OBJECTIVE： To establish a method for simultaneous determination of matrine， linarin， berberine hydrochloride and osthole in Shenju lotion. METHODS： HPLC was performed on the column of Dionex C18 with mobile phase of acetonitrle - 0.1％ triethylamine （pH was adjusted to 7.2 with phosphoric） （gradient elution） at a flow rate of 1.0 ml/min， detection wavelength was 210 nm， column temperature was 30 ℃， and injection volume was 20 μl. RESULTS： The linear range was 1.400-8.400 μg for matrine （r＝0.999 4）， 0.152-0.912 μg for linarin （r＝0.999 6）， 0.248-1.488 μg for berberine hydrochloride （r＝0.999 9） and 0.128-0.768 μg for osthole （r＝0.999 4）； limit of quantitation was 84.85， 3.15， 15.03， 7.76 ng， limit of detection was 24.13， 0.89， 4.27， 2.20 ng； RSDs of precision， stability and reproducibility tests were lower than 2％； recoveries were 96.29％-99.06％ （RSD＝1.10％，n＝9）， 96.49％-98.27％ （RSD＝0.69％，n＝9）， 99.26％-101.13％（RSD＝0.62％，n＝9） and 96.27％-98.64％ （RSD＝0.89％，n＝9）. CONCLUSIONS： The method is simple， accurate and reproducible， and can be used for the simultaneous determination of matrine， linarin， berberine hydrochloride and osthole in Shenju lotion.