Gas chromatography/quadrupole mass spectrometry (GC/qMS), gas chromatography/Fourier transform infrared spectrometry (GC/FTIR) and gas chromatography/orthogonal acceleration time-of-flight mass spectrometry (GC/oaTOFMS) were applied to analyze the volatile oil from the stem of Acanthopanax senticosus (Rupr. et Maxim.) Harms. Based on the spectra search function of GC/qMS with the aid of the discriminability of the geometrical isomer by GC/FTIR and the ability to determine the accurate mass charge ratio (m/z) by GC/oaTOFMS, 68 GC eluants were identified successfully. Compared with the results obtained by GC/qMS only, the analytical results obtained by these hyphenated methods of gas chromatography are more reliable.