HPLC法测定兔眼角膜中艾沙康唑的浓度

Concentration Determination of Isavuconazole in Rabbit Cornea by HPLC

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 C_(18),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10μL。取日本大耳白兔15只,双眼分别滴加0.2%艾沙康唑混悬液50μL,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 2~2.24μg/mL质量浓度范围内线性关系良好(r^2=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。

OBJECTIVE：To establish a method for the concentration determination of isavuconazole in rabbit cornea. METHODS：HPLC was performed on column of Waters X-Bridge C（18） 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 10μL. 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.