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Development of a generic flow-injection analysis method for compounds with a secondary amine or amide function, using an experimental design approach: Part II. Selection and evaluation of the chemical reaction parameters
Authors:C VanneckeMS Bloomfield  Y Vander HeydenDL Massart
Institution:a ChemoAc, Vrije Universiteit Brussel, Pharmaceutical Institute, Laarbeeklaan 103, 1090 Brussels, Belgium
b GlaxoSmithKline, Consumer Healthcare, St. Georges Avenue, Weybridge, Surrey KT13 0DE, UK
Abstract:A second part in the development of a generic flow injection analysis (FIA) method to determine compounds with a secondary amine or amide in their structure is described. This part consists in the selection and evaluation of the chemical reaction conditions. Sodium hypochlorite first converts the secondary amine or the amide to a primary amine. The latter reacts with o-phthalaldehyde (OPA) and a thiol (N-acetylcysteine (NAC)) to form a derivative which can be measured fluorimetrically. To investigate the influence of the different chemical reaction parameters on the peak height for a set of 31 pharmaceutical compounds, a quarter-fraction factorial design for six factors at two levels (26-2-resolution IV, 16 experiments) was executed. Effects on the responses were calculated for each compound. Parallel coordinate geometry (PCG) plots and principal component analysis (PCA) were also applied on the measured responses as aids in the interpretation of the results.
Keywords:Secondary amines  Generic FIA-system  Fractional factorial design  Parallel coordinate geometry  Principal component analysis
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