荧光标记法测定大鼠血浆中海胆黄多糖研究

通过FITC荧光标记海胆黄多糖SEP,研究大鼠尾静脉注射给药SEP的血浆代谢。结果表明,荧光标记的海胆黄多糖SEP-Tyr-FITC纯度可达99%。与空白对照组及SEP组相比,SEP-Tyr-FITC组小鼠脾细胞增殖无显著差异。其在大鼠体内的血浆清除半衰期为0.24 h,消除速率常数(Ke)为2.867,给药后的最大血药浓度(cmax)为860μg/mL,消除半衰期(t1/2)为0.24 h,药-时曲线下面积(AUCt)为366.4 h·μg/mL。所用方法的定量下限为0.02 mg/mL,标准曲线线性范围为0.02～1.0 mg/mL。最低定量限(LLOQ)、低、中、高4个浓度批内变异系数和批间变异系数均小于15%,相对回收率均在85%～115%之间。同时,各浓度SEP血浆样品处理前或处理后室温放置6 h、-20℃冷冻融解1次稳定性良好,相对标准偏差均小于10%。

The Metabolism of A Strongylocentrotus Nudus Eggs Polysaccharide in Rat Plasma

The metabolism of a polysaccharide from Strongylocentrotus nudus eggs(SEP) in rat plasma was detected by labeling SEP with fluorescein isothiocyanate(FITC).The results showed that the purity of fluorescence labeled polysaccharide SEP-Tyr-FITC was 99%.FITC-labeling(200μg/mL) didn′t significantly affect the activity of SEP to splenocytes.The elimination half-time of SEP-Tyr-FITC in rat plasma was 0.24 h by intravenous injection of SEP-Tyr-FITC(50 mg/kg).The area under the concentration-time curve was 366.4 h·μg/mL.The elimination rate constant(ke) was 2.867 1/h.The maximum plasma concentration(Cmax) was 860 μg/mL.In this method,the lower limit of quantification(LLOQ) was 0.02mg/ml.The assay was linear over the tested concentration range(0.02~1.0 mg/mL).The recoveries of the method were 85%~115%,with CV of the intra-and inter-day being less than 15%.The concentration-time curve of SEP-Tyr-FITC was in accordance with a two-compartment model.The concentrations of the SEP inplasma samples were stability under the condition of stored at room temperature for 6h before or after sample preparation,or frozen-thawed once.