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目的 建立特定培养基中莫西沙星的HPLC测定法。方法 取样品100μL,以加替沙星作内标,采用乙腈沉淀蛋白;常温下氮气吹干,用0.01 mol·L-1盐酸200μL复溶后进样。色谱柱:Alltech Nucleosil 100(C18)柱(10μm,4.6 mm×250mm);流动相:缓冲液-甲醇-乙腈(66.8:15.1:18.1),缓冲液为三乙胺-磷酸-水(0.37:0.30:99.3);流速1.3 mL·min-1,荧光检测器的激发波长和发射波长分别为295和418nm。结果在1.0-50.0 μg·mL-1内线性关系良好,相关系数r=0.999 9(n=7)。高低2个浓度质控样品的日内RSD在0.67％-0.81％之间,日间RSD在0.48％-0.89％之间,方法学回收率为98.43％-103.51％。结论本法灵敏、可靠、耐用、操作简便易行、分析时间短,并成功地应用于莫西沙星体外模型的细菌耐药性发展研究。
Objective To establish an HPLC assay for moxifloxacin in specific media. Methods 100 μL of sample was taken with gatifloxacin as internal standard, and the protein was precipitated by acetonitrile. After drying under nitrogen at room temperature, the sample was reconstituted with 200 μL of 0.01 mol·L-1 hydrochloric acid. The mobile phase consisted of buffer-methanol-acetonitrile (66.8: 15.1: 18.1) and the buffer was triethylamine-phosphoric acid-water (0.37: 0.30 : 99.3). The flow rate was 1.3 mL · min-1. The excitation and emission wavelengths of the fluorescence detector were 295 and 418 nm, respectively. The results showed good linearity in the range of 1.0-50.0 μg · mL-1 with a correlation coefficient of 0.999 9 (n = 7). The RSDs of the two control samples at high and low concentrations ranged from 0.67% to 0.81%, the intraday RSD ranged from 0.48% to 0.89%, and the methodological recoveries ranged from 98.43% to 103.51%. Conclusion This method is sensitive, reliable, durable, simple and easy to operate, short analysis time, and successfully applied to the development of bacterial drug resistance in vitro model of moxifloxacin.