de Oliveira Juliana Kovalczuk, Ronik Débora Fernanda Veres, Ascari Jociani, Mainardes Rubiana Mara, Khalil Najeh Maissar
Department of Pharmacy, Laboratory of Pharmaceutical Nanotechnology, Universidade Estadual do Centro-Oeste, Rua Simeão Camargo Varela de Sá, 85040-080 Guarapuava, Brazil.
Department of Biological Sciences, Universidade Tecnológica Federal do Paraná, Rua Cerejeiras S/N, 85892-000 Santa Helena, PR, Brazil.
J Pharm Anal. 2017 Apr;7(2):129-133. doi: 10.1016/j.jpha.2016.08.001. Epub 2016 Aug 3.
In this study, we developed and validated a fast, specific, sensitive, precise and stability-indicating high performance liquid chromatography (HPLC) method to determine the drug apocynin in bovine serum albumin (BSA) nanoparticles. Chromatographic analyses were performed on an RP C column and using a photodiode array detector at a wavelength of 276 nm. Mobile phase consisted of a mixture of acetonitrile and 1% acetic acid (60:40, v/v), and it was eluted isocratically at a flow rate of 0.8 mL/min. The retention time of apocynin chromatographic peak was 1.65 min. The method was linear, precise, accurate and specific in the range of 5-100 μg/mL. The intra- and inter-day precisions presented relative standard deviation (RSD) values lower than 2%. The method was robust regarding changes in mobile phase proportion, but not for flow rate. Limits of detection and quantitation were 78 ng/mL and 238 ng/mL, respectively. Apocynin was exposed to acid and alkali hydrolysis, oxidation and visible light. The drug suffered mild degradation under acid and oxidation conditions and great degradation under alkali conditions. Light exposure did not degrade the drug. The method was successfully applied to determine the encapsulation efficiency of apocynin in BSA nanoparticles.
在本研究中，我们开发并验证了一种快速、特异、灵敏、精确且具有稳定性指示功能的高效液相色谱（HPLC）方法，用于测定牛血清白蛋白（BSA）纳米粒中的药物白杨素。色谱分析在反相C柱上进行，使用光电二极管阵列检测器，检测波长为276 nm。流动相由乙腈和1%乙酸的混合物（60:40，v/v）组成，以0.8 mL/min的流速等度洗脱。白杨素色谱峰的保留时间为1.65 min。该方法在5 - 100 μg/mL范围内呈线性、精确、准确且特异。日内和日间精密度的相对标准偏差（RSD）值均低于2%。该方法在流动相比例变化方面具有稳健性，但在流速变化时不具有稳健性。检测限和定量限分别为78 ng/mL和238 ng/mL。白杨素经过了酸、碱水解、氧化和可见光照射处理。该药物在酸和氧化条件下发生轻度降解，在碱条件下发生严重降解。光照未使该药物降解。该方法成功应用于测定白杨素在BSA纳米粒中的包封率。
