Abstract
HPLC analysis for organic acids in tobacco was optimized with the aid of statistical experimental design, a central composite face-centered design. In the design, only thirteen HPLC analyses were needed for identifying two optimal separation parameters. A Bio-Rad Aminex HPX-87H column was used for the analyses. An optimal separation for seven acids in tobacco was found at a temperature of 57°C and a mobile phase of 0.032 N sulfuric acid solution, or at a temperature of 70°C and a mobile phase of 0.024 N sulfuric acid solution, with a flow rate of 0.6 ml min −1.
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