A new approach for similarity analysis of chemical profiles of Scrophularia buergeriana was developed. The roots of S. buergeriana collected in Andong, Korea were analyzed using HPLC-DAD and the peak areas of the chromatograms were used to construct the chemical profiles of S. buergeriana. The analytical conditions were validated for specificity, precision and repeatability. The standard chemical profiles were established from the chromatograms of nine standard analytical extracts from three batches. The correlation coefficients of peak areas of characteristic compounds between the chromatograms of the standard and test samples were calculated for quantitative expression of the chemical profiles. The results showed that the HPLC chemical profiles of the samples of same geographic origin were relatively consistent. This new approach could be applied to the quality assessment of herbal medicines with complex chemical compositions.