Method for detecting genotoxic impurities in moxifloxacin hydrochloride raw material or preparation thereof

Abstract

The invention relates to the technical field of pharmaceutical analysis, and particularly discloses a method for detecting genotoxic impurities in a moxifloxacin hydrochloride raw material or a preparation thereof. According to the method, a high performance liquid chromatography and mass spectrometry combined method is adopted for detection, and liquid chromatography conditions are as follows: a C18 chromatographic column is used, a formic acid aqueous solution with the volume concentration of 0.05%-0.2% is used as a mobile phase A, an acetonitrile solution of formic acid with the volume concentration of 0%-0.2% is used as a mobile phase B, and gradient elution is performed; the mass spectrum adopts an ESI ion source and positive ion detection mode, the parent ion is 431.15 m / z, the daughter ion is 401.18 m / z, and the collision voltage is 10V-25V. The method provided by the invention is strong in specificity and high in sensitivity, the detection limit is 0.625 ppm, the blank that N-nitroso moxifloxacin impurities in a moxifloxacin hydrochloride raw material or a preparation cannot be effectively detected in the prior art is filled up, and the medication safety risk of a moxifloxacin hydrochloride medicine is further reduced.

Description

Technical Field

[0001] The invention relates to the technical field of drug analysis, and in particular to a method for detecting genotoxic impurities in a moxifloxacin hydrochloride raw material or a preparation thereof. Background Art

[0002] The chemical name of moxifloxacin is: 1-cyclopropyl-7-﹛(S,S)-2,8-diazo-bicyclo[4.3.0]non-8-yl﹜-6-fluoro-8-methoxy-1,4-dihydro-4-oxo-3-quinolinecarboxylic acid, CAS number: 151096-09-2. This drug is a synthetic quinolone antibacterial drug used to treat symptoms caused by respiratory tract infections. The structural formula is as follows.

[0003]

[0004] The existing process for synthesizing moxifloxacin hydrochloride and the existing process for producing moxifloxacin hydrochloride tablets may cause the nitrosating reagents remaining in the raw materials or in the pipeline to react with the raw material moxifloxacin to generate the genotoxic nitrosamine impurity N-nitroso-moxifloxacin (N-nitrosomoxifloxacin), the chemical structure of which is shown below.

[0005]

[0006] The above-mentioned N-nitrosomoxifloxacin impurities are nitrosamine genotoxic impurities, which need to be strictly controlled in raw materials and tablets. According to the regulatory requirements of the FDA or the European Union, the acceptable intake of N-nitrosomoxifloxacin is 1500ng / day. According to the ICH M7 guideline, the acceptable limit of N-nitrosomoxifloxacin in moxifloxacin API and tablets is calculated as AI (ng / day) / MDD (mg / day)=1500ng / day / 400mg / day=3.75ppm; wherein AI represents the acceptable daily intake, and MDD represents the maximum daily dose of the drug. The acceptable limit of N-nitrosomoxifloxacin in moxifloxacin API and tablets is 3.75ppm (in terms of moxifloxacin), the impurity detection limit (LOD) is 1 / 6 of the acceptable limit, the detection limit (LOD) is generally required to be 0.625ppm, and the quantitative limit (LOQ) is generally required to be 1.875ppm.

[0007] Since the acceptable limit of N-nitrosomoxifloxacin in moxifloxacin hydrochloride bulk drug and tablets is only 3.75ppm, the detection sensitivity of conventional high performance liquid chromatography (HPLC) test methods cannot meet the requirements, and there are technical difficulties in sensitively and accurately detecting trace amounts of N-nitrosomoxifloxacin in moxifloxacin bulk drug and tablets. No reports on detection methods for N-nitrosomoxifloxacin that can achieve a detection limit lower than 0.625ppm have been found in the prior art. Therefore, it is urgent to provide a method that can sensitively and accurately test N-nitrosomoxifloxacin in moxifloxacin hydrochloride and moxifloxacin hydrochloride tablets, so as to accurately control the content of N-nitrosomoxifloxacin impurities in moxifloxacin hydrochloride bulk drug and tablets. Summary of the invention

[0008] Aiming at the problem that the prior art cannot effectively detect trace amounts of N-nitroso moxifloxacin in a moxifloxacin hydrochloride raw material or preparation, the present invention provides a method for detecting genotoxic impurities in a moxifloxacin hydrochloride raw material or preparation thereof.

[0009] In order to solve the above technical problems, the technical solution provided by the present invention is:

[0010] A method for detecting genotoxic impurities in a moxifloxacin hydrochloride raw material or a preparation thereof, wherein the genotoxic impurity is N-nitrosomoxifloxacin, and high performance liquid chromatography-mass spectrometry is used for detection, comprising the following steps:

[0011] (1) Preparation of test solution and reference solution:

[0012] Take the moxifloxacin hydrochloride raw material or its preparation and prepare a test solution with a solvent;

[0013] Take N-nitrosomoxifloxacin reference substance and prepare reference substance solutions with a series of concentrations using solvent;

[0014] (2) Testing the reference solution and the test solution of the series of concentrations, wherein the conditions of the high performance liquid chromatography are:

[0015] A C18 chromatographic column was used, with a formic acid aqueous solution having a volume concentration of 0.05%-0.2% as the mobile phase A, and an acetonitrile solution having a volume concentration of 0%-0.2% formic acid as the mobile phase B, and gradient elution was performed;

[0016] The mass spectrometer adopts an ESI ion source and a positive ion detection mode, wherein the quantitative ion of the N-nitrosomoxifloxacin is: a parent ion of 431.15 m / z, a daughter ion of 401.18 m / z, and a collision voltage of 10V-25V.

[0017] Compared with the prior art, the method for detecting genotoxic impurities in a moxifloxacin hydrochloride raw material or its preparation provided by the present invention adopts a C18 chromatographic column, a specific mobile phase, a gradient elution mode, and a specific mass spectrometry condition, so as to achieve accurate detection of trace amounts of N-nitrosomoxifloxacin in a moxifloxacin hydrochloride raw material or its preparation, and the detection limit of N-nitrosomoxifloxacin is 0.625ppm, and the quantitative limit is 1.875ppm, which meets the detection requirements of the guiding principles of ICH M7. In addition, the method provided by the present invention has strong specificity, high sensitivity, and good linear relationship. At the same time, it has good accuracy and durability, and can achieve trace detection of potential process impurities N-nitrosomoxifloxacin in a moxifloxacin hydrochloride raw material or its preparation, which can be used as the basis for quality monitoring of moxifloxacin hydrochloride drugs, and makes up for the gap that the prior art cannot effectively detect genotoxic impurities of N-nitrosomoxifloxacin in moxifloxacin hydrochloride, thereby helping to reduce the drug safety risk of moxifloxacin hydrochloride drugs.

[0018] Preferably, the mobile phase A is 0.05% formic acid aqueous solution, and the mobile phase B is acetonitrile.

[0019] The preferred mobile phase can better separate moxifloxacin hydrochloride and N-nitrosomoxifloxacin without generating baseline interference, and can effectively improve the peak shape, thereby making the detection results more accurate and precise.

[0020] Preferably, the elution procedure of the gradient elution is as follows:

[0021] 0 min, 78%-82% mobile phase A, 22%-18% mobile phase B;

[0022] 3 min, 78%-82% mobile phase A, 22%-18% mobile phase B;

[0023] 5 min, 0%-2% mobile phase A, 100%-98% mobile phase B;

[0024] 15 min, 0%-2% mobile phase A, 100%-98% mobile phase B;

[0025] 15.1 min, 78%-82% mobile phase A, 22%-18% mobile phase B;

[0026] 20min, 78%-82% mobile phase A, 22%-18% mobile phase B.

[0027] Further preferably, the elution procedure of the gradient elution is as follows:

[0028] 0 min, 80% mobile phase A, 20% mobile phase B;

[0029] 3 min, 80% mobile phase A, 20% mobile phase B;

[0030] 5 min, 0% mobile phase A, 100% mobile phase B;

[0031] 15 min, 0% mobile phase A, 100% mobile phase B;

[0032] 15.1 min, 80% mobile phase A, 20% mobile phase B;

[0033] 20 min, 80% mobile phase A, 20% mobile phase B.

[0034] The preferred gradient elution sequence can improve the separation between the main component and N-nitrosomoxifloxacin and the detection sensitivity, making the detection result quantitatively accurate and highly precise.

[0035] Preferably, the chromatographic column is C18, 4.0mm*100mm, 5μm.

[0036] The preferred chromatographic column can reduce baseline interference and improve the sensitivity and accuracy of N-nitrosomoxifloxacin detection.

[0037] Preferably, the column temperature is 33°C-37°C, and the flow rate is 0.25mL / min-0.35mL / min.

[0038] More preferably, the column temperature is 35° C. and the flow rate is 0.3 mL / min.

[0039] Preferably, the injection volume is 20 μL.

[0040] The preferred detection conditions are conducive to the effective detection of the genotoxic impurity N-nitrosomoxifloxacin in moxifloxacin hydrochloride and improve the sensitivity of the detection, thereby achieving the purpose of effectively and accurately detecting the trace amount of N-nitrosomoxifloxacin in moxifloxacin hydrochloride.

[0041] Preferably, the scanning mode of the mass spectrometer is multiple reaction monitoring, the desolvation gas temperature is 220℃-300℃, the ion source temperature is 100℃-120℃, the desolvation gas flow rate is 550L / h-750L / h, the cone gas flow rate is 0L / h-50L / h, the capillary voltage is 2.8kV-3.5kV, the cone voltage is 15V-40V, the secondary cone voltage is 1V-5V, and the sextupole lens voltage is 0.1V-0.5V.

[0042] Furthermore, the desolvation gas temperature is 300°C, the ion source temperature is 120°C, the desolvation gas flow rate is 700L / h, the cone gas flow rate is 35L / h, the capillary voltage is 3.0kV, the cone voltage is 30V, the secondary cone voltage is 3V, and the sextupole lens voltage is 0.3V.

[0043] The optimal mass spectrometry detection conditions can maximize the accuracy of the determination of trace N-nitrosomoxifloxacin in moxifloxacin hydrochloride.

[0044] Preferably, the concentration of the test solution is 4 mg / mL.

[0045] The preferred test sample concentration is conducive to better peak shapes of the main component and N-nitrosomoxifloxacin impurities, higher column efficiency, and more accurate integration, thereby facilitating more accurate calculation of the content of N-nitrosomoxifloxacin in the test sample.

[0046] Preferably, the concentration of N-nitrosomoxifloxacin in the reference solution is 12 ng / mL, 18 ng / mL, 24 ng / mL, 30 ng / mL, 36 ng / mL, and 45 ng / mL.

[0047] Preferably, the solvent for preparing the test solution and the reference solution is a mixed solution of acetonitrile and water in a volume ratio of 50:50.

[0048] The detection method provided by the present invention can realize accurate quantitative detection of trace amounts of N-nitrosomoxifloxacin impurities in a moxifloxacin hydrochloride raw material or a preparation, thereby being beneficial to monitoring the preparation process of the moxifloxacin hydrochloride raw material or the preparation thereof, and being more beneficial to truly reflecting the quality of the moxifloxacin hydrochloride raw material or the preparation thereof, thereby ensuring the clinical medication safety of the moxifloxacin hydrochloride product, and having high practical value. BRIEF DESCRIPTION OF THE DRAWINGS

[0049] Figure 1 This is the LC-MS graph of the blank solution under 2.1 in Example 2;

[0050] Figure 2 The LC-MS graph of the moxifloxacin hydrochloride raw material under 2.1 in Example 2;

[0051] Figure 3 The LC-MS diagram of the moxifloxacin hydrochloride tablets under item 2.1 in Example 2;

[0052] Figure 4 This is the LC-MS graph of the sensitivity solution under 2.1 in Example 2;

[0053] Figure 5 The LC-MS graph of the accuracy solution 2 of the Moxixa hydrochloride raw material under Item 2.3 in Example 2;

[0054] Figure 6 The LC-MS graph of the accuracy solution 2 of the Moxixa hydrochloride tablets under Item 2.3 in Example 2;

[0055] Figure 7 This is the LC-MS graph of the sensitive solution in Comparative Example 1. DETAILED DESCRIPTION

[0056] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0057] Example 1

[0058] 1.1 Solution preparation

[0059] (1) Blank solution / dilution solution: acetonitrile and water in a 50:50 volume ratio.

[0060] (2) Reference substance stock solution: Take about 10 mg of N-nitrosomoxifloxacin reference substance and place it in a 100-mL volumetric flask. Dissolve it with diluent and dilute it to the mark. Shake well to prepare N-nitrosomoxifloxacin stock solution A. Accurately pipette 1 mL of N-nitrosomoxifloxacin stock solution A into a 100-mL volumetric flask, dilute it to the mark with diluent, and shake well to prepare stock solution B. Accurately pipette 5 mL of stock solution B into a 10-mL volumetric flask, dilute it to the mark with diluent, and shake well to prepare a reference substance stock solution with a concentration of 500 ng / mL.

[0061] (3) Reference substance solution: Accurately pipette 240 μL, 360 μL, 480 μL, 600 μL, 720 μL, and 900 μL of the above reference substance stock solution into six 10 mL volumetric flasks, dilute to the mark with diluent, and shake well to obtain reference substance solutions with concentrations of 12 ng / mL, 18 ng / mL, 24 ng / mL, 30 ng / mL, 36 ng / mL, and 45 ng / mL, respectively, as a series of reference substance solutions.

[0062] (4) Sensitivity solution: Accurately pipette 2083 μL of the reference solution with a concentration of 12 ng / mL into a 10 mL volumetric flask, dissolve it with diluent and dilute to the mark, shake well, and use it as the sensitivity solution (the concentration of N-nitrosomoxifloxacin is 2.5 ng / mL, equivalent to the test sample concentration of 0.625 ppm).

[0063] (5) Raw material test solution: Take 40 mg of moxifloxacin hydrochloride raw material in a 10 mL volumetric flask, dissolve and dilute to the scale with diluent, shake well, and prepare two parallel portions to obtain a test solution with a concentration of 4 mg / mL.

[0064] Tablet test solution: Take 3 moxifloxacin hydrochloride tablets, grind into powder, accurately weigh 72.5 mg into a 10 mL volumetric flask, add 5 mL of diluent and ultrasonically dissolve for 5 min, add diluent to the scale, shake well, centrifuge at 12000 r / min for 5 min, take the supernatant, prepare two portions in parallel, and prepare a test solution with a concentration of 4 mg / mL (calculated as moxifloxacin hydrochloride).

[0065] 1.2 Detection method of N-nitrosomoxifloxacin in moxifloxacin hydrochloride raw materials or preparations:

[0066] Chromatographic column: C18, 4.0mm*100mm, 5μm;

[0067] Mobile phase A: 0.05% formic acid in water;

[0068] Mobile phase B: acetonitrile;

[0069] Flow rate: 0.3 mL / min;

[0070] Column temperature: 35 °C;

[0071] Sample pan temperature: 15°C;

[0072] Injection volume: 20 μL;

[0073] Injector needle cleaning fluid: acetonitrile;

[0074] Gradient elution, the elution program is as follows:

[0075]

[0076]

[0077] Mass spectrometry conditions:

[0078] Ion source type: electrospray ionization (ESI);

[0079] Mass spectrometry time period settings:

[0080]

[0081] ESI source parameter settings:

[0082]

[0083] Desolvation temperature: 300°C;

[0084] Source temperature: 120°C;

[0085] Desolvation gas flow rate: 700L / h

[0086] Cone hole gas flow: 35L / h;

[0087] Capillary voltage: 3.0kV;

[0088] Cone voltage: 30V;

[0089] Secondary cone voltage: 3V;

[0090] Sextupole lens voltage: 0.3V.

[0091] After the detection system is stable, inject the blank solution to ensure that there is no interference from abnormal peaks, and then perform sample testing.

[0092] Example 2

[0093] Methodological validation:

[0094] 2.1 Exclusivity

[0095] Take 20 μL of each of the blank solution, reference solution, sensitivity solution, raw material test solution and tablet test solution prepared in Example 1, and perform HPLC-MS / MS detection according to the above conditions. Each solution is injected once, and the sensitivity solution is injected 6 times. The results are shown in Table 1. The chromatograms are shown in Table 1. Figure 1-Figure 4 shown.

[0096] Table 1 Specificity results of sensitivity solutions

[0097]

[0098] The test results showed that the baseline was stable, there was no interference in the quantitative ion channel of N-nitrosomoxifloxacin, and the RSD of the peak area of ​​the 6-pin sensitivity solution was 2.14%, indicating that the method had good specificity.

[0099] 2.2 Limit of detection and limit of quantification

[0100] Limit of detection solution (LOD): the same as the sensitivity solution in Example 1, with a concentration of 2.5 ng / mL.

[0101] Limit of quantitation solution (LOQ): Take 2.5 mL of the reference solution with a concentration of 30 ng / mL prepared in Example 1, place it in a 10 mL volumetric flask, dilute to the mark with diluent, and shake well to use as the limit of quantitation solution.

[0102] After the system is stable, inject the blank solution to ensure that there is no abnormal peak interference, then inject the detection limit solution and quantification limit solution, record the chromatogram, and the results are shown in Table 2.

[0103] Table 2 Detection limit and quantification limit test results

[0104]

[0105] The results show that the detection limit of the method of the present invention is 2.5 ng / mL, equivalent to 0.625 ppm of the test sample concentration; the quantification limit is 7.5 ng / mL, equivalent to 1.875 ppm of the test sample concentration, the detection limit signal-to-noise ratio S / N is greater than 3, the quantification limit signal-to-noise ratio S / N is greater than 10, and the 5-injection quantification limit peak area RSD% is less than 10%.

[0106] 2.3 Accuracy

[0107] According to Example 1, prepare N-nitrosomoxifloxacin stock solution, reference solution of a series of concentrations, sensitivity solution, raw material test solution and tablet test solution.

[0108] Raw material accuracy solution: Accurately weigh 40.05 mg, 40.11 mg, and 40.02 mg of moxifloxacin hydrochloride raw material test sample and place them in three 10 mL volumetric flasks respectively. First add an appropriate amount of diluent to dissolve, then accurately pipette 240 μL, 300 μL, and 360 μL of N-nitrosomoxifloxacin stock solution into three 10 mL volumetric flasks respectively, then add diluent to dilute to the scale, shake well, and use them as accuracy solution-1, accuracy solution-2, and accuracy solution-3.

[0109] Tablet accuracy solution: Take 3 moxifloxacin hydrochloride tablets, grind into powder, accurately weigh 72.55 mg, 72.58 mg, and 72.60 mg of the powder and place them in 3 10 mL volumetric flasks respectively, add appropriate amount of diluent to dissolve, accurately pipette 240 μL, 300 μL, and 360 μL of N-nitrosomoxifloxacin stock solution into 3 10 mL volumetric flasks respectively, then add diluent to dilute to the scale, shake well, and use them as accuracy solution-1, accuracy solution-2, and accuracy solution-3.

[0110] After the system is stable, inject the blank solution to ensure that there is no abnormal peak interference, then inject the sensitivity solution (1 injection) and the reference solution of a series of concentrations. The impurity signal-to-noise ratio in the sensitivity solution must not be less than 3, and the linear coefficient of the reference solution is R 2 It should not be less than 0.99. After meeting the requirements, inject the blank solution (1 injection), and then inject the test solution and accuracy solution (3 injections for each concentration of accuracy solution). The results are shown in Tables 3-4 and Figure 5-6 shown.

[0111] Table 3 Content of N-nitrosomoxifloxacin in moxifloxacin hydrochloride raw material and tablets

[0112]

[0113] Table 4 Accuracy results of moxifloxacin hydrochloride raw materials and tablets

[0114]

[0115]

[0116] The results showed that the accuracy of the content of N-nitrosomoxifloxacin was good under the detection conditions, the test results were accurate and reliable, the recovery rate of the N-nitrosomoxifloxacin content accuracy experiment was between 80% and 120%, and the RSD met the standard requirements.

[0117] 2.4 Linear range

[0118] Reference solution: the same as the reference solution of the same concentration as in Example 1.

[0119] Operation steps: After the system is stable, inject the blank solution to ensure that there is no abnormal peak interference, then inject the sensitivity solution (1 injection), the impurity signal-to-noise ratio in the sensitivity solution must not be less than 3, and then inject the reference solution respectively, and record the LC-MS graph. With concentration (ng / mL) as the X-axis and peak area as the Y-axis, perform linear regression, and the results are shown in Table 5.

[0120] Table 5 Linear range

[0121] Compound Name Calibration curve Linear range <![CDATA[R 2 ]]> N-nitrosomoxifloxacin y=283.91x-741.21 12.17ng / mL-45.63ng / mL 0.9984

[0122] The results showed that N-nitrosomoxifloxacin showed a good linear relationship in the range of 12.17 ng / mL-45.63 ng / mL.

[0123] 2.5 Precision

[0124] Moxifloxacin hydrochloride raw material precision solution: Prepare raw material accuracy solution-2 in the same way as in 2.4 Accuracy;

[0125] Moxifloxacin hydrochloride tablet precision solution: Prepare tablet accuracy solution-2 in the same way as in 2.4 Accuracy.

[0126] Operation steps: After the system is stable, inject the blank solution to ensure that there is no abnormal peak interference, then inject the sensitivity solution (1 injection), the impurity signal-to-noise ratio in the sensitivity solution must not be less than 3, and then inject the raw material precision solution (6 injections) and tablet precision solution (6 injections) respectively, record the LC-MS graph, take the peak area of ​​each N-nitromoxacin, calculate the average value and relative standard deviation (RSD), and the results are shown in Table 6.

[0127] Table 6 Precision test results (peak area)

[0128] serial number Raw material precision solution Tablet Precision Solution 1 9082.51 4325.05 2 9279.22 4250.11 3 9110.77 4214.25 4 9154.27 4278.88 5 9003.68 4202.25 6 9192.96 4290.08 average value 9137.235 4260.103 RSD(%) 1.04 1.10

[0129] The results show that under the detection conditions of the present invention, the precision of the quantitative analysis results of N-nitrosomoxifloxacin is good.

[0130] 2.6 Durability

[0131] Prepare blank solution, reference solution, raw material test solution, and tablet test solution according to Example 1.

[0132] The above prepared solution was taken for LC-MS detection, and the LC-MS graph was recorded. The durability of N-nitrosomoxifloxacin was investigated by fine-tuning the chromatographic conditions: flow rate 0.25 mL / min and 0.35 mL / min, column temperature 33°C and 37°C, and mobile phase B in the gradient was adjusted by ±2%. The results are shown in Table 7.

[0133] Table 7 Durability test results (content of N-nitrosomoxifloxacin)

[0134]

[0135] The results showed that under the detection conditions of the present invention, fine-tuning the chromatographic conditions had no effect on the detection of N-nitrosomoxifloxacin, indicating that the method had good durability.

[0136] Comparative Example 1

[0137] The detection method provided in this comparative example is different from that in Example 1 in that the mobile phase A is replaced with a 0.05% ammonium acetate aqueous solution, and the other detection conditions are exactly the same. The sensitivity solution in Example 1 is injected for detection, and the results are as follows: Figure 7 shown.

[0138] As can be seen from the figure, the response intensity of N-nitrosomoxifloxacin in the sensitivity solution is significantly reduced and cannot meet the detection limit requirement of N-nitrosomoxifloxacin.

[0139] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modification, equivalent substitution or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for detecting genotoxic impurities in a moxifloxacin hydrochloride raw material or a preparation thereof, characterized in that: The genotoxic impurity is N-nitrosomoxifloxacin, and the detection is performed by high performance liquid chromatography-mass spectrometry, comprising the following steps: (1) Preparation of test solution and reference solution: Take the moxifloxacin hydrochloride raw material or its preparation and prepare a test solution with a solvent; Take N-nitrosomoxifloxacin reference substance and prepare reference substance solutions with a series of concentrations using solvent; (2) Testing the reference solution and the test solution of the series of concentrations, wherein the conditions of the high performance liquid chromatography are: A C18 chromatographic column was used, with a formic acid aqueous solution having a volume concentration of 0.05%-0.2% as the mobile phase A, and an acetonitrile solution having a volume concentration of 0%-0.2% formic acid as the mobile phase B, and gradient elution was performed; The mass spectrometer adopts an ESI ion source and a positive ion detection mode, wherein the quantitative ion of the N-nitrosomoxifloxacin is: a parent ion of 431.15 m / z, a daughter ion of 401.18 m / z, and a collision voltage of 10V-25V.

2. The method for detecting genotoxic impurities in a moxifloxacin hydrochloride raw material or a preparation thereof according to claim 1, characterized in that: The elution procedure of the gradient elution is as follows: 0 min, 78%-82% mobile phase A, 22%-18% mobile phase B; 3 min, 78%-82% mobile phase A, 22%-18% mobile phase B; 5 min, 0%-2% mobile phase A, 100%-98% mobile phase B; 15 min, 0%-2% mobile phase A, 100%-98% mobile phase B; 15.1 min, 78%-82% mobile phase A, 22%-18% mobile phase B; 20min, 78%-82% mobile phase A, 22%-18% mobile phase B.

3. The method for detecting genotoxic impurities in a moxifloxacin hydrochloride raw material or a preparation thereof according to claim 2, characterized in that: The elution procedure of the gradient elution is as follows: 0 min, 80% mobile phase A, 20% mobile phase B; 3 min, 80% mobile phase A, 20% mobile phase B; 5 min, 0% mobile phase A, 100% mobile phase B; 15 min, 0% mobile phase A, 100% mobile phase B; 15.1 min, 80% mobile phase A, 20% mobile phase B; 20 min, 80% mobile phase A, 20% mobile phase B.

4. The method for detecting genotoxic impurities in a moxifloxacin hydrochloride raw material or a preparation thereof according to claim 1, characterized in that: The chromatographic column is C18, 4.0mm*100mm, 5μm.

5. The method for detecting genotoxic impurities in a moxifloxacin hydrochloride raw material or a preparation thereof according to claim 1, characterized in that: The mobile phase A is 0.05% formic acid aqueous solution, and the mobile phase B is acetonitrile.

6. The method for detecting genotoxic impurities in a moxifloxacin hydrochloride raw material or a preparation thereof according to claim 1, characterized in that: The column temperature was 33°C-37°C, and the flow rate was 0.25mL / min-0.35mL / min.

7. The method for detecting genotoxic impurities in a moxifloxacin hydrochloride raw material or a preparation thereof according to claim 6, characterized in that: The column temperature was 35 °C and the flow rate was 0.3 mL / min.

8. The method for detecting genotoxic impurities in a moxifloxacin hydrochloride raw material or a preparation thereof according to claim 1, characterized in that: The injection volume was 20 μL.

9. The method for detecting genotoxic impurities in a moxifloxacin hydrochloride raw material or a preparation thereof according to any one of claims 1 to 3, characterized in that: The scanning mode of the mass spectrometer is multiple reaction monitoring, the desolvation gas temperature is 220°C-300°C, the ion source temperature is 100°C-120°C, the desolvation gas flow rate is 550L / h-750L / h, the cone gas flow rate is 0L / h-50L / h, the capillary voltage is 2.8kV-3.5kV, the cone voltage is 15V-40V, the secondary cone voltage is 1V-5V, and the sextupole lens voltage is 0.1V-0.5V.

10. The method for detecting genotoxic impurities in a moxifloxacin hydrochloride raw material or a preparation thereof according to claim 9, characterized in that: The desolvation temperature was 300°C, the ion source temperature was 120°C, the desolvation flow rate was 700L / h, the cone gas flow rate was 35L / h, the capillary voltage was 3.0kV, the cone voltage was 30V, the secondary cone voltage was 3V, and the sextupole lens voltage was 0.3V.

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