Nanoencapsulation of apocynin and vanillic acid extracted from Picrorhiza kurroa Royle ex Benth plant roots and its characterisation.

Picrorhiza kurroa Royle ex Benth (P.kurroa) is an important medicinal plant in the ayurvedic system for treating various liver and inflammatory conditions. The present study aimed to extract the phytocompounds from various extracts (Acetone, Chloroform, Ethanol, Ethyl acetate, Hexane, and Methanol) of P. kurroa. Further, the major phytocompounds were nano-encapsulated by PLGA (Poly-lactic-co-glycolic acid) method and characterized to enhance activity towards the target. The highest polyphenolic value was found to be 323.2 ± 16.6 and 316.3 ± 12.1 µg GAEq./mg in ethanolic and methanolic extracts. The highest flavonoid value was found to be 280.3 ± 19.8 and 300.8 ± 15.2 in ethanolic and methanolic extracts µg QEq./mg. P. kurroa exhibited DPPH radical scavenging with IC50 of 38.2 ± 1.1 and 43.7 ± 1.8 µg/mL and also showed potent ferric reducing power and total antioxidant activities. The major phytocompounds, such as apocynin (AP) and vanillic acid (VA), were confirmed using HPLC. Further, the nano-encapsulation of apocynin and vanillic acid successfully achieved by PLGA methods. The average particle size of nano-encapsulated apocynin, vanillic acid is 350 nm, 204.4 nm, and zeta potential were -25.3 mv and -11.2 mv. Nanoformulations showed an apocynin and vanillic acid encapsulation efficiency of 93.6% and 93.3%, respectively. SEM and AFM confirmed the round and smooth morphology of the nanoparticles. The results of XRD confirmed the amorphous nature of nanoformulations. FTIR technique confirm the presence of biomolecules inside the polymer. The thermal stability of nanoformulations determined by DSC analysis showed endothermic peak. The prepared and characterization apocynin, vanillic acid nanoparticles revealed their good quality index, suggesting that potential use in pharmacy and phytotherapy as a source of natural antioxidant.

Apocynin exerts neuroprotective effects in fumonisin b1-induced neurotoxicity via attenuation of oxidative stress and apoptosis in an animal model.

The Fusarium verticillioides produces a mycotoxin, that is, fumonisin b1 (Fb1), which commonly infects corn and agricultural commodities. The Fb1 showed hepatotoxicity, neurotoxicity, and carcinogenicity in animals. Hence, the present investigation aimed to evaluate the effect of apocynin (AP) on Fb1-induced neurotoxic effects and its mechanism in the mice model and cell line. The male Balb/c mice, with the 6.75 mg/kg bwt of Fb1 were injected subcutaneously for 5 days to induce neurotoxicity. A significant elevation of serotonin (5-HT) was observed in mice treated with Fb1 in the whole brain showing biogenic amines may reflect Fb1 neurotoxicity, but the negatively regulated mechanisms were attenuated by the pretreatment of AP. In addition, AP pretreatment normalized apoptotic changes in histology and immunohistochemistry studies. In Western blotting studies, apoptotic genes were upregulated and oxidative stress genes were downregulated due to Fb1 treatment; while treating with AP, these gene expressions were rectified. Further cell cytotoxicity was investigated by MTT and lactate dehydrogenase (LDH) assays in SH-SY5Y cell line. MTT and LDH assays indicated the IC50 value to be 150 µM of Fb1, which was protected by 100 µg of AP. The electron microscopy evaluated the Fb1-induced apoptotic conditions and its cell morphology recovery by AP. These results suggest that nicotinamide adenine dinucleotide phosphate hydrogen oxidase-mediated reactive oxygen species is the primary upstream signal leading to increased Fb1-mediated neurotoxicity in mice. The use of the antioxidant AP reversed the toxin-induced oxidative stress and apoptosis by its antioxidant potency.

Indigenous fungal strains isolation and molecular identification from coffee pulp for the production of pectic oligosaccharides.

Coffee pulp (CP) is a significant agro-industrial waste generated during coffee bean processing, which possess substantial environmental contamination and is rich in pectin. The primary objective of this study was to investigate the conversion of pectin extracted from coffee pulp into pectic oligosaccharides (POS) using native microbial strains. The study aimed to optimize the growing conditions, including temperature, time, and pectin concentration, to assess the productivity of pectinase. Two fungal strains that exhibited the highest growth on CP were isolated and subsequently identified as Aspergillus fumigatus P-1007 and A. fumigatus HA1, employing 5.8S rRNA gene sequencing. The optimization of temperature for the organism was carried out between 25 and 45 °C; compared to the other temperatures at 45 °C the productivity of pectinase was high; the exact temperature was used for the time experiment where we found that compared to the A. fumigatus P-1007, A. fumigates HA1 was showed high enzyme productivity on 6th day. Hence, the highest productivity of endo-pectinase was seen at a temperature of 45 °C on the 6th day using isolated A. fumigates HA1 in the CP with 1% of coffee pectin. Additionally, the produced POS were screened and confirmed through TLC and HPLC analysis. The antioxidant activity of the POS derived from the separated CP demonstrated an effective concentration (EC50) of 400 µg/ml. The study indicates that the efficient utilization of CP waste for producing potentially valuable functional food ingredients, such as POS, holds promise for commercial development. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03811-9.

Profiling of Lipids, Nutraceuticals, and Bioactive Compounds Extracted from an Oilseed Rich in PUFA.

Polyunsaturated fatty acid (PUFA) rich vegetable oils are nutritionally and economically important agriculture based commodities. The lipid profile, nutraceutical content, and antioxidant activity of Indian chia seed oil (CSO) were analysed and compared with PUFA rich vegetable oils. The total oil content was 28% (w/w) with α-linolenic acid (ALA; 65%) as the predominant fatty acid and a n-3/n-6 ratio of 3.5. The tocopherol content was 144 mg/kg of oil, with γ + ß being the most abundant. The squalene content was 178.47 mg/100 g of oil, and the total phenolic content was 0.014 mg GAE/g of oil. The identity of major polyphenols in the methanolic extract of CSO were established by LC-HRMS. FTIR spectra of CSO exhibited characteristic features that were identical to other PUFA rich oils. Results demonstrate that the Indian CSO is an excellent source of essential fatty acids and key nutraceuticals.

Physicochemical characterization and nutraceutical compounds of the selected spice fixed oils.

Spices and herbs are well appreciated for their medicinal properties since ancient times. Till date, spices are being explored for volatile oils (essential), flavour and for addressing many chronic diseases. In the present study, we investigated the physicochemical properties, fatty acid composition, differential scanning calorimetry (DSC), elemental composition and nutraceutical compounds of fixed oils (non-volatile) from five selected spices viz., Alpinia galanga, Cinnamomum zeylanicum, Trigonella foenum-graecum, Foeniculum vulgare, and Myristica fragrans. The fixed oil (FO) content of volatiles-free powders of the five selected spices ranged from 1.58% (C. zeylanicum) to 26.43% (M. fragrans). The studied FO showed a good quality index which was analysed by estimation of free fatty acids, iodine value and unsaponifiable matter. The fatty acid analysis showed high palmitic acid in the FO of A. galanga and C. zeylanicum. High linoleic, oleic, and myristic acid levels were observed in T. foenum-graecum, F. vulgare and M. fragrans FOs, respectively. The nutraceutical compounds such as total phenolics were high in C. zeylanicum FO (0.53%). Hence the studied FO could be an excellent alternative to oil nutraceutical compounds. It may be used as a functional ingredient in foods which needs further validation for value addition.

Utilization of coffee pulp waste for rapid recovery of pectin and polyphenols for sustainable material recycle.

Coffee pulp is one of the major underutilized byproduct of coffee processing in farm level. Disposal of this agro-industrial waste has become one of the most challenging tasks for coffee planters. However, most of the efforts are towards the management of coffee pulp as an effluent, and not-on re-use. The problem is compounded due to the large volumes produced in diluted forms, which makes it expensive to reuse. The preliminary proximate analysis of coffee pulp indicated it to be rich in pectin and polyphenols. The efficacy of various chemicals like ethanol, sulfuric acid, hydrochloric acid, nitric acid, ammonium oxalate and metal salts for effective precipitation of pectin from coffee pulp was evaluated. HPLC characterization of the extracted and concentrated polyphenols fractions was analyzed. The maximum extraction of pectin was achieved by using metal salts and ethanol with 6.0% and 6.7% on wet weight basis respectively. The equivalent weight of extracted pectin (1180.5 mg/g) was found to be higher than that of commercial pectin (724.8 mg/g). The methoxyl content of the commercial pectin and crude pectin were 9.3 and 5.6% respectively. Gallic, vanillin, catechin, ethyl catechol, coumaric, Caffeic, and ferulic acid were the major polyphenols as quantified by the HPLC. The polyphenol fraction showed a good antioxidant activity with phosphomolybdate, FRAP, DPPH, and ABTS radicals respectively. The sustainable utilization of coffee pulp as a source of pectin and polyphenols with good antioxidant activities could help to solve the problem of waste generated in coffee processing in farm level.

Spice fixed oils as a new source of γ-oryzanol: nutraceutical characterization of fixed oils from selected spices.

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 ß-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 ß-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.

Chemical composition, nutraceuticals characterization, NMR confirmation of squalene and antioxidant activities of Basella rubra L. seed oil.

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.