The invention relates to a pharmaceutical composition, and in particular relates to a composition containing various microelements, a preparation and a preparation method thereof. The composition comprises ferrous gluconate dihydrate, a zinc gluconate trihydrate compound, copper gluconate, manganese gluconate, sodium fluoride, cobalt gluconate dihydrate, sodium iodide, sodium iodide pentahydrate, ammonium heptamolybdate tetrahydrate, chromic chloride hexahydrate and a medicine auxiliary material. The form of the pharmaceutical composition can be injection or freeze-dried powder injection. According to the invention, organic iron, organic zinc, organic copper, organic manganese and organic cobalt are adopted to replace chloride, so that not only is the property of the composition more stable, but also the adverse reaction is reduced, and the bioavailability of the medicine is increased. Therefore, the preparation provided by the invention is better in safety and more suitable for clinical application.