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HPLC-荧光法测定血样和组织样品中羟基喜树碱的研究
Determination of Hydroxycamptothecine in rats by HPLC with fluorescence detection
【摘要】 [目的]建立了HPLC-荧光法检测血浆与组织样品中羟基喜树碱的方法。[方法]Waters Symmetry C18色谱柱(150×4.6 mm,5μm);流动相为乙腈和0.3%冰醋酸(用三乙胺调pH至5.0)(体积比:20∶80);检测器激发波长363 nm;发射波长550 nm;柱温35℃;流速1.0 mL·min-1。[结果]血浆与组织样品中羟基喜树碱与内标(喜树碱)峰分离良好,内源性杂质不干扰样品峰。血浆及各组织HCPT浓度在2~1 000 ng·m L-1的浓度范围内具有良好的线性关系。大鼠血浆的线性回归方程为:Y=0.223 279X+0.104 877,r2=0.981 7。小鼠肝、瘤组织的线性回归方程分别为:肝Y=0.227 993X+0.088 526,r2=0.977 3;瘤Y=0.232 519X+0.099 206,r2=0.981 6。日内、日间精密度RSD值小于8%,提取回收率在98.87%~117.28%之间。[结论]本实验方法操作简单,专属性好,准确度高,分析时间短,可作为生物样品中羟基喜树碱浓度的测定及药代动力学研究的分析方法。
【Abstract】 [Objective]To establish a method for determining Hydroxycamptothecin in rat blood plasma and mice tissues by HPLCwith fluorescence detector.[Methods]The chromatographic conditions were as follows: A Waters Symmetry C18 column( 4.6 mm×150 mm,5 μm), a mobile phase consisted of acetonitrile-0.3% acetic acid( pH tuned to 5 with triethylamine)( Volume ralio: 20∶ 80), the column temperature at 35 ℃, the flow rate with 1.0 m L·min-1 and the fluorescence detection at λex= 363 nm and λem= 530 nm.[Results]In the plasma and tissue samples, the Hydroxycamptothecin was well separated from the internal standard( Camptothecin) peak, and the peaks of Hydroxycamptothecin and Camptothecin were interfered by the endogenous impurity. The peak area and the concentration of HCPT in plasma and tissue has a good linear relationship in the range of 2 ~ 1 000 ng·m L-1.The linear regression equation of plasma samples was: Y = 0.223 279 X + 0.104 877, r2= 0.981 7. The linear regression equationof liver and tumor tissues samples were: liver, Y = 0.227 993 X + 0.088 526, r2= 0.977 3; tumor, Y = 0.232 519 X + 0.099206, r2= 0.981 6, respectively. The recoveries of HCPT was between 98.87% ~ 117.28%. The inter-day and intra-day relative standard deviations was smaller than 8%.[Conclusion]The method is simple, has good specificity and accuracy, and can be used to determine hydroxycamptothecin in biological samples and investigate the pharmacokinetics.
【Key words】 Hydroxycamptothecine; HPLC; fluorimetric method; methodological investigation;
- 【文献出处】 河南大学学报(医学版) ,Journal of Henan University(Medical Science) , 编辑部邮箱 ,2018年03期
- 【分类号】R969
- 【下载频次】169