Enhanced cellular uptake, transport and oral bioavailability of optimized folic acid-loaded chitosan nanoparticles.

Folic acid is a synthetic form of folate widely used for food fortification. However, its bioavailability is limited due to its inherent instability at several conditions. Therefore, a suitable encapsulation system is highly required. In the present study, the fabrication condition for folic acid-loaded chitosan nanoparticle (FA-Chi-NP) was optimized and then subjected to characterization. The optimized formulation had the particle size, zeta potential, and encapsulation efficiency of 180 nm, +52 mV, and 90%, respectively. In vitro release profile showed a controlled release of folic acid from the nanoparticles. Treatment of Caco-2 cells with the formulation showed no adverse effects based on MTT and LDH assays, and also, the cellular uptake was significantly higher after 2 h compared to free folic acid. Further, the oral administration of rats with FA-Chi-NPs (1 mg/kg BW) increased the plasma level of both folic acid (3.2-fold) and its metabolites such as tetrahydrofolate (2.3-fold) and 5-methyltetrahydrofolate (1.6-fold) significantly compared to free folic acid. In a bio-distribution study, duodenum and jejunum were found to be the primary sites for absorption. These findings suggest that chitosan may be a promising carrier for the delivery of folic acid and, therefore, could be exploited for various food applications.

Development of nano-encapsulated green tea catechins: Studies on optimization, characterization, release dynamics, and in-vitro toxicity.

Green tea (Camelia sinesis) has been acknowledged for plethora of pharmacological activities attributed by green tea catechins (GTC), however, poor bioavailability, short half-life and stability issues hamper its use as a therapeutic agent. Chitosan and sodium -tripolyphosphate were used to encapsulate GTC, which reduced its degradation in gastro-intestinal tract. The average size, zeta potential, and encapsulation efficiency of nano-encapsulated epigallocatechin-3-gallate (EGCG) rich GTC were observed as 250 nm, +49.8 mV, and 87%, respectively. The morphological and physico-chemical characterizations affirmed the size, stability, and encapsulation efficacy of nano-encapsulated green tea catechins. The In-vitro simulated release model showed the release of GTC in the intestinal phase via zero order kinetics. Cell viability studies were conducted on PC12 cell line to validate safety efficacy of nanoencapsulated GTC. Moreover, the study concluded that stability, bioavailability and bioactivity of EGCG was improved by nanoencapsulation of GTC, thereby rendering it a potent nanoceutical for clinical implications. PRACTICAL APPLICATIONS: Green tea catechin has enormous health endorsing activities. One of the major potentials of GTC is its antioxidant activity that plays a promising role in the prevention of various lethal disorders. In the present study, nanoencapsulation is used as a potential approach to improve the low bioavailability of green tea catechin. The results enlightened that nanoencapsulation of green tea catechin could be useful for improving the stability of green tea catechin in the GI tract as well as its bioaccessibility. Henceforth, this strategy restores the stability and bioavailability of green tea catechin that could be practically implied as a nutraceutical in the food and pharmaceutical industry as it can enhance the biological activity of catechins in catechin rich green tea-related products.

Influence of cooking methods on antinutritional factors, oligosaccharides and protein quality of underutilized legume Macrotyloma uniflorum.

Macrotyloma uniflorum is a salubrious but underutilized legume mainly consumed in semi-arid zones of Africa, Australia and India. Various antinutritional factors- phytates, oxalates and oligosaccharides- has limited its consumption. Current work describes the influence of various thermal processing technologies - autoclaving, microwave, micronization and extrusion- on antinutritional profile, phenolic acid profile and protein digestibility of two selected varieties. Significant (p ≤ 0.05) reduction in antinutritional content was observed for horse gram grains processed using various thermal technologies. Among all processing technologies extrusion caused marked degradation in antinutritional components. The reduction in oxalic acid, trypsin inhibitor, phytates and tannins ranged from 33 to 87 %, 77-82%, 33-60% and 51-66% respectively. Further, the decline in content of various oligosaccharides viz. raffinose, stachyose and verbascose varied from 36 to 61 %, 25-49% and 30-74% respectively for both the varieties. Although extrusion caused significant (p ≤ 0.05) decrease in the essential amino acid index and protein efficiency ratio, simultaneous increased was observed in biological value. The processing had significant (p ≤ 0.05) impact on grain antinutritional content and also retained its substantial functional properties. This establishes the utility of grain and promote the introduction of these new grains and enlarge the market of novel healthy foods.

Toxic metabolite profiling of Inocybe virosa.

Wild mushroom foraging involves a high risk of unintentional consumption of poisonous mushrooms which is a serious health concern. This problem arises due to the close morphological resemblances of toxic mushrooms with edible ones. The genus Inocybe comprises both edible and poisonous species and it is therefore important to differentiate them. Knowledge about their chemical nature will unambiguously determine their edibility and aid in an effective treatment in case of poisonings. In the present study, the presence of volatile toxic metabolites was verified in Inocybe virosa by gas chromatography. Methyl palmitate, phenol, 3,5-bis (1,1-dimethyl ethyl) and phytol were the identified compounds with suspected toxicity. The presence of the toxin muscarine was confirmed by liquid chromatography. The in vitro study showed that there was negligible effect of the digestion process on muscarine content or its toxicity. Therefore, the role of muscarine in the toxicity of Inocybe virosa was studied using a bioassay wherein metameters such as hypersalivation, immobility, excessive defecation, heart rate and micturition were measured. Administration of muscarine resulted in an earlier onset of symptoms and the extract showed a slightly stronger muscarinic effect in comparison to an equivalent dose of muscarine estimated in it. Further, the biological fate of muscarine was studied by pharmacokinetics and gamma scintigraphy in New Zealand white rabbits. Significant amount of the toxin was rapidly and effectively concentrated in the thorax and head region. This study closely explains the early muscarinic response such as miosis and salivation in mice. By the end of 24 h, a relatively major proportion of muscarine administered was accumulated in the liver which stands as an explanation to the hepatotoxicity of Inocybe virosa. This is one of the rare studies that has attempted to understand the toxic potential of muscarine which has previously been explored extensively for its pharmaceutical applications.

Influence of processing methodology on phenolic acid content, fatty acid profile and oxidative stability of Macrotyloma uniflorum.

Study was conducted to determine the influence of different cooking methods namely pressure, flaking, infra-red, microwave and extrusion processing on rate of oxidative stability, fatty acid profile and functional properties of horse gram. The rate of auto-oxidation was significantly (p < 0.05) affected during storage and found maximum at 0.00 aw and minimum at 0.33 aw for both varieties namely GPM-6 and PAIYUR-2. The extrusion processed grains were more susceptible to auto-oxidation. The iron content of grain increased significantly (p < 0.05) during extrusion (1.22 ± 0.50 to 1.65 ± 0.15 mg 100 g-1 for PAIYUR-2 and 1.19 ± 0.45 to 1.59 ± 0.12 mg 100 g-1 for GPM-6). Whereas, tocopherol content decreased during extrusion (8.05 ± 0.15 to 2.28 ± 0.23 mg 100 g-1 for PAIYUR-2 and 6.48 ± 0.46 to 1.68 ± 0.15 mg 100 g-1 for GPM-6). Ellagic (12.36 ± 0.35 and 10.71 ± 0.29 mg 100 g-1), vanillic (15.20 ± 0.23 and 12.48 ± 0.18 mg 100 g-1), and coumaric acid (14.68 ± 0.71 and 8.97 ± 0.66 mg 100 g-1) were the major phenolic acids whereas, linoleic (35.53 ± 0.30 and 35.46 ± 0.19%), palmitic (26.08 ± 0.26 and 25.97 ± 0.33%), and linolenic acid (13.44 ± 0.18 and 10.13 ± 0.21%) were the major fatty acids present in raw grain for PAIYUR-2 and GPM-6 respectively. Phenolic and fatty acids were significantly (p < 0.05) affected during processing. The oxidative stability of microwave processed grains was maximum whereas, extrusion processed grains showed minimum oxidative stability during storage. Study explicitly describes that native grains and the grains which did not undergo destruction of naturally present cellular structure, were less prone to oxidation. The oxidation rate of grains was found dependent on composition, processing and storage environment of grains. Hence, all these factors need to be considered to ensure the stability of processed food during storage.

Antioxidant, Biomolecule Oxidation Protective Activities of Nardostachys jatamansi DC and Its Phytochemical Analysis by RP-HPLC and GC-MS.

The study aimed at analyzing the metabolite profile of Nardostachys jatamansi using RP-HPLC, GC-MS and also its antioxidant, biomolecule protective and cytoprotective properties. The 70% ethanolic extract of Nardostachys jatamansi (NJE) showed the presence of polyphenols and flavonoids (gallic acid, catechin, chlorogenic acid, homovanillin, epicatechin, rutin hydrate and quercetin-3-rhamnoside) analyzed by RP-HPLC, whereas hexane extract revealed an array of metabolites (fatty acids, sesquiterpenes, alkane hydrocarbons and esters) by GC-MS analysis. The antioxidant assays showed the enhanced potency of NJE with a half maximal inhibitory concentration (IC50) value of 222.22 ± 7.4 µg/mL for 2,2-diphenyl-1-picrylhydrazyl (DPPH), 13.90 ± 0.5 µg/mL for 2,2'-azino-bis(3-ethyl benzothiazoline-6-sulfonic acid) diammonium salt (ABTS), 113.81 ± 4.2 µg/mL for superoxide, 948 ± 21.1 µg/mL for metal chelating and 12.3 ± 0.43 mg FeSO4 equivalent/g of extract for ferric reducing antioxidant power assays and was more potent than hexane extract. NJE effectively inhibited 2,2'-azobis(2-methylpropionamidine) dihydrochloride (AAPH)-induced oxidation of biomolecules analyzed by pBR322 plasmid DNA damage, protein oxidation of bovine serum albumin and lipid peroxidation assays. The observed effects might be due to the high content of polyphenols, 53.06 ± 2.2 mg gallic acid equivalents/g, and flavonoids, 25.303 ± 0.9 mg catechin equivalents/g, of NJE compared to the hexane fraction. Additionally, the extract abrogated the protein, carbonyl, and ROS formation, and NJE showed cytotoxicity in SH-SY5Y neuronal cells above 75 µg/mL. Thus, the study suggests that the herb unequivocally is a potential source of antioxidants and could aid in alleviating oxidative stress-mediated disorders.