The invention relates to a preparation method of a compound amino acid injection 18AA. The method includes the following steps that firstly, infusion bottles are washed and then conveyed to a filling room; secondly, arginine is stirred to be dissolved, and then valine, phenylalanine, alanine, aspartic acid, proline, serine, glutamic acid, glycine, tyrosine, isoleucine, leucine, methionine, threonine and histidine are added in sequence and stirred to be dissolved; thirdly, the solution is cooled to 30-50 DEG C, lysine acetate, tryptophan, cysteine and a sodium bisulfite solution are added, and after stirring is carried out for dissolution, water for injection is supplemented to a full dose; fourthly, after capping and sealing are carried out, sterilization is carried out. In this way, full dissolution of the components is promoted, and the light transmittance of the compound amino acid injection 18AA is guaranteed; besides, the property of the compound amino acid injection is stable, long-time storage is easy, and absorption is promoted.