The invention belongs to the technical field of electrochemical sensors, and in particular relates to an Au(111)-like nano-particle modified non-enzyme glucose sensor electrode, and a preparation method and application of the same. The preparation method comprises the following steps of: firstly, wiping a conductive substrate with alcohol, then, respectively performing ultrasonic washing with acetone, isopropyl alcohol and ultrapure water, adding the conductive substrate in the mixed liquor of deionized water, ammonia water and hydrogen peroxide, heating until boiling, finally, cleaning the conductive substrate with the ultrapure water and blowing the conductive substrate to dry with nitrogen; in the electrolyte solution composed of sodium perchlorate and chloroauric acid solution, firstly, performing electrolytic deposition on the conductive substrate by means of chronoamperometry, then, further performing electrolytic deposition on the conductive substrate by means of cyclic voltammetry, and cleaning the conductive substrate with the deionized water. The non-enzyme glucose sensor electrode is very sensitive to the concentration variation of the glucose and presents better linear relation, moreover, the non-enzyme glucose sensor electrode has better selectivity to the glucose, only needs 1.5s response time to the glucose and can achieve the detection limit of 5mu moL/L.