荧光分光光度法检测大鼠膀胱组织中表阿霉素浓度

Determination of Epirubicin concentration in rat bladder tissues by fluorescence spectrometry

目的建立大鼠膀胱组织中表阿霉素(Epirubicin,EPI)的荧光分光光度法检测方法。了解聚维酮(polyvidone,PVP)是否有促进EPI进入膀胱壁的作用。方法建立雌性Sprague-Dawley大鼠膀胱灌注模型,经大鼠尿道单独灌注1mg·mL-1表阿霉素或联合1%聚维酮溶液灌注后,保留药物2h后,处死大鼠,分离及匀浆膀胱组织,用含0.3mol·L-1HCl的50%乙醇抽提,离心取上清液,用荧光分光光度计在λex/em为470/554nm测定其荧光值,通过标准曲线方程计算表阿霉素浓度。结果样本扫描未见杂峰,日内精密度<2.75%,日间精密度<4.01%,表阿霉素于肿瘤组织匀浆中抽提回收率>86.0%,表阿霉素联合1%聚维酮灌注组的膀胱组织内表阿霉素浓度明显高于单灌表阿霉素组,P<0.01。结论本方法特异性高,快速、简便,结果准确可靠,适用于大鼠膀胱组织中EPI药代动力学研究。1%PVP联合膀胱灌注能促使表阿霉素进入膀胱壁。

OBJECTIVE To study the fluorospectrophotometer method for determining Epirubicin（EPI） concentration in rat bladder tissues and learn whether 1% polyvidone（PVP） can increase the transport of EPI into the rat urinary bladder wall.METHODS The model of bladder irrigation of the female Sprague-Dawley rat was established.Two hours after irrigating 1mg·mL-1 EPI with or without 1% polyvidone by rat urethra the rats were killed and the bladder tissues were extracted,mashed and resuspended in HCl 0.3mol·L-1 in 50% ethanol.Following centrifugation,the supernatant was removed and assayed spectrofluorometrically at λex 470 nm and λem 554 nm.The concentration of EPI was calculated by the standard curve of EPI.RESULTS The intra-day and inter-day RSD were 2.75% and 4.01% respectively.The recovery of EPI from bladder tissues was 86.0%.The concentration of EPI was higher in associated medication group（use EPI and PVP） than in single medication group（only use EPI）,P0.01.CONCLUSION This method is simple,rapid,sensitive and of excellent precision,so it is suitable for EPI pharmacokinetic studies in bladder tissues.1%PVP increased the transport of EPI into the urinary bladder wall.