GC-MS, HPLC, UV-VIS, AND FTIR CHARACTERIZATION OF BIOACTIVE PHYTOCONSTITUENTS IN THE HYDROETHANOLIC LEAF EXTRACT OF DIGERA MURICATA (L.) MART: A MULTI-ANALYTICAL APPROACH

Digera muricata (L.) Mart (Amaranthaceae) has a well-documented history of therapeutic application in South
Asian folk medicine, yet the identity of its constituent bioactive molecules has remained largely uncharacterised at the
molecular level. Our previous study established that the hydro-ethanolic leaf extract harbours exceptional quantities of
total phenolics, flavonoids, saponins, tannins, and steroids, pointing to substantial pharmacological potential. The present
work follows from that quantitative baseline by applying multi-instrumental characterization techniques, viz., Gas
Chromatography-Mass Spectrometry (GC-MS), High-Performance Liquid Chromatography (HPLC), Ultraviolet-Visible
(UV-Vis) spectroscopy, and Fourier-Transform Infrared (FTIR) spectroscopy, to characterise the specific molecular
constituents responsible for those activities. GC-MS analysis resolved 30 chromatographic peaks, with n-hexadecanoic
acid (palmitic acid, 57.08%) and octadecanoic acid (10.97%) as dominant constituents, accompanied by phytol,
tetradecanoic acid, and muskolactone among the biologically relevant minor components. HPLC profiling at 280 nm
tentatively identified 12 polyphenolic constituents, with rutin (39.73%), ellagic acid (14.12%), and catechin (11.49%)
accounting for the majority of the peak area. The UV-Vis analysis showed that the major absorptions were found at 202.6
nm and 250.1 nm, which were associated with aromatic conjugation and flavonoid Band-II chromophores. FTIR
established the presence of hydroxyl, aliphatic, alkene, aromatic, and ether functionalities, which were in agreement with
the GC-MS and HPLC profiles. The data collectively have offered a molecular basis of the antioxidant, anti-inflammatory,
and anticancer potentials of this plant, and have identified rutin, ellagic acid, palmitic acid and phytol as priority
compounds to be isolated and their bioactivity validated.