Pellett Jackson, Lukulay Patrick, Mao Yun, Bowen William, Reed Robert, Ma M, Munger R C, Dolan J W, Wrisley Loren, Medwid K, Toltl N P, Chan C C, Skibic M, Biswas Kallol, Wells Kevin A, Snyder L R
Pfizer Global Research & Development, Pharmaceutical Sciences, Analytical R&D, Michigan Laboratories, Ann Arbor, MI, USA.
J Chromatogr A. 2006 Jan 6;1101(1-2):122-35. doi: 10.1016/j.chroma.2005.09.080. Epub 2005 Oct 19.
A general procedure is proposed for the rapid development of a reversed-phase liquid chromatographic (RP-LC) separation that is "orthogonal" to a pre-existing ("primary") method for the RP-LC separation of a given sample. The procedure involves a change of the mobile-phase organic solvent (B-solvent), the replacement of the primary column by one of very different selectivity, and (only if necessary) a change in mobile phase pH or the use of a third column. Following the selection of the "orthogonal" B-solvent, column and mobile phase pH, further optimization of peak spacing and resolution can be achieved by varying separation temperature and either isocratic %B or gradient time. The relative "orthogonality" of the primary and "orthogonal" RP-LC methods is then evaluated from plots of retention for one method versus the other. The present procedure was used to develop "orthogonal" methods for nine routine RP-LC methods from six pharmaceutical analysis laboratories. The relative success of this approach can be judged from the results reported here.
本文提出了一种通用方法，用于快速开发与给定样品的反相液相色谱（RP-LC）分离的现有（“主要”）方法“正交”的反相液相色谱分离方法。该方法包括改变流动相有机溶剂（B溶剂），用选择性差异很大的柱子替换主要柱子，以及（仅在必要时）改变流动相pH值或使用第三根柱子。在选择“正交”B溶剂、柱子和流动相pH值之后，可以通过改变分离温度和等度%B或梯度时间来进一步优化峰间距和分离度。然后从一种方法的保留值与另一种方法的保留值的图中评估主要RP-LC方法和“正交”RP-LC方法的相对“正交性”。本方法用于为六个药物分析实验室的九种常规RP-LC方法开发“正交”方法。从这里报告的结果可以判断这种方法的相对成功程度。
