The invention discloses a method and a kit for rapidly detecting homovanillic acid, vanillylmandelic acid, 5-hydroxyindoleacetic acid and creatinine in human urine. The method comprises the following steps of 1) preparing a sample dilution working solution, 2) under a dark condition, taking a urine sample, adding EDTA and Na2S2O3 into the urine sample, then adding acetic acid for acidification, and uniformly mixing to obtain a to-be-detected urine sample, and (3) mixing a to-be-detected urine sample, a calibration product, a urine quality control product and the sample diluent working solution, sealing a film, vorticing, centrifuging, taking supernate, and detecting by adopting liquid chromatography-mass spectrometry. When a urine sample prepared from random urine is detected, homovanillic acid, vanillylmandelic acid, 5-hydroxyindoleacetic acid and creatinine can be quantified at a time only by 10 microliters of urine sample, the pretreatment method is simple and rapid, the operation time is saved, neuroblastoma and carcinoid in a human body can be effectively screened and diagnosed according to the indexes, and the pheochromocytoma can be screened in an auxiliary manner.