Department of Pharmacy, Birla Institute of Technology and Science Pilani, Pilani Campus, Jhunjhunu, Rajasthan, 333031, India.
Department of Pharmacy, Birla Institute of Technology and Science Pilani, Pilani Campus, Jhunjhunu, Rajasthan, 333031, India.
J Pharm Biomed Anal. 2019 Oct 25;175:112765. doi: 10.1016/j.jpba.2019.07.013. Epub 2019 Jul 12.
The objective of present work was to develop rapid, sensitive, selective, accurate and precise RP-HPLC method for analysis of Efavirenz from combination anti-HIV drug (Efavirenz-Enfuvirtide) incorporated into polymer-lipid hybrid nanoparticles (PLN). Chromatographic separation of Efavirenz was performed on Waters Spherisorb 5 μm ODS (C18) column (4.6 x 250 mm) with acetonitrile and 10 mM phosphate buffer pH 6.8 (70:30, v/v) as mobile phase. The UV detection wavelength was 246 nm. The method was found to be linear between the concentration range of 500-20000 ng/ml with 160 ng/ml and 480 ng/ml as limit of detection and limit of quantitation respectively. Heteroscedasticity of calibration curve responses was minimized using weighted least square regression analysis. The method was found to be specific for analysis of Efavirenz in presence of Enfuvirtide (a fusion inhibitor peptide), formulation excipients and release media, accurate (average recovery rate: 99.9 ± 9.39%) and precise (%RSD < 2%). The validated RP-HPLC method could be effectively utilized to determine % entrapment efficiency (%EE), % drug loading (%DL), % cumulative drug release and drug content of Efavirenz from Efavirenz-Enfuvirtide co-loaded PLN.
本工作的目的是开发一种快速、灵敏、选择性好、准确和精密的反相高效液相色谱法（RP-HPLC），用于分析组合抗 HIV 药物（依非韦伦-恩夫韦肽）中聚合物脂质杂化纳米粒（PLN）载带的依非韦伦。依非韦伦的色谱分离在 Waters Spherisorb 5μm ODS（C18）柱（4.6×250mm）上进行，流动相为乙腈和 10mM 磷酸盐缓冲液 pH 6.8（70:30，v/v）。检测波长为 246nm。该方法在 500-20000ng/ml 的浓度范围内呈线性关系，检测限和定量限分别为 160ng/ml 和 480ng/ml。采用加权最小二乘法回归分析最小化了校准曲线响应的异方差性。该方法特异性强，可用于分析依非韦伦存在时的依非韦伦（融合抑制剂肽）、制剂赋形剂和释放介质，准确度高（平均回收率：99.9±9.39%），精密度好（%RSD<2%）。验证后的 RP-HPLC 方法可有效用于测定依非韦伦-恩夫韦肽共载 PLN 中的包封效率（%EE）、载药量（%DL）、累积药物释放度和药物含量。
