高效液相色谱法测定兔组织中羟基喜树碱的浓度

Determination of Hydroxycamptothecin Concentration in Rabbit Tissues by HPLC

目的:建立以高效液相色谱-紫外检测法测定兔组织中羟基喜树碱浓度的方法。方法:组织样品制成匀浆后,加入内标喜树碱,用甲醇-乙腈(1:1)沉淀离心后取上清液进行色谱分析。色谱柱:Lichrospher C18柱(250mm×4.6mm,5μm),C18预柱(10mm×4.6mm,5μm);流动相:乙腈-0.075mol/L醋酸铵缓冲液(30:70);流速:1.0ml/min;检测波长:384nm;柱温:30℃。结果:羟基喜树碱保留时间为4.5min,内标喜树碱保留时间为10.0min,定量线性范围为40-1 600ng/ml。组织中羟基喜树碱高、中、低3种浓度的回收率范围为95.54％～107.58％,日内精密度≤7.70％,日间精密度≤6.69％。 结论:本法简便实用,定量准确,可满足羟基喜树碱药动学研究的需要。

OBJECTIVE: To establish a HPLC - UV method for determination of hydroxycamptothecin(HCPT) in rabbit tissues (liver, kidney, stomach, lung, spleen, heart, intestine) .METHODS: After homogenization, tissue samples and internal standard, eamptothecine, were precipitated with CH3OH-CH3CN( 1 : 1) and then centrifugalized.20μl of supernatant was injected and measured by HPLC - UV method.The chromatographic column was Lichrospher C18 column: (250mm × 4.6mm, 5μm), CH3CN - 0.075mol/L NH4AC buffer(pH6.4) (30 : 70, contain 5mol/L tyiethylamine) served as mobile phase with flow rate of 1.0ml/ min .Detection wavelength was 384nm .RESULTS: The retention time of hydroxyeamptothecin was 4.5min.A good linearity was shown in the concentration range of (40- 1 600)ng/ml .The recovery was between 95.24% and 107.58% . The intra day RSD was less than 7.70% and the inter - day RSD was less than 6.69% .CONCLUSION: This method is simple,pratical and accurate.It could be applied to pharmacokinetic study of hydroxyeamptothecin.