Abstract: Objective To identify the structure of the alkaloid extract of Gelsemium from east Guangdong province of China. Methods Fourier transform infrared (FTIR) absorption spectrometry, hydrogen and carbon spectra of nuclear magnetic resonance (NMR) and distortionless enhancement by polarization transfer (DEPT) analysis were used for the structural identification of the alkaloid exstract of Gelsemium. Results The frequency, intensity and shape of the extract’s characteristic peaks in infrared absorption spectra (4 000.0-400.0 cm^-1) and ¹H NMR, ¹³C NMR were recognized and compared. The molecular structure of the sample was consistent with the theoretically derived model. Conclusion The extract is structurally identical to koumine, which may provide evidence for its safe clinical application and establishment of Chinese medicine fingerprint database of Gelsemium.