平行SFC-MS快速手性和手性分析及半制备分析

手性和非手性SFC分析需要溶剂和柱筛选，以确定最佳的溶剂-柱组合，并为兴趣峰提供所需的分辨率。随着手性柱和非手性柱数量的增加以及分析样品数量的增加，需要更高的通量。然而，反相并不是手性分离的理想分离技术，SFC也为非手性化合物的反相提供了正交分离。此外，更小的颗粒柱导致更高的压力，需要更昂贵的设备。UHPLC的规模扩大已经建立，但不如SFC的规模扩大那么简单，而且使用CO2醇流动相，修复性SFC中的馏分回收要容易得多。在这个应用中，我们展示了在溶剂和手性和非手性分析的柱筛选中具有更高通量的平行SFC。平行SFC提供多达10种溶剂和24个柱，但在确定最佳溶剂-柱组合后，还能够进一步优化分离。在优化之后，该系统允许放大到制备纯化。本应用中结论：由于SFC对手性和非手性分离都需要柱状筛选和溶剂筛选，因此平行筛选显著减少了总分析时间。这个分析时间可以减少5倍，但仍然允许在初始筛选后进一步开发单独的列方法。此外，该系统还提供了在初始筛选和分离优化后扩大净化规模的能力。质谱仪的加入不仅有助于手性纯化，而且对非手性分离也势在必行。质谱仪通过质量区分化合物的能力，仍然允许质谱仪与平行SFC一起工作，为手性和非手性分析提供最快和最强大的分离技术。 Parallel SFC-MS for Rapid Chiral and Achiral Analytical and Semi-Prep Analysi s DJ Tognarelli1， John Burchell1， Satoe lijima2， Tomohiro Moteki?， Atsushi Tsukamoto2， Masao Bounoshita2， Yasuyo Sato2， Miki Kuwajima2. 1 JASCO Incorporated， 28600 Mary's Court，Easton， MD 21601 USA. 2 JASCO Corporation， 2967-5 Ishikawa-machi， Hachioji， Tokyo， 192-8537 Japan.email ： dtognarelli@jascoinc.com Chiral and achiral SFC analysis requires solvent and column screening i n order to determine the best solvent-column combination and provide the desired resolution for the peaks of interest. With the increasing number of chiral and achiral columns and the increase in the number of samples to analyze， higher throughput is required. The implementat i on of UHPLC using shorter length and smal l er particle columns has increased to address this throughput need. However， reversed-phase is not the desired separation technique for chiral separat i ons and SFC also offers an orthogonal separation to reversed-phase for achiral compounds . In addition， smaller particle columns lead to higher pressures requiring more expensive equipment. Scale-up from UHPLC has been established， but is not as straightforward as SFC scale-up and the fraction recovery in preparative SFC i s much easier with t he CO，-alcohol mobile phase. In t his application we show the parallel SFC for higher throughput i n solvent and column screening for chiral and achiral analysis. The parallel SFC offers up to 10 solvents and 24columns， but also the ability to further optimize the separation after the best solvent-column combination is determined. After the optimization， the system allows for scale-up to preparative puri f ication. Methods and Materials Flavanone， ibuprofen， ketoprofen， sulfamethazine and methanol， were purchased from Sigma-Aldr i ch (St. Louis， MO， USA). The terpenes sample containing 19 terpenes was obtained from Restek. The CO， cyl i nder was purchased from Airgas (PA， USA). Columns were acquired from multiple suppliers： Princeton Chromatography (Princeton， NJ， USA)，Phenomenex (T orrance， CA， USA) and Chiral Technologies (PA， USA). The JASCO Parallel SFC system consisted of a CO2 pump， co-solvent pump， autosampler， column oven， UV detectors， Advion CMS-L， and back pressure regulator. Results Ibuprofen Screen - Run ti me 15 minutes Flavanone Screen 0.01.02.0 3.0 4.05.00.01.02.0 3.04.05.0 0.01.02.00 3.0 4.05.00.01.02.0 3.04.05.0 The results of the 4 column parallel screen for ibuprofen and flavanone are shown above which required only 15 minutes each to screen 12 col u mns. This results in the reduction i n screening t ime by 45 minutes compared to run each column individually. The best separation is circled in red and the system also allows for further optimizat i on on the best column without any changes. Parallel screening in SFC not only applies to chiral separations as shown in the achiral screening of t erpenes on 12 columns. Both chiral and achiral columns were used for this screen as often chiral columns have provided achiral separations. This show the abili t y for a simultaneous ch i ral and achiral screening. Terpenes Screen The addition o f a mass spectrometer is useful for ch i ral separat i ons for identity confirmat i on especially with the presence of impurities， but certainly offers a significant advantage for achiral separations . As shown be l ow the 2 column parallel screening was run i nto the CMS mass spectrometer. With the presence of both chiral and achiral compounds， the peaks are more easily conf i rmed and identified with the mass spectrometer. The mass spectrometer is able to detect the compounds coming from both columns and when peaks co-elute wi t hin a column or co-elute when recombined after running parallel ， the mass spectrometer was able to different i ate and identify the compounds. This capabil i ty provided a powerful identi fi cation tool while stil l t aking advantage of the speed provided by parallel screening. Conclusions As SFC requires column and solvent screening for both chiral and achiral separation， parallel screening significant l y reduces the total analysis time . This analysis time can be red u ce as much as 5 fold， but sti l l allows for further individual column method develop after the initial screen. In addition the system also offers the abili t y to scale up for pur i f i cation af t er t he initial screen and separat i on optimization. The addition of the mass spectrometer not only aids in chiral puri f ications， but is imperative for achiral separations. The power of the mass spectrometer to di f ferentiate compounds by mass， still allows for the mass spectrometer to work with paral l el SFC providing the fastest and most powerful separation technique for both chira l and achiral analysis.