用于组合文库初步纯化的自动化高通量液-液萃取

Automated high-throughput liquid-liquid extraction for initial purification of combinatorial libraries.

作者信息

Peng S X, Henson C, Strojnowski M J, Golebiowski A, Klopfenstein S R

机构信息

Health Care Research Center, Procter & Gamble Company, Mason, Ohio 45040, USA.

出版信息

Anal Chem. 2000 Jan 15;72(2):261-6. doi: 10.1021/ac990946v.

DOI:10.1021/ac990946v

PMID:10658318

Abstract

An automated high-throughput liquid-liquid extraction (LLE) methodology has been developed and utilized for the initial purification of the combinatorial library samples containing unreacted amines and other water-soluble byproducts or impurities. Various extraction solvents were evaluated along with different extraction devices. The LLE method was automated using 96-well-format plates and a robotic liquid-handling workstation. In the optimized LLE method, crude combinatorial library samples were dissolved in a water-immiscible organic solvent, butyl acetate, and added to each well in a 96-well-format plate packed with an inert support material coated with hydrochloric acid. Separation occurs based on the partitioning of the compounds between two liquid phases. Product recovery, purity, and amine removal efficiency were determined by HPLC with and without precolumn derivatization. The automated method was successfully applied to the cleanup of some representative combinatorial library samples with greater than 98% amine removal and an average product purity of 90%. The application of the automated high-throughput LLE method should greatly reduce the labor, time, and cost associated with the purification of combinatorial libraries.

摘要

已开发并利用一种自动化高通量液-液萃取（LLE）方法，用于初步纯化含有未反应胺以及其他水溶性副产物或杂质的组合文库样品。对各种萃取溶剂以及不同的萃取装置进行了评估。LLE方法使用96孔板和机器人液体处理工作站实现了自动化。在优化的LLE方法中，将粗组合文库样品溶解于与水不混溶的有机溶剂乙酸丁酯中，并添加到装有涂有盐酸的惰性支撑材料的96孔板的每个孔中。分离基于化合物在两个液相之间的分配。通过有无柱前衍生化的高效液相色谱法（HPLC）测定产物回收率、纯度和胺去除效率。该自动化方法成功应用于一些代表性组合文库样品的净化，胺去除率大于98%，平均产物纯度为90%。自动化高通量LLE方法的应用应能大大减少与组合文库纯化相关的劳动力、时间和成本。