A method for detecting genotoxic impurities in moxifloxacin hydrochloride raw material or its preparation

Abstract

The application relates to the technical field of pharmaceutical analysis, and particularly discloses a method for detecting genotoxic impurities in moxifloxacin hydrochloride raw materials or preparations. The method is detected by high performance liquid chromatography-mass spectrometry, liquid chromatography conditions are as follows: a C18 chromatographic column is used, a 0.05%-0.2% formic acid aqueous solution is used as a mobile phase A, a 0%-0.2% formic acid acetonitrile solution is used as a mobile phase B, and gradient elution is carried out; the mass spectrometry adopts an ESI ion source, a positive ion detection mode, a parent ion is 431.15 m / z, a daughter ion is 401.18 m / z, and a collision voltage is 10V-25V. The method provided by the application has strong specificity and high sensitivity, the detection limit is 0.625ppm, the blank that the prior art cannot effectively detect N-nitrosomoxifloxacin impurities in moxifloxacin hydrochloride raw materials or preparations is made up, and then the safety risk of moxifloxacin hydrochloride drugs is reduced.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of pharmaceutical analysis, and particularly relates to a detection method of genotoxic impurities in moxifloxacin hydrochloride raw materials or preparations thereof. BACKGROUND

[0002] Moxifloxacin is chemically named as 1-cyclopropyl-7-﹛(S,S)-2,8-diazido- bicyclo[4.3.0]non-8-yl﹛-6-fluoro-8-methoxy-1,4-dihydro-4-oxo-3-quinolinecarboxylic acid, and its CAS number is 151096-09-2. The drug is an artificially synthesized quinolone antibacterial drug, which is used for treating related symptoms caused by respiratory tract infections, and its structural formula is as follows.

[0003]

[0004] The existing synthesis process of moxifloxacin hydrochloride and the existing process of moxifloxacin hydrochloride tablets can cause the reaction of nitrosation reagent residues in raw materials or pipeline residues and moxifloxacin raw materials to generate genotoxic nitrosamine impurities N-nitroso-moxifloxacin, and its chemical structure is as follows.

[0005]

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

[0007] Since the acceptable limit of N-nitroso-moxifloxacin in moxifloxacin hydrochloride bulk drug and tablet is only 3.75ppm, the detection sensitivity of the conventional high performance liquid chromatography (HPLC) test method cannot meet the requirements, and there is a technical difficulty in sensitively and accurately detecting trace amounts of N-nitroso-moxifloxacin in moxifloxacin hydrochloride bulk drug and tablet. There is no report on the detection method of N-nitroso-moxifloxacin with a detection limit lower than 0.625ppm in the prior art. Therefore, it is urgent to provide a method for sensitively and accurately detecting N-nitroso-moxifloxacin in moxifloxacin hydrochloride and moxifloxacin hydrochloride tablets, so as to accurately control the content of N-nitroso-moxifloxacin impurity in moxifloxacin hydrochloride bulk drug and tablet. SUMMARY

[0008] In view of the problem that the trace amount of N-nitroso-moxifloxacin in moxifloxacin hydrochloride bulk drug or preparation cannot be effectively detected in the prior art, the present application provides a method for detecting genotoxic impurities in moxifloxacin hydrochloride bulk drug or preparation thereof.

[0009] To solve the above technical problems, the technical scheme provided by the present application is:

[0010] A method for detecting genotoxic impurities in moxifloxacin hydrochloride bulk drug or preparation thereof, the genotoxic impurity being N-nitroso-moxifloxacin, which is detected by high performance liquid chromatography-mass spectrometry, comprising the following steps:

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

[0012] Take moxifloxacin hydrochloride bulk drug or preparation thereof, and prepare a test solution with a solvent;

[0013] Take N-nitroso-moxifloxacin control product, and prepare a series of control solution with different concentrations with a solvent;

[0014] (2) Detecting the series of control solution with different concentrations and the test solution, wherein the conditions of the high performance liquid chromatography are as follows:

[0015] A C18 chromatographic column is used, 0.05%-0.2% formic acid aqueous solution is used as mobile phase A, and 0%-0.2% formic acid acetonitrile solution is used as mobile phase B, and gradient elution is performed;

[0016] The mass spectrometry uses an ESI ion source and a positive ion detection mode, wherein the quantitative ions of N-nitroso-moxifloxacin are as follows: the parent ion is 431.15m / z, the daughter ion is 401.18m / z, and the collision voltage is 10V-25V.

[0017] Compared with the prior art, the method for detecting genotoxic impurities in moxifloxacin hydrochloride raw materials or preparations thereof provided by the application realizes accurate detection of trace N-nitroso moxifloxacin in moxifloxacin hydrochloride raw materials or preparations thereof by using a C18 chromatographic column, through gradient elution with specific mobile phases, and under specific mass spectrometry conditions, the detection limit of N-nitroso moxifloxacin is 0.625 ppm, the quantification limit is 1.875 ppm, which meets the detection requirements of the ICH M7 guideline. The method provided by the application has strong specificity, high sensitivity, good linear relationship, good accuracy and durability, and can realize trace detection of potential process impurities N-nitroso moxifloxacin in moxifloxacin hydrochloride raw materials or preparations thereof, and can be used as a basis for quality control of moxifloxacin hydrochloride drugs, which makes up the blank that the prior art cannot effectively detect N-nitroso moxifloxacin genotoxic impurities in moxifloxacin hydrochloride, and is conducive to reducing 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] Preferably, the mobile phase can better separate moxifloxacin hydrochloride and N-nitroso moxifloxacin under the premise of not causing baseline interference, and effectively improve the peak shape, so that the accuracy and precision of the detection result are higher.

[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] 20 min, 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 degree between the main component and N-nitroso moxifloxacin and the sensitivity of detection, so that the detection result is quantitative accurate and has high precision.

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

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

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

[0038] Further preferably, the column temperature is 35℃, and the flow rate is 0.3 mL / min.

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

[0040] The preferred detection condition is conducive to the effective detection of the genotoxic impurity N-nitroso moxifloxacin in moxifloxacin hydrochloride, improves the sensitivity of detection, so as to achieve the purpose of effectively and accurately detecting the trace amount of N-nitroso moxifloxacin in moxifloxacin hydrochloride.

[0041] Preferably, the scan 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 is 550-750 L / h, the cone gas flow is 0-50 L / h, the capillary voltage is 2.8-3.5 kV, the cone voltage is 15-40 V, the secondary cone voltage is 1-5 V, and the hexapole lens voltage is 0.1-0.5 V.

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

[0043] The preferred mass spectrometry detection condition can maximize the accuracy of determination of trace N-nitroso-moxifloxacin in moxifloxacin hydrochloride.

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

[0045] The preferred test sample concentration is advantageous for better peak shape of the main component and N-nitroso-moxifloxacin impurity, high column efficiency and accurate integration, thereby facilitating more accurate calculation of the content of N-nitroso-moxifloxacin in the test sample.

[0046] Preferably, the concentration of N-nitroso-moxifloxacin in the control solution is 12 ng / mL, 18 ng / mL, 24 ng / mL, 30 ng / mL, 36 ng / mL or 45 ng / mL.

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

[0048] The detection method provided by the present application can realize accurate quantitative detection of trace N-nitroso-moxifloxacin impurity in moxifloxacin hydrochloride raw material or preparation, thereby facilitating monitoring of the preparation process of moxifloxacin hydrochloride raw material or preparation, more favorably reflecting the quality of moxifloxacin hydrochloride raw material or preparation, and ensuring the clinical medication safety of moxifloxacin hydrochloride product, which has high practical value. BRIEF DESCRIPTION OF DRAWINGS

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

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

[0051] Figure 3 LC-MS graph of moxifloxacin hydrochloride tablet under item 2.1 in Example 2;

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

[0053] Figure 5 LC-MS graph of accuracy solution 2 of moxifloxacin hydrochloride raw material under item 2.3 in Example 2;

[0054] Figure 6 LC-MS graph of accuracy solution 2 of moxifloxacin hydrochloride tablet under item 2.3 in Example 2;

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

[0056] In order to make the objectives, technical solutions and advantages of the present application clearer, the present application will be further described in detail below with reference to examples. It should be understood that the specific examples described herein are only used to explain the present application and should not be used to limit the present application.

[0057] Example 1

[0058] 1.1 Preparation of solution

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

[0060] (2) Reference substance stock solution: about 10 mg of N-nitroso-moxifloxacin reference substance was taken into a 100 mL volumetric flask, dissolved and diluted to the mark with the diluent, and shaken to obtain N-nitroso-moxifloxacin stock solution A; 1 mL of N-nitroso-moxifloxacin stock solution A was accurately transferred into a 100 mL volumetric flask, diluted to the mark with the diluent, and shaken to obtain stock solution B; 5 mL of stock solution B was accurately transferred into a 10 mL volumetric flask, diluted to the mark with the diluent, and shaken to obtain a reference substance stock solution with a concentration of 500 ng / mL.

[0061] (3) Reference substance solution: 240 μL, 360 μL, 480 μL, 600 μL, 720 μL, and 900 μL of the above reference substance stock solution were accurately transferred into six 10 mL volumetric flasks, respectively, diluted to the mark with the diluent, and shaken 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: 2083 μL of the reference substance solution with a concentration of 12 ng / mL was accurately transferred into a 10 mL volumetric flask, dissolved and diluted to the mark with the diluent, and shaken to obtain a sensitivity solution (the concentration of N-nitroso-moxifloxacin was 2.5 ng / mL, which was equivalent to a test substance concentration of 0.625 ppm).

[0063] (5) Raw material test substance solution: 40 mg of moxifloxacin hydrochloride raw material was taken into a 10 mL volumetric flask, dissolved and diluted to the mark with the diluent, and shaken to obtain a test substance solution with a concentration of 4 mg / mL. Two samples were prepared in parallel.

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

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

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

[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℃;

[0071] Sample disc temperature: 15℃;

[0072] Injection volume: 20 μL;

[0073] Injector needle cleaning solution: acetonitrile;

[0074] Gradient elution, elution program as follows:

[0075]

[0076]

[0077] Mass spectrometry conditions:

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

[0079] Mass spectrometry time period setting:

[0080]

[0081] ESI source parameter settings:

[0082]

[0083] Desolvation gas temperature: 300℃;

[0084] Source temperature: 120℃;

[0085] Desolvation gas flow: 700 L / h

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

[0087] Capillary voltage: 3.0 kV;

[0088] Cones hole voltage: 30 V;

[0089] Secondary cones hole voltage: 3 V;

[0090] Hexapole rod lens voltage: 0.3 V.

[0091] After the system is stable, inject the blank solution, and ensure that there is no abnormal peak interference before injecting the sample for detection.

[0092] Example 2

[0093] Method validation:

[0094] 2.1 Specificity

[0095] Take 20 μL of the blank solution, the reference solution, the sensitivity solution, the raw material test solution, and the tablet test solution prepared in Example 1, respectively, and perform HPLC-MS / MS detection according to the above conditions. Inject the sample 1 needle for each solution, and inject the sensitivity solution 6 times. The results are shown in Table 1, and the chromatograms are shown in Figures 1-4 .

[0096] Table 1 Specificity results of the sensitivity solution

[0097]

[0098] The test results show that the baseline is stable, there is no interference in the quantitative ion channel of N-nitrosomoxifloxacin, the RSD of the peak area of the sensitivity solution is 2.14% for 6 needles, indicating that the method has good specificity.

[0099] 2.2 Detection limit and quantification limit

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

[0101] The quantification limit solution (LOQ) is prepared by taking 2.5 mL of the reference solution with a concentration of 30 ng / mL prepared in Example 1, placing it in a 10 mL volumetric flask, diluting to the mark with the dilution solution, and shaking well.

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

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

[0104]

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

[0106] 2.3 Accuracy

[0107] An N-nitroso moxifloxacin stock solution, a series of concentration reference solutions, a sensitivity solution, a raw material test sample solution and a tablet test sample solution were prepared according to the preparation method in Example 1.

[0108] The raw material accuracy solution: 40.05 mg, 40.11 mg and 40.02 mg of the moxifloxacin hydrochloride raw material test sample were accurately weighed into three 10 mL volumetric flasks, and then 240 μL, 300 μL and 360 μL of the N-nitroso moxifloxacin stock solution were accurately added into the three 10 mL volumetric flasks, respectively, after being dissolved with an appropriate amount of diluent. Then, the diluent was added to the mark, and the mixture was shaken to obtain the accuracy solution-1, accuracy solution-2 and accuracy solution-3.

[0109] The tablet accuracy solution: three moxifloxacin hydrochloride tablets were ground into powder, and 72.55 mg, 72.58 mg and 72.60 mg of the powder were accurately weighed into three 10 mL volumetric flasks, respectively. Then, 240 μL, 300 μL and 360 μL of the N-nitroso moxifloxacin stock solution were accurately added into the three 10 mL volumetric flasks, respectively, after being dissolved with an appropriate amount of diluent. Then, the diluent was added to the mark, and the mixture was shaken to obtain the accuracy solution-1, accuracy solution-2 and accuracy solution-3.

[0110] After the system was stabilized, the blank solution was injected to ensure that there was no abnormal peak interference. Then, the sensitivity solution (1 needle) and the series of concentration reference solutions were injected. The signal-to-noise ratio of the impurities in the sensitivity solution should be greater than 3, and the linear coefficient R of the reference solution should be greater than 0.99. 2 After the above conditions were met, the blank solution (1 needle) was injected, and then the test sample solution and the accuracy solution (3 injections for each concentration of the accuracy solution) were injected. Figures 5-6

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

[0112]

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

[0114] ​

[0115]

[0116] The results show that the accuracy of the content of N-nitrosomoxifloxacin is good under the detection condition, the detection result is accurate and reliable, the recovery rate of the accuracy experiment of N-nitrosomoxifloxacin content is between 80%-120%, and the RSD meets the standard requirement.

[0117] 2.4 Linear range

[0118] Control solution: the series concentration control solution under the item of example 1.

[0119] Operation steps: after the system is stable, inject the blank solution, ensure that there is no abnormal peak interference, first inject the sensitivity solution (1 needle), the signal-to-noise ratio of impurities in the sensitivity solution should not be less than 3, then inject the control solution respectively, record the LC-MS diagram. With concentration (ng / mL) as X axis, peak area as Y axis, linear regression is carried out, and the results are shown in table 5.

[0120] Table 5 Linear range

[0121] Compound Name Calibration Curve Linear Range [R 2 ]] N-nitroso moxifloxacin y = 283.91x - 741.21 12.17 ng / mL - 45.63 ng / mL 0.9984

[0122] The results show that N-nitrosomoxifloxacin presents 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: the raw material accuracy solution-2 prepared under the item of 2.4 accuracy;

[0125] Moxifloxacin hydrochloride tablet precision solution: the tablet accuracy solution-2 prepared under the item of 2.4 accuracy.

[0126] Operation steps: after the system is stable, inject the blank solution, ensure that there is no abnormal peak interference, first inject the sensitivity solution (1 needle), the signal-to-noise ratio of impurities in the sensitivity solution should not be less than 3, then inject the raw material precision solution (6 needles) and the tablet precision solution (6 needles) respectively, record the LC-MS diagram, take the peak area of N-nitrosomoxifloxacin respectively, 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] Number Stock 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 9137.235 4260.103 RSD (%) 1.04 1.10

[0129] The results show that under the detection condition of the application, the precision of the quantitative analysis result of N-nitrosomoxifloxacin is good.

[0130] 2.6 Durability

[0131] The blank solution, the reference solution, the raw material test sample solution and the tablet test sample solution were prepared according to the method in Example 1.

[0132] The above-prepared solution was subjected to LC-MS detection, and the LC-MS graph was recorded. The durability of N-nitroso-moxifloxacin 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, mobile phase B adjustment ± 2% in gradient, and the results are shown in Table 7.

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

[0134]

[0135] The results show that under the detection conditions of the present application, fine-tuning the chromatographic conditions has no effect on the detection of N-nitroso-moxifloxacin, indicating that the method has good durability.

[0136] Comparative Example 1

[0137] The detection method provided in this comparative example is different from Example 1 only in that the mobile phase A is replaced with 0.05% ammonium acetate aqueous solution, and the rest of the detection conditions are exactly the same. The sensitivity solution under Example 1 was injected for detection, and the results are shown in Table 2. Figure 7

[0138] As can be seen from the graph, the response intensity of N-nitroso-moxifloxacin in the sensitivity solution is significantly reduced, which cannot meet the detection limit requirements of N-nitroso-moxifloxacin.

[0139] The above only describes the preferred embodiments of the present application and is not intended to limit the present application. Any modification, equivalent replacement or improvement made within the spirit and principle of the present application shall be included in the protection scope of the present application.​

Claims

1. A method for detecting genotoxic impurities in moxifloxacin hydrochloride raw material or its preparations, characterized in that, The genotoxic impurity is N-nitrosomoxifloxacin, which was detected using high-performance liquid chromatography-mass spectrometry (HPLC-MS / MS), including the following steps: (1) Preparation of test solution and reference solution: Take moxifloxacin hydrochloride raw material or its preparation, and prepare a test solution with a solvent; Take N-nitrosomoxifloxacin reference standard and prepare a series of reference standard solutions with solvent; (2) The series of concentrations of reference solutions and test solutions are analyzed, wherein the conditions for high-performance liquid chromatography are: A C18 column was used, with mobile phase A being a 0.05%–0.2% (v / v) formic acid aqueous solution and mobile phase B being a 0%–0.2% (v / v) formic acid acetonitrile solution, and gradient elution was performed. The gradient elution program 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; 20 min, 78%-82% mobile phase A, 22%-18% mobile phase B; The mass spectrometer uses an ESI ion source and a positive ion detection mode. The quantitative ions for N-nitrosomoxifloxacin are: the parent ion is 431.15 m / z, the daughter ion is 401.18 m / z, and the collision voltage is 10V-25V.

2. The method for detecting genotoxic impurities in moxifloxacin hydrochloride raw material or its formulation as described in claim 1, characterized in that, The gradient elution procedure 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.

3. The method for detecting genotoxic impurities in moxifloxacin hydrochloride raw material or its formulation as described in claim 1, characterized in that, The chromatographic column was Ultimate® C18, 4.0 mm * 100 mm, 5 µm.

4. The method for detecting genotoxic impurities in moxifloxacin hydrochloride raw material or its formulation as described in claim 1, characterized in that, The mobile phase A is a 0.05% formic acid aqueous solution, and the mobile phase B is acetonitrile.

5. The method for detecting genotoxic impurities in moxifloxacin hydrochloride raw material or its formulation as described in claim 1, characterized in that, The column temperature was 33℃-37℃, and the flow rate was 0.25mL / min-0.35mL / min.

6. The method for detecting genotoxic impurities in moxifloxacin hydrochloride raw material or its formulation as described in claim 5, characterized in that, The column temperature was 35℃ and the flow rate was 0.3 mL / min.

7. The method for detecting genotoxic impurities in moxifloxacin hydrochloride raw material or its formulation as described in claim 1, characterized in that, The injection volume was 20 µL.

8. The method for detecting genotoxic impurities in moxifloxacin hydrochloride raw material or its preparations as described in claim 1 or 2, characterized in that, The mass spectrometer was used in multiple reaction monitoring (MRM) mode. The desolvation gas temperature was 220℃-300℃, the ion source temperature was 100℃-120℃, the desolvation gas flow rate was 550L / h-750L / h, the cone gas flow rate was 0L / h-50L / h, the capillary voltage was 2.8kV-3.5kV, the cone voltage was 15V-40V, the second-stage cone voltage was 1V-5V, and the hexagonal lens voltage was 0.1V-0.5V.

9. The method for detecting genotoxic impurities in moxifloxacin hydrochloride raw material or its formulation as described in claim 8, characterized in that, The desolvation gas temperature is 300℃, the ion source temperature is 120℃, the desolvation gas flow rate is 700L / h, the conical orifice gas flow rate is 35L / h, the capillary voltage is 3.0kV, the conical orifice voltage is 30V, the secondary conical orifice voltage is 3V, and the hexagonal lens voltage is 0.3V.

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