Currently, there is an increasing interest on the development of topical formulations containing rosmarinic acid (RA) due to its well-documented antioxidant activity. This study aimed to develop and validate a stability-indicating ultra-fast liquid chromatography (UFLC) method for the determination of RA in nanoemulsions, porcine skin and nasal mucosa intended to be applied in permeation/retention studies and for development of topical nanoemulsions. Chromatographic separation was carried out using a C18 column packed with 2.6 μm particle size in isocratic conditions using as mobile phase water:acetonitrile (83:17, v/v), acidified with 0.1% trifluoracetic acid (v/v), with a total time of analysis of 3.5 min and detection at 330 nm. RA analysis was specific in the presence of both non-biological (blank nanoemulsion and receptor fluid) and biological matrices (porcine ear skin and porcine nasal mucosa). No interference of degradation products of RA was verified after different stress conditions such as acidic, alkaline, oxidative, light exposure (UV-A and UV-C) and thermal demonstrating the method stability-indicating property. The analytical (0.1–10.0 μg·mL⁻¹) and bioanalytical (0.5–10.0 μg·mL⁻¹) linearity was proved by analysis of the calibration curves of RA and no matrix effect was observed. The method was sensitive, precise and accurate, and showed recovery higher than 85%. The method was considered robust as evaluated by a Plackett-Burman experimental design. In the validated conditions, the RA was determined in the nanoemulsions obtained by spontaneous emulsification procedure (1.007 ± 0.040 mg·mL⁻¹), porcine ear skin (1.13 ± 0.19 μg·cm⁻²) and nasal mucosa (22.46 ± 3.99 μg·cm⁻²) after retention/permeation studies. Thus, a highly sensitive, simple, fast and stability-indicating method was developed for RA analysis during the development of topical nanoemulsions and bioanalytical assays in complex matrices.

目前，由于含迷迭香酸（RA）的抗氧化剂活性得到充分证明，因此对局部制剂的开发越来越感兴趣。这项研究旨在开发和验证一种稳定性指示超快速液相色谱（UFLC）方法，用于测定打算用于渗透/保留研究和用于局部纳米乳剂的纳米乳剂，猪皮肤和鼻黏膜中的RA。使用填充有2.6μm粒径的C18色谱柱在等度条件下进行色谱分离，使用流动相为水：乙腈（83:17，v / v），用0.1％三氟乙酸（v / v）酸化，总计分析时间为3.5分钟，并在330 nm处检测。在非生物（空白纳米乳剂和受体液）和生物基质（猪耳皮肤和猪鼻黏膜）均存在的情况下，RA分析具有特异性。在不同的应力条件下（如酸性，碱性，氧化性，光照（UV-A和UV-C））和热证明了方法的稳定性指示性能后，未证实RA的降解产物受到干扰。解析度（0.1–10.0μg·mL通过分析RA的校准曲线证明了^-1）和生物分析（0.5-10.0μg·mL ^-1）线性，没有观察到基质效应。该方法灵敏，准确，准确，回收率高于85％。通过Plackett-Burman实验设计评估，该方法被认为是可靠的。在验证的条件下，通过自发乳化程序（1.007±0.040 mg·mL ^-1），猪耳皮肤（1.13±0.19μg·cm ^-2）和鼻黏膜（22.46±3.99μg·厘米^-2），经过保留/渗透研究。因此，在局部基质纳米乳剂和复杂基质中生物分析方法的开发过程中，开发了一种高度灵敏，简单，快速且稳定的指示方法用于RA分析。