-85% to 65-75% at a constant rate, and the volume fraction of mobile phase B was changed from 15-25% to 25-35% at a constant rate; At 40-45 min, the volume fraction of mobile phase A was 65-75%, and the volume fraction of mobile phase B was 25-35%; From 45 to 45.1 minutes, the volume fraction of mobile phase A was changed at a constant rate from 65-75% to 75-85%, and the volume fraction of mobile phase B was changed at a constant rate from 25-35% to 15-25%; At 45.1-50 min, the volume fraction of mobile phase A was 75-85%, and the volume fraction of mobile phase B was 15-25%.

6. The detection method according to any one of claims 1 to 5, characterized in that The ion source of the mass spectrometer includes an electrospray ion source; The mass spectrometry modes include positive ion mode and negative ion mode.

7. The detection method according to any one of claims 1 to 6, characterized in that The retention time of the polymer impurities is 15-35 min.

8. The detection method according to any one of claims 1 to 7, characterized in that The detection method adopts an external standard method and uses cefuroxime as a reference substance to perform quantitative analysis on polymer impurities.

9. The detection method according to any one of claims 1 to 8, characterized in that The cefuroxime sodium polymer impurities include a molecular formula of C 31 H 29 N7O 14 S2, C 25 H 24 N6O 12 S.C. 30 H 28 N6O 13 S2, C 30 H 28 N6O 14 S2, C 31 H 29 N7O 14 S2, C 30 H 29 N7O 12 S2, C 25 H 24 N6O 12 S or C 31 H 32 N8O 14 Any one or a combination of at least two of S2.

10. Use of the method for detecting polymer impurities in a product containing cefuroxime sodium according to any one of claims 1 to 9 in detecting a product containing cefuroxime sodium; Preferably, the product comprises an injection containing cefuroxime sodium.

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