Fourier transform infrared spectra (FTIR) of royal jelly (RJ) stored at different temperature and after different storage periods were measured, a series of correlation analysis among the spectra were carried out by using the spectra of new-harvested RJ as a standard. The results showed that the correlationcoefficient of amideband and the relative intensity ratios of I₁₆₄₇/I₁₅₄₁, I₁₆₄₇/I₁₄₀₉,I₁₆₄₇/I₁₂₄₇and I₁₆₄₇/I₁₀₅₄of RJ samples’spectra decreased with extension of storage time and temperature, and existed good linear correlations with the storage time, with the order of their change extent being28℃>16℃>4℃>-18℃.According to the spectra change laws and practical experiences of RJ storage, the correlation coefficient of amide band Ⅰand four relative intensity ratios I₁₆₄₇/I₁₅₄₁, I₁₆₄₇/I₁₄₀₉, I₁₆₄₇/I₁₂₄₇and I₁₆₄₇/I₁₀₅₄ were selected as assessment indexes of RJ freshness. The threshold value of correlation coefficient was set to be 0.9100, and the threshold values of the four relativeintensity ratios were set to be 1.744, 2.430, 3.345 and 1.412 respectively. Once one or more indexes are lower than the corre-sponding threshold values, the RJ sample can be considered as a stale sample. So, FTIR spectroscopy combined with several data-processing methods would be an effective method for overall assessing the freshness of RJ.