Improved HPLC method for doxorubicin quantification in rat plasma to study the pharmacokinetics of micelle-encapsulated and liposome-encapsulated doxorubicin formulations |
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Authors: | Wei Guangli Xiao Shuhua Si Duanyun Liu Changxiao |
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Institution: | Tianjin Key Laboratory of Pharmacokinetics and Pharmacodynamics, Tianjin Institute of Pharmaceutical Research, Tianjin 300193, People's Republic of China. weigl510@163.com |
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Abstract: | An improved simple, rapid and accurate HPLC method for quantification of doxorubicin derived from micelle-encapsulated or liposome-encapsulated doxorubicin formulation in rat plasma was described. The mobile phase consisting of a mixture of methanol-water containing 0.1% formic acid anhydrous and 0.1% ammonia solution (25%), pH 3.0], 60:40, was delivered at a flow rate of 1.0 mL/min. Sample preparation for micelle- or liposome-encapsulated doxorubicin in rat plasma were achieved directly by protein precipitation with acetonitrile. Doxorubicin and daunorubicin (internal standard, IS) were separated on a C(18) reversed-phase HPLC column and quantified by a fluoresence detection with an excitation wavelength of 475 nm and an emission wavelength of 580 nm. The linearity was obtained over the range of 5.0-1000.0 ng/mL and 1.0-200.0 microg/mL for doxorubicin and the lower limit of quantitation was 5.0 ng/mL. For each level of quality control samples, inter- and intra-assay precision was less than 9.6 and 5.1% (relative standard deviation), respectively, and percentage error was within +/-2.6%. The extraction recoveries of doxorubicin in the range of 10 ng/mL to 100 microg/mL in rat plasma were between 94.1 and 105.6%. This method was successfully applied to the pharmacokinetic study of doxorubicin formulations after i.v. administration to rats. |
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Keywords: | pharmacokinetics doxorubicin daunorubicin high‐performance liquid chromatography (HPLC) |
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