Non‐Targeted and Targeted Analyses Using Gas Chromatography–Mass Spectrometry for Detecting Fungal Spoilage in Red Pepper Powder

Red pepper powder (RPP), derived from Capsicum annuum L., is vulnerable to adulteration with fungal-spoiled RPP for profit. This study aimed to detect fungal spoilage in RPP by integrating non-targeted analysis (NTA) and targeted analysis (TA) of non-polar metabolites using gas chromatography-mass spectrometry. NTA revealed significant differences in free fatty acid profiles between unspoiled and fungal-spoiled RPP (p < 0.05), specifically due to Alternaria alternata and Colletotrichum acutatum infections. Partial least squares discriminant analysis distinguished the groups and identified palmitic, linoleic, and oleic acids as key markers. TA confirmed significantly elevated levels of these fatty acids in spoiled samples (p < 0.05). A blind test demonstrated spoilage detection at a minimum of 2.5 w/w%, even when mixed with unspoiled samples. These findings establish a reliable approach for identifying fungal spoilage in RPP, offering valuable insights for food safety monitoring and quality control. PRACTICAL APPLICATIONS: This approach offers a reliable tool for food manufacturers, quality control laboratories, and regulatory agencies to verify the authenticity and safety of RPP products. By enabling early and sensitive detection of fungal spoilage, it helps prevent economic fraud and supports consumer health protection. Adoption of this method could strengthen quality assurance protocols and guide regulatory standards in the RPP industry.