Abstract: In this paper, three methods for determination of jatrorrhizine are reported and compared with each other by spectrophotometry, TLC-scanning and RP-HPLC. For the Berberis julianae Schneid cell cultural extracts, jatrorrhizine can be determined directly by spectrophotometry at wavelength 430 nm. The sample can also be determined by reflective absorption scanning at wavelength 450nm after developed on TLC plate, or it can be injected into HPLC on ODS revers phase column and adopted phosphoric acid-acetonitrile system as mobile phase, detection at 346nm. The excellect results obtained with any one of these methods were in good accord with each other.