The gaseous hormone ethylene regulates different processes in plant life. Ethylene is generally considered as a stress hormone that is stimulated by biotic and abiotic stress. To understand its role in different arrays of plant life, in-vivo quantification of ethylene is essential to understand the physiological aspects of plant metabolism. Several techniques are employed for its accurate estimation; one such popular technique is gas chromatography. Gas chromatography is commonly employed for the estimation of different types of volatile compounds. The quantification of ethylene by gas chromatography is reliable and efficient as a large number of samples can be estimated simultaneously. The measurement of ethylene is accomplished by utilizing a standard curve that is prepared from certified ethylene gas used as the standard. Here, we describe a gas chromatography–flame ionization detection (GC-FID)-based method for the quantification of ethylene from live plants.