The invention provides moxifloxacin hydrochloride glucose injection and a preparation method and use thereof. The method for preparing the injection comprises the following steps of: adding water for injection accounting for 20 to 98 percent of the batch volume into an ingredient tank, and adding glucose, a metal complexing agent and the moxifloxacin hydrochloride in a ratio; after stirring to fully dissolve the components, regulating the pH value to between 4.0 and 4.5 by using 1mol/L hydrochloric acid solution or 1mol/L sodium hydroxide, adding medicinal carbon accounting for 0.05 percent (W/V) of the total volume, uniformly stirring, maintaining the temperature of between 70 and 80 DEG C for 20 minutes, and performing circular filtering for over 20 minutes; replenishing the water for injection to the batch scale, stirring for 5 to 10 minutes, and detecting the pH value of the prepared solution (controlling to between 4.0 and 4.5); after determining that no residual water is present in an elevated tank and a pipeline, opening a valve of the elevated tank, and sampling liquid medicament at a self-circulation pipeline sampling port after the liquid medicament circulates for 20 minutes through a filter element and the elevated tank; detecting according to the intermediate quality standard, requiring that the content of the moxifloxacin hydrochloride is between 1.52 and 1.68 mg/ml, the glucose content is between 47.5 and 52.5 mg/ml, and the pH value is between 4.0 and 4.5; after the intermediate is detected to be qualified, beginning to fill; and conveying the filled semi-finished products into a sterilizing cabinet for sterilization, wherein the sterilization condition is to sterilize for 8 to 30 minutes at 121 DEG C through thermal pressure steam.