Natural flavors and fragrances are generally mixtures in a background matrix, with many similar structures. As a result of their physicochemical characteristics and a prevailing sensory response, these compounds are crucial to the food, essential oil, and cosmetic fields. The use of flavor and fragrance substances for enjoyment, religious, or medicinal reasons is as old as mankind. Today, the production of flavors and fragrances has been transformed, evolving from physical and chemical isolation from natural plant resources as in ancient times and still of immense importance today, to chemical synthesis or biological (enzymatic or microbial) processes to generate desired fragrance and flavor compounds. The large number of fragrance and flavor compounds in natural samples with similar structures and physicochemical characteristics, different polarities, and a wide range of volatilities demand the use of highly efficient separation techniques. Developments in analytical technology for volatile compounds are critically relevant, because poor resolution of target volatiles from matrix compounds and the lack of precision in identification that limits structural assessment of compounds in complex matrices raises challenges at the frontier of instrument capabilities. The difficulty is compounded at lower compound abundance. Flavor/fragrance compounds are especially prone to poor specificity in mass spectrometry, arising from their many isomers.