In summary, using untargeted lipidomics analysis as a dis-
covery tool, this study identified lipids in the sphingolipid, ether lipid, glycerophospholipid and
glycerolipid metabolism pathways as novel biomarkers for estrogen-deficient conditions induced by
ovariectomy. Subsequent evaluation of the therapeutic potential of
C. comosa in the OVX rats revealed that C. comosa extract and powder gave a superior performance
over E2 and DPHD in terms of both the comparative table of individual lipid species and the OPLS-DA
score plot of negative-mode data. Moreover, because Cer were the only lipids affected by the E2
treatment, it could also suggest the increased Cer levels as a potential cause of metabolic
disturbance under estrogen deprivation, an effect which was re- versed by C. comosa treatment.
Future work will aim to dissect the mechanisms underlying alterations in the levels of these
lipids, especially Cer, and to determine the potential additive or sy- nergistic effects from other
compounds in C. comosa. We believe that the LC–MS-based lipidomics platforms and biomarkers pre-
sented in this work can be utilized as tools for evaluating other modern and traditional medicines
in this model organism and in general in the future.