The invention discloses a preparation method of an aluminum-containing adjuvant hepatitis B vaccine, belonging to the biotechnology field. The preparation method is characterized in that an aluminum adjuvant Al(OH)3 is produced by an on-line reaction, i.e. after a phosphate buffer solution (PBS), a KAl(SO4)2 solution and a hepatitis B surface antigen stock solution are mixed, an NaOH solution is added to the mixed solution, an Al(OH)3 adjuvant is continuously produced, and simultaneously, hepatitis B surface antigens are continuously coated and adsorbed; and the process is called 'in-situ adsorption'. In the invention, the Al(OH)3 adjuvant is produced by an in-situ reaction to greatly improve the adsorption rate of the hepatitis B surface antigens, thereby improving the immunogenicity of the antigens, being capable of more effectively causing organisms to generate an immune response, and producing more protective antibodies. The practice proves that the aluminum adjuvant hepatitis B vaccine produced by the method disclosed by the invention has the advantages of small inoculation amount, few adverse responses, high antibody positive conversion rate and the like, and can induce high-level antibody response after being immunized. Simultaneously, the processing steps are also simplified, and the production cost is greatly lowered.