Rapid method development for chiral separation in drug discovery using sample pooling and supercritical fluid chromatography-mass spectrometry.
A novel strategy for rapid chiral method development has been implemented using sample pooling and supercritical fluid chromatography-mass spectrometry (SFC-MS) on four chiral stationary phases, namely Chiralpak AD and AS, and Chiralcel OJ and OD, and eight different modifier concentrations (5 to 40% methanol-0.2% isopropylamine). The screening is performed under an outlet pressure of 110 bar at 35 degrees C, and at a flow-rate of 2.5 ml/min for the initial 20 min and then ramped up to 4 ml/min and held for 4.5 min to elute all solutes from the column. The entire process is fully automated from injection to data processing, and operates unattended for 15 h overnight to obtain optimal chiral separation for multiple compounds. A unique feature of using SFC-MS to monitor chiral synthesis is the negligible interferences from achiral impurities. In addition, with SFC-MS, enantiomeric excess can be determined with much lower detection limits than UV and much shorter analysis times compared to normal-phase/reversed-phase liquid chromatography.
Zhao
Yining
Y
Discovery Analytical Science, Amgen Inc., One Amgen Center Drive, Mail Stop 29-M-B, Thousand Oaks, CA 91320, USA. yzhao@amgen.com
Woo
Gregory
G
Thomas
Samuel
S
Semin
David
D
Sandra
Pat
P
D000688:Amylose; D001322:Autoanalysis / Q000379:methods; D002482:Cellulose; D002851:Chromatography, High Pressure Liquid; D025924:Chromatography, Supercritical Fluid / Q000295:instrumentation / Q000379:methods*; D007202:Indicators and Reagents; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods*; D004364:Pharmaceutical Preparations / Q000302:isolation & purification*; D013237:Stereoisomerism; D013678:Technology, Pharmaceutical
D007202:Indicators and Reagents; D004364:Pharmaceutical Preparations; D002482:Cellulose; D000688:Amylose