Abstract: 3-pyridinemethanolwas extracted from Anoectochilus roxburghiiby the ultrasonic-microwave assisted extraction(UMAE). After pretreatment using solid phase extraction(SPE) technology, 3-pyridinemethanolwas identified and determined by high performance liquid chromatography-electrospray ion traps mass spectrometry. The chromatographic separation was achieved on an Agilent TC-C₁₈column（5 μm，4.6×250 mm）using methanol -0.02 mol/L ammonium acetate（5∶95,V/V）as the mobile phase at a flow rate of 1.0 mL/min, and 260 nm as the detection wavelength. The analyte showed good linear regression relationship between peak area and concentrations in the range of 1~10 μg/mL (r=0.9996).The recovery rate was 92.0%~96.2% with an RSD of 2.36%. The proposed method is simple, accurate and rapid.