The invention provided a method for preparing high-purity Mezlocillin sodium, and a method for preparing the Mezlocillin sodium powder injection. The method includes (1) dissolving Mezlocillin sodiumcrude product in purified water, and adjusting the pH value to less than 7 with the pH regulator; (2) extracting with organic solvent, separating the organic phase, drying with desiccant, and vacuum recovering to dry to obtain the product; (3) dissolving the product with eluting agent of mixed solution of hydrocarbon solvent and chlorine-containing solvent, eluting with alumina as filler of medium-pressure chromatography column, and collecting the fraction in a sectional manner; (4) mixing fractions with the Mezlocillin acid content of no smaller than 80%, vacuum concentrating, dissolving into ethanol, and basifying with base to separate precipitate; and (5) re-crystallizing the precipitate with ethanol, and freeze drying to obtain the high-purity Mezlocillin sodium.