Nowadays, the spectroscopic databases used for the modeling of Earth and planetary atmospheres provide only electric-dipole transitions for polyatomic molecules (H2O, CO2, N2O, CH4, O3…). Very recently, electric-quadrupole transitions have been detected in the high sensitivity cavity ring down spectrum (CRDS) of water vapour near 1.3 μm [A. Campargueet al.,Phys. Rev. Res., 2020,2, 023091, D. This discovery paved the way to systematic searches of quadrupole transitions in water vapor and other polyatomic molecules. In the present work, on the basis of high accuracyab initiopredictions, H216O quadrupole lines are detected for the first time in the 5.4 μm and 2.5 μm regions where they are predicted to have their largest intensities (up to 10−26cm per molecule). A total of twelve quadrupole lines are identified in two high sensitivity Fourier transform spectra recorded with a 1064 m path length. Ten lines in the 4030-4150 cm−1region are assigned to theν3band while the lines near 1820 and 1926 cm−1belong to theν2band. The derived line intensities which are largely above the dipole intensity cut-off of the standard spectroscopic databases, agree nicely with the theoretical predictions. We thus conclude that the calculated line list of quadrupole transitions, validated by the present measurements, should be incorporated in the spectroscopic databases.