The study of fruit-ripening mechanism is vital as it plays a key role in the maintenance of fruit quality. Use of various xenobiotics for quick ripening has been shown to impact the quality of fruit, which in turn affect human health. In the present study, we made an attempt to understand the metabolic perturbations in Carica papaya Linn. (papaya), which has been ripened either by the ripening practice (room temperature process as control) and/or ripening agents (calcium carbide and ethylene) using gas chromatography-mass spectrometry (GC-MS) based metabolomics. The partial least squares-discriminant analysis has revealed significant alternations in 13 metabolites mainly sugars, amino acids, fatty acids, and organic acids as well as disturbances in five metabolic pathways due to different ripening practice/agents. The individual comparison of calcium carbide with control and ethylene with control has found 13 and 11 metabolites, respectively, which are common to the PLS-DA of three ripening groups. The GC-MS-based metabolomics has been able to predict the metabolic perturbations in papaya resulting from the ripening practice/agents. The findings from the present analysis has a wide application in food quality and will help to address safety concerns. K E Y W O R D S calcium carbide, ethylene, metabolomics, Papaya (Carica papaya Linn.), ripening agents 1 INTRODUCTION At present, various ripening agents are being used in the process of fruit ripening. The variations in the ripening process of fruit not only affect the fruit quality but also associated with several health concerns such as hormonal imbalance, cerebral edema, reduced immunity, hypoxia, hematological and biochemical parameter changes, and increased susceptibility to cancers. 1-4 Different ripening practices/agents are used mainly for fruits like mango, papaya, banana, plums, etc. 5 Ripening practices are used mainly to improve the appearance by maintaining uniformity in ripening and delivery of fruit for human consumption. However, these accelerated ripening processes may alter the fruit metabolism and fruit quality due to Abbreviations: AUROC, area under receiver operating characteristics; MetPA, metabolic pathway analysis; PCA, principal component analysis; PLS-DA, partial least squares-discriminant analysis; VIP, variable importance in projection This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.