The current research investigates the spray behavior of lemon peel oil (LPO) and butanol in a controlled environment under various engine-like conditions. The liquid spray morphology of both fuel blends is captured using a standard Mie scattering technique, and the liquid spray penetration length is compared to a baseline fuel isooctane. In order to simulate and create engine-like conditions, these experiments are carried out in a constant volume chamber under various pressure and temperature conditions. Furthermore, the combustion quality of binary and ternary blends is studied using an optical gasoline direct injection engine at three different injection timings. According to the constant volume spray study, isooctane has the shortest penetration. Because of its higher boiling point, LPO has a longer liquid spray penetration length. Despite its lower boiling point, butanol penetrates better than isooctane. The temperature was also discovered to influence liquid spray tip penetration length more than pressure significantly. In-cylinder combustion imaging results also revealed that injection timing significantly impacts combustion. Although butanol improves combustion, LPO-dominant blends demonstrated more diffusion burning due to poor evaporation characteristics. The blends prepared for the study were similar to gasoline in combustion conditions. It was discovered that these blends ran optimally without requiring any modifications to existing engines, even though late injection is recommended to improve combustion quality and peak performance.