Box–Behnken Design-Based Optimized UHPLC Method for the Analysis of Lipid-Based SNEDDS Ferrying Hydrocortisone and Thymoquinone
Abstract
High Resolution Image Download MS PowerPoint Slide This study aimed to develop and validate a robust ultrahigh-performance liquid chromatography (UHPLC) method for the simultaneous quantification of hydrocortisone (HCT) and thymoquinone (TMQ) from self-nanoemulsifying drug delivery systems (SNEDDS). A Box–Behnken experimental design was applied to identify the impact of critical parameters, including the column temperature (20–40 °C), flow rate (0.2–0.4 mL/min), and buffer ratio (60–75%), on chromatographic responses. The optimized conditions comprised a column temperature of 20 °C, a flow rate of 0.2 mL/min, and a buffer ratio of 70.7%, achieving excellent peak resolution and asymmetry. The method demonstrated linearity (R 2 > 0.999) across the concentration range of 1–30 μg/mL for both analytes. Precision studies revealed relative standard deviations below 9.39% and 2.55% for HCT and TMQ, respectively, while accuracy ranged from 96.41% to 102.69% for HCT and from 97.83% to 106.21% for TMQ. The method exhibited high sensitivity with limits of detection of 0.203 and 0.129 μg/mL and limits of quantification of 0.615 and 0.392 μg/mL for HCT and TMQ, respectively. The method was robust, sensitive, and reproducible. The validated method was successfully applied to analyze HCT and TMQ contents with high recovery rates of 96.8 ± 2.53% and 101.5 ± 2.37%, respectively. The retention time for HCT was relatively shorter compared to that for TMQ, which may be attributed to various…
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