Regalado Erik L, Helmy Roy, Green Mitchell D, Welch Christopher J
Merck Research Laboratories, Rahway, NJ, USA.
J Sep Sci. 2014 May;37(9-10):1094-102. doi: 10.1002/jssc.201400038. Epub 2014 Apr 10.
In this study, we investigate the separation of a variety of mixtures of drugs, metabolites, and related analogs including representatives of the carbamazepine, methylated xanthine, steroid hormone, nicotine, and morphine families using several automated chromatographic method development screening systems including ultra high performance liquid chromatography, core-shell HPLC, achiral supercritical fluid chromatography (SFC), and chiral SFC. Of the 138 column and mobile phase combinations examined for each mixture, a few chromatographic conditions afford the best overall performance, with a single achiral SFC method (4.6 × 250 mm, 3.0 μm GreenSep Ethyl Pyridine, 25 mM isobutylamine in methanol/CO2) affording good separation for all samples. Four of these mixtures were also resolved by achiral SFC on the Luna HILIC and chiral SFC Chiralpak IB columns using methanol or ethanol with 25 mM isobutylamine as polar modifiers. Modifications of standard chromatography screening conditions afforded fast separation methods (from 1 to 5 min) for baseline resolution of all components of each of these challenging sets of closely related compounds.
在本研究中，我们使用几种自动色谱方法开发筛选系统，包括超高效液相色谱、核壳高效液相色谱、非手性超临界流体色谱（SFC）和手性SFC，研究了多种药物、代谢物及相关类似物混合物的分离，这些混合物包括卡马西平、甲基化黄嘌呤、甾体激素、尼古丁和吗啡家族的代表物。对于每种混合物，在考察的138种色谱柱和流动相组合中，有几种色谱条件具有最佳的整体性能，其中一种非手性SFC方法（4.6×250 mm，3.0μm GreenSep乙基吡啶，甲醇/二氧化碳中含25 mM异丁胺）对所有样品都有良好的分离效果。其中四种混合物也通过在Luna HILIC柱和手性SFC Chiralpak IB柱上进行非手性SFC分离，使用甲醇或乙醇并添加25 mM异丁胺作为极性改性剂。对标准色谱筛选条件进行修改后，得到了快速分离方法（1至5分钟），可对这些具有挑战性的紧密相关化合物组中的每种化合物的所有组分进行基线分离。
