Metabolomic profiling of pumpkin (Cucurbita spp.) male flowers using integrated LC-HRMS, GC-MS and 1H NMR.

Pumpkin (Cucurbita spp.) male flowers are edible tissues traditionally consumed in several regions worldwide; however, information regarding their metabolite composition remains limited. In this study, four agronomically important pumpkin varieties cultivated in northeastern Brazil were investigated using an integrated metabolomics workflow combining LC-HRMS, GC-MS and 1H NMR. The complementary analytical platforms enabled the annotation of 151 metabolites spanning amino acids, carbohydrates, phenylpropanoids, flavonoids, terpenoids, fatty acids and other specialized metabolites, while increasing annotation confidence through orthogonal analytical information. Multivariate and univariate analyses revealed clear genotype-dependent metabolic differences, particularly in carbohydrate, amino acid and phenolic metabolism, whereas PCA showed no clear separation according to planting-bed position. The integrated analytical strategy broadened chemical coverage beyond that achievable with any individual platform and provided new insights into the molecular composition of pumpkin flowers. These findings strengthen current knowledge of Cucurbitaceae floral metabolism and support the nutritional, technological and economic valorization of pumpkin flowers as an underexplored edible resource.