This study was conducted to investigate the potential of hyperspectral imaging technique (900-1700 nm) for nondestructive determination of inosinic acid (IMP) in chicken. Hyperspectral images of chicken flesh samples were acquired, and their mean spectra within the images were extracted. The quantitative relationship between the mean spectra and reference IMP value was fitted by partial least squares (PLS) regression algorithm. A PLS model (MAS-PLS) built with moving average smoothing (MAS) spectra showed better performance in predicting IMP content, leading to correlation coefficients (R P ) of 0.951, root mean square error (RMSEP) of 0.046 mg/g, and residual predictive deviation (RPD) of 3.152. Regression coefficient (RC), successive projections algorithm (SPA), stepwise, competitive adaptive reweighted sampling (CARS), and uninformative variable elimination (UVE) were used to select the optimal wavelengths to optimize the MAS-PLS model.