Early experiments:

One of the first known experiments on the relationship between combustion and air was conducted by the second century BCE Greek writer on mechanics, Philo of Byzantium. In his work Pneumatica, Philo observed that inverting a vessel over a burning candle and surrounding the vessel's neck with water resulted in some water rising into the neck. Philo incorrectly surmised that parts of the air in the vessel were converted into the classical element fire and thus were able to escape through pores in the glass. Many centuries later Leonardo da Vinci built on Philo's work by observing that a portion of air is consumed during combustion and respiration.

In the late 17th century, Robert Boyle proved that air is necessary for combustion. English chemist John Mayow refined this work by showing that fire requires only a part of air that he called spiritus nitroaereus or just nitroaereus. In one experiment he found that placing either a mouse or a lit candle in a closed container over water caused the water to rise and replace one-fourteenth of the air's volume before extinguishing the subjects. From this he surmised that nitroaereus is consumed in both respiration and combustion.