Sperm capacitation, fertilization and embryo development take place in the oviduct during the periovulatory period of the estrous cycle. Phospholipids are crucial metabolites for sperm capacitation and early embryo development. The aim of this study was to monitor the abundance of phospholipids in the bovine oviductal fluid (OF) according to the stage of the estrous cycle and the side relative to ovulation. Pairs of bovine oviducts were collected in a slaughterhouse and classified into four stages of the estrous cycle: post-ovulatory (Post-ov), mid-luteal (Mid-lut), late-luteal (Late-lut) and pre-ovulatory (Pre-ov) phases (n = 17 cows/stage). Cell-free OF from oviducts ipsilateral and contralateral to the site of ovulation were analyzed using MALDI-TOF mass spectrometry. Lipid identification was achieved by high resolution mass spectrometry. A total of 274 lipid masses were detected in the mass range of 400-1000 Da, corresponding mostly to phosphatidylcholines (PC), lysoPC, phosphatidylethanolamine (PE), lysoPE and sphingomyelins (SM). Ipsilateral and contralateral OF did not differ in their lipid profiles at any stage of the cycle. However, 127 and 96 masses were differentially abundant between stages in ipsilateral and contralateral OF, respectively. Highest differences in lipid profiles were observed in the Pre-ov vs. Mid-lut and Pre-ov vs. Late-lut comparisons in both sides relative to ovulation. Differential abundance of specific molecules of PC, PE, SM and l-carnitine were observed at Pre-ov and Post-ov compared with the luteal phase. This work proposes new candidates potentially able to regulate sperm capacitation and early embryo development.