Abstract: This research aims to establish a RP-HPLC method for analysis of bioactive constituents such as tryptanthrin,2(3H)-benzoxazolinone,indirubin,indigo,acteoside,martynoside of different parts of Baphicacanthus cusia (Nees) Bremek in different habitats.The chromatographic separation was performed on ZORBAX SB-C₁₈ column（4.6×250 mm,5 μm）with gradient elution consisting of acetonitrile and water at a flow rate 1 mL/min.The UV detection wavelength was 280 nm and the column temperature was set at 30 ℃.The six kinds of compounds in B.cusia separated well,and the linear relation was obtained (r ≥ 0.999 1).The average recoveries were 100.25%,99.93%,100.46%,99.94%,9992% and 100.02%,with RSD 1.46,0.13,0.52,0.09,0.16 and 0.40.The method is simple,quick,and good reproducibility,which can provide the basis for the rational use of the active parts of B.cusia.

Key words: content determination analysis, Baphicacanthus cusia (Nees) Bremek, RP-HPLC, different habitats, different parts