Poor temperature management along a cold chain leads to fruit quality deterioration and loss. In order to determine the threshold value of temperature fluctuation in a cold chain, peach fruits were stored in four different virtual cold chains applying different temperature–time scenarios. Core temperature profiling, the physicochemical qualities, and the activities of the peaches’ antioxidant enzymes were monitored during cold storage and shelf life. Abusive temperature management (temperature increased to 20 and 15 °C three times) resulted in a significant increase in a peach’s core temperature to the highest temperature measured: 17.6 °C. The ethylene production rate at the end of the shelf life of peaches under these temperatures was 21.03–28.16% higher than the constant-temperature group and accompanied by significantly lower levels of flesh firmness, titratable acid content, total phenol and flavonoid content, and peroxidase (POD) and catalase (CAT) activities (p< 0.05). The results of a principal component analysis (PCA) and heatmap confirmed the results. Limited temperature increases (10 °C) in a cold chain had little impact on the quality of the peaches, while temperature increases higher than 15 °C three times would negatively affect the quality of the peaches significantly. The temperature of a cold chain needs to be controlled precisely to reduce the loss of peaches.