DED describes a group of tear film disorders that cause irritation and ocular surface damage. The most common subtypes of DED include aqueous and lipid deficiency, although most patients with DED have abnormalities in both tear components . Considering dry eye prevalence and cost, known risk, and underlying pathogenesis of DED seems to be determinant to achieve effective prevention and treatments. Age (menopause) and female sex are considered as valid risk factors for DED development as stated in the literature . In recent years, LC-MS/MS has had a pivotal role in routine clinical chemistry thanks to a wide range of applicability, easy sample preparation, and high analytical specificity, which allows for the identification, characterization and quantification of chemical compounds as target analytes based on their respective molecular masses and fragmentation patterns . This technique allows the simultaneous analysis of a large number of metabolites from many different biological matrices in order to investigate multifactorial disease, as in the case of DED.