Mass-Directed制备SFC——回收率和纯度测定

超临界流体色谱（SFC）是一种发展迅速的技术。最初几乎只用于热不稳定和手性化合物，现在它被认为是一种广泛应用于各种化合物的通用分析和纯化技术。它比常规HPLC和Prep-lc的产量更高，并具有溶剂消耗、耗材和处理成本低得多的“绿色”优势。SFC与多种检测方法兼容，包括紫外电喷雾（ESI）和大气压化学电离（APCI）质谱法。本应用中结论：CMS可以简单地连接到分析天平和制备天平SFC，在紫外线检测器之前使用简单的被动拆分UV和MS信号之间的定时延迟被确定为3.6秒对每种分析物收集的所有级分进行了分析测试，结果表明每种级分的纯度均为99.9%或更高UV引发的回收率与MS-TIC引发的组分之间的差异，黄酮和磺胺二甲嘧啶峰的回收率小于2%，卡马西平的回收率低于6%Results Mass-Directed Fraction Collection for Preparative SFC Recove r y and P u r i ty D e t er min ation usi n g a J asco P r ep SFC coupled with the expression Compa ct Mass Spe c tromet e r using a simple passive split Mark Hardink *， Frank R iley & Qi Yan， P fizer ； Simon Prosser， Adv i on；John Burchell & DJ Tognarelli ， JASCO Introduction Supercri t ical fluid chromatography (SFC) is a rapidly growing technique.Initially used almost exclusively only for thermally labile and chiral DIe compounds， i t is now accepted as a versatile analytical and pur i fication technique applied a cross a broad range of compounds. It is higher-throughput than regular HPLC and Prep-LC and offers the green’ advantages of much lower solvent consumption， consumable and disposal costs. SFC is compatible with a wide range of detection methods inc l uding UV，electrospray (ESl) and atmospheric pressure chemical ionization (APCI ) mass spectrometry. Methods The initial tests explored a wide range of flow rates， both isocratic and gradient ， to test performance and rel i ability of the epassive splitter， coincident t i ming of MS and UV signal and the clearance of the source with highly concentrated samples. The system was further evaluated by testing the recovery and purity of compounds i n a three-component mi x ture. Test #1：：Three component standard mixture，700 儿 injections， gradient elution，mass-directed collection using TIC of each peak per i njection starting with Peak A， then B &C (Flavone， Carbamazepine， and Sulfamethazine). Repeat for aa total of 5injections. Measure the purity and recovery of the three collected fractions . Test #2：：Three component standard m i xture，700 pL i njections， gradient elution，UV-directed collection using UV of each peaks per injection start i ng with Peak A， then B & C (Fl a vone，Carbamazepine， and Sulfamethazine). Repeat for a total of 5 injections. Measure the purity and recovery of the three col l ected fractions. Experimental TIC triggered fraction collection (5 injections) 477.8mg 50 mL 9.56 mg/m L 700 u L 7 46.8244 mg 44.4mg 94.8% C a rbamaz e pi ne 478.2 50 9.56 700 5 33.474 31.4 93.8 Sulf a metha zi ne 473.9 50 9.48 700 5 33.173 31.8 95.9 * Two a dditional f r a ctions of Flavone were collected duri n g ins t rument set -u p and not disposed，showi n g a l arger collect e d s ample . UV triggered Fractio n collection (5 injections) JASCO /ADVION S F C PRE P rec o ve r y Dif fere n c e Weight of Sample Sample injection # of Theory sample Sample Vs. UV trigger analyte Volume concentration volume Injections injected on Column Recovered Recoverv Flavone 477.8mg 50 mL 9.56 mg/mL 700uL 5 33.446 mg 31.7mg 94.8% Carbamazepine 478.2 9.56 700 5 33.474 33.5 100.1 6.3 Sulfamethazine 473.9 9.48 700 5 33.173 31.3 94.4 -1.6 Conclusions · The CMS c a n be simply interfaced to both analytical scale and prep scale SFC using a simple passive split before the UV detector · Timing delay between UV and MS signals was determined to be 3.6 seconds · All fractions collected of each analyte were tested analytically and showed that each was 99.9% pure or greater · Di f ference between the recoveries of that were t riggered by UV verses the fractions triggered by MS-TIC were less than 2% for the flavone and sulfamethazine peak and 6% for the carbamazepine