The invention relates to the technical field of serum phospholipid detection and particularly relates to a serum phospholipid detecting reagent. A reagent R1 comprises a buffer solution, phospholipase D, DAOS, ascorbic acid oxidase, bilirubin oxidase, polyethylene glycol 6000, cane sugar, xanthan gum, mannitol, trehalose, BSA, glycerol propoxylate-block-ethoxylate (Pluranic L64) and an aseptic. A reagent R2 comprises the buffer solution, 4-aminoantipyrine, peroxidase, choline oxidase, the polyethylene glycol 6000, the cane sugar, the xanthan gum, the mannitol, the trehalose, the BSA, the glycerol propoxylate-block-ethoxylate (Pluranic L64) and the aseptic. The HEPES buffer solutions and a novel Trinder reaction chromogen DAOS are adopted. A plurality of stabilizers are added to obviously improve stability of the detecting reagent. The bilirubin oxidase and the ascorbic acid oxidase are added, thus effectively avoiding interference caused by bilirubin and ascorbic acid and greatly improving interference-resisting capability of the detecting reagent. In addition, addition of the glycerol propoxylate-block-ethoxylate (Pluranic L64) which is an optional nonionic surfactant prevents reaction system turbidity, enhances substrate stability and improves the interference-resisting capability of the detecting reagent.