Yan Bing, Collins Nathan, Wheatley Jeffrey, Irving Mark, Leopold Kyle, Chan Cecilia, Shornikov Alex, Fang Liling, Lee Andrew, Stock Mike, Zhao Jiang
Discovery Chemistry Division, Discovery Partners International, Inc., 385 Oyster Point Boulevard, South San Francisco, CA 94080, USA.
J Comb Chem. 2004 Mar-Apr;6(2):255-61. doi: 10.1021/cc0340527.
We have developed a high-throughput purification system to purify combinatorial libraries at a 50-100-mg scale with a throughput of 250 samples/instrument/day. We applied an accelerated retention window method to shorten the purification time and targeted one fraction per injection to simplify data tracking, lower QC workload, and simplify the postpurification processing. First, we determined the accurate retention time and peak height for all compounds using an eight-channel parallel LC/UV/MS system, and calculated the specific preparative HPLC conditions for individual compounds. The preparative HPLC conditions include the compound-specific gradient segment for individual compounds with a fixed gradient slope and the compound-specific UV or ELSD threshold for triggering a fraction collection device. A unique solvent composition or solvent strength was programmed for each compound in the preparative HPLC in order to elute all compounds at the same target time. Considering the possible deviation of the predicted retention time, a 1-min window around the target time was set to collect peaks above a threshold based on UV or ELSD detection. Dual column preparative instruments were used to maximize throughput. We have purified more than 500 000 druglike compounds using this system in the past 3 years. We report various components of this high-throughput purification system and some of our purification results.
我们开发了一种高通量纯化系统，可在50-100毫克规模下纯化组合文库，通量为250个样品/仪器/天。我们应用了加速保留窗口方法来缩短纯化时间，并每次进样靶向一个馏分，以简化数据跟踪、降低质量控制工作量并简化纯化后处理。首先，我们使用八通道平行LC/UV/MS系统确定所有化合物的准确保留时间和峰高，并计算单个化合物的特定制备型HPLC条件。制备型HPLC条件包括具有固定梯度斜率的单个化合物的化合物特异性梯度段以及触发馏分收集装置的化合物特异性UV或蒸发光散射检测器（ELSD）阈值。为制备型HPLC中的每种化合物设定独特的溶剂组成或溶剂强度，以便在相同的目标时间洗脱所有化合物。考虑到预测保留时间可能存在偏差，在目标时间周围设置1分钟的窗口，基于UV或ELSD检测收集高于阈值的峰。使用双柱制备仪器以最大化通量。在过去3年中，我们使用该系统纯化了超过500,000种类药物化合物。我们报告了这种高通量纯化系统的各个组件以及我们的一些纯化结果。
