HPLC法测定兔眼角膜中艾沙康唑的浓度
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篇名: HPLC法测定兔眼角膜中艾沙康唑的浓度
TITLE:
摘要: 目的:建立测定兔眼角膜中艾沙康唑浓度的方法。方法:采用高效液相色谱(HPLC)法。色谱柱为Waters X-Bridge C18,流动相为0.2%冰乙酸-乙腈(49 ∶ 51,V/V),流速为0.7 mL/min,柱温为40 ℃,检测波长为284 nm,进样量为10 μL。取日本大耳白兔15只,双眼分别滴加0.2%艾沙康唑混悬液50 μL,于给药后5、15、30、60、90 min分别处死3只兔并剖取其角膜样品,处理后进样测定艾沙康唑浓度。结果:角膜样品中的艾沙康唑在0.031 2~2.24 μg/mL质量浓度范围内线性关系良好(r2=0.999 9),方法回收率为91.7%~105.6%(RSD≤6.88%,n=5),提取回收率为100.50%~106.10%(RSD≤2.16%,n=3),日内(n=5)、日间(n=3)精密度和稳定性(n=5)试验的RSD均小于10%。结论:本方法专属性强、准确度高、重现性好,可快速、准确地测定兔眼角膜中艾沙康唑的浓度。
ABSTRACT: OBJECTIVE: To establish a method for the concentration determination of isavuconazole in rabbit cornea. METHODS: HPLC was performed on column of Waters X-Bridge C18 with mobile phase of 0.2% glacial acetic acid-acetonitrile (49 ∶ 51, V/V) at flow rate of 0.7 mL/min, column temperature was 40 ℃, detection wavelength was 284 nm, and injection volume was 10 μL. Fifteen Japanese white rabbits with big ears were selected, 50 μL of 0.2% isavuconazole suspension was dropped into eyes, then every 3 rabbits were executed after 5, 15, 30, 60, 90 min of administration and cornea samples were taken respectively. The determination of isavuconazole were determined after treating. RESULTS: The isavuconazole in leaching solution showed good linear range in 0.031 2-2.24 μg/mL (r2=0.999 9); oethod recovery rate was 91.7%-105.6% (RSD≤6.88%, n=5); extraction recovery rate was 100.50%-106.10% (RSD≤2.16%, n=3); and RSDs of inter-day (n=3) and intra-day (n=3) precision and stability (n=5) tests were lower than 10%. CONCLUSIONS: The method shows strong specificity, high accuracy, good reproducibility, and can rapidly and accurately determine the concentration of isavuconazole in rabbit cornea.
期刊: 2017年第28卷第31期
作者: 周静静,周天洋,张俊杰
AUTHORS: ZHOU Jingjing,ZHOU Tianyang,ZHANG Junjie
关键字: 高效液相色谱法;艾沙康唑;角膜;兔
KEYWORDS: HPLC; Isavuconazole; Cornea; Rabbits
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