Detection method of avibactam sodium related substances

Abstract

The invention discloses a detection method of avibactam sodium related substances, and relates to the technical field of medicine detection. According to the method, a high performance liquid chromatography method is adopted for detection, and chromatographic conditions are as follows: a C18 column is adopted as a chromatographic column; a mobile phase A is a 0.15 mol / L ammonium dihydrogen phosphate solution, a mobile phase B is 0.15 mol / L ammonium dihydrogen phosphate solution-acetonitrile (70: 30), and gradient elution is carried out; the sample injection volume is 5-20 [mu] l, the flow velocity is 0.8 ml / min, and the column temperature is 15-25 DEG C. The method for detecting the avibactam sodium related substances can effectively detect the avibactam sodium related substances, has the advantages of good specificity, high sensitivity, high accuracy and strong interference resistance, and can meet the avibactam sodium detection requirements.

Description

Technical Field

[0001] The present invention relates to the technical field of medical detection, and in particular to a method for detecting avibactam sodium-related substances. Background Art

[0002] Avibactam sodium, also known as [(2S,5R)-2-carbamoyl-7-carbonyl-1,6-diazabicyclo[3.2.1]octan-6-yl]sulfate monosodium salt, belongs to the diazabicyclooctanone class of compounds and is a promising new β-lactamase inhibitor. While it lacks significant antibacterial activity on its own, it exhibits broad-spectrum antibacterial activity when used in combination with various cephalosporins and carbapenems. Furthermore, avibactam sodium itself lacks a β-lactam structure and does not induce the production of β-lactamases. Therefore, avibactam sodium is a highly promising new β-lactamase inhibitor that can effectively alleviate the problem of antibiotic resistance. With the widespread research and application of avibactam sodium, the requirements for its quality control are becoming increasingly stringent. The detection of related substances is a key component of avibactam sodium's quality control.

[0003] Currently, existing methods for detecting related substances of avibactam sodium have some shortcomings. For example, some detection methods do not take into account the response differences between avibactam sodium and impurities in high-performance liquid chromatography, resulting in low detection accuracy. In addition, some methods are not highly specific. There may be related substances with similar structures to avibactam sodium, and existing detection methods may not be able to accurately distinguish these substances from avibactam sodium itself and other normal impurities. In addition, the impurities in avibactam sodium and its related substances have similar structures and close polarity, and their retention times are too short, making it difficult to separate avibactam sodium from its impurities.

[0004] Therefore, it is of great significance to develop a method for detecting related substances of avibactam sodium with high sensitivity, strong specificity and high accuracy to meet the requirements of drug quality control and ensure the quality and safety of avibactam sodium. Summary of the Invention

[0005] To address the above shortcomings, the present invention provides a method for detecting avibactam sodium-related substances. This method has the characteristics of high detection sensitivity, strong specificity, and high accuracy, and can effectively separate and detect five related substances that may be produced in avibactam sodium. The specific technical solution is as follows: A method for detecting avibactam sodium-related substances, wherein the method comprises detecting the avibactam sodium-related substances by high performance liquid chromatography; The relevant substances are shown in the following table:

[0006] The detection method comprises the following steps: Step 1: Preparation of test solution Take avibactam sodium, place it in a volumetric flask, add mobile phase A to dissolve and quantitatively dilute to the scale, shake well to obtain a test solution with a concentration of 1~4 mg / mL; Step 2: Control solution preparation Take the test solution prepared in step 1, place it in a volumetric flask, quantitatively dilute it to the mark with mobile phase A, and shake well to obtain a control solution with a concentration of 1-4 μg / mL; Step 3: Chromatographic conditions The chromatographic column used was a C18 column; mobile phase A was 0.1-0.2 mol / L ammonium dihydrogen phosphate solution, and mobile phase B was 0.1-0.2 mol / L ammonium dihydrogen phosphate solution-acetonitrile, with gradient elution; the injection volume was 5-20 μl; the flow rate was 0.8-0.9 ml / min; the column temperature was 15-30°C; and the detection wavelength was 193-197 nm. Step 4: Determination The test solution of step 1 and the control solution of step 2 were respectively aspirated, injected into the high performance liquid chromatograph, and the data were read; based on the control solution and the corresponding chromatographic peak area, the chromatographic peak area of ​​the related substances in the test solution was determined by locating the impurities by relative retention time, and the content of the related substances in the avibactam sodium test sample was calculated by the self-reference method with correction factor.

[0007] Preferably, the chromatographic column C18 column is Waters HSS T3, with a specification of 4.6×250 mm and 3.5 μm; the chromatographic column further comprises a trapping column, which is Shimadzu SHIMADZU Ghost trap DS, with a specification of 7.6 mm ID×30 mm.

[0008] Preferably, the volume ratio of ammonium dihydrogen phosphate solution to acetonitrile in the mobile phase B is 70:30.

[0009] Preferably, the gradient elution procedure is as follows: by volume percentage, the initial ratio is: the amount of mobile phase A is 100%, and the amount of mobile phase B is 0%; 0-16 min, the amount of mobile phase A is 100%, and the amount of mobile phase B is 0%; 16-40 min, the amount of mobile phase A changes from 100% to 60%, and the amount of mobile phase B changes from 0% to 40%; 40-40.1 min, the amount of mobile phase A changes from 60% to 100%, and the amount of mobile phase B changes from 40% to 0%; 40.1-50 min, the amount of mobile phase A is 100%, and the amount of mobile phase B is 0%; the final ratio is: the amount of mobile phase A is 100%, and the amount of mobile phase B is 0%.

[0010] Preferably, the injection volume is 20 μl, the flow rate is 0.8 ml / min, the column temperature is 25° C., and the wavelength is 195 nm.

[0011] Preferably, the mobile phase A is a 0.15 mol / L ammonium dihydrogen phosphate solution; and the mobile phase B is a 0.15 mol / L ammonium dihydrogen phosphate solution-acetonitrile.

[0012] Preferably, the content of related substances is calculated by peak area according to the correction factor self-control method, and the calculation formula is as follows:

[0013] Where: A 供 is the peak area of ​​each impurity in the test solution; A 对 is the peak area of ​​the self-reference solution; F is the correction factor for each impurity.

[0014] The present invention also provides an application of the above-mentioned method for detecting avibactam sodium-related substances in the quality control of avibactam sodium.

[0015] Compared with the prior art, the present invention has the following beneficial effects: 1. The present invention uses ammonium dihydrogen phosphate solution and acetonitrile as mobile phases, controls the concentration and ratio of the two, and optimizes the elution procedure to achieve the separation of (2S,5R)-7-oxo-2-sulfonylformamide-1,6-diazabicyclo[3.2.1]octane-6-yl sulfate monoester disodium salt, [(2R,5R)-2-formyl-7-oxo-1,6-diazabicyclo[3.2.1]-octane-6-yl]-sulfuric acid monoester sodium salt, (2S,5R)-6-hydroxy-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxamide, ((3R,6S)-6-formamidopiperidin-3-yl(ethoxycarbonyl)amino)sulfonate and ((((3R,6S)-6-formamido-1-(ethoxycarbonyl)amino)sulfonate) in avibactam sodium. The effective separation and detection of five impurities, including (piperidin-3-yl)oxy)sulfonic acid, with good separation effect and high accuracy, can effectively control drug quality, reduce possible adverse reactions in drug clinical trials and treatment, and ensure the effectiveness and safety of patients' medication.

[0016] 2. The present invention's method for detecting avibactam sodium-related impurities is highly specific, with no mutual interference between the impurities and the blank solvent. The resolution between each component peak is greater than 1.5, enabling efficient separation and detection of five impurities at one time. Furthermore, the present invention's chromatographic conditions provide high sensitivity for detecting impurities. Furthermore, the present invention's method is robust, with minor changes in chromatographic conditions not affecting the separation of the component peaks. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for describing the embodiments.

[0018] Figure 1 This is the full wavelength scanning spectrum in Example 1 of the present invention; Figure 2 This is the specificity investigation spectrum in Example 1 of the present invention; Figure 3 is the linear equation curve in Example 1 of the present invention; Figure 4 This is the accuracy inspection spectrum in Example 1 of the present invention; Figure 5 The graphs of the sample solution and the control solution under the basic conditions of the durability investigation in Example 1 of the present invention are as follows; Figure 6 This is a sample solution diagram for durability testing in Example 1 of the present invention; Figure 7 This is a reference solution spectrum for the durability study in Example 1 of the present invention; Figure 8 This is the sample solution spectrum in Comparative Example 1 of the present invention; Figure 9 These are the impurity location solution atlas and the test sample solution atlas in Comparative Example 2 of the present invention. DETAILED DESCRIPTION

[0019] The specific embodiments of the present invention are described in detail below, but it should be understood that the scope of protection of the present invention is not limited by the specific embodiments. Unless otherwise defined, all technical terms used hereinafter have the same meaning as those generally understood by those skilled in the art. The technical terms used herein are only for the purpose of describing specific embodiments and are not intended to limit the scope of protection of the present invention. Unless otherwise specified, the various raw materials, reagents, instruments and equipment used in the present invention can be purchased from the market or can be prepared by existing methods.

[0020] The sources of the instruments and equipment and some reagents used in the following specific embodiments are shown in Table 1.

[0021] Table 1 Instruments, consumables, and reagents models and sources Example 1

[0022] 1. A method for detecting related substance impurities of avibactam sodium, comprising the following steps: (1) Preparation of test solution: Take 40±0.5 mg of the avibactam sodium test sample to be tested, accurately weigh it, place it in a 10 ml volumetric flask, add mobile phase A to dissolve it and dilute it to the scale, shake well, and obtain the solution; (2) Preparation of control solution: Accurately measure 1 ml of the test solution, place it in a 100 ml volumetric flask, add mobile phase A to dilute to the mark, and shake well; accurately measure 1 ml of the test solution, place it in a 10 ml volumetric flask, add mobile phase A to dilute to the mark, and shake well. (3) Chromatographic conditions were as follows: chromatographic column: Waters HSS T3 4.6×250 mm, 3.5 μm; trapping column: Shimadzu SHIMADZU Ghost trap DS 7.6 mm ID×30 mm; mobile phase A was 0.15 mol / L ammonium dihydrogen phosphate solution, mobile phase B was 0.15 mol / L ammonium dihydrogen phosphate solution-acetonitrile (70:30), injection volume was 20 μl, flow rate was 0.8 ml / min, column temperature was 25°C; the gradient elution program by volume percentage is shown in Table 2; Table 2 Gradient elution program (4) Detection: Accurately measure 20 μl of the test solution and the reference solution, inject them into the liquid chromatograph, and record the chromatogram. If a chromatographic peak with the same relative retention time as the reference substance impurity-related substance appears in the chromatogram of the test solution, it indicates that the impurity is present. Calculate the impurity content by peak area using the self-reference method with a correction factor. The calculation formula is as follows:

[0023] Where: F is the correction factor for each impurity; A 供 is the peak area of ​​each impurity in the test solution; A 对 The peak area of ​​the self-reference solution. Acceptable criteria: The relative retention time, correction factor, and limit of each impurity peak are shown in Table 3: Table 3 Relative retention time, correction factor and limit of each impurity peak

[0024] Acceptable standard: The content of impurity 1, impurity 2, impurity 3, impurity 4 and impurity 5 shall not exceed 0.10%.

[0025] 2. Determination of detection wavelength A full wavelength scan was performed using an ultraviolet detector to detect avibactam sodium, impurity 1, impurity 2, impurity 3, impurity 4, and impurity 5, and the target peak absorption wavelength was examined. The full wavelength scan results are shown in FIG. Figure 1 .

[0026] pass Figure 1 It can be seen that all impurities and avibactam sodium have terminal absorption, so the wavelength of the measurement method is selected as 195 nm.

[0027] 3. Method Validation 3.1 Specificity Investigation According to the test solution preparation method under "1. A method for detecting related substances of avibactam sodium", 39.98 mg of the test sample was accurately weighed and placed in a 10 ml volumetric flask. Mobile phase A was added to dissolve and dilute to the scale, and shaken to obtain the test solution.

[0028] Impurity 1, impurity 2, impurity 3, impurity 4, impurity 5 reference substance stock solutions: Weigh 2.222 mg of impurity 1 reference substance, 2.315 mg of impurity 2 reference substance, 2.649 mg of impurity 3 reference substance, 2.693 mg of impurity 4 reference substance, and 2.900 mg of impurity 5 reference substance into 10 ml volumetric flasks respectively, add mobile phase A to dissolve and dilute to the scale, shake well, and obtain the stock solutions of each impurity reference substance.

[0029] Impurity mixed reference substance stock solution: Accurately measure 1 ml each of impurity 1, impurity 2, impurity 3, impurity 4, and impurity 5 reference substance stock solutions, place them in the same 10 ml volumetric flask, add mobile phase A to dilute to the scale, shake well, and obtain.

[0030] Impurity localization solution: Accurately measure 1 ml of each of the above impurity reference stock solutions and place them in 10 ml volumetric flasks respectively. Add mobile phase A to dilute to the scale and shake well.

[0031] Sample solution: Weigh 41.38 mg of avibactam sodium test sample into a 10 ml volumetric flask, add 1 ml of impurity mixed reference stock solution, add mobile phase A to dissolve and dilute to the scale, and shake well.

[0032] Take blank solution (mobile phase A), impurity location solution, and sample solution for testing. The results are shown in Table 4 and Figure 2 The results showed that the blank solution, test solution and other impurities did not interfere with the detection of impurity-related substances, and the method had good specificity.

[0033] Table 4 Results of investigation on specificity of relevant substances

[0034] 3.2 Investigation of limit of quantification and limit of detection Prepare the impurity reference substance stock solution according to the preparation method of impurity reference substance stock solution under “3.1 Specificity Investigation”.

[0035] Avibactam sodium reference substance stock solution: Weigh 2.576 mg of avibactam sodium reference substance, place it in a 10 ml volumetric flask, add mobile phase A to dissolve and dilute to the scale, shake well; accurately measure 1 ml, place it in a 10 ml volumetric flask, add mobile phase A to dilute to the scale, shake well, and obtain.

[0036] Take appropriate amount of impurity reference stock solution and avibactam sodium reference stock solution, quantitatively dilute with mobile phase A, the signal-to-noise ratio is greater than 10, the quantification limit results are: avibactam sodium is 0.046134μg / ml, impurity 1 is 0.037557μg / ml, impurity 2 is 0.11518μg / ml, impurity 3 is 0.12290μg / ml, impurity 4 is 0.16144μg / ml, impurity 5 is 0.06416μg / ml, the signal-to-noise ratio is greater than 3, the detection limit results are: avibactam sodium is 0.023067μg / ml, impurity 1 is 0.018779μg / ml, impurity 2 is 0.05759μg / ml, impurity 3 is 0.06145μg / ml, impurity 4 is 0.08072μg / ml, impurity 5 is 0.03208μg / ml.

[0037] 3.3 Linearity and range Prepare the impurity reference substance stock solution according to the preparation method of impurity reference substance stock solution under “3.1 Specificity Investigation”.

[0038] Prepare the avibactam sodium reference stock solution according to the preparation method of avibactam sodium reference stock solution under "3.2 Investigation of Quantitation Limit and Detection Limit".

[0039] Linear solution: Take appropriate amount of impurity reference substance stock solution and avibactam sodium reference substance stock solution, dilute them step by step with mobile phase A to different concentrations, and use them as linear solutions. Take each solution and analyze it from low concentration to high concentration. Perform linear regression with concentration as the horizontal axis and peak area as the vertical axis. The results are shown in Table 5 and Figure 3 .

[0040] Table 5 Linearity test results of related substances

[0041] 3.4 Accuracy Investigation Sample recovery experiments were performed at 50%, 100%, and 150% of the impurity limits.

[0042] Prepare the impurity reference substance stock solution according to the preparation method of impurity reference substance stock solution under “3.1 Specificity Investigation”.

[0043] Recovery solution 50%, 100%, 150%: Weigh 40 mg of the test sample and place it in a 10 ml volumetric flask. Add 0.5, 1.0, and 1.5 ml of the impurity reference stock solution, respectively. Dissolve it in mobile phase A and dilute to the scale. Shake well.

[0044] The recovered solution was sampled and analyzed. The results are shown in Table 6 and Figure 4The results showed that the RSD% recovery of each impurity in the test solution at each concentration level was between 90% and 108%, and the RSD values ​​of the recovery rates of each impurity were all less than 10%, indicating that the method of the present invention has high accuracy.

[0045] Table 6 Accuracy test results

[0046] 3.5 Precision investigation Prepare the recovery solution according to the preparation method of 100% recovery solution under “3.4 Accuracy Test”.

[0047] The precision of the recovered solution was tested six times on different days, by different analysts, and using different equipment. The precision of the impurity content of the test solution was calculated 12 times. The results are shown in Table 7.

[0048] The results showed that the RSD values ​​of the impurity contents in the 12 recovered solutions were all less than 20%, and the injection precision was good, meeting the requirements.

[0049] Table 7 Precision inspection results

[0050] 3.6 Durability Assessment Prepare the sample solution according to the preparation method of recovery solution-100% solution under "3.4 Accuracy Test".

[0051] Control solution: Accurately measure 1 ml of the sample solution, place it in a 100 ml volumetric flask, add mobile phase A to dilute to the scale, and shake well; accurately measure 1 ml, place it in a 10 ml volumetric flask, add mobile phase A to dilute to the scale, and shake well.

[0052] Under the basic chromatographic conditions specified in "1. A Detection Method for Related Substances of Avibactam Sodium", fine-tune the chromatographic conditions (see Table 8). After the system stabilizes, sample each solution (control solution and sample solution) for analysis.

[0053] The results are shown in Table 9 and the patterns are shown in Figures 5-7 The results show that the chromatographic conditions were fine-tuned, the solvent blank had no interference, and the RSD values ​​of the impurity contents in the sample solution were all less than 15%, indicating that the durability of the method of the present invention meets the requirements.

[0054] Table 8 Durability test conditions

[0055] Table 9 Durability test results

[0056] Comparative Example 1 This comparative example provides a method for detecting related substances of avibactam sodium, comprising the following steps: (1) The preparation of the sample solution is the same as that of the sample solution under “3.1 Specificity Investigation” in Example 1.

[0057] (2) Chromatographic conditions: Chromatographic column: Waters HSS T3 4.6×250 mm, 3.5 μm; trapping column: Shimadzu SHIMADZUGhost trap DS 7.6 mm ID×30 mm; mobile phase A was 0.15 mol / L ammonium dihydrogen phosphate solution, mobile phase B was 0.15 mol / L ammonium dihydrogen phosphate solution-acetonitrile (70:30), injection volume was 20 μl, flow rate was 0.8 ml / min, column temperature was 25°C; the gradient elution program by volume percentage is shown in Table 10: Table 10 Gradient elution program

[0058] (3) Take the sample solution and analyze it. The results are shown in Figure 8 .

[0059] The results showed that no impurities were detected after a column retention time of 40 min. Therefore, the gradient elution program could be adjusted to shorten the run time.

[0060] Comparative Example 2 This comparative example provides a method for detecting related substances of avibactam sodium, comprising the following steps: (1) The preparation of the impurity localization solution is the same as that of the impurity localization solution under “3.1 Specificity Investigation” in Example 1.

[0061] (2) The preparation of the test solution is the same as that of the test solution under “3.1 Specificity Investigation” in Example 1.

[0062] (3) Chromatographic conditions: Column: Thermo scientific Hypercarb TM 4.6 × 100 mm, 5 μm; mobile phase A was 0.1 mol / L potassium dihydrogen phosphate solution, mobile phase B was 0.2 mol / L potassium dihydrogen phosphate solution-acetonitrile (50:50), injection volume was 10 μl, flow rate was 1.5 ml / min, column temperature was 60°C; the gradient elution program by volume percentage is shown in Table 11; Table 11 Gradient elution program

[0063] (4) Take the impurity location solution and the test solution for analysis. The results are shown in Table 12 and Figure 9 .

[0064] Table 12 Results of Comparative Example 2 The results showed that the retention times of impurities 1 and 2 were close to those of avibactam sodium, and the retention times of impurities 4 and 5 were close to each other. Avibactam sodium and the impurities could not be effectively separated.

[0065] Example 2 The related substances of three batches of avibactam sodium were tested according to the detection method of Example 1. The experimental results are shown in Table 13. No impurities were detected in the samples, which met the product requirements.

[0066] Table 13 Test results of multiple batches of samples of related substances

[0067] In summary, the method of the present invention realizes the preparation of (2S,5R)-7-oxo-2-sulfonylformamide-1,6-diazabicyclo[3.2.1]octane-6-yl sulfate monoester disodium salt, [(2R,5R)-2-formyl-7-oxo-1,6-diazabicyclo[3.2.1]-octane-6-yl]-sulfuric acid monoester sodium salt, (2S,5R)-6-hydroxy-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxamide, ((3R,6S)-6-formamidopiperidin-3-yl(ethoxycarbonyl)amino)sulfonic acid ammonium and ((((3R,6S)-6-formamido-1-(ethoxycarbonyl)amino)sulfonic acid ammonium) in avibactam sodium. The effective separation and detection of five impurities, including (piperidin-3-yl)oxy)sulfonic acid, with good separation effect, high accuracy, strong specificity, high sensitivity, and good durability, can effectively control drug quality, reduce possible adverse reactions in drug clinical trials and treatment, and ensure the effectiveness and safety of patients' medication.

[0068] The foregoing descriptions of specific exemplary embodiments of the present invention are for purposes of illustration and description. These descriptions are not intended to limit the invention to the precise forms disclosed, and it is apparent that many variations and modifications are possible in light of the foregoing teachings. The exemplary embodiments have been selected and described for the purpose of explaining the specific principles of the invention and their practical application, thereby enabling those skilled in the art to realize and utilize a variety of exemplary embodiments of the invention and various options and modifications. The scope of the invention is intended to be defined by the claims and their equivalents.

Claims

1. A method for detecting avibactam sodium-related substances, characterized in that: The detection method is to detect the related substances of avibactam sodium by high performance liquid chromatography; The related substances include: (2S,5R)-7-oxo-2-sulfonylformamide-1,6-diazabicyclo[3.2.1]octane-6-yl sulfate monoester disodium salt, [(2R,5R)-2-formyl-7-oxo-1,6-diazabicyclo[3.2.1]-octane-6-yl]-sulfuric acid monoester sodium salt, (2S,5R)-6-hydroxy-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxamide, ((3R,6S)-6-formamidopiperidin-3-yl(ethoxycarbonyl)amino)sulfonic acid ammonium and ((((3R,6S)-6-formamido-1-(ethoxycarbonyl))amino)piperidin-3-yl)oxy)sulfonic acid; The detection method comprises the following steps: Step 1: Preparation of test solution Take avibactam sodium, place it in a volumetric flask, add mobile phase A to dissolve and quantitatively dilute to the scale, shake well to obtain a test solution with a concentration of 1~4 mg / mL; Step 2: Control solution preparation Take the test solution prepared in step 1, place it in a volumetric flask, quantitatively dilute it to the mark with mobile phase A, and shake well to obtain a control solution with a concentration of 1-4 μg / mL; Step 3: Chromatographic conditions The chromatographic column used was a C18 column; mobile phase A was 0.1-0.2 mol / L ammonium dihydrogen phosphate solution, and mobile phase B was 0.1-0.2 mol / L ammonium dihydrogen phosphate solution-acetonitrile, with gradient elution; the injection volume was 5-20 μl; the flow rate was 0.8-0.9 ml / min; the column temperature was 15-30°C; and the detection wavelength was 193-197 nm. Step 4: Determination The test solution of step 1 and the control solution of step 2 were respectively aspirated, injected into the high performance liquid chromatograph, and the data were read; based on the control solution and the corresponding chromatographic peak area, the chromatographic peak area of ​​the related substances in the test solution was determined by locating the impurities by relative retention time, and the content of the related substances in the avibactam sodium test sample was calculated by the self-reference method with correction factor.

2. The method for detecting an avibactam sodium-related substance according to claim 1, wherein The chromatographic column C18 column is a Waters HSS T3 column with a specification of 4.6×250 mm and 3.5 μm. The chromatographic column also includes a trapping column, which is a Shimadzu SHIMADZU Ghost trap DS column with a specification of 7.6 mm ID×30 mm.

3. The method for detecting an avibactam sodium-related substance according to claim 1, wherein The volume ratio of ammonium dihydrogen phosphate solution to acetonitrile in the mobile phase B is 70:

30.

4. The method for detecting avibactam sodium-related substances according to claim 1, wherein The gradient elution procedure is as follows: by volume percentage, from 0 to 16 min, the amount of mobile phase A is 100%, and the amount of mobile phase B is 0%; from 16 to 40 min, the amount of mobile phase A changes from 100% to 60%, and the amount of mobile phase B changes from 0% to 40%; from 40 to 40.1 min, the amount of mobile phase A changes from 60% to 100%, and the amount of mobile phase B changes from 40% to 0%; 40.1-50min, the amount of mobile phase A is 100%, and the amount of mobile phase B is 0%.

5. The method for detecting avibactam sodium-related substances according to claim 1, wherein The injection volume was 20 μl, the flow rate was 0.8 ml / min, the column temperature was 25° C., and the wavelength was 195 nm.

6. The method for detecting avibactam sodium-related substances according to claim 1, wherein The mobile phase A is 0.15 mol / L ammonium dihydrogen phosphate solution; the mobile phase B is 0.15 mol / L ammonium dihydrogen phosphate solution-acetonitrile.

7. The method for detecting avibactam sodium-related substances according to claim 1, wherein The content of related substances was calculated by peak area using the correction factor self-control method. The calculation formula is as follows: Where: A 供 is the peak area of ​​each impurity in the test solution; A 对 is the peak area of ​​the self-reference solution; F is the correction factor for each impurity.

8. Use of the method for detecting avibactam sodium-related substances according to any one of claims 1 to 7 in the quality control of avibactam sodium.

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