CN104931632A - Method for detecting residual tartaric acid amount in moxifloxacin hydrochloride - Google Patents

Abstract

The invention discloses a method for detecting residual tartaric acid in moxifloxacin hydrochloride, which is detected by high-performance liquid chromatography, adopts AD-H chiral chromatographic column, and uses n-hexane solution containing trifluoroacetic acid and diethylamine-containing The mixed solution of ethanol solution is the mobile phase, and the detection wavelength is 205nm. The method has the advantages of good linearity, good injection precision, stable solution within 12 hours, and accurate and reliable recovery results. A comprehensive study provides an effective method.

Description

Determination method of tartaric acid residue in moxifloxacin hydrochloride

technical field

The invention relates to a method for detecting drug quality, in particular to a method for detecting residual tartaric acid in moxifloxacin hydrochloride.

Background technique

Moxifloxacin hydrochloride is a fourth-generation quinolone broad-spectrum antibacterial drug used to treat respiratory and skin infections.

During the synthesis of moxifloxacin hydrochloride, there may be tartaric acid residues. Because tartaric acid only absorbs at the end, there is absorption at 205nm, and there is no absorption at 293nm, the maximum absorption wavelength of moxifloxacin hydrochloride. Under the chromatographic conditions for the detection of related substances of cifloxacin, tartaric acid is almost not retained, so it is necessary to establish a detection method for the residual tartaric acid in moxifloxacin hydrochloride to ensure the quality of the drug.

Contents of the invention

The object of the present invention is to provide a method for detecting residual tartaric acid in moxifloxacin hydrochloride, so as to accurately measure the residual tartaric acid in moxifloxacin hydrochloride and ensure the quality of medicines.

The tartaric acid residue detection method in moxifloxacin hydrochloride of the present invention is detected by high performance liquid chromatography, the chromatographic column adopts AD-H chiral chromatographic column, and the mobile phase is n-hexane solution containing trifluoroacetic acid and diethyl ether The mixed liquid of amine in ethanol solution, the detection wavelength is 205nm.

Further, the above-mentioned n-hexane solution containing trifluoroacetic acid preferably has a trifluoroacetic acid content of 0.1-0.5% (volume percentage), most preferably 0.3%.

The ethanol solution containing diethylamine preferably has a diethylamine content of 0.1-0.4% (volume percentage), most preferably 0.2%.

In the above mobile phase, the volume ratio of the n-hexane solution containing trifluoroacetic acid and the ethanol solution containing diethylamine is preferably 75-85:15-25, most preferably 80:20.

The size of the above AD-H chiral chromatographic column is 250 mm×4.6 mm, 5 μm; the flow rate of the mobile phase during detection is 0.3-1.0 ml/min, preferably 0.5 ml/min.

In a specific embodiment of the present invention, the high performance liquid chromatography conditions are: chromatographic column AD-H column, chromatographic column specification: 250mm×4.6mm, 5μm; %Diethylamine ethanol=80:20 (volume ratio) is the mobile phase, the detection wavelength is 205nm, and the flow rate is 0.5ml/min.

The specific determination method can be: take an appropriate amount of moxifloxacin hydrochloride to be tested, accurately weigh it, add an appropriate amount of methanol to dissolve, dilute with mobile phase to make a solution containing 2mg-3mg (such as 3mg) of moxifloxacin hydrochloride per 1ml, shake well , as the test solution; take another appropriate amount of tartaric acid reference substance, accurately weigh it, dissolve it with an appropriate amount of methanol, dilute it with mobile phase to make a solution containing 0.15mg～0.30mg (such as 0.15mg) of tartaric acid per 1ml, shake well, and use as Reference substance solution; precision measures need testing solution and each 10 μ l of reference substance solution, injects liquid chromatograph respectively, record chromatogram; If there is the chromatographic peak of tartaric acid in the chromatogram of need testing solution, by external standard method, with Calculate the peak area to obtain the residual amount of tartaric acid in the moxifloxacin hydrochloride test product.

The tartaric acid residue detection method in moxifloxacin hydrochloride provided by the invention has the advantages of good linearity, good sampling precision, stable solution within 12 hours, accurate and reliable recovery results, etc., and the method can accurately determine moxifloxacin hydrochloride The residual amount of tartaric acid provides an effective method for the comprehensive research on related substances of moxifloxacin hydrochloride.

Description of drawings

Fig. 1 is the chromatogram of solvent methanol and mobile phase (MP);

Fig. 2 is the chromatogram of tartaric acid;

Fig. 3 is moxifloxacin hydrochloride chromatogram;

Fig. 4 is the HPLC figure of system applicability of tartaric acid and moxifloxacin hydrochloride.

detailed description

The present invention will be further described by specific examples below, but the scope of the present invention is not limited in any way.

Chromatographic conditions

Chiral chromatographic column (AD-H column 250mm×4.6mm, 5μm) with amylose-tris-(3,5-xylylcarbamate) silica gel as the stationary phase; n-hexane (containing 0.3 % trifluoroacetic acid) - ethanol (containing 0.2% diethylamine) = 80:20 (volume ratio) as the mobile phase (MP), the detection wavelength is 205nm, and the flow rate is 0.5ml/min.

Assay

Take an appropriate amount of moxifloxacin hydrochloride to be tested, accurately weighed, add an appropriate amount of methanol to dissolve, dilute with mobile phase to make a solution containing 3mg of moxifloxacin hydrochloride per 1ml, shake well, and use it as the test solution; take another tartaric acid reference substance Appropriate amount, accurately weighed, dissolved with appropriate amount of methanol, diluted with mobile phase to make a solution containing 0.15 mg of tartaric acid per 1 ml, shaken up, as the reference substance solution. Precisely measure 10 μl each of the test solution and the reference solution, inject them into the liquid chromatograph, and record the chromatograms. If there is a chromatographic peak of tartaric acid in the chromatogram of the test solution, it shall be calculated by the peak area according to the external standard method.

Positioning of each sample solution

1) Take the solvent methanol and the mobile phase to directly inject samples for determination. The chromatogram is shown in Figure 1;

2) Tartaric acid: Take 20.23mg of tartaric acid, put it in a 10ml measuring bottle, dissolve it with an appropriate amount of methanol, dilute the mobile phase to the mark, and see the chromatogram in Figure 2;

3) Moxifloxacin hydrochloride: take 10.5g of moxifloxacin hydrochloride, weigh it accurately, put it in a 10ml measuring bottle, dissolve it with an appropriate amount of methanol, and dilute the mobile phase to the mark. The chromatogram is shown in Figure 3;

4) Mixed solution of tartaric acid and moxifloxacin hydrochloride: see Figure 4 for the chromatogram.

It can be seen from Figures 1 to 4 that tartaric acid and moxifloxacin hydrochloride can be separated by baseline, and solvent methanol and mobile phase do not interfere with the detection of tartaric acid in moxifloxacin hydrochloride.

System Suitability Test

Take about 11mg of tartaric acid and about 75mg of moxifloxacin hydrochloride, weigh them accurately, put them in the same 25ml measuring bottle, add an appropriate amount of methanol to dissolve, dilute the mobile phase to the mark, shake well, and use it as a system suitability solution. Take 10 μl of the system suitability solution, inject it into the liquid chromatograph, and record the chromatogram, see Figure 4, the separation degree of tartaric acid and moxifloxacin hydrochloride should be greater than 1.0. Continuously inject 6 needles, record the RSD% and resolution of the peak area of the two chromatographic peaks, see the table below.

Applicability results of tartaric acid detection system in moxifloxacin hydrochloride

sample tartaric acid retention time Moxifloxacin hydrochloride retention time Tartaric acid peak area Separation 1 15.943 19.056 737687 2.80 2 15.976 19.003 735296 2.72 3 15.986 18.965 743716 2.66 4 15.98 18.95 734083 2.65 5 15.998 18.931 737567 2.62 6 16.009 18.904 740236 2.57 average 15.98 18.97 738097.50 2.67 S 0.02 0.05 3480.09 0.08 RSD% 0.14 0.29 0.47 2.95

Conclusion: From the above results, it can be seen that the separation degree of tartaric acid and moxifloxacin hydrochloride is 2.65, and the RSD% of the peak area of tartaric acid is 0.47%. Indicates that the system suitability meets the test requirements.

Linear range:

Tartaric acid had good linearity in the range of 91.20μg/ml-304.20μg/ml; moxifloxacin hydrochloride had good linearity in the range of 31.50μg/ml-105.00μg/ml.

Minimum detection limit:

The lower quantification limit of tartaric acid was less than 1000ng.

Injection precision:

After six consecutive injections, the RSD% of the retention time and peak area of moxifloxacin hydrochloride was less than 2%, indicating good injection precision.

Solution Stability:

Take the test solution in the injection precision test, place it at room temperature, and inject samples at different times to investigate the stability of the solution. Conclusion: When the moxifloxacin hydrochloride sample solution was placed at room temperature, the RSD% of the peak area within 12 hours was 2.70%, and the solution was basically stable within 12 hours.

Recovery test:

The RSDs of recoveries of 9 samples with three concentrations of high, medium and low were all in the range of 98.0-102.0%, meeting the determination requirements.

Determination of tartaric acid residues in three batches of moxifloxacin hydrochloride samples

Three batches of samples were inspected according to the above-mentioned inspection method for tartaric acid residue in moxifloxacin hydrochloride, and the results are as follows.

Determination results of tartaric acid residues in three batches of moxifloxacin hydrochloride samples

batch number 20110301 20110302 20110303 tartaric acid% not detected not detected not detected

Conclusion: The tartaric acid residues in the three batches of samples were all less than 1.0%.

Conclusion: Through the verification of the method of tartaric acid residue in moxifloxacin hydrochloride, the method has good linearity, good injection precision, the solution is stable within 12 hours, and the recovery rate is accurate and reliable. Therefore, it can be seen that the established method can accurately determine hydrochloric acid. The residual amount of tartaric acid in moxifloxacin provides an effective method for comprehensive research on related substances of moxifloxacin hydrochloride.

Claims (9)

1. a detection method of tartaric acid residue in moxifloxacin hydrochloride, detect by high performance liquid chromatography, chromatographic column adopts AD-H chiral chromatographic column, and mobile phase is the n-hexane solution containing trifluoroacetic acid and containing two The mixture of ethylamine and ethanol solution has a detection wavelength of 205nm. 2. The detection method according to claim 1, wherein the content of trifluoroacetic acid in the n-hexane solution containing trifluoroacetic acid is 0.1% to 0.5%. 3. The detection method according to claim 1, wherein the content of diethylamine in the ethanol solution containing diethylamine is 0.1-0.4%. 4. The detection method according to claim 1, wherein the volume ratio of the n-hexane solution containing trifluoroacetic acid and the ethanol solution containing diethylamine is 75～85:15～25. 5. The detection method according to claim 1, characterized in that, the specification of the AD-H chiral chromatographic column is 250 mm×4.6 mm, 5 μm. 6. The detection method according to claim 1, characterized in that the flow rate of the mobile phase is 0.3-1.0 ml/min. 7. The detection method as claimed in claim 1, characterized in that, the high performance liquid chromatography condition is: adopt AD-H chiral column chromatography column, column specification: 250mm×4.6mm, 5μm; to contain 0.2～0.5% The n-hexane solution of trifluoroacetic acid: ethanol solution containing 0.1-0.4% diethylamine = 75-85:15-25 volume ratio mixture is the mobile phase; the detection wavelength is 205nm; the flow rate is 0.3-1.0ml/min. 8. The detection method as claimed in claim 7, characterized in that, the high performance liquid chromatography condition is: adopt AD-H chiral chromatographic column, chromatographic column specification: 250mm * 4.6mm, 5 μm; To contain 0.3% trifluoroacetic acid n-hexane solution: ethanol solution containing 0.2% diethylamine = 80:20 volume ratio mixture is the mobile phase; the detection wavelength is 205nm; the flow rate is 0.5ml/min. 9. The detection method according to any one of claims 1 to 8, wherein the specific detection steps include: 1) Take an appropriate amount of moxifloxacin hydrochloride to be tested, accurately weigh it, add appropriate amount of methanol to dissolve, dilute with mobile phase to make a solution containing 2mg-3mg of moxifloxacin hydrochloride per 1ml, shake well, and use it as the test solution; 2) Take an appropriate amount of tartaric acid reference substance, accurately weigh it, dissolve it with an appropriate amount of methanol, dilute it with mobile phase to make a solution containing 0.15-0.30 mg of tartaric acid per 1 ml, shake well, and use it as the reference substance solution; 3) Precisely measure 10 μl each of the test solution and the reference solution, inject them into the liquid chromatograph respectively, and record the chromatogram; 4) If there is a chromatographic peak of tartaric acid in the chromatogram of the test solution, use the external standard method to calculate the peak area to obtain the residual tartaric acid in the moxifloxacin hydrochloride test product.