In order to ensure the long-term reliability and safety of power transformers, it is important to continuously monitor the characteristics of insulating oil, which not only helps in understanding its behavior over time but also ensures the safety of the equipment. The current study analyzes in-service insulating oil with the aim of relating deterioration and changes in the oil with service aging. Insulating oil samples were collected from three power transformers, with a voltage level of 220 kV and 132 kV, installed at a 220 kV substation. Electrical and chemical characteristics were obtained, and the impact of service aging and the relationships among load variation, oil, and winding temperatures with the characteristics were evaluated. Variations in the dielectric dissipation factor and breakdown voltage with service aging were recorded for all transformers, while the moisture content increased with each service year. Among the concentrations of gases present in the insulating oil, carbon monoxide, oxygen, and nitrogen concentrations increased after each service year. The impact of load variation on the breakdown voltage of the 132 kV transformer oil was more prominent than for the 220 kV transformers. The analysis of gas ratios and moisture content identified the degradation of cellulose insulation in all transformers, which was due to the presence of electrical faults.