Abstract: A determination method of anthocyanins in Xuzishu NO.6 by high performance liquid chromatography (HPLC) was established. 13 kinds of components with R>1 and over 1000 of USP theoretical plate number have been identified by this method which the resolution were 1.5% phosphoric acid and 35% acetonitrile (1% phosphoric acid), and the RSD was 0.42%~8.16%.The samples from XZ₁₅ (34°28'N 117°30'E), SN₁₅ (33°97'N 117°98'E), and YC₁₅ (33°00'N 120°82'E) were analyzed. The results revealed that the main component was cyanidin 3-caffeoyl-p-hydroxybenzoyl-sophoroside-5-glucoside.Whereas XZ₁₅ sample contained 13 kinds of components with highest content of total anthocyanins, YC₁₅ sample only contained 8 kinds of components but the highest main component contents. And SN₁₅ sample contained 13 kinds of components but low-content anthocyanins. The results of cluster analysis also showed that the anthocyanin composition differences among the 3 samples were relative to their origins, whereas YC₁₅ showed more differences because of its components reduce and main component contents increases. The method was well separated and proved to be sensitive and accurate. The results revealed that the anthocyanin composition was mostly affected by their origins. Therefore, this method is applied to geographical classification of Xuzishu No.6.