HPLC quantification of doxorubicin in plasma and tissues of rats treated with doxorubicin loaded poly(alkylcyanoacrylate) nanoparticles
HPLC quantification of doxorubicin in plasma and tissues of rats treated with doxorubicin loaded poly(alkylcyanoacrylate) nanoparticles
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DOI：
10.1016/j.jchromb.2012.01.025
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发表时间：
2012-03-01
影响因子：
3
通讯作者：
Moussa, Fathi
中科院分区：
文献类型：
--
作者：
Alhareth, Khairallah;Vauthier, Christine;Moussa, Fathi
多柔比星（Dox）是一种重要的抗肿瘤药物，但其剂量相关性的急性心脏毒性、骨髓抑制和肿瘤细胞的多药耐药性限制了其长期临床应用。为了提高阿霉素的抗肿瘤疗效和降低毒性，人们开发了许多给药系统，包括聚氰基丙烯酸烷基酯（PACA）纳米粒。本实验室采用氧化还原自由基乳液聚合法制备了一种具有潜在隐身性能的新型PACA纳米粒子，并将其与Dox结合。为比较研究不同剂型的阿霉素（Dox）PACA纳米粒（Dox-PACA）在大鼠体内的药代动力学和生物分布，建立了一种简便、快速的高效液相色谱法（HPLC）测定阿霉素（Dox）PACA纳米粒（Dox-PACA）大鼠血浆和组织中阿霉素（Dox）浓度。Dox在4.4 min洗脱，并与其主要代谢物多柔比星酚（Doxl）和多柔比星酮（Doxon）以及用作内标物（IS）的依达替尼（Ida）很好地分离。通过液-液萃取实现从生物介质中萃取Dox。在大鼠血浆中，对于0.05和1 μ g/mL的Dox-PACA，从血浆和组织（肝、脾和心脏）中回收的总Dox（即游离Dox和与纳米颗粒结合的Dox）分别为71%和78%，在组织中，对于0.5和10 μ g/g的Dox分别为73%和80%。该方法在血浆中Dox浓度为0.05 - 1.5 μ g/mL范围内呈线性。该方法的检测限为每次注射0.5 ng Dox（50 μ L）。在0.05~1 μ g/mL浓度范围内，日间和日内精密度分别为97.1-102.9%和97.3-101.7%。初步数据表明，该方法可以应用于测定静脉给药后，与PACA纳米粒子的阿霉素的药代动力学和生物分布。(c)2012爱思唯尔有限公司版权所有。
The long-term clinical use of doxorubicin (Dox), one of the most important anticancer agent in use, is limited by dose-related acute cardiotoxicity, myelo-suppression and multidrug resistance developed by cancer cells. To improve the antitumor efficacy and reduce the toxicity of Dox, many drug delivery systems have been developed, including poly(alkylcyanoacrylate) (PACA) nanoparticles. A new formulation of PACA nanoparticles with potential stealth properties were prepared by redox radical emulsion polymerization and associated to Dox in our laboratory. To comparatively investigate the pharmacokinetics and the biodistribution of different formulations of Dox associated PACA nanoparticles, a simple and rapid high performance liquid chromatographic method (HPLC) was developed for the quantification of Dox in plasma and tissues of rats treated with Dox loaded PACA nanoparticle (Dox-PACA). Dox was eluted at 4.4 min and it was well separated from its main metabolites doxorubicinol (Doxl) and doxorubicinon (Doxon) and idarubicin (Ida) used as internal standard (IS). Extraction of Dox from biological media was achieved by liquid-liquid extraction. The recovery of total Dox (i.e. free Dox and Dox associated with nanoparticles) from plasma and tissues (liver, spleen and heart) spiked with Dox-PACA were 71 and 78% for 0.05 and 1 mu g/mL in rat plasma, respectively, and 73% and 80% for 0.5 and 10 mu g/g in tissues, respectively. The method is linear from 0.05 to 1.5 mu g/mL of Dox in plasma. The limit of detection of the method is 0.5 ng of Dox per injection (50 mu L). The between-day and within-day precisions of the method were 97.1-102.9% and 97.3-101.7% for concentrations ranging from 0.05 to 1 mu g/mL, respectively. Preliminary data suggested that this method can be applied to determine the pharmacokinetic and biodistribution of Dox associated with PACA nanoparticles after intravenous administration to rats. (c) 2012 Elsevier B.V. All rights reserved.