CN-122017096-A - Resolution detection method of nadifloxacin enantiomer

Abstract

The invention relates to the technical field of medicine analysis, in particular to a resolution detection method of a nadifloxacin enantiomer. The method adopts a high performance liquid chromatography to split a sample solution containing a nadifloxacin enantiomer, and uses a diode array detector to detect the sample solution, wherein the high performance liquid chromatography adopts Agilent ZORBAX Eclipse XDB-C 8 chromatographic columns, and L-isoleucine copper acetate is used as a chiral mobile phase additive. The method realizes the resolution of the nadifloxacin enantiomer, the analysis time is shortened by about half by only 22min, the resolution is far greater than 1.5, and the resolution determination requirement is met. The method is simple to operate, economical and practical, has good linear relation, precision and sensitivity, and can be used for resolution and quality control of the nadifloxacin enantiomer.

Inventors

• HONG TING
• CHEN XI
• YANG YISHENG
• LI DAN
• XIAO XIAOWU
• ZHOU ZHIQIANG
• LIAO YONGGEN

Assignees

• 江西省药品检验检测研究院

Dates

Publication Date

20260512

Application Date

20260403

Claims (7)

1. 1. A method for detecting the resolution of a nadifloxacin enantiomer is characterized in that a high performance liquid chromatography is adopted to split a sample solution containing the nadifloxacin enantiomer, and a diode array detector is used for detection, wherein the high performance liquid chromatography adopts Agilent ZORBAX Eclipse XDB-C 8 chromatographic columns, and L-isoleucine copper acetate is used as a chiral mobile phase additive.
2. 2. The method for detecting the resolution of the nadifloxacin enantiomer according to claim 1, wherein the preparation method of the mobile phase in the high performance liquid chromatography is characterized in that L-isoleucine and copper acetate are used as raw materials and dissolved in water to prepare an L-isoleucine copper acetate solution, ammonia water is used for adjusting the pH value to 5.6-7.0 to be used as the mobile phase A, and methanol is used as the mobile phase B.
3. 3. The method for detecting the resolution of the nadifloxacin enantiomer according to claim 2, wherein the volume ratio of the mobile phase A to the mobile phase B is 57-60:43-40, and the elution method is isocratic elution.
4. 4. The method for resolution detection of nadifloxacin enantiomer according to claim 2, wherein the molar concentration of L-isoleucine in the L-isoleucine copper acetate solution is 4-8 mmol/L, the molar concentration of copper acetate is 4-8 mmol/L, and the concentration ratio of L-isoleucine to copper acetate is 1.5-2.5:1.
5. 5. The method for resolution detection of nadifloxacin enantiomer according to claim 1, wherein the chromatographic column is Agilent ZORBAX Eclipse XDB-C 8 μm, 4.6x250 mm.
6. 6. The method for resolution detection of nadifloxacin enantiomer according to claim 1, wherein the preparation method of the sample solution containing nadifloxacin enantiomer is that a sample of nadifloxacin is dissolved by acetonitrile to prepare a sample solution with a concentration of nadifloxacin of 500-1500 mug/ml.
7. 7. The method for resolution detection of nadifloxacin enantiomer according to claim 1, wherein the chromatographic column temperature is 40 ℃, the detection wavelength is 345nm, the sample injection amount is 20 μl, and the flow rate is 1.0ml/min.

Description

Resolution detection method of nadifloxacin enantiomer Technical Field The invention relates to the technical field of medicine analysis, in particular to a resolution detection method of a nadifloxacin enantiomer. Background The nadifloxacin (Nadifloxacin) is a third-generation quinolone antibacterial agent developed by the Japanese large scale company in the beginning of the 80 th century, has a unique benzoquinolizine structure from the structural point of view, has a chiral carbon in a chemical structural formula, is connected with a parent ring and is a C-C bond, the unique structure enables the whole molecular configuration, rigidity and hydrophobicity of the nadifloxacin to be obviously different from those of the former-generation sarcin, the antibacterial activity, pharmacokinetics and toxicity of two enantiomers of the nadifloxacin are obviously different from those of the two enantiomers of the nadifloxacin in vivo and in vitro, and separation and detection of the enantiomers of the nadifloxacin are realized through high performance liquid chromatography, so that the nadifloxacin is an effective method for measuring the purity of single enantiomer. The applicant has prepared a mobile phase by adding potassium dihydrogen phosphate/ethanol to a C 18 chromatographic column and using hydroxypropyl-beta-cyclodextrin as a chiral mobile phase additive, and resolved the nadifloxacin enantiomer. In addition, the method has the advantages that the monopotassium phosphate in the mobile phase is easy to crystallize and separate out, the instrument is easy to be blocked and damaged, the system maintenance cost is high, and the method needs to find different resolution detection methods applicable to the nadifloxacin enantiomer. Disclosure of Invention Aiming at the defects of the prior art, the invention aims to provide a method for detecting the resolution of the nadifloxacin enantiomer, which can realize the complete separation of two enantiomers of nadifloxacin, obviously shorten the time consumption of single resolution and realize the resolution and quality control of the nadifloxacin enantiomer. In order to achieve the above purpose, the invention adopts the following technical scheme: The application provides a method for detecting the resolution of a nadifloxacin enantiomer, which adopts a high performance liquid chromatography to resolve a sample solution containing the nadifloxacin enantiomer, and uses a diode array detector to detect the sample solution, wherein the high performance liquid chromatography adopts Agilent ZORBAX Eclipse XDB-C8 chromatographic columns and uses L-isoleucine copper acetate as a chiral mobile phase additive. Further, the preparation method of the mobile phase in the high performance liquid chromatography comprises the steps of dissolving L-isoleucine and copper acetate serving as raw materials in water to prepare an L-isoleucine copper acetate solution, adjusting the pH value to 5.6-7.0 by using ammonia water to serve as the mobile phase A, and using methanol as the mobile phase B. Further, the volume ratio of the mobile phase A to the mobile phase B is 57-60:40-43. Preferably, the preparation method of the mobile phase in the high performance liquid chromatography comprises the steps of dissolving L-isoleucine and copper acetate serving as raw materials in water to prepare an L-isoleucine copper acetate solution, adjusting the pH value to 7.0 by using ammonia water to serve as a mobile phase A, and using methanol as a mobile phase B, wherein the volume ratio of the mobile phase A to the mobile phase B is 60:40. Further, the molar concentration of L-isoleucine in the L-isoleucine copper acetate solution is 4-8 mmol/L, the molar concentration of copper acetate is 4-8 mmol/L, and the concentration ratio of L-isoleucine to copper acetate is 1.5-2.5:1. Preferably, the molar concentration of L-isoleucine in the L-isoleucine copper acetate solution is 8mmol/L, the molar concentration of copper acetate is 4mmol/L, and the ratio of the concentrations of L-isoleucine and copper acetate is 2:1. Further, the column was Agilent ZORBAX Eclipse XDB-C 8 μm, 4.6X1250 mm. Further, the preparation method of the sample solution containing the nadifloxacin enantiomer comprises the steps of dissolving a nadifloxacin sample by acetonitrile to prepare the sample solution with the nadifloxacin concentration of 500-1500 mug/ml. Further, the column temperature of the chromatographic column is 40 ℃, the detection wavelength is 345nm, the sample injection amount is 20 mu L, and the flow rate is 1.0ml/min. Compared with the prior art, the method has the beneficial effects that L-isoleucine copper acetate is adopted as a chiral mobile phase additive, a chromatographic column is screened, and through optimizing various experimental parameters, the separation of the nadifloxacin enantiomer is finally realized on the Agilent ZORBAX Eclipse XDB-C 8 chromatographic column, the separation degree between the