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中国临床药理学与治疗学 ›› 2004, Vol. 9 ›› Issue (10): 1179-1182.

• 研究原著 • 上一篇    下一篇

高效液相色谱法测定犬全血中两种基喜树碱活性成份

张徐宁1, 丁建花, 刘苏怡, 刘强, 范益, 胡刚   

  1. 南京医科大学药理学系, 南京210029, 江苏
  • 收稿日期:2004-09-14 修回日期:2004-09-16 出版日期:2004-10-26 发布日期:2020-11-23
  • 通讯作者: 胡刚, 男, 博士, 教授, 博士生导师, 研究方向主要为临床药理与实验治疗学。Tel:025-86863169 E-mail:ghu @njmu. edu. cn
  • 作者简介:张徐宁, 女, 在职研究生, 淮阴卫校教师。
  • 基金资助:
    江苏省教育厅产学研研究基金项目(№JH02026)

Quantitative determination of two active constituents of 10-hydroxycamptothecin(HCPT)in whole blood of dog by HPLC

ZHANG Xu-Ning, DING Jian-Hua, LIU Su-Yi, LIU Qiang, FAN Yi, HU Gang   

  1. Department of Pharmacology, Nanjing Medical University, Nanjing, 210029, Jiangsu, China
  • Received:2004-09-14 Revised:2004-09-16 Online:2004-10-26 Published:2020-11-23

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(L-HCPT)含量的方法。方法: 采用HPLC-荧光法, 检测器激发波长为363 nm, 发射波长为550 nm, Discovery C18 色谱柱,流动相为甲醇-乙腈-三乙胺-0. 05 mol·L-1 磷酸氢二钾缓冲液(34 ∶5 ∶ 1 ∶60, pH:6. 8), 流速1. 0 ml·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C-HCPT 、L-HCPT峰分离良好, 内源性杂质不干扰样品峰, 最低检测浓度为2. 5 μg·L-1。C-HCPT 、L-HCPT 浓度范围在2. 5 ~ 1500 μg·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7 d 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C-HCPT 和LHCPT在体内的药代动力学特征。

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

Abstract: 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-acetonitriletriethylamine-0. 05 mol·L-1 dipotassium hydrogen phosphate buffer solution(34 ∶5 ∶1 ∶60, 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.

Key words: lactone form of 10-hydroxycamptothecin, carboxylate form of 10-hydroxycamptothecin, whole blood, HPLC

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