Statistics aided optimization for high-performance liquid chromatographic analysis of organic acids in tobacco

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 degrees C and a mobile phase of 0.032 N sulfuric acid solution, or at a temperature of 70 degrees C and a mobile phase of 0.024 N sulfuric acid solution, with a flow rate of 0.6 ml min(-1).