The present invention discloses a carbon base long-acting nitrogen fertilizer biochemistry inhibitor preparation method, which concretely comprises: preparing a nitrogen fertilizer biochemistry inhibitor such as a urease inhibitor or a nitrification inhibitor into an aqueous solution with a certain concentration, adding the obtained aqueous solution to a carbonaceous absorption agent, uniformly mixing, drying, grinding and screening with a sieve, wherein the used carbonaceous absorption agent has wide sources, and can be active carbon, biological carbon or black carbon, the inhibitor content in the resulting product is 0.4-33.3%, and the obtained product can be directly applied into the soil in the powder form during a nitrogen-containing fertilizer applying process or before or after applying of a nitrogen-containing fertilizer, and can further be mixed into a nitrogen-containing fertilizer after granulation or be adopted as a coating layer to be coated on the outer portion of a particulate nitrogen-containing fertilizer. According to the present invention, the solid-liquid mixing absorption method is applied to ensure absorb the inhibitor to the surface and the internal of the carbonaceous material to the maximal degree, such that a slow-release effect is excellent, and an inhibition effect on soil enzyme activity is long-lasting. In addition, the used carbonaceous material has the wide sources, the cost is low, and the carbon base long-acting nitrogen fertilizer biochemistry inhibitor can be adopted as a carbon fixation manner to increase soil carbon sequestration and improve a soil environment, and is a veritable green fertilizer additive.