Aljohar Haya I, Al-Hossaini Abdullah M, Alzammay Seham A, Alhabardi Samiah, Maher Hadir M, Ahmed Aya R
Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia.
Department of Pharmaceutics, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia.
BMC Chem. 2025 Jan 13;19(1):14. doi: 10.1186/s13065-024-01372-3.
A simple, rapid, and reproducible high-performance liquid chromatography (HPLC) method has been developed and validated for the determination of β-sitosterol in the pharmaceutical dosage form of moist exposed burn ointment (MEBO). This method involved an effective sample procedure for extraction of β-sitosterol from MEBO using an alkali saponification agent composed of 0.8 N ethanolic NaOH and diethyl ether. The chromatographic separation was achieved on a C18 column (50 × 3.0 mm, 2.5 μm), using a mobile phase composed of methanol and acetonitrile (70:30 v/v) pumped in an isocratic mode at a flow rate of 0.7 mL/min. The column temperature was maintained at 40 °C, the injection volume was 10 µL, and the detection wavelength was 203 nm. Employing these conditions, the retention time was found to be 2.10 min. The developed method was validated for its specificity, linearity, accuracy, precision, the limit of detection, the limit of quantification, robustness, and solution stability based on International Council for Harmonisation (ICH) guidelines Q2 (R1). Our proposed method demonstrated superior performance compared to other reported methods. It exhibited a linearity range of 30 to 500 µg/mL and improved detectability with a limit of detection (LOD) of 4.65 µg/mL, highlighting its high sensitivity. Additionally, the separation was achieved in a remarkably short analysis time of just 2.1 min, which not only enhanced throughput but also significantly minimized waste and solvent consumption, thereby making it a more sustainable and effective alternative for β-sitosterol extraction. Moreover, in the light of green and white analytical chemistry, a comprehensive ecological and sustainable tri-color coded assessment protocol was established. The proposed method has been successfully applied to quantify β-sitosterol in commercial products (MEBO, Avomeb and BISTROl) demonstrating its suitability for routine quality control analysis of β-sitosterol in pharmaceutical ointment dosage forms.
已开发并验证了一种简单、快速且可重复的高效液相色谱（HPLC）方法，用于测定湿润暴露烧伤膏（MEBO）药物剂型中的β-谷甾醇。该方法采用由0.8 N乙醇氢氧化钠和乙醚组成的碱性皂化剂，从MEBO中有效提取β-谷甾醇。色谱分离在C18柱（50×3.0 mm，2.5μm）上进行，流动相由甲醇和乙腈（70:30 v/v）组成，以等度模式泵送，流速为0.7 mL/min。柱温保持在40℃，进样体积为10 μL，检测波长为203 nm。采用这些条件，保留时间为2.10分钟。根据国际协调理事会（ICH）指南Q2（R1），对所开发方法的特异性、线性、准确性、精密度、检测限、定量限、稳健性和溶液稳定性进行了验证。我们提出的方法与其他报道的方法相比表现出优异的性能。它的线性范围为30至500 μg/mL，检测限（LOD）为4.65 μg/mL，提高了可检测性，突出了其高灵敏度。此外，分离在仅2.1分钟的极短分析时间内完成，这不仅提高了通量，还显著减少了废物和溶剂消耗，从而使其成为β-谷甾醇提取更可持续、更有效的替代方法。此外，根据绿色和白色分析化学，建立了全面的生态和可持续三色编码评估方案。所提出的方法已成功应用于商业产品（MEBO、Avomeb和BISTROl）中β-谷甾醇的定量，证明了其适用于药膏剂型中β-谷甾醇的常规质量控制分析。
