摘要
目的建立高效液相色谱法(HPLC)测定犬全血中羟喜树碱两种活性成份羧酸型羟喜树碱(CHCPT)和内酯型羟喜树碱(LHCPT)含量的方法。方法采用HPLC荧光法,检测器激发波长为363nm,发射波长为550nm,DiscoveryC18色谱柱,流动相为甲醇乙腈三乙胺-0.05mol·L-1磷酸氢二钾缓冲液(345160,pH6.8),流速1.0ml·min-1,N2000色谱工作站采集数据,外标法计算药物浓度。结果全血样品中CHCPT、LHCPT峰分离良好,内源性杂质不干扰样品峰,最低检测浓度为2.5μg·L-1。CHCPT、LHCPT浓度范围在2.5～1500μg·L-1间呈线性相关,回归方程分别为y=11196x-921.56和y=11840x-5188.4,相关系数分别为r=0.9998和0.9991,绝对回收率分别为85.5%～98.6%和78.4%～89.1%,相对回收率分别为100.7%～103.9%和96.8%～102.0%,日内和日间变异均小于15.0%,提取后样品-75℃冻存7d稳定。结论本实验方法具有专属性,且灵敏、可靠,适用于HCPT类制剂给药后血药浓度的检测,有助于深入研究HCPT两种活性成份CHCPT和LHCPT在体内的药代动力学特征。
AIM: To quantify the lactone form and carboxylate form of 10 hydroxycamptothecin (HCPT) in whole blood of dog by develop a simple, rapid, and sensitive high performance liquid chromatography (HPLC) method. METHODS: The concentration of hydroxycamptothecinum was assayed on a DiscoveryC18 column with a mobile phase consisting of methanol acetonitrile triethylamine - 0.05 mol·L -1 dipotassium hydrogen phosphate buffer solution(345160,pH= 6.8 )at a flow rate of 1.0 ml·min -1 , and HCPT was monitored with fluorescence detector, excitation and emission wavelengths being 363 nm and 550 nm, respectively. RESULTS: There was a good linear response range of 2.5 -1500 μg·L -1 while the limit of quantification was 2.5 μg·L -1 . The relative recovery rate C HCPT of was 100.7 %- 103.9 %,L HCPT of was 96.8 %- 102.0 %; the absolute recovery rate of C HCPT was 85.5 %- 98.6 %, and L HCPT was 78.4 %- 89.1 %. The intra day and inter day variations were all less than 15%. The HCPT in supernatant in refrigerator was stable at -75 ℃ for 7 days. CONCLUSSION: The method is simple, sensitive, reliable, and suitable for the quantitative determination of hydroxycamptothecin following i.v. administration, and it is also helpful to improve the study on pharmacokinetics of two active constituents C HCPT and L HCPT of HCPT.
出处
《中国临床药理学与治疗学》
CAS
CSCD
2004年第10期1179-1182,共4页
Chinese Journal of Clinical Pharmacology and Therapeutics
基金
江苏省教育厅产学研研究基金项目(№JH02026)
关键词
羧酸型羟喜树碱
内酯型羟喜树碱
全血
高效液相色谱法
lactone form of 10 hydroxycamptothecin
carboxylate form of 10-hydroxycamptothecin
whole blood
HPLC
