<jats:title>Abstract</jats:title><jats:p>During<jats:italic>Leishmania</jats:italic>transmission sand flies inoculate parasites and saliva into the skin of vertebrates. Saliva has anti-haemostatic and anti-inflammatory activities that evolved to facilitate bloodfeeding, but also modulate the host’s immune responses. Sand fly salivary proteins have been extensively studied, but the nature and biological roles of protein-linked glycans remain overlooked. Here, we characterised the profile of<jats:italic>N</jats:italic>-glycans from the salivary glycoproteins of<jats:italic>Lutzomyia longipalpis</jats:italic>, vector of visceral leishmaniasis in the Americas.<jats:italic>In silico</jats:italic>predictions suggest half of<jats:italic>Lu. longipalpis</jats:italic>salivary proteins may be<jats:italic>N</jats:italic>-glycosylated. SDS-PAGE coupled to LC-MS analysis of sand fly saliva, before and after enzymatic deglycosylation, revealed several candidate glycoproteins. To determine the diversity of<jats:italic>N</jats:italic>-glycan structures in sand fly saliva, enzymatically released sugars were fluorescently tagged and analysed by HPLC, combined with highly sensitive LC-MS/MS, MALDI-TOF-MS, and exoglycosidase treatments. We found that the<jats:italic>N</jats:italic>-glycan composition of<jats:italic>Lu. longipalpis</jats:italic>saliva mostly consists of oligomannose sugars, with Man<jats:sub>5</jats:sub>GlcNAc<jats:sub>2</jats:sub>being the most abundant, and a few hybrid-type species. Interestingly, some glycans appear modified with a group of 144 Da, whose identity has yet to be confirmed. Our work presents the first detailed structural analysis of sand fly salivary glycans.</jats:p>