Real-time assessment of enantiomeric purity via a polarimetric/absorption detector response function.

The development of methodology appropriate for the rapid and precise assessment of enantiomeric purity is a critical need in the life sciences with impact in a number of areas including biomedical research, biotechnology, and pharmaceutical science. Real-time assessment of enantiomeric purity is critical to decisions related to possible product purity and/or the need for, and the type of additional processing. Recently, we have shown that laser-based polarimetric detection, in combination with ultraviolet detection, can be used to assess enantiomeric purity in real-time as an adjunct to the separation process. A mass-independent response function is obtained from the ratio of the normalized polarimetric signal relative to the normalized UV signal. This response ratio will be shown to be equivalent to the enantiomeric excess and independent of concentration and chromatographic resolution. The methodology will be evaluated as a function of injected mass, enantiomeric excess, chromatographic resolution, and peak asymmetry.