The H1N1pdm09 influenza strain's resistance to Tamiflu® (oseltamivir) due to the H275Y mutation has highlighted the importance of neuraminidase (NA) in viral pathogenicity, prompting increased interest in including NA antigens in seasonal influenza vaccines. While NA's glycosylation is crucial for viral function, most studies have focused on the catalytic head domain, leaving the stalk region's glycosylation patterns largely unexplored despite its potential antigenic significance. This study characterizes glycan variations and occupancy in both head and stalk regions of wild-type and oseltamivir-resistant recombinant NA (rNA) from the H1N1pdm09 strain. We introduced the resistance-associated H275Y mutation and fitness-restoring mutations (V241I and N369K) through site-directed mutagenesis, expressing rNA in HEK293 cells. Glycopeptide analysis was performed in ZenoTOF7600 mass spectrometer following proteolytic digestion of purified proteins. Glycan formulas were identified using Byonic software, with manual curation of glycopeptide spectra for structure-specific fragments. Our comprehensive analysis revealed distinct glycosylation patterns across the NA protein, with all identified glycosites showing complete occupancy. The four glycosites in the head domain (N88, N146, N235, N386) were predominantly modified with complex glycans, while the conserved sites in the stalk region (N50, N58, N63, N68) exhibited a mixed profile of both oligomannose and complex glycans, suggesting domain-specific glycosylation preferences. Notably, we did not detect sialylated or sulphated glycopeptides. This study represents one of the first comprehensive glycosylation profiles of a pandemic-emerged NA strain, encompassing both head and stalk domains. Our findings provide valuable insights into NA glycosylation patterns that could inform future vaccine development strategies, particularly in addressing drug resistance in pandemic influenza strains.