Ankara University, Faculty of Pharmacy, Department of Pharmaceutical Technology, Ankara, Turkey.
Ankara University, Institute of Forensic Sciences, Department of Forensic Toxicology, Ankara, Turkey; Hitit University Faculty of Arts & Sciences, Department of Analytical Chemistry, Corum, Turkey.
J Pharm Biomed Anal. 2018 May 30;154:285-293. doi: 10.1016/j.jpba.2018.03.021. Epub 2018 Mar 12.
This is the first report in literature using hydrophilic interaction liquid chromatography (HILIC) in combination with diode array detector (DAD) for stability indicating determination of 5-Fluorouracil (5-FU) from its bulk form, pharmaceutical preparations, developed solid lipid nanoparticle (SLN) and nano structured lipid carrier (NLC) drug delivery systems as well as the rat skin extracts. The separation was performed at 45 °C, on Sequant Zic HILIC (250 mm × 4.60 mm ID, 5 μm, 200 A), peek HPLC column. Mobile phase is consisting of a mixture of acetonitrile: buffer containing 5 mM ammonium acetate (95:5; v/v). The pH of the mobile phase was adjusted to 7.0 using 1 M NaOH. The analysis was carried out at 0.75 mL min flow rate with a detection wavelength of 265 nm and the injection volume was arranged as 10 μL. The developed method was fully validated in accordance with the International Council on Harmonization (ICH) Guidelines. Specificity of this method was demonstrated by forced degradation studies. As a result of calibration studies, the calibration curve was found linear in the concentration range of 1-250 μg mL (R = 0.999). The precision of this technique calculated within the frame of intra-day and inter-day based on a percentage of relative standard deviation (RSD%) values (<2%). The limits of detection and quantification were 11 and 37 ng mL respectively. On the other hand, 5-FU loaded SLN and NLC formulations with average particle size of 370 nm were also developed and compared in order to increase the permeation of drug into the rat skin. Ex-vivo Penetration/Permeation Studies indicated that higher dermal accumulation of 5-FU was obtained with NLC formulation. As a conclusion, the present work expressed the optimization and the validation of a selective, simple, precise and accurate fully validated HILIC method with sufficient sensitivity for the estimation of 5-FU in raw materials, marketed formulation and rat skin extract after applying both of the commercial product and newly developed nanoparticulate drug delivery systems on to the rat skins with high percentage recoveries.
这是文献中首次使用亲水相互作用色谱（HILIC）结合二极管阵列检测器（DAD）来对 5-氟尿嘧啶（5-FU）进行稳定性指示测定，包括其原料药、开发的固体脂质纳米粒（SLN）和纳米结构脂质载体（NLC）药物递送系统以及大鼠皮肤提取物。分离在 45°C 下进行，使用 Sequant Zic HILIC（250mm×4.60mm ID，5μm，200A），Peek HPLC 柱。流动相由乙腈：含 5mM 乙酸铵的缓冲液（95:5，v/v）组成。使用 1M NaOH 将流动相的 pH 值调节至 7.0。分析在 0.75mL/min 的流速下进行，检测波长为 265nm，进样量为 10μL。该方法按照国际协调会议（ICH）指南进行了全面验证。通过强制降解研究证明了该方法的特异性。作为校准研究的结果，发现校准曲线在 1-250μg/mL 的浓度范围内呈线性（R=0.999）。根据相对标准偏差（RSD%）值（<2%）计算，该技术的精密度在日内和日间范围内均得到验证。检测限和定量限分别为 11 和 37ng/mL。另一方面，还开发并比较了平均粒径为 370nm 的负载 5-FU 的 SLN 和 NLC 制剂，以增加药物向大鼠皮肤的渗透。离体渗透/渗透研究表明，NLC 制剂可获得更高的 5-FU 皮肤蓄积量。总之，本工作表达了一种选择性、简单、精确和准确的完全验证的 HILIC 方法的优化和验证，该方法具有足够的灵敏度，可用于估计原料药、市售制剂和大鼠皮肤提取物中的 5-FU，并且在将商业产品和新开发的纳米药物递送系统应用于大鼠皮肤后，可获得高回收率。
