Tu Ying, Yan Bing
Department of Chemical Biology and Therapeutics, St. Jude Children's Research Hospital, Memphis, TN, USA.
Methods Mol Biol. 2012;918:117-26. doi: 10.1007/978-1-61779-995-2_9.
An automated high-throughput method applied to the production and analysis of libraries of natural -products for high-throughput biological screening is described. The production of the library includes solid-phase extraction of crude extracts to remove polyphenols, followed by automated preparative high-performance liquid chromatography (HPLC) fractionation. Libraries of fractions are analyzed by an ultra-performance liquid chromatography-UV diode-array detection-evaporative light scattering detection-mass spectrometry system (UPLC/PDA/ELSD/MS) to provide information that facilitates characterization of compounds in active fractions. This system fractionates 2,600 unique natural product samples per year, providing fractions in 0.5-10 mg scale for creation of libraries that could be used for the screening of multiple targets to identify hits for various applications including drug discovery.
本文描述了一种应用于天然产物库生产和分析的自动化高通量方法，用于高通量生物筛选。该库的生产包括对粗提物进行固相萃取以去除多酚，随后进行自动化制备型高效液相色谱（HPLC）分级分离。通过超高效液相色谱-紫外二极管阵列检测-蒸发光散射检测-质谱系统（UPLC/PDA/ELSD/MS）对分级分离后的库进行分析，以提供有助于表征活性组分中化合物的信息。该系统每年可对2600个独特的天然产物样品进行分级分离，提供0.5-10毫克规模的组分，用于创建可用于筛选多个靶点以识别各种应用（包括药物发现）的命中物的库。
