The aim of this work was to develop and validate an analytical method using HPLC for the determination of propranolol in the different layers of the skin to be used in kinetic studies of skin permeation. The development of the method was based on the suitability of the chromatogram, and the validation followed the international health regulation for bioanalytical methods. In addition, the method was tested in an in vitro permeation assay using porcine skin. The drug was determined using an RP‐C18 column at 30°C, a mobile phase comprising acidic aqueous phase:acetonitrile (75:25 v/v), at a flow rate of 1.0 mL min⁻¹, and UV detection at 290 nm. The method was demonstrated to be selective against skin contaminants, linear in a wide range of concentrations (3–20 μg mL⁻¹), sensitive enough to quantify less than 0.1% of the drug dosage in skin matrices, and precise regardless of analysis variations such as day of analysis, analyst, or equipment. In addition, the method presented a high drug extraction capacity greater than 90% for all skin layers (stratum corneum, hair follicle, and remaining skin). Finally, the method was successfully tested in skin permeation assays, proving its value in the development of topical formulations containing propranolol.

中文翻译：

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建立反相高效液相色谱法测定不同皮肤层中普萘洛尔的方法

这项工作的目的是开发和验证一种使用HPLC测定皮肤不同层中普萘洛尔的分析方法，该方法可用于皮肤渗透动力学研究。该方法的开发基于色谱图的适用性，其验证遵循了生物分析方法的国际卫生法规。另外，该方法在使用猪皮的体外渗透测定中进行了测试。使用RP-C18色谱柱在30°C，流动相包括酸性水相：乙腈（75:25 v / v），流速1.0 mL min ^-1和290 nm的UV检测下测定药物。证明该方法对皮肤污染物具有选择性，在各种浓度（3–20μgmL ^-1），灵敏度足以量化皮肤基质中少于0.1％的药物剂量，并且无论分析变化（例如分析日，分析人员或设备）如何都精确。此外，该方法对所有皮肤层（角质层，毛囊和剩余皮肤）的提取率均超过90％。最后，该方法在皮肤渗透试验中成功进行了测试，证明了其在开发含有普萘洛尔的局部制剂中的价值。