Atmospheric CO2 column concentration (column-averaged dry air mole fractions of atmospheric carbon dioxide) data obtained by ground-based hyperspectral observation is an important source of data for the verification and improvement of the results of CO2 retrieval based on satellite hyperspectral observation. However, few studies have been conducted on atmospheric CO2 column concentration retrieval based on ground-based spectral hyperspectral observation in China. In the present study, we carried out the ground-based hyperspectral observation in Xilingol Grassland, Inner Mongolia of China by using an observation system which is consisted of an optical spectral analyzer, a sun tracker, and some other elements. The atmospheric CO2 column concentration was retrieved using the observed hyperspectral data. The effect of a wavelength shift of the observation spectra and the meteorological parameters on the retrieval precision of the atmospheric CO2 concentration was evaluated and analyzed. The results show that the mean value of atmospheric CO2 concentration was 390.9 μg·mL-1 in the study area during the observing period from July to September. The shift of wavelength in the range between -0.012 and 0.042 nm will generally lead to 1 μg·mL-1 deviation in the CO2 retrievals. This study also revealed that the spectral transmittance was sensitive to meteorological parameters in the wavelength range of 6 357~6 358, 6 360~6 361, and 6 363~6 364 cm-1. By comparing the CO2 retrievals derived from the meteorological parameters observed in synchronous and non-synchronous time, respectively, with the spectral observation, it was showed that the concentration deviation caused by using the non-synchronously observed meteorological parameters is ranged from 0.11 to 4 μg·mL-1. These results can be used as references for the further improvement of retrieving CO2 column concentration based on spectral observation.