Al-Sayah Mohammad A, Rizos Panagiota, Antonucci Vincent, Wu Naijun
Merck & Co. Inc., Rahway, NJ 07065, USA.
J Sep Sci. 2008 Jul;31(12):2167-72. doi: 10.1002/jssc.200700594.
Ultra performance LC (UPLC) was evaluated as an efficient screening approach to facilitate method development for drug candidates. Three stationary phases were screened: C-18, phenyl, and Shield RP 18 with column dimensions of 150 mm x 2.1 mm, 1.7 microm, which should theoretically generate 35,000 plates or 175% of the typical column plate count of a conventional 250 mm x 4.6 mm, 5 microm particle column. Thirteen different active pharmaceutical ingredients (APIs) were screened using this column set with a standardized mobile-phase gradient. The UPLC method selectivity results were compared to those obtained for these compounds via methods developed through laborious trial and error screening experiments using numerous conventional HPLC mobile and stationary phases. Peak capacity was compared for columns packed with 5 microm particles and columns packed with 1.7 microm particles. The impurities screened by UPLC were confirmed by LC/MS. The results demonstrate that simple, high efficiency UPLC gradients are a feasible and productive alternative to more conventional multiparametric chromatographic screening approaches for many compounds in the early stages of drug development.
超高效液相色谱（UPLC）被评估为一种有效的筛选方法，以促进候选药物的方法开发。筛选了三种固定相：C-18、苯基和Shield RP 18，柱尺寸为150 mm×2.1 mm，1.7微米，理论上应产生35000个塔板数，或传统250 mm×4.6 mm、5微米粒径柱典型塔板数的175%。使用该柱组和标准化流动相梯度筛选了13种不同的活性药物成分（API）。将UPLC方法的选择性结果与通过使用多种常规HPLC流动相和固定相进行费力的反复筛选实验开发的方法对这些化合物获得的结果进行了比较。比较了填充5微米粒径颗粒的柱和填充1.7微米粒径颗粒的柱的峰容量。通过UPLC筛选出的杂质通过LC/MS进行了确认。结果表明，对于药物开发早期的许多化合物，简单、高效的UPLC梯度是比更传统的多参数色谱筛选方法更可行、更有效的替代方法。
