The invention discloses half antigen and antigen of tenuazonic acid and a preparation method and application thereof. The antigen can serve as a compound of the tenuazonic acid half antigen and is provided with the structure as the formula I. The antigen of the tenuazonic acid is obtained by mixing TeAH and carrier protein, dissolving carrier protein into phosphate buffer solution (PBS), droppingglutaraldehyde solution, conducting stirring and reaction and then conducting dialysis through physiological saline, or the ntigen of the tenuazonic acid is obtained by dissolving TeAHGA into N, N-dimethylformamide, adding bicyclocaproyl carbodiimide and N-hydroxy succinimide into the mixture, stirring the liquid, conducting centrifugation to obtain supernatant as A liquid, dissolving carrier protein in the PBS, stirring and dissolving the PBS to obtain B liquid, dropping the A liquid into the B liquid for reaction and conducting dialysis through physiological saline. The antiserum valence obtained by antigen immune animals can reach 1: 3.2*104, the linear range of TeAH ranges from 0.130ng/mL to 8.789ng/mL(IC20-IC80), the half inhibition concentration is 1.095ng/mL, and the produced antigen is high in specificity, high in sensitivity and high in accuracy. Formula I.