Abstract: OBJECTIVE To establish characteristic HPLC chromatograms for Jubei mixture, and to simultaneous determine its six components by HPLC-ELSD. METHODS An HPLC specific chromatogram of the Jubei mixture was established. The chromatographic column was Agilent ZORBAX-C₁₈(250 mm×4.6 mm, 5 μm), gradient elution was conducted with acetonitrile(A)-0.1% phosphoric acid aqueous solution(B) as the mobile phase, the flow rate was 1.0 mL·min^-1, and the column temperature was 30℃, the detection wavelength was 250 nm. An HPLC-ELSD method was established and the chromatography was accomplished on a Waters T3 C₁₈(250 mm×4.6 mm, 5 μm) column, the mobile phase consisted of acetonitrile(A)-1% acetic acid aqueous solution(B) in a gradient mode, the flow rate was 1.0 mL·min^-1, the column temperature was set at 30℃. RESULTS The characteristic chromatograms of the HPLC with baicalin as the reference peak was established, and eight common peaks were defined. The linear ranges of amygdalin, liquiritin, peimine, peiminine, platycodin D and ophiopogonin D were good within their own ranges(r ≥ 0.999). The average recovery were 99.1%, 98.6%, 100.9%, 97.7%, 98.9% and 98.0%, RSD were 0.4%, 1.0%, 1.1%, 1.2%, 0.7% and 1.3%, respectively. CONCLUSION The established methods can be used to qualitative identification and quantitative determination of Jubei mixture, can effectively used for evaluation and quality control of Jubei mixture.