Detection method of moxifloxacin hydrochloride residual solvent

Abstract

The invention discloses a method for detecting a moxifloxacin hydrochloride residual solvent. The method comprises the following steps: (1) respectively preparing a blank solution, a reference solution and a test solution; (2) sample injection detection; chromatographic conditions are as follows: an instrument used is a gas chromatograph with a hydrogen flame ionization detector, a chromatographic column is a capillary column taking polyethylene glycol as a stationary liquid, headspace sample injection is adopted to inject into the gas chromatograph, hydrogen is taken as a carrier gas, sample injection detection is performed respectively to obtain a chromatogram of a reference substance solution and a chromatogram of a test sample solution, and the chromatogram of the reference substance solution and the chromatogram of the test sample solution are calculated. And detecting and analyzing by adopting an external standard method to complete the determination of the moxifloxacin hydrochloride residual solvent. According to the method, a plurality of residual solvents are placed in the same system for detection, the organic solvents are completely separated, the linear relation is good in the investigated range, and the detection efficiency is greatly improved.

Description

Technical Field

[0001] The invention relates to the technical field of drug analysis, and in particular to a method for detecting residual solvents of moxifloxacin hydrochloride. Background Art

[0002] Moxifloxacin hydrochloride is a fourth-generation fluoroquinolone antibiotic developed by Bayer, Germany. Its chemical name is 1-cyclopropyl-6-fluoro-7-([S, S]-2,8-diazabicyclo[4.3.0]nonan-8-yl)-8-methoxy-1,4-dihydro-4-oxo-3-quinolinecarboxylic acid hydrochloride. Its trade name is Avelox. Its structural formula is as follows: .

[0003] Moxifloxacin hydrochloride is a broad-spectrum antibiotic that has antibacterial activity not only against Gram-negative bacteria, but also against Gram-positive cocci, mycoplasmas, chlamydia, anaerobic bacteria, etc. It is clinically used to treat socially acquired pneumonia caused by Staphylococcus aureus, Haemophilus influenzae, Pneumococcus, Moraxella catarrhalis, Klebsiella pneumoniae, etc., acute exacerbation of chronic bronchitis, acute sinusitis, etc. Moxifloxacin hydrochloride has the advantages of high efficiency, low toxicity, low level of drug resistance, and almost no obvious phototoxicity. It is one of the fluoroquinolone antibiotics with more clinical applications.

[0004] Residual solvents in drugs refer to organic volatile compounds produced or used in the production of raw materials or excipients, as well as in the preparation of preparations. Since residual solvents not only have no therapeutic effect, but may also increase the toxic side effects of drugs and affect the stability of drugs, all organic solvents should be removed as much as possible. In order to protect patients from the harm of residual organic solvents in drugs, it is necessary to measure the residual amount of organic solvents introduced into drugs during the production process.

[0005] For example, CN 104230924 B (application number 201410405137.5) discloses a method for synthesizing moxifloxacin hydrochloride, comprising the following steps: using a main ring chelate compound having a structure as shown in formula (I) and (S,S)-2,8-diazabicyclo[4.3.0]nonane as raw materials and triethylamine as an acid absorbent, and after sufficient condensation reaction in acetonitrile, concentrating, dissolving after treatment, acidolysis to form salt, crystallization, filtering, washing, and drying to obtain moxifloxacin hydrochloride; wherein the acetonitrile is evaporated and then acidified in ethanol to obtain moxifloxacin hydrochloride; the method for synthesizing the main ring chelate compound shown in formula (I) is as follows: acetic anhydride is heated to 70°C, boric acid is slowly added at 70-90°C, the temperature is raised to reflux for 1h, then cooled to 70°C, 1-cyclopropyl-6,7-difluoro-1 , 4-dihydro-8-methoxy-4-oxo-3-quinolinecarboxylic acid ethyl ester, heated to 100-105°C for 1h, cooled to 0°C, washed with water, filtered, washed with crystals, and dried to obtain a chelate. In the preparation process, ethanol, acetonitrile and acetic acid were used.

[0006] According to the requirements of the ICH guidelines: the residual limits of ethanol, acetonitrile and acetic acid are 0.5%, 0.041% and 0.5%. Therefore, the determination of ethanol, acetonitrile and acetic acid in moxifloxacin hydrochloride is of great significance to the drug quality of moxifloxacin hydrochloride and the health of patients.

[0007] Regarding the detection method of residual solvents, Chinese patent document CN 104297365 B (application number 201410476437.2) discloses a method for determining the residual solvent acetic acid in moxifloxacin hydrochloride, which comprises the following steps: A. Preparation of test solution: Take a moxifloxacin sample, dissolve it in dimethyl sulfoxide and dilute it to a 0.1g / ml solution as the test solution.

[0008] B. Preparation of reference solution: Take an appropriate amount of glacial acetic acid, dissolve it in dimethyl sulfoxide and dilute it into a 0.5 mg / ml solution as the reference solution.

[0009] C. Determination: Using gas chromatography, accurately measure 1.0 to 2.0 μl of the test solution and the reference solution, inject them into the gas chromatograph, and record the chromatogram; D. Calculation: Calculate the acetic acid content by the external standard method; wherein the gas chromatography conditions are: Chromatographic column: capillary column with 6% cyanopropylphenyl-94% dimethylpolysiloxane as stationary liquid, column length 30m, inner diameter 0.32mm, liquid film thickness 1.8μm; detector: hydrogen flame ionization detector; injection port temperature: 200℃; Detector temperature: 200-250°C; carrier gas: nitrogen; carrier gas flow rate: 0.8-1.2 ml / min; column temperature: programmed temperature rise; wherein the specific method of the programmed temperature rise is: initial temperature 30-50°C, maintained for 3-7 minutes, heated to 100°C at a rate of 30-50°C per minute, maintained for 3-7 minutes, then heated to 200-250°C at a rate of 40-60°C per minute, maintained for 3-7 minutes.

[0010] Chinese patent document CN 111751459 A (application number 202010461550.9) discloses a method for simultaneously detecting multiple residual solvents in sitafloxacin, wherein the multiple residual solvents include methanol, ethanol, acetonitrile, tert-butyl alcohol, dichloromethane, methyl tert-butyl ether, n-hexane, ethyl acetate, tetrahydrofuran, dioxane and toluene, and the method comprises the following steps: 1) Determine the gas chromatography conditions: chromatographic column: capillary column with weakly polar polysiloxane as the stationary liquid; carrier gas: nitrogen; injection: headspace injection, headspace equilibrium temperature is 80°C-100°C, and headspace equilibrium time is 20min-40min; Column temperature: Program temperature rise, starting at 38°C-42°C, maintain for 20 min, then increase to 200°C at a rate of 20°C / min ℃, maintain for 5 minutes; Inlet: Inlet temperature is 200℃; Detector: FID detector, detector temperature is 280℃; Column flow rate: 3.8-4.2mL / min; Split ratio: 8:1-12:1.

[0011] 2) Preparation of reference solution and test solution: The reference solution was prepared as follows: accurately weigh appropriate amounts of methanol, ethanol, acetonitrile, tert-butanol, dichloromethane, methyl tert-butyl ether, n-hexane, ethyl acetate, tetrahydrofuran, dioxane and toluene, respectively, placed in a volumetric flask, dissolve with N, N-dimethylacetamide and quantitatively dilute to the mark, shake well, accurately measure an appropriate amount, and place in an empty bottle at the top as the reference solution; the test solution was prepared as follows: accurately weigh an appropriate amount of sitafloxacin sample, place in a volumetric flask, dissolve with N, N-dimethylacetamide and quantitatively dilute to the mark, shake well, accurately measure an appropriate amount, and place in an empty bottle at the top as the test solution.

[0012] 3) Perform methodological validation: The methodological validation includes specificity, sensitivity, linearity and range, system precision, repeatability and recovery.

[0013] 4) Determination of residual solvents in sitafloxacin samples: The determination is carried out according to the following method: accurately measure the test solution and the reference solution, respectively perform determination according to the gas chromatography conditions in step 1), record the chromatogram, and calculate the content by peak area according to the external standard method. Summary of the invention

[0014] The technical problem to be solved by the present invention is to provide a method with high sensitivity and accuracy suitable for detecting residual solvents of moxifloxacin hydrochloride.

[0015] The technical solution to achieve the purpose of the present invention is a method for detecting residual solvents in moxifloxacin hydrochloride, comprising the following steps: ① Prepare blank solution, reference solution and test solution respectively.

[0016] The blank solution is a diluent, and the diluent is a mixed solution of 1-methyl-2-pyrrolidone and water.

[0017] The reference substance solution is a mixed solution of ethanol, acetonitrile, acetic acid and a diluent.

[0018] The test solution is obtained by dissolving moxifloxacin hydrochloride to be tested in a diluent.

[0019] ② Sampling test; The chromatographic conditions are as follows: the instrument used is a gas chromatograph with a hydrogen flame ionization detector, the chromatographic column is a capillary column with polyethylene glycol as the stationary liquid, headspace injection is used for injection into the gas chromatograph, hydrogen is used as the carrier gas, the injection port temperature is 180°C to 220°C, the column temperature gradient is set to increase the temperature, and the hydrogen flame ionization detector temperature is 220°C to 250°C.

[0020] The chromatograms of the reference solution and the test solution were obtained by injection testing respectively, and the external standard method was used for detection and analysis to complete the determination of the residual solvent in moxifloxacin hydrochloride.

[0021] The diluent is a mixed solution of 1-methyl-2-pyrrolidone and water in a volume ratio of 1 to 2:1.

[0022] The carrier gas flow rate is 1.8 mL / min to 2.3 mL / min.

[0023] Preferably, the carrier gas flow rate is 2.0 mL / min to 2.1 mL / min.

[0024] When the column temperature was increased in a gradient manner, the starting temperature was 45-50°C, maintained for 4 minutes, and then increased to 230°C at a rate of 10°C per minute and maintained for 5 minutes.

[0025] As an option, the injection order of each solution into the gas chromatograph for detection is 1 injection of blank solution, 5 injections of reference solution, and 2 injections of test solution, and the injection volume is 1 mL.

[0026] The split ratio of the split direct injection mode was 20:1.

[0027] Optionally, the gas chromatography column model is selected from Agilent INNOWAX 19095N-123 and Agilent DB-264; the specifications of the gas chromatography column are 30m*0.53mm, 1μm; 30m×0.53mm, 3μm; 60m×0.53mm, 3μm; or 60m×0.32mm, 1μm.

[0028] The above detection method can simultaneously detect the residual amounts of ethanol, acetonitrile and acetic acid in moxifloxacin hydrochloride.

[0029] The present invention has positive effects: the present invention optimizes the detection conditions for the detection of residual solvents of moxifloxacin hydrochloride, such as ethanol, acetonitrile and acetic acid. Under the condition of ensuring good theoretical pedal and separation, multiple residual solvents are placed in the same system for detection, and these organic solvents are completely separated, and the linear relationship is good within the investigated range, which greatly improves the detection efficiency. At the same time, the sensitivity and accuracy of the present invention meet the detection requirements of the residual amounts of ethanol, acetonitrile and acetic acid, and ensure the quality control of the preparation process of moxifloxacin hydrochloride. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 It is the gas chromatogram of the blank solution in Example 1.

[0031] Figure 2 It is the gas chromatogram of the reference substance solution in Example 1.

[0032] Figure 3 It is the gas chromatogram of the test solution in Example 1.

[0033] Figure 4 It is the gas chromatogram of the reference substance solution in Example 2.

[0034] Figure 5 It is the gas chromatogram of the test solution in Example 2. DETAILED DESCRIPTION

[0035] (Example 1) The detection method of the residual solvent of moxifloxacin hydrochloride in this embodiment comprises the following steps: ① Prepare blank solution, reference solution and test solution respectively.

[0036] The blank solution is a diluent, and the diluent is a mixed solution of 1-methyl-2-pyrrolidone-water with a volume ratio of 2:1.

[0037] The reference substance solution is obtained by diluting the reference stock solution, wherein the reference stock solution is prepared as follows: Accurately weigh 208.55 mg of acetonitrile into a 20 mL volumetric flask, dilute to the mark with the diluent, and mix well. Accurately measure 1 mL of the above solution into a 50 mL volumetric flask, then accurately weigh 125.6 mg of ethanol and 125.3 mg of acetic acid into the aforementioned 50 mL volumetric flask, dilute to the mark with the diluent, and mix well to serve as the control stock solution.

[0038] The reference solution was prepared as follows: 5 mL of the reference stock solution was accurately measured into a 50 mL volumetric flask, diluted to the mark with a diluent, and mixed to obtain the reference solution.

[0039] The test solution was prepared as follows: 256.59 mg of moxifloxacin hydrochloride raw material was accurately weighed, added to a headspace bottle with 5 mL of diluent, shaken, dissolved, and sealed to prepare the test solution.

[0040] ② Sample injection test.

[0041] The chromatographic conditions are as follows: the instrument used is a gas chromatograph with a hydrogen flame ionization detector, which is an Agilent-7890B gas chromatograph in this embodiment; the headspace injector is an Agilent-7697A; the chromatographic column is a capillary column with polyethylene glycol as the stationary phase, which is an Agilent INNOWAX 19095N-123 in this embodiment; the specification is 30m*0.53mm, 1μm; hydrogen is used as the carrier gas, and the flow rate is 2.1mL / min; the split ratio is 20:1; headspace injection is used, the injection port temperature is 200°C, and the column temperature gradient is set to rise, the starting temperature is 50°C, maintained for 4 minutes, and the temperature is raised to 230°C at a rate of 10°C per minute, and maintained for 5 minutes; the detector temperature is 250°C, and the injection volume is 1mL.

[0042] In addition to the above-mentioned 30m*0.53mm, 1μm, the gas chromatography column can also be of the following specifications: 30m×0.53mm, 3μm; 60m×0.53mm, 3μm; or 60m×0.32mm, 1μm.

[0043] Accurately pipette 1 mL of blank solution, reference solution, and test solution, and inject into the gas chromatograph, including 1 injection of blank solution, 5 injections of reference solution, and 1 injection of test solution, for determination, and record the chromatogram. According to the chromatogram, the residual amounts of ethanol, acetonitrile, and acetic acid in moxifloxacin hydrochloride were calculated by the external standard method.

[0044] Figure 1is the gas chromatogram of the blank solution. Figure 1 It can be seen that the blank solution does not interfere with the detection of the solvent peak.

[0045] Figure 2 This is the gas chromatogram of the reference solution. The separation degrees of the solvent peaks in the reference solution are all greater than 1.5, and several organic solvents have achieved complete separation, which meets the requirements. The peak shape is good, the peak purity is good, and the theoretical plate number is greater than 3000, which meets the standards.

[0046] Figure 3 The gas chromatogram of the test solution shows that the test sample contains ethanol, and the calculated content is 0.0134%, which is lower than the limit specified in the ICH guidelines, indicating that the method of the present invention is a detection method with high sensitivity and accuracy.

[0047] (Example 2) The detection method of the residual solvent of moxifloxacin hydrochloride in this embodiment comprises the following steps: ① Prepare blank solution, reference solution and test solution respectively.

[0048] The blank solution is a diluent, and the diluent is a mixed solution of 1-methyl-2-pyrrolidone-water with a volume ratio of 1:1.

[0049] The reference solution is obtained by diluting the reference stock solution, wherein the reference stock solution is prepared as follows: accurately weigh 147.4 mg of ethanol and 153.5 mg of acetic acid in a 10 mL volumetric flask, dilute to scale with the diluent, and mix well. In addition, accurately weigh 139.3 mg of acetonitrile in a 100 mL volumetric flask, dilute to scale with the diluent, and mix well. Each is used as the reference stock solution.

[0050] The reference solution was prepared as follows: 1 mL of the reference stock solution was accurately measured into a 100 mL volumetric flask, diluted to the mark with a diluent, and mixed to obtain the reference solution.

[0051] The test solution was prepared as follows: 150 mg of moxifloxacin hydrochloride raw material was accurately weighed, added to a headspace bottle with 5 mL of diluent, shaken, dissolved, and sealed to obtain the test solution.

[0052] ② Sample injection test.

[0053] The chromatographic conditions were as follows: The instrument used was an Agilent-7890B gas chromatograph; the headspace injector was an Agilent-7697A; the chromatographic column was an Agilent DB-264 with specifications of 30m*0.53mm, 3μm; hydrogen was used as the carrier gas with a flow rate of 2.0mL / min; the split ratio was 20:1; headspace injection was adopted, the injection port temperature was 210℃, the column temperature gradient was set, the starting temperature was 45℃, maintained for 4 minutes, and then increased to 230℃ at a rate of 10℃ per minute and maintained for 5 minutes; the detector temperature was 220℃, and the injection volume was 1mL.

[0054] Accurately pipette 1 mL of blank solution, reference solution, and test solution, and inject into the gas chromatograph, including 1 injection of blank solution, 5 injections of reference solution, and 1 injection of test solution, for determination, and record the chromatogram. According to the chromatogram, the residual amounts of ethanol, acetonitrile, and acetic acid in moxifloxacin hydrochloride are calculated by the external standard method.

[0055] Figure 4 This is the gas chromatogram of the reference solution. The separation degrees of the solvent peaks in the reference solution are all greater than 1.5, and several organic solvents have achieved complete separation, which meets the requirements. The peak shape is good, the peak purity is good, and the theoretical plate number is greater than 3000, which meets the standards.

[0056] See Figure 5 The gas chromatogram of the test solution showed that the test sample contained ethanol, and the calculated content was 0.014%, which was lower than the limit specified in the ICH guidelines and met the requirements.

Claims

1. A method for detecting residual solvents of moxifloxacin hydrochloride, characterized in that The following steps are involved: ① Prepare blank solution, reference solution and test solution respectively; The blank solution is a diluent, and the diluent is a mixed solution of 1-methyl-2-pyrrolidone and water; The reference substance solution is a mixed solution of ethanol, acetonitrile, acetic acid and a diluent; The test solution is obtained by dissolving moxifloxacin hydrochloride to be tested in a diluent; ② Sampling test; The chromatographic conditions were as follows: the instrument used was a gas chromatograph with a hydrogen flame ionization detector, the chromatographic column was a capillary column with polyethylene glycol as the stationary phase, headspace injection was used to inject into the gas chromatograph, hydrogen was used as the carrier gas, the injection port temperature was 180°C to 220°C, the column temperature was set to a gradient temperature increase, and the hydrogen flame ionization detector temperature was 220°C to 250°C; The chromatograms of the reference solution and the test solution were obtained by injection testing respectively, and the external standard method was used for detection and analysis to complete the determination of the residual solvent in moxifloxacin hydrochloride.

2. The method for detecting residual solvent of moxifloxacin hydrochloride according to claim 1, characterized in that: The diluent is a mixed solution of 1-methyl-2-pyrrolidone and water in a volume ratio of 1 to 2:

1.

3. The method for detecting residual solvent of moxifloxacin hydrochloride according to claim 1, characterized in that: The carrier gas flow rate is 1.8 mL / min to 2.3 mL / min.

4. The method for detecting residual solvent of moxifloxacin hydrochloride according to claim 1, characterized in that: The carrier gas flow rate is 2.0 mL / min to 2.1 mL / min.

5. The method for detecting residual solvent of moxifloxacin hydrochloride according to claim 1, characterized in that: When the column temperature was increased in a gradient manner, the initial temperature was 45-50°C, maintained for 4 minutes, and then increased to 230°C at a rate of 10°C per minute and maintained for 5 minutes.

6. The method for detecting residual solvent of moxifloxacin hydrochloride according to claim 1, characterized in that: The injection order of each solution into the gas chromatograph for detection is 1 injection of blank solution, 5 injections of reference solution, and 2 injections of test solution, and the injection volume is 1 mL.

7. The method for detecting residual solvent of moxifloxacin hydrochloride according to claim 1, characterized in that: The split ratio of the split direct injection mode was 20:

1.

8. The method for detecting residual solvent of moxifloxacin hydrochloride according to claim 1, characterized in that: The gas chromatography column model is selected from Agilent INNOWAX 19095N-123 and Agilent DB-264; the specifications of the gas chromatography column are 30m*0.53mm, 1μm; 30m×0.53mm, 3μm; 60m×0.53mm, 3μm; or 60m×0.32mm, 1μm.

9. The method for detecting residual solvent of moxifloxacin hydrochloride according to claim 1, characterized in that: The residual amounts of ethanol, acetonitrile and acetic acid in moxifloxacin hydrochloride can be detected simultaneously.

Images