Based on parsed colchicine biosynthetic pathway in Colchicum autumnale L.(C.autumnale), the intermediate metabolites of colchicine biosynthetic pathway (IMCBP) were analyzed by HPLC–Q/TOF MS in Hemerocallis citrina Baroni (H. citrina ) and C.autumnale samples. The result showed that IMCBP was found in C.autumnale but not detected in H. citrine samples. Demecolcine (Cp5)，deacetylcolchicine(Cp6) and colchicine(Cp7)were confirmed by authentic standard substances. The results support that there are no colchicine biosynthetic pathway and colchicine in H. citrina.