Abstract: OBJECTIVE To establish a liquid chromatography-tandem mass spectrometry(HPLC-MS/MS) method to assay bornenol-7-O-β-D-apiofuranosyl(1®6)-β-D-glucopyranoside(BAG) and borneol-7-O-β-D-glucopyranoside(BG) in Ophiopogonis Radix and Shenmai injection. METHODS Loganin was used as internal standard. The separation was performed on Agilent Poroshell 120 EC-C₁₈ column(4.6 mm×100 mm, 2.7 μm) with gradient elution of acetonitrile(A)-0.1% formic acid solution(B) at a flow rate of 0.5 mL·min^-1. The column temperature was 30℃. All the samples were determined using positive electrospray ionization with multiple reaction minitoring mode. The ion pairs of BAG, BG and internal standard were m/z 449.5→137.1, 317.1→137.1, 391.2→179.1, respectively. RESULTS The linear ranges of BAG and BG were 0.20-10.32 μg·mL^-1(r=0.995 3, n=5), 0.02-1.51 μg·mL^-1(r=0.998 0, n=6), and the quantification limits were 8.1, 3.5 μg·mL^-1, respectively. Intra-day and inter-day precision and repeatability tests were good(RSD<10%), and BAG and BG were stable within 24 h(RSD<10%). The average recoveries(n=3) were 99.9%(RSD=1.93%) and 99.6%(RSD=3.09%), respectively. CONCLUSION The established method is accurate, simple and reproducible, which can be used in separation and determination of BAG and BG in Ophiopogonis Radix and Shenmai injection.