Department of Pharmaceutical Technology, Faculty of Pharmacy, University of Porto, Rua Aníbal Cunha 164, 4050-047 Porto, Portugal.
J Pharm Biomed Anal. 2010 Jun 5;52(2):167-72. doi: 10.1016/j.jpba.2010.01.007. Epub 2010 Jan 15.
The objective of this work was to develop and validate a rapid reversed-phase (RP) high-performance liquid chromatography (HPLC) method for the in vitro pharmaceutical characterization of dapivirine-loaded polymeric nanoparticles. Chromatographic runs were performed on a RP C18 column with a mobile phase comprising acetonitrile-0.5% (w/v) triethanolamine solution in isocratic mode (80:20, v/v) at a flow rate of 1 ml/min. Dapivirine was detected at a wavelength of 290 nm. The method was shown to be specific, linear in the range of 1-50 microg/ml (R(2)=0.9998), precise at the intra-day and inter-day levels as reflected by the relative standard deviation values (less than 0.85%), accurate (recovery rate of 100.17+/-0.35%), and robust to changes in the mobile phase and column brand. The detection and quantitation limits were 0.08 and 0.24 microg/ml, respectively. The method was successfully used to determine the loading capacity and association efficiency of dapivirine in poly(lactic-co-glycolic acid)-based nanoparticles and its in vitro release.
本工作旨在开发并验证一种快速反相（RP）高效液相色谱（HPLC）法，用于载有双夫定的聚合物纳米粒的体外药物特征研究。色谱运行在 RP C18 柱上，采用等度洗脱模式，流动相由乙腈-0.5%（w/v）三乙醇胺溶液（80:20，v/v）组成，流速为 1ml/min。双夫定于 290nm 波长处检测。该方法具有专属性，在 1-50μg/ml 范围内线性良好（R²=0.9998），日内和日间精密度的相对标准偏差值均小于 0.85%，准确度（回收率为 100.17±0.35%）高，且对流动相和色谱柱品牌的变化具有稳健性。检测限和定量限分别为 0.08 和 0.24μg/ml。该方法成功用于测定聚（乳酸-共-乙醇酸）基纳米粒中双夫定的载药量和结合效率及其体外释放。
