Accurate quantitation of low dose, multi-active dissolution samples poses unique challenges in the pharmaceutical industry, often resulting in separate HPLC methods for each active or the use of multiple detectors for increased sensitivity. In this study, we report a fast, isocratic HPLC method utilizing only UV detection for dissolution testing of low dose desogestrel and ethinylestradiol tablets. Rapid separation is completed in 5 min using isocratic elution at a flow rate of 0.45 mL/min, with a column temperature at 30 °C, an injection volume of 50 μL and the detection wavelength at 200 nm. After extensive method development and optimization, the cyano stationary phase was used to overcome the large difference in hydrophobicity for desogestrel and ethinylestradiol, providing balanced retention for both analytes under isocratic elution. Chromatography modeling software was used to provide a rapid analysis of multiple columns and chromatography conditions. The optimized method boasts fast and efficient separation through use of a short, small I.D. column and a large injection volume of dissolution solution to achieve high sensitivity. The stable baseline from an isocratic separation allows low detection wavelengths to be used, resulting in accurate and precise quantitation of both desogestrel and ethinylestradiol. The method has been successfully validated for specificity, linearity, accuracy and precision in the range of 75 – 600 ng/mL for desogestrel and 10 – 80 ng/mL for ethinylestradiol using both HPLC and UHPLC systems. The method robustness was characterized using a design of experiment approach, and the operational design region of the method was established.

低剂量、多活性溶出度样品的准确定量给制药行业带来了独特的挑战，通常导致对每种活性物质采用单独的 HPLC 方法或使用多个检测器来提高灵敏度。在本研究中，我们报告了一种快速、等度的 HPLC 方法，该方法仅利用 UV 检测对低剂量去氧孕烯和炔雌醇片剂进行溶出度测试。使用等度洗脱，流速为 0.45 mL/min，柱温为 30 °C，进样量为 50 μL，检测波长为 200 nm，在 5 分钟内完成快速分离。经过广泛的方法开发和优化，氰基固定相用于克服去氧孕烯和炔雌醇疏水性的巨大差异，在等度洗脱下为两种分析物提供平衡的保留。色谱建模软件用于提供多色谱柱和色谱条件的快速分析。优化后的方法通过使用短小内径柱和大溶出液进样量实现了高灵敏度的快速高效分离。等度分离的稳定基线允许使用低检测波长，从而对去氧孕烯和炔雌醇进行准确和精确的定量。使用 HPLC 和 UHPLC 系统成功验证了该方法的特异性、线性、准确度和精密度，去氧孕烯的范围为 75 – 600 ng/mL，炔雌醇的范围为 10 – 80 ng/mL。使用实验设计方法表征方法的稳健性，并建立了该方法的操作设计区域。