如何快速建立手性药物分析的LC及LC-MS-MS方法.ppt

Chiral Method Development Techniques for Analytical Prep HPLC Proposed Structure of Vancomycin Proposed Structures of Glycopeptide CSPs Proposed Structures of Glycopeptide CSPs Features of CHIROBIOTIC Phases Reversed phase Largest and most versatile number of separations Polar Organic phase Fast kinetics, second major group of separations Best suited for LC/MS systems Normal phase Generally provides ca 10-15% of the total methods Column coupling possible Rapid, broad screening protocols Each column can be used in all 3 modes Broad Selectivity Based on the Same Stereogenic Center: CHIROBIOTIC T: Amino Alcohols What is the Polar Organic Mode? The Polar Organic Mode was developed originally by Dr D W Armstrong and Astec in ’85 for use with CYCLOBOND cyclodextrin technology 100 % MeOH with added acid and base (typically 0.1%, each with a range of 1.0 to 0.001%), or equivalent salts Method development is very simple and fast What is the Polar Organic Mode? The Polar Organic Mode terminology is now frequently applied to the Daicel phases, but in this case it means 100% organic only, usually MeOH, ACN or MeOH/EtOH The mechanism (predominantly hydrogen bonding) for the POM on Daicel is different from that on the CHIROBIOTIC phases and the optimization process is different What is the Polar Organic Mode? The mechanism for the POM on the CHIROBIOTIC phases is predominantly ionic, making it essential for acid and base to be added. Thus, Polar Ionic Mode? Advantages of Polar Ionic Mode? on CHIROBIOTIC V, T and R No problem with salt containing compounds Compatible with column switching from C18 column to chiral column Scale-up is easy due to the volatility of mobile phase components Complementary to hexane/ethanol separations on polysaccharide CSPs For LC/MS, can use ammonium trifluoroacetate, acetate or formate (0.001 to 0.5% w/v in methanol) Applicable to the use of gradients for method development Normal phase vs Polar Ionic? Phase CHIROBIOTIC columns appear to be