Quantitation of coeluting components via HPLC: Using UV absorption to separate coeluting species

ANYL 140

Maria Tsiouris, maria_tsiouris@merck.com1, Lei Qi1, John J. Donnelly1, Eric S. Ahuja2, and Holly J. Perpall1. (1) Regulatory and Analytical Sciences - Analytical, Merck Manufacturing Division, Merck and Co, Inc, WP82-30, 770 Sumneytown Pike, PO Box 4, West Point, PA 19486-004, (2) Quality Operations Europe, Middle East and Africa, Merck Manufacturing Division, Merck and Co, Inc, Waardereg 39, Haarlem, 2301BN, Netherlands
An HPLC procedure for simultaneous quantitation of a process impurity coeluting with an oxidative degradate is developed without changing the chromatographic separation conditions specified in the filed stability indicating method. The degradate contribution to the coeluted peak area is quantitated by capitalizing on the shift in the UV spectrum of the oxidation product vs. the process impurity. The absorption ratio of the oxidative degradate at its maximum versus the method specified wavelength is determined and a second detection channel, at the absorption maximum of the oxidation product, is added to the existing HPLC method. The contribution of the process impurity is determined through subtraction. This work represents an elegant technique for separation of co-eluting components in cases where changes to the HPLC chromatographic separation conditions are not desired.
 

General Papers
7:00 PM-9:00 PM, Sunday, 10 September 2006 Moscone Center -- Hall D, Poster

Division of Analytical Chemistry

The 232nd ACS National Meeting, San Francisco, CA, September 10-14, 2006