Metabolomic Analysis of Aging in Drosophila

The whole flies at 30 days of age were collected, and the sample preparation methods of metabolomics analysis were assessed as reported previously [25 (link)]. Briefly, samples were separated using Agilent 1290 UHPLC (Agilent Technologies, Santa Clara, CA, USA) equipped with the ACQUITY UPLC BEH Amide column (1.7 μm, 2.1 mm × 100 mm), and analyzed by Triple 6600 TOF mass spectrometer (AB Sciex, Concord, Toronto, ON, Canada). Metabolites were identified based on the exact mass of their MS and tandem MS spectra, which were then searched and compared using a laboratory database (Shanghai Applied Protein Technology Co., Ltd., Shanghai, China). The initially processed data were enumerated with SIMCA software (V14.1, Umetrics, Ume, Västerbotten, Sweden) for mode identification following normalization to total peak intensity (by weight of the complete flies). Following the collection of valid data, principal component analysis (PCA) and orthogonal partial least-squares discriminant analysis (OPLS-DA) were applied to differentiate STP from the CT group. Variable importance in projection (VIP) > 1 and p < 0.05 were employed as criteria for screening potential biomarkers, and the KEGG metabolomics pathway analysis was constructed to reveal the most relevant pathway for STP to exert anti-aging effects.