Abstract A rapid and specific method was developed for simultaneous quantification of hydrocortisone 21 acetate (HCA), dexamethasone (DEX), and fluocinolone acetonide (FCA) in whitening cream formulations using reversed-phase high-performance liquid chromatography. The effect of the composition of the mobile phase, analysis temperature, and detection wavelength was investigated to optimize the separation of studied components. The analytes were finally well separated using ACE Excel 2, C18 AR column having 150 mm length, 3 mm internal diameter, and 2 µm particle size at 35°C using methanol with 1% formic acid and double-distilled deionized water in the ratio of 60:40 (v/v), respectively, as the mobile phase in isocratic mode. Ten microliters of sample were injected with a flow rate of 0.5 mL/min. The specificity, linearity, accuracy, precision, recovery, limit of detection (LOD), limit of quantification (LOQ), and robustness were determined to validate the method as per International Conference on Harmonization guidelines. All the analytes were simultaneously separated within 8 min, and observed retention times of HCA, DEX, and FCA were 4.5, 5.5, and 6.9 min, respectively. The proposed method showed good linearity with the correlation coefficient, R 2 = 0.999 over the range of 1–150 µg/mL for all standards. The linear regression equations were y = 12.7x + 118.7 (r = 0.999) for HCA, y = 12.9x + 106.8 (r = 0.999) for DEX, and y = 12.9x + 96.8 (r = 0.999) for FCA. The LOD was 0.25, 0.20, and 0.08 µg/mL for HCA, FCA, and DEX and LOQ was 2.06, 1.83, and 1.55 µg/mL for HCA, FCA, and DEX, respectively. The recovery values of HCA, DEX, and FCA ranged from 100.7–101.3, 102.0–102.6, and 100.2–102.0%, respectively, and the relative standard deviation for precision (intra- and interday) was less than 2, which indicated repeatability and reproducibility. The novelty of the method was described by forced degradation experimentation of all analytes in the combined form under acidic, basic, oxidative, and thermal stress. The proposed method was found to be simple, rapid, and reliable for the simultaneous determination of HCA, DEX, and FCA in cosmetics.

中文翻译：

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用于同时定量化妆品中醋酸氢化可的松 21、地塞米松和醋酸氟轻松的稳定性指示 HPLC-DAD 分析

摘要 开发了一种使用反相高效液相色谱同时定量美白霜配方中醋酸氢化可的松 (HCA)、地塞米松 (DEX) 和氟轻松 (FCA) 的快速且特异性的方法。研究了流动相组成、分析温度和检测波长的影响，以优化研究组分的分离。最终使用 ACE Excel 2、长度为 150 mm、内径为 3 mm、粒径为 2 µm 的 C18 AR 色谱柱在 35°C 下使用含有 1% 甲酸和双蒸去离子水的甲醇对分析物进行很好的分离60:40 (v/v)，分别作为等度模式的流动相。以 0.5 mL/min 的流速注入 10 微升样品。特异性、线性、准确度、精密度、根据国际协调会议指南确定了回收率、检测限 (LOD)、定量限 (LOQ) 和稳健性，以验证该方法。所有分析物在 8 分钟内同时分离，观察到的 HCA、DEX 和 FCA 的保留时间分别为 4.5、5.5 和 6.9 分钟。所提出的方法在所有标准品的 1–150 µg/mL 范围内与相关系数显示出良好的线性关系，R 2 = 0.999。HCA 的线性回归方程为 y = 12.7x + 118.7 (r = 0.999)，DEX 为 y = 12.9x + 106.8 (r = 0.999)，FCA 为 y = 12.9x + 96.8 (r = 0.999)。HCA、FCA 和 DEX 的 LOD 为 0.25、0.20 和 0.08 µg/mL，HCA、FCA 和 DEX 的 LOQ 分别为 2.06、1.83 和 1.55 µg/mL。HCA、DEX 和 FCA 的回收率范围为 100.7-101.3、102.0-102.6 和 100.2-102。分别为 0%，精密度（日内和日间）的相对标准偏差小于 2，表明具有重复性和再现性。通过在酸性、碱性、氧化和热应力下以组合形式对所有分析物进行强制降解实验，描述了该方法的新颖性。发现该方法简单、快速且可靠，可同时测定化妆品中的 HCA、DEX 和 FCA。