School of Pharmacy, College of Pharmacy, Taipei Medical University, Taipei, 11042, Taiwan.
Graduate Institute of Pharmacognosy, College of Pharmacy, Taipei Medical University, Taipei, 11042, Taiwan.
Anal Methods. 2024 Sep 26;16(37):6420-6428. doi: 10.1039/d4ay00687a.
The rapid isolation of natural products and efficient drug screening are pivotal in expediting drug development. Techniques ranging from traditional open column chromatography to medium-pressure liquid chromatography (MPLC), and the latest Sepbox technology, have been developed to accelerate separation processes and streamline drug development timelines. The Sepbox system combines two-dimensional high-performance liquid chromatography (2D-HPLC) and solid-phase extraction (SPE) technologies, coupled with UV and evaporative light scattering detection (ELSD) systems, offering various column options to cater to diverse sample requirements. Furthermore, the Sepbox system automates and expedites sample fractionation into numerous fractions, facilitating subsequent high-throughput screening and analysis. Despite previous emphasis on 2D-HPLC development, optimizing separation conditions with the Sepbox system poses challenges due to the requirement for substantial sample and solvent quantities, limiting its practicality compared to conventional methods. Hence, this study employed eight standard compounds to explore the correlation between retention factor () values obtained from high-performance thin-layer chromatography (HPTLC) plates and retention times on the Sepbox main column. Mass spectrometry was utilized to confirm the retention times of the standard compounds. The findings yielded a conversion equation between HPTLC values and Sepbox main column retention times, thereby enhancing the separation efficiency of Sepbox 2D-2000 system. Finally, the efficacy of this method was validated using leaf crude extracts and its purified compounds, demonstrating the rapid optimization of suitable elution conditions for the Sepbox 2D-2000 system using HPTLC.
快速分离天然产物和高效药物筛选对于加速药物开发至关重要。已经开发出从传统的开放柱层析到中压液相色谱（MPLC）以及最新的 Sepbox 技术等技术，以加速分离过程并简化药物开发时间表。Sepbox 系统结合了二维高效液相色谱（2D-HPLC）和固相萃取（SPE）技术，与 UV 和蒸发光散射检测（ELSD）系统相结合，提供了各种柱选项以满足不同的样品要求。此外，Sepbox 系统可以自动化并加速将样品分离成多个馏分，从而方便随后进行高通量筛选和分析。尽管以前强调 2D-HPLC 的发展，但由于 Sepbox 系统需要大量的样品和溶剂，因此优化分离条件具有挑战性，与传统方法相比，其实用性有限。因此，本研究使用了八个标准化合物来探索从高效薄层色谱（HPTLC）板获得的保留因子（）值与 Sepbox 主柱上的保留时间之间的相关性。使用质谱法确认了标准化合物的保留时间。得出了 HPTLC 值和 Sepbox 主柱保留时间之间的转换方程，从而提高了 Sepbox 2D-2000 系统的分离效率。最后，使用叶粗提物及其纯化化合物验证了该方法的有效性，证明了使用 HPTLC 可以快速优化 Sepbox 2D-2000 系统的合适洗脱条件。
