Osterici Radix, the dried roots of Ostericum koreanum Maximowicz, has long been used to treat cold, headache and arthralgia in Chinese medicine. Notopterygii Rhizoma et Radix, the dried rhizomes and roots of Notopterygium incisum Ting et H.T. Chang or Notopterygium forbesii Boiss (family: Umbelliferae), are well-known traditional Chinese medicines. These all herbal medicines were named kangwhoal in China, Japan and Korea. However no discrimination methods for the origin and no simultaneous analysis methods for bioactive components are currently available. We developed simultaneous analysis of six biomarkers including nodakenin (1), oxypeucedanin (2), bisabolangelone (3), notopterol (4), imperatorin (5), and isoimperatorin (6) using HPLC method. HPLC chromatographic separation was conducted with C18 column and mobile phase of acetonitrile (A) - water (B) (gradient, (A) 35%-65% (30 min)). The analytical condition was validated with specificity, selectivity, linearity (correlation coefficients of all bio marker were more than 0.999.), recovery (94.9-106.8%), precision and accuracy (intra and inter RSD < 2.99%). In addition, principal component analysis and cluster analysis was performed on analytical data of 38 commercial kangwhoal samples from different places. We confirmed to classify kangwhoal origins and this study was applied to quality evaluation and chemotaxonomy of medicinal herbs.