A coix seed oil comprising 13 triglycerides extracted from coix seeds, formulation and application thereof. Specifically, the coix seed oil comprises the following 11 triglyceride components (calculated by mass percentage): glyceryl trilinoleate (4.87-6.99%), glyceryl 1-oleate-2,3-dilinoleate (13.00-18.69%), glyceryl 1-palmitate-2,3-dilinoleate (5.25-7.54%), glyceryl1,3-dioleate-2-linoleate (13.23-19.02%), glyceryl 1-palmitate-2-linoleate-3-oleate (10.26-14.75%), glyceryl 1,3-dipalmitate-2-linoleate (2.28-3.28%), glyceryl trioleate (14.44-20.76%), glyceryl 1-palmitate-2,3-dioleate (8.06-11.58%), glyceryl 1-oleate-2-linoleate-3-stearate (1.37-1.97%), glyceryl 1,3-dipalmitate-2-oleate (1.52-2.19%), and 1,2-dioleate-3-stearate (1.29-1.86%). The coix seed oil can be used in the preparation of anti-tumour medicine.