The invention relates to a method for quick and non-destructive determination of content of amylase in corn single grains and application thereof. The method mainly comprises a step (1) of collecting corn single grain materials; a step (2) of collecting near infrared spectra of a sample; a step (3) of using a conventional chemical method to determine the content of amylase of the sample; a step (4) of preprocessing the obtained near infrared spectra and eliminating interference factors; a step (5) of establishing a calibration model between a chemical value of the content of amylase in the corn single grains and the near infrared spectra, and checking the model; a step (6) of externally verifying the model; and a step (7) of collecting near infrared spectra of a to-be-measured sample, and using the established near infrared model to perform quick and non-destructive determination of the content of amylase in the to-be-measured sample.