豚鼠耳蜗内淋巴液阿霉素含量高效液相色谱法测定方法的建立

Assay of Adriamycin in Endolymph of Guinea Pig Cochlea By High Performance Liquid Chromatography

目的建立测定豚鼠耳蜗内淋巴液中阿霉素(adriamycin,ADM)含量的方法,为进一步探讨耳毒性物质在内耳中积聚浓度提供方法学基础。方法选取12只健康豚鼠,静脉注射ADM,24小时后快速处死动物,采用高效液相色谱法(highperformanceliquidchromatography,HPLC)检测豚鼠耳蜗内淋巴液阿霉素含量。结果豚鼠耳蜗内淋巴液中ADM浓度在2.56～1398.00ngml范围内线性关系良好,线性相关系数r=0.999(n=10);内淋巴液中ADM的平均方法回收率为96.3%,相对标准差(relativestandarddeviation,RSD)为2.61%;最低检测限为:1.9ngml。结论HPLC法灵敏度高,可以准确地测出豚鼠耳蜗内淋巴液中ADM的含量,适用于静脉给药后ADM在耳蜗内淋巴液中积聚浓度的检测。

Objective To create a method to assay adriamycin in endolymph of guinea pig cochlea,and provide methodology background to study the accumulation of ototoxicity substance in inner ear.Methods 12 normal guinea pig were killed quickly after 24 hours following adriamycin injection.The concentrate of adriamycin in endolymph of guinea pig cochlea was assayed by high performance liquid chromatography(HPLC).Results The linear relation of concentration of adriamycin in endolymph was well when the concentration was from 2.56 ng/mL to 1398.00 ng/mL.Correlation coefficient of linearity was 0.999 (n=10).The percentage of retrieve for adriamycin in endolymph was 96.3%.Relative standard deviation (RSD) was 2.61%.The lowest limitation was 1.9 ng/ml .Conclusion The sensibility of HPLC is high,and it can accurately detect the concentration of adriamycin in endolymph of guinea pig cochlea,especially the adriamycin is administrated in vein.