Eleven consecutive N '-n-acylnornicotines from cherry-red tobacco were structurally elucidated and quantitively analyzed using chromatography and mass spectrometry. All of these N '-n-acylnornicotines are first reported in cherry-red tobacco, whereas N '-propionylnornicotine, N '-n-valerylnornicotine, N '-n-nonanoylnornicotine and N '-n-undecanoylnornicotine are first reported in natural products. The concentration distribution of the identified N '-n-acylnornicotines was studied and it was found that N '-n-octanoylnornicotine and N '-n-hexanoylnornicotine showed the highest concentration, accounting for 94% of the detected N '-n-acylnornicotines. The cherry-red color density of the related tobacco leaves was found to be positively correlated with the concentration of the N '-n-acylnornicotines, whereas the ultraviolet-visible spectra of the N '-n-acylnornicotines showed no absorption larger than 300 nm, indicating the discovered compounds are not responsible for the cherry-red color appearance.