In vitro fertilization (IVF) is one of the most common and effective type of assisted reproductive technology (ART) to treat infertility. The success rate of ART relies heavily on the quality of the embryo’ development. Therefore, analysis of embryo culture media for both research and development as well as quality control is crucial. Q-TOF was popular in identifying unknown compounds because of its high resolution mass and MS/MS spectrum. In this poster, three kind of human embryo culture media were investigated. The energy substrates, amino acids, antioxidant and antibiotic were identified using accurate mass and MS/MS library.