高效液相色谱荧光检测法测定人血浆中9-氨基喜树碱的浓度

HPLC determination of 9-aminocamptothecin in human plasma with fluorescence detector

目的:建立测定人血浆中9-氨基喜树碱浓度的高效液相色谱-荧光检测法。方法;以喜树碱为内标,以C18固相萃取小柱处理血浆样品,采用荧光检测器检测。色谱柱为KromasilC18色谱分析柱(250mm×4.6mm,5μm),甲醇-磷酸盐缓冲液(48∶52,pH=2.5)为流动相,流速为1.0mL.min-1,柱温30℃,荧光检测器激发波长366nm,发射波长456nm,进样量50μL。结果:9-氨基喜树碱血浆浓度线性范围为0.5～100ng·mL-1(r=0.9998),最低定量限为0.5ng·mL-1,方法回收率为96.8%～102.5%,日内精密度RSD为5.8%～7.3%,日间精密度RSD为1.3%～6.6%。结论:该方法操作简便、专属性强且灵敏度高,适用于血浆9-氨基喜树碱浓度测定和人体药动学研究。

Objective：To establish an HPLC method with fluorescence detection for the quantitation of 9-aminocamptothecin in human plasma.Methods：Camptothecin was adopted as the internal standard.9-aminocamptothecin was extracted from plasma using solid phase extraction （SPE）.The Kromasil C18 column （250 mm×4.6 mm,5 μm） was adopted as analytical column.The mobile phase consisted of methanol and phosphate buffer solution （48：52,pH=2.5）.The flow rate was 1.0 mL·min-1,and the column temperature was at 30 ℃.Excitation wavelength and emission wavelength of fluorescence detection were set at 366 nm and 456 nm,respectively.The injection volume was 50 μL.Results：A good linear relationship was obtained in the concentration range of 0.5 to 100 ng·mL-1.The limit of quantitative concentration was 0.5 ng·mL-1.The relative recovery was between 96.8% and 102.5%.The intra-day precision was between 5.8% and 7.3% and The inter-day precision was between 1.3% and 6.6%.Conclusions：The presented method is simple,accurate and sensitive.It was successfully applied to the determination of 9-aminocamptothecin in human plasma and clinical pharmacokinetic study.