Australia is in an extinction crisis with many of our iconic marsupial species threatened. Assisted reproductive technologies should be a vital part of conservation efforts, yet the fundamental method of in vitro fertilisation (IVF) has not been achieved for marsupials. The reason for this is a lack of understanding surrounding the signalling environment within the oviduct at the time sperm meets egg. Marsupial sperm are highly regulated, with research showing sperm will bind to the oocyte only when cultured in media that is conditioned with oviductal epithelial cells. Investigating this further, we used liquid chromatography mass spectrometry (LC-MS) to obtain both the metabolome and proteome for the marsupial oviduct, using the Fat-tailed dunnart as a model. Samples were taken when dunnarts were in oestrous and compared to non-oestrous samples. Interestingly, results suggest fructose is a candidate for the dunnart seminal sugar, with n-acetyl glucosamine (NAG) only lowly detected. Carbohydrate levels reflect what has been observed in previous marsupial embryo work. Serum from each animal was also subjected to LC-MS to obtain a profile for sex steroid hormones, including estradiol and progesterone. We have shown that there is a difference in the oviductal environment during oestrous and plan to use these results to synthesise a media that will better support marsupial sperm capacitation and oocyte binding. The high levels of hypotaurine and L-glutamine during oestrous, as well as the detection of all essential and non-essential amino acids but cysteine, suggest commercial IVF media may be a good starting point, to which additional factors may be added. This is the first study to extensively characterise the marsupial oviductal environment and will be a fundamental resource for the field of marsupial IVF moving forward.