Basella rubra (Malabar spinach) is a commonly consumed green leafy vegetable in southern parts of India. The chemical composition, nutraceuticals characterization, squalene Nuclear Magnetic Resonance (NMR), in vitro antioxidant activities and cytotoxicity of B. rubra seed oil (33.08%) was investigated. Gas chromatography-mass spectrometry (GC/MS) analysis revealed the presence of palmitic (27.21 μmol%), oleic (33.83 μmol%) and linoleic acid (26.02 μmol%) with a total of 64.38 μmol% unsaturated fatty acids respectively. HPLC nutraceutical characterization showed a major constituent of gallic acid (11.23 mg%), γ-tocopherols (17.74 mg%), cycloartenylferulate (1.7 mg%), and squalene (1 g%). Squalene was further recovered (98%), purified (99.9%), and confirmed through 1H and 13C NMR. The in vitro antioxidant activities recorded by using 2,2-diphenyl-1-picrylhydrazyl (EC50 = 6 mg mL-1), ferric reducing antioxidant power (361.85 mM of Trolox Eq./100 g) and 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (EC50 = 56.19 mg mL-1) scavenging activity. In vitro cytotoxicity assessed on 3T3-L1 showed good cell survival without any toxicity (upto 400 μg mL-1). B. rubra seed oil has proven nutraceuticals and antioxidant potentials with least toxicity which can be recommended for functional foods applications.

γ-Oryzanol is an important group of nutraceuticals that play a key role in addressing metabolic disorders. This study, for the first time, examined volatile-free spice fixed oils (FOs) as an alternate plant source for γ-oryzanol and other nutraceuticals (phenolics, flavonoids, phytosterols, and tocopherols) using HPLC, HR-MS and NMR. The in vitro antioxidant activities of FOs were also analysed. The selected spices were Alpinia galanga, Cinnamomum zeylanicum, Trigonella foenum-graecum, Foeniculum vulgare and Myristica fragrans. The major polyphenols and flavonoids quantified were gallic, protocatechuic, vanillic, syringic, para-coumaric, ferulic, rutin, trans-cinnamic, and quercetin. T. foenum-graecum FOs recorded high levels of ergosterol (48.56 mg/100 g) and stigmasterol (247.36 mg/100 g). The fucosterol levels were high in A. galanga (268.31 mg/100 g) FOs, whereas C. zeylanicum FOs showed high content of β-sitosterols (7037.77 mg/100 g). C. zeylanicum and T. foenum-graecum FOs recorded high α-tocopherol content (47.55 and 15.96 mg/100 g respectively). C. zeylanicum FOs showed high levels of three ferulates, namely, cycloartenyl ferulate, 24-methylene cycloartenyl ferulate and β-sitosteryl ferulate, whose contents were 89.42, 170.23 and 50.23 mg/100 g respectively which was confirmed by HRMS with a molecular mass (m/z) of 601.45, 615.47, and 589.45 respectively. Further, γ-oryzanol ferulates in C. zeylanicum FOs were confirmed by 1H-NMR analysis. The acidified methanolic extractives of FOs showed high free radical scavenging activity and antioxidant potential. These spice FOs have excellent antioxidant activities, and are novel potential functional ingredients against lifestyle disorders.