This study presents an alternative preliminary strategy to identify and quantify serum hFSH glycoforms based on immunopurification assay and mass spectrometry (MS) using parallel reaction monitoring (PRM) analysis. In this study, we provide MS-PRM data acquisition method for hFSH glycopeptides identification with high specificity and their quantification by extracting the chromatographic traces of selected fragments of glycopeptides. Once set up in all its features, the proposed method could be transferred to the clinic to improve the fertility treatments and follow-ups in men and women.