【摘要】 目的:建立高效液相色谱(HPLC)法测定新西兰大白兔眼玻璃体腔两性霉素B。方法:兔眼玻璃体溶液两性霉素B标本100μL经过冷冻干燥后,加入乙腈水(70∶30)20μL去蛋白,并溶解浓缩回收目标药物,13 000 r·min-1离心5 min,取10μL进样分析。色谱条件为流动相:乙腈10 mmol·L-1磷酸二氢钠溶液(35∶65),流速:0.20 mL·min-1,C18色谱柱,柱温:25 ℃,检测波长:410nm。结果:两性霉素B在3~500μg·L-1内线性关系良好(r=0.998 8),高、中、低3 种浓度的方法回收率为99%~108%,绝对回收率为83%~93%;日内、日间RSD分别为3.1%~12.6%,3.8%~12.9%;检测灵敏度为1μg·L-1。结论:本方法适用于兔眼玻璃体腔两性霉素B的定量分析,过程简单,方法稳定,结果可靠。更多还原

【Abstract】 OBJECTIVE To establish a method for determination of amphotericin B in vitreous humor of New Zealand rabbit by HPLC.METHODS Preparations of bio-samples include condensation by cryodesiccation, deproteinization and recovery by adding 20μL acetonitrile-water(70∶30). Mobile phase of acetonitrile-10 mmol·L -1 NaH_ 2 PO_ 4 (35∶65) and ZOBARX C_ 18 colum were used, the flow rate was 0.20 mL·min -1 and detection wavelength 410 nm. RESULTS The linear range of amphotericin B was 3~500 μg·L -1 ( r =\{0.998 8\}).The absolute and relative recoveries of three levels of bio-samples were 99%-108%、83%-93%. The RSD of intra-day and inter-day were 3.1% - 12.6% , 3.8% - 12.9% respectively. The detection limit was 1 μg·L -1 .CONCLUSION This method was proved to be accurate,simple and reliable for determination of amphotericin B in vitreous humor of New Zealand rabbit eyes.更多还原