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Fried pepper oil (FPO) is a widely used flavoring oil in China, but it suffers from complex chemical reactions that give an unpleasant odor. The aim of this paper is to study the main causes of off-flavor in FPO, as well as identify key off-flavor compounds in FPO. Fifty-eight volatiles were identified using HS-SPME–GC–MS (headspace solid-phase microextraction–gas chromatography–mass spectrometry) during storage. Fifteen off-flavor compounds were initially targeted by odor activity values (OAVs) calculations and partial least squares regression (PLSR) analysis. Eventually, five key off-flavor compounds were identified by recombination experiments and omission test, which were hexanal, (E) - 2-octenal, terpinyl acetate, α -terpineol, and hexanoic acid. The off-flavors could derive from the oxidative degradation of long-chain fatty acids and the acid-catalyzed rearrangement of terpenes. The results of the study provide a theoretical basis for the regulation of off-flavor in FPO and a practical guide for the industrial production of FPO. [Display omitted] • Five odorants with off-flavor have been identified as markers of flavor damage. • Off-flavor of FPO was most related to aldehydes and alcohols. • Oxidative cleavage of long-chain fatty acids could be a main source of off-flavor. • Terpene degradation could be lead to flavor attenuation. [ABSTRACT FROM AUTHOR]