The invention belongs to the fields of analytical chemistry and clinical medicine, and discloses a urine fatty acid metabolite marker related to idiopathic male infertility, as well as a detection method and application thereof. The marker is urine fatty acid metabolites, namely, tetradecanedioic acid and/or hexadecanedioic acid, and is detected by adopting a UPLC-Q exactive MS method; the marker can be used for auxiliary diagnosis and monitoring of the idiopathic male infertility, has high sensitivity and specificity, and has a clinical popularization value.