Currently, it must be acknowledged that little is known about the quantity and make-up of oligosaccharides (OS) found in breast-fed babies' feces as well as their metabolic fate. In the present work, UPLC-QE-HF-MS was successfully adopted to identify the profiles of human milk oligosaccharides (HMOs) in the breast milk of four mothers and fecal OS in the feces of their breast-fed infant. There were significant variations and differences in both number and composition between HMOs and fecal OS. The early-life gastrointestinal microbiota metabolism may be triggered into the advanced breakdown, synthesis, bioconversion, or redesign of HMOs. The fate of HMOs during passage through the gastrointestinal tract may be profoundly informed by the comparison of OS between breast milk and fecal OS profiles. The characterization of fecal OS could be applied as a valuable tool for monitoring the gastrointestinal fate of HMOs and reflecting infant development at different stages of lactation. Further research on the gastrointestinal bioconversion of HMOs profiles is required, including secretor type and the lactation time of milk, as well as baby feeding.