Abstract: OBJECTIVE To establish a specific HPLC chromatogram of Ganoderma lucidum spore and a method for determination of triolein. METHODS The chromatographic separation was performed on an ACCHROM Unitary-C₁₈ column (250 mm×4.6 mm, 5 μm) with the mobile phase of acetonitrile-isopropanol(51∶49). The flow rate was 1.0 mL·min^-1 at 30 ℃ with the detection by ELSD. RESULTS The results showed that the chromatographic fingerprint was completed with 10 recognizable peaks, of which 8 peaks kaempferol-3-O-β-D (trilinolein, 1,2-linolein-3-olein, 1,2-linolein-3-palmitin, 1,2-olein-3-linolein, 1-palmitin-2-olein-3-linolein, triolein, 1,2-olein-3-palmitin and 1,2-olein-3-stearin) were determined. The linear range of triolein was 0.613 2-18.40 μg(r=0.998 9), the average recovery was 104.1%(RSD=1.8%). CONCLUSION The combination of specific chromatograms and determination of triolein is suitable for use in the quality control of Ganoderma lucidum spore.