Food web research is rapidly expanding through study of natural fractional abundance of ¹⁵N in individual amino acids. This paper overviews the principles of this isotope approach, and from my perspective, reanalyzes applications, and further extends the discussion. It applies kinetic isotope effects that enriches ¹⁵N in certain amino acids associated with the metabolic processes, which was clearly demonstrated by observations of both natural ecosystem and laboratory experiments. In trophic processes ‘trophic amino acids’ such as glutamic acid that significantly enrich ¹⁵N, whereas ‘source amino acids’ such as phenylalanine and methionine show little ¹⁵N enrichment. Through various applications conducted over the years, the principles of the method have shown to operate well and disentangle complex food webs and relevant problems. Applications include food chain length estimate, nitrogen resource assessment, tracking fish migration, and reconstruction of paleodiet. With this approach, considerations of a wide range of classical issues have been reinvigorated, while in the same time, new challenging frontiers are emerging.