Ferric Reducing Antioxidant Potential Assay Research Articles

Evaluating the in vitro antidiabetic, antibacterial and antioxidant properties of copper(II) Schiff base complexes: An experimental and computational studies

In recent years, metal-based compounds have been explored as potential remedies for majorhealth issues such as diabetes, bacterial infections, and oxidative stress, which impact the general well-being of humans globally. Diabetes, bacterial infections, and oxidative stress are major health concerns affecting millions worldwide. Researchers have been exploring the use of metal-based compounds as potential treatments for these conditions. In our quest towards identifying biologically viable metal-based compounds, three novel Cu(II) complexes were synthesized from halogen-substituted ON donor Schiff base ligands. These ligands were obtained by condensing 2-hydroxybenzldehyde with three different halogen-substituted anilines. The ligands and their complexes were characterized using various spectroscopic techniques such as 1H and 13C nuclear magnetic resonance, Fourier Transform-Infrared, Ultraviolet–visible spectra, elemental analysis, mass spectrometry, and single-crystal X-ray spectroscopy to elucidate their structures. The compounds were screenedfor antidiabetic (α-glucose and α-amylase enzymes inhibition assay), antioxidant (2,2-diphenyl-1-picrylhydrazyl(DPPH), ferric reducing antioxidant potential (FRAP) assay, and nitric oxide(NO) radical scavenging), and antimicrobial activities. The pharmacological activity revealed that the efficacy of the ligands was enhanced upon coordination, and CuL3 is more potent among the complexes, having higher inhibition activity on α-glucosidase (IC50 = 100.1 μM) than the control, whose IC50 is 112.4 μM on the same enzyme. For radical scavenging activity, the results showed that the complexes have higher scavenging activity on DPPH radicals than their ligands and the control. For the FRAP assay, the complexes show higher activity than their ligands. Similarly, NO radical scavenging activity results indicate the ligands have higher activity than their complexes, while the complexes outperformed the control at all concentrations tested. The In vitro antibacterial results on some selected Gram-positive and Gram-negative bacteria revealed the complexes have higher antibacterial activity than their corresponding ligands, with CuL3 taking the lead, showing a MIC value of 4 g/mL on some of the bacteria tested. Also, density functional theory calculations at different levels of DFT and basis sets were used to investigate the optimized geometry, electronic structure (energy of HOMO and LUMO, Mulliken atomic charge, dipole moment, hardness, and softness, electrophilicity, and spectral properties) of the synthesized compounds. Molecular docking studies were carried out to investigate the binding affinity of the receptor proteins to other proteins and to establish the mechanism of action at the active sites.

Evaluation of Antioxidant Potential of Asystasia Chelonoides var.chelonoides Nees.(Acanthaceae)

The ethanolic leaf extract of Asystasia chelonoides var. chelonoides Nees. was evaluated using various in vitro and ex vivo assays. Evaluations were made for DPPH radical scavenging activity, NO radical scavenging activity, Super oxide radical scavenging activity ,Ferric Reducing Antioxidant Potential (FRAP) assay, Total antioxidant activity , Reducing Power activity and Lipid Peroxidation inhibition. Over all findings revealed that A.chelonoides leaf exhibited four extracts namely (AC HEX, AC CHL, AC ETH and AC CRD) and were selected for the study, AC ETH and AC CRD showed potent antioxidant activity by inhibiting DPPH radical in a concentration dependent manner . The nitric oxide radical scavenging activity of all the samples increased as their concentration increased from 20 µg/mL to 320 µg/ mL. AC CRD showed its maximum superoxide radical scavenging of 78.56 % at 320 µg/mL and the EC50 was found to be 93.26 µg/mL. The extracts exhibited a dose dependent reducing power activity within the applied concentrations (20 µg/mL to 100 µg/mL) and AC CRD at 100 µg/mL concentration showed highest activity, which was almost comparable to standard ascorbic acid. In FRAP assay the crude extract of A. chelonoides (AC CRD ) showed 138.14 µg trolox equivalent/g of dry extract respectively. Total antioxidant activity of AC CRD, AC HEX, AC CHL and AC ETH were found to be 98.27, 14.86, 20.16 and 72.56 µg of ascorbic acid equivalent/mg of dry extract respectively. A.chelonoides crude extract and ethanolic fraction showed potent inhibition of FeCl2AA stimulated rat liver lipid peroxidation in concentration dependent manner.

Phytochemical Screening, gc-ms Analysis and Antioxidant Activity of Methanol Stem Bark Extract of Cassia Sieberiana DC

The over production of free radicals in the human body is known to cause oxidative stress which plays an important role in the pathogenesis of many diseases. Therefore, the search for natural antioxidants would continue to be a major area of research for many decades. This study was to investigate the in vitro antioxidant activities and to determine the phytochemicals present in Cassia sieberiana. The methanol stem bark extract of C. sieberiana was subjected to phytochemical screening and Gas Chromatography-Mass Spectrometry (GC-MS) analysis. Its antioxidant activities were assayed in vitro using nitric oxide scavenging assay, ferric reducing antioxidant potential assay, lipid peroxidation inhibitory assay and 2-diphenyl-1-picrylhydrazyl scavenging assay. The phytochemical screening of the stem bark extract of C. sieberiana showed saponins, alkaloids, flavonoids, steroids, tannins, phlobatannins, terpernoids, cardiac glycosides, phenolics and reducing sugar. The GC-MS Table revealed that the methanol extract has 51 bioactive compounds with 3-0-Methyl-d-glucose having the highest percentage of 12.16 % while 5.alpha-Androstan-3-beta-ol,4,4-dimethyl- having the lowest percentage of 0.09 %. The antioxidant study showed concentration-dependent radical scavenging activity of the methanol stem bark extract of C. sieberiana. This concentration-dependent result was also observed when compared with ascorbic acid used as the standard drug. The presence of different phytochemicals such as alkaloids, saponins, tannins, phenols, flavonoids, among other things, might be responsible for C. sieberiana antioxidant activity. The study concluded that the methanol stem bark extract of C. sieberiana contains 51 bioactive compounds and showed good antioxidant activity which could be therapeutic in several human diseases. Key words: Cassia sieberiana, Phytochemical, Antioxidant, Pharmaceuticals

Evaluation of acute oral toxicity, cytotoxicity, antidepressant and antioxidant activities of Japanese mint (Mentha arvensis L.) oil

Menthol-rich essential oil of Mentha arvensis L . (Japanese mint) have been identified as a valuable and new source of natural bioactive compounds with applications not only in industry but also in our daily life. The present study aimed to assess the cytotoxicity, acute toxicity, antidepressant and antioxidant activities of Japanese mint oil (JMO). Brine shrimp lethality assay was performed to investigate cytotoxicity of JMO. The acute toxicity was evaluated by Lorke's method. Forced swimming test (FST) was performed for determining antidepressant property. The antioxidant potentials were performed by DPPH scavenging assay, ferric reducing antioxidant potential (FRAP) and Fe 2+ ion chelating ability methods. In acute toxicity study, JMO showed toxic effects within 5 minutes and estimated LD 50 value was 2070.4 µl/kg body weight after 10 days oral treatment. The brine shrimp lethality assay exhibited concentration–dependent approach and the LC 50 value was determined to be 139.73 l/ml, with lower and upper limits of 82.2428 l/ml and 202.7082 l/ml, respectively. In the FST method, intraperitoneal injections of JMO emulsion at doses of 78l/kg, 156l/kg, and 225l/kg of body weight resulted in a significant (p<0.005) increase in struggling time and a reduction in immobility. This observation had a nearly identical effect to the reference drug. Besides, JMO showed weak antioxidant activity in DPPH assay with an IC 50 of 72.58±6.16387 μl/ml. But in other two methods, it exhibited strong antioxidant abilities whereas 1.4010 μl/ml reducing power for FRAP assay and 66.66% chelating rate for Fe2+ ion chelating ability test at 100 μl/ml concentration. The investigation clearly showed that JMO have significant antidepressant and cytotoxic properties whereas acute toxicity was moderate. Besides, JMO also possess considerable antioxidant activities.

Polysaccharides, Total Phenolic, and Flavonoid Content from Different Kenaf (Hibiscus cannabinus L.) Genotypes and Their Antioxidants and Antibacterial Properties.

Kenaf (Hibiscus cannabinus L.) is a valuable plant with a potential health benefit because of its extensive bioactive compounds. Leaf extracts of 33 kenaf genotypes were investigated for their polysaccharide, total phenolic, and flavonoid content. The antioxidant properties were evaluated by 2,2-Diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS), and ferric reducing antioxidant potential (FRAP) assays. Antimicrobial capacity was also assessed against Staphylococcus aureus and Escherichia coli using a disc diffusion assay. The polysaccharide content varied from 6.45–16.12 mg glucose per g DW. Total phenolic and flavonoid content ranged from 6.03–21.15 mg GAE/g DW and 1.55–9.24 mg RE/g DW, respectively. Similarly, varied values in the range 20.55–79.99% of inhibition by DPPH, 56.28–88.30% of inhibition by ABTS and 1.26–5.08 mmol Fe2+/g DW by FRAP assays were obtained for antioxidants of the genotype extracts. Extracts from CS4 and CS2 genotypes had the highest antioxidant activities. Kenaf leaves exhibited antibacterial activity against Staphylococcus aureus and Escherichia coli. Strong correlation was found between antioxidant activity with polysaccharide (DPPH, r = 0.893; ABTS, r = 0.819; FRAP, r = 0.864) and total phenolic content (DPPH, r = 0.850; ABTS, r = 0.959; FRAP, r = 0.953). The results suggested that the kenaf leaves could be used as a natural antioxidants and antimicrobial in food industries.

Self-Crosslinked Ellipsoidal Poly(Tannic Acid) Particles for Bio-Medical Applications

Self-crosslinking of Tannic acid (TA) was accomplished to obtain poly(tannic acid) (p(TA)) particles in single step, surfactant free media using sodium periodate (NaIO4) as an oxidizing agent. Almost monodisperse p(TA) particles with 981 ± 76 nm sizes and −22 ± 4 mV zeta potential value with ellipsoidal shape was obtained. Only slight degradation of p(TA) particles with 6.8 ± 0.2% was observed at pH 7.4 in PBS up to 15 days because of the irreversible covalent formation between TA units, suggesting that hydrolytic degradation is independent from the used amounts of oxidation agents. p(TA) particles were found to be non-hemolytic up to 0.5 mg/mL concentration and found not to affect blood clotting mechanism up to 2 mg/mL concentration. Antioxidant activity of p(TA) particles was investigated by total phenol content (TPC), ferric reducing antioxidant potential (FRAP), trolox equivalent antioxidant capacity (TEAC), total flavanoid content (TFC), and Fe (II) chelating activity. p(TA) particles showed strong antioxidant capability in comparison to TA molecules, except FRAP assay. The antibacterial activity of p(TA) particles was investigated by micro-dilution technique on E. coli as Gram‑negative and S. aureus as Gram-positive bacteria and found that p(TA) particles are more effective on S. aureus with over 50% inhibition at 20 mg/mL concentration attained.

Unleashing the Biological Potential of Fomes fomentarius via Dry and Wet Milling.

Fomes fomentarius, usually referred to as tinder conk, is a common wood-based fungus rich in many interesting phytochemicals and with an unique porous structure. Dry or wet ball milling of this sponge on a planetary mill results in small particles with sizes in the range of 10 µm or below. Suspended in water and without preservatives or other stabilizers, the resulting micro-suspensions are sterile for around six days, probably due to the increased temperatures of around 80 °C especially during the wet milling process. The suspensions also exhibit excellent antioxidant activities as determined in the DPPH, ferric reducing antioxidant potential (FRAP) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assays. In the DPPH assay, IC50 values of 0.02–0.04% w/v and 0.04% w/v were observed for dry and wet milled samples, respectively. In the FRAP assay, IC50 values of <0.02% w/v and 0.04% w/v were observed for dry and wet milled samples, respectively. In contrast, the ABTS assay provided IC50 values of 0.04% w/v and 0.005% w/v, respectively. Notably, this activity is mostly—albeit not exclusively—associated with the highly porous particles and their large surfaces, although some active ingredients also diffuse into the surrounding aqueous medium. Such suspensions of natural particles carrying otherwise insoluble antioxidants on their surfaces provide an interesting avenue to unleash the antioxidant potential of materials such as sponges and barks. As dry milling also enables longer storage and transport, applications in the fields of medicine, nutrition, agriculture, materials and cosmetics are feasible.

Simultaneous Detection and Validation of Analytical Markers of Swertia chirata by HPLC-DAD to Evaluate the Potency of Extracts and Fractions against Antioxidant Potential

The present study is based on the selection of extract and fraction of Swertia chirata plant for the antioxidant potential with HPLC fingerprinting, which includes the simultaneous detection and quantification of four analytical markers protocatechuic acid (PCA), swertiamarin (SM), mangiferin (MF) and amarogentin (AG) by HPLC-DAD. The yield of water extract (SWA), hydroalcoholic extract (SHA) and fractions of hydroalcoholic extracts were evaluated for their antioxidant potential against 2,2-diphenyl-1-picrylhydrazyl-hydrate free radical assay (DPPH assay), 2,2′-azino-bis(3- ethylbenzothiazoline-6-sulfonic acid radical scavenging assay (ABTS assay), total reducing assay (TRA), ferric reducing antioxidant potential assay (FRAP assay), total antioxidant capacity assay (TAC assay). The hydroalcoholic extracts (SHA) can be a better choice as compared to water extract (SWA) due to higher yield of extract (13.680 ± 0.548%) and higher antioxidant activity against DPPH assay, ABTS assay, TRA assay, FRAP assay and TAC assay. In hydroalcoholic extract (SHA), ethyl acetate fraction (SEA) showed most potent activity against DPPH (IC50 = 0.008 ± 0.002 mg/mL) and ABTS (0.025 ± 0.001 mg/mL). n-Hexane fraction of SHA showed higher FRAP (28.664 ± 3.153 μmol/mL) and TAC (3.263 ± 0.325 μmol/mL) value (equivalent to ascorbic acid in μmol/mL) but showed very low yield (0.468 ± 0.018%), SBU showed higher TRA value (0.413 ± 0.309 mg/mL). The ethyl acetate fraction (SEA) can be a choice for antioxidant as it showed second highest FRAP (19.547 ± 2.119 μmol/mL) and TAC (2.750 ± 0.466 μmol/mL) with better yield (2.473 ± 0.594%) as compared to n-hexane (SH) fractions (0.468 ± 0.018%).

Quality Assessment and Antioxidant Activities of the Blossoms of Inula Nervosa Wall.

Currently, although Inula nervosa Wall is substantially investigated, little is understood about blossoms of Inula nervosa Wall (BINW). In this work, we systematically investigated the antioxidant activity of the extract from BINW by various standard assays including 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical ability, 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) di-ammonium salt radical cation (ABTS), and ferric reducing antioxidant potential (FRAP). Chemical compounds were tentatively identified through an UHPLC-QTOF-MS system. Furthermore, the contents of nine compounds were detected with UHPLC method coupled with photodiode array (PDA) detector. By carefully analyzing the quantitative data via clusters analysis and principal component analysis (PCA). Forty-six compounds were tentatively identified, and our results showed that nine compound samples in 21 batches of BINW collected from different areas could be differentiated and analyzed by a heatmap visualization. In addition, the contents of nine compounds (flavonoids, phenolic acids) exhibited a total of higher amounts and better antioxidant activities from Yunnan than those from the other three origins. Our study not only developed a powerful platform to explain the difference between traditional Chinese medicines species that are closely related through the chemometric and chemical profiling, but also presented a useful method to establish quality criteria of BINW with multiple origins. To characterize the BINW in detail, we not only performed DPPH, FRAP, and ABTS assays to investigate its antioxidant activity, but also established UHPLC-QTOF-MS/MS- and UHPLC-PDA-based methods to comprehensively identify and qualitatively analyze its components.

Analyses of In-vitro antioxidant and anticancer activity of Cissus quadrangularis stem extract in osteoblastic cell line -UMR- 106

Cancer is one of the leading causes of death worldwide with an estimated 7.8 lakh deaths in 2018 in India. Osteosarcoma, a primary tumour of the bone is the second highest cause of bone cancer related death in young people. It is treated with chemotherapy, radiation, and surgery which cause side effects. Certain traditional plants may have antitumour activities. This research aimed to evaluate the invitro antioxidant and apoptosis induced activity of stem extract of cissus quadrangularis (CQ) on osteoblastic cell line (UMR-106). Various extracts of CQ was subjected to phytochemical screening and antioxidant studies. Ethanolic extract of CQ showed maximum phytoconstituents and better antioxidant activity for both DPPH(2,2-diphenyl-1-picryl-hydrazyl-hydrate)and FRAP(Ferric reducing antioxidant potential) assay. The UMR-106 cell line was treated with different concentrations of the extract ranging from 25μg to 400μg for varying time periods. The dose and time optimization of the ethanolic extract was evaluated in the osteoblastic cell line (UMR-106) using MTT assay (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) which showed IC50 value with 200μg/ml at 24 hours. There was a reduction in the percentage of cells by 50% in the treated group by SRB assay showing the cytotoxic potency of the extract. Flow cytometry analysis indicated that the extract of CQ induced apoptosis in UMR-106 cell line. Ethanolic extract of CQ has antioxidant effects, also inducing G0/G1 arrest and apoptosis in osteoblasts, suggesting that this might represent a novel and effective agent against bone cancer.

The Impact of Maltodextrin and Inulin on the Protection of Natural Antioxidants in Powders Made of Saskatoon Berry Fruit, Juice, and Pomace as Functional Food Ingredients.

The objective of this study was to examine the effect of inulin and maltodextrin applied during vacuum drying of Saskatoon berry fruit, juice, and pomace on the retention of bioactive compounds and antioxidant capacity (radical scavenging capacity (ABTS), ferric reducing antioxidant potential (FRAP)) of powders obtained. Ultra-high performance liquid chromatography (UPLC-PDA-ESI-MS/MS) was used to identify major groups of polyphenolic compounds, such as: flavan-3-ols (35% of all polyphenols for fruit powder, 33% for juice powder, and 39% for pomace powders of all polyphenols), anthocyanins (26% for fruit powder, 5% for juice powder, and 34% for pomace), phenolic acids (33% for fruit powder, 55% for juice powder, and 20% for pomace powder), and flavanols (6% for fruit powder, 6% for juice powder, and 7% for pomace powder). In general, the content of polyphenols was more dependent on the content than on the type of carrier used for drying, regardless of the matrix tested. The average sum of polyphenols and the antioxidant activity (for ABTS and FRAP assay) of the powders with 30% of carrier addition were 5054.2 mg/100 g dry matter (d.m.) as well as 5.3 and 3.6 mmol Trolox/100 g d.m. in the ABTS and FRAP tests, respectively. The increase in carrier concentration by 20% caused a decrease of 1.5-fold in the content of polyphenols and a 1.6-fold and 1.5-fold in the antioxidant potential, regardless of the matrix tested. The principal component analysis (PCA) analysis indicated that the freeze-drying process led to the lowest degradation of the identified compounds, regardless of the matrix tested, with the exception of juice and pomace powders dried by vacuum drying at 60 °C. In this case, the release of (−)-epicatechin was observed, causing an increase in the flavanol contents. Thus, this work demonstrated the effect of processing and matrix composition on the preservation of antioxidant bioactives in Saskatoon berry powders. Properly designed high-quality Saskatoon berry powders with the mentioned carriers may be used as nutraceutical additives to fortify food products and to improve their functional properties.

The ramification of fermentation time on antioxidant properties of Napier grass herbal tea by black tea processing method

Tea fermentation relates to the enzymatic oxidation of the leaves of the tea and the formation of dark pigments. This method is usually conducted in a setting where temperature and humidity can be regulated. However, the impact of fermentation time on the antioxidant characteristics of Napier grass herbal tea is still unanswered. This study aimed to determine the effect of fermentation time on antioxidant properties of Napier grass black tea. Napier grass was subjected to fermentation for 3, 6, 9, 12 and 24 hrs. Fresh and dried samples were extracted in water (95°C, 30 mins) and the extracts were then analysed by total phenolic content (TPC) assay, total flavonoid content (TFC) assay, diphenyl-picryl-hydrazyl (DPPH) assay, Ferric reducing antioxidant potential (FRAP) assay, ferric thiocyanate (FTC) method and thiobarbituric acid (TBA) method. Sample fermented for 9 hrs showed the highest results in TPC (18.32±0.38) and FRAP assay (91.00±2.78). Whereas, the highest flavonoids and antioxidant activity were found in dried sample without fermentation (control) in TFC (97.82±13.00) and DPPH (90.02±1.11). However, samples fermented for 3 to 9 hrs showed no significant difference with that of the fresh sample in TFC and DPPH indicating 3 to 9 hrs fermentation time did not affect the antioxidant properties of Napier grass. In FTC and TBA, all the fermented samples showed lower antioxidant activity than that of fresh samples and control. The results suggested that 9 h of fermentation time is suitable for the preparation of Napier grass black tea as it exhibited high antioxidant properties in TPC, TFC, DPPH and FRAP assays.

Effect of steaming time on antioxidant properties of Napier grass herbal tea by green tea processing method

This study aimed to determine the effect of steaming time on antioxidant properties of Napier grass green tea. Napier grass was subjected to steaming for 1, 2, 3, 4, 6, 8 and 10 mins. Fresh and dried samples were extracted in water (95°C, 30 mins) and the extracts were then analysed by total phenolic content (TPC) assay, total flavonoid content (TFC) assay, diphenyl-picryl-hydrazyl (DPPH) assay, Ferric reducing antioxidant potential (FRAP) assay, ferric thiocyanate (FTC) method and thiobarbituric acid (TBA) method. Sample steamed for 8 mins showed the highest TPC (18.32±0.26), TFC (152.71±5.74) and 109.88±5.44 in FRAP assay. High antioxidant activity was found in sample steamed for 3 to 10 min (81.63±1.19 to 83.50±1.10) in DPPH which were not significantly different with the fresh sample indicating steaming can retain the phytochemical compounds. Samples undergone 6 to 10 mins steaming time were found to have high lipid peroxidation in ferric thiocyanate (75.02±2.96 to 81.01±6.68) and thiobarbituric acid (85.99±1.56 to 86.21±1.44) assays. The results suggested that 8 mins of steaming time is suitable for Napier grass green tea as it exhibited the greatest antioxidant properties

Health-Promoting Properties of Brazilian Unconventional Food Plants

This study lies on the unexplored and unprecedented opportunities for the discovery of Unconventional Food Plants (UFPs) rich in nutrients and bioactive compounds. UFPs grow spontaneously and have been consider an important food sources and demonstrate health-promoting properties. This study aimed to investigate the phytochemical profiles together with the anti-inflammatory and antioxidant in vitro activities of the Brazilian UFPs Hypochaeris chillensis, Emilia fosbergii and Emilia sonchifolia. Preliminary secondary metabolites were identified by LC–MS/MS analysis. Antioxidant activity was determined by the ABTS cation radical scavenging capacity, ferric reducing/antioxidant potential (FRAP), and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assays. The effects of UFPs on nitric oxide (NO), superoxide anion generation (O2•−), and pro-inflammatory cytokine (TNF-α and IL-6) production and on nuclear factor kappa B (NF-κB) activity was determined using Griess reagent, immunoenzymatic assay kits (ELISA) and chemiluminescence measurements in cell-based assays, respectively. Phytochemical analysis revealed significant amount of total phenolic content in all of the studied plants. E. fosbergii showed the highest antioxidant capacity in all of the chemical assays, exhibiting IC50 values of 32.9 ± 4.8, 50.2 ± 2.3 and 24.0 ± 2.9 µg mL−1 in the ABTS, DPPH and FRAP assays, respectively. The studied UFPs showed a significant intracellular reduction in NO and O2•− production in LPS-stimulated RAW 264.7 macrophages. Additionally, UFPs suppressed the production of the pro-inflammatory cytokines TNF-α and IL-6 in a dose-dependent manner. Moreover, E. sonchifolia suppressed NF-κB activity. Altogether, the investigated UFPs exhibited promising bioactive compounds that were capable of neutralizing free radicals, controlling oxidative stress and modulating the inflammatory process.

(2E)-2-Benzylidene-4,7-dimethyl-2,3-dihydro-1H-inden-1-one (MLT-401), a novel arylidene indanone derivative, scavenges free radicals and exhibits antiproliferative activity of Jurkat cells

Abstract Background The arylidene indanone scaffold has contributed many lead molecules in chemotherapeutic anticancer agent research. Objectives To determine the oxidant-scavenging activities and antiproliferative activity of (2E)-2-benzylidene-4,7-dimethyl-2,3-dihydro-1H-inden-1-one (MLT-401), an arylidene indanone derivative. Methods Jurkat cells, primary lymphocytes, and Vero cells were treated with MLT-401. Antioxidant properties of MLT-401 were determined using 2,2-diphenyl-1-picrylhydrazyl (DPPH)-based, 2,2′-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS)-based, and ferric-reducing antioxidant potential (FRAP) assays. Inhibition of cell proliferation was determined using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide-based assay. Nuclear status was determined using a DNA fragmentation assay, and cell cycle stage was analyzed by flow cytometry. Mitochondrial membrane enzyme activities were measured using colorimetric methods. Results The antioxidant assays gave MLT-401 half maximal inhibitory concentration (IC50) values of 1611 nM (DPPH-based assay), 2115 nM (ABTS-based assay), and 1586 nM (FRAP assay). MLT-401 inhibited proliferation of Jurkat cells with a concentration for 50% of maximal inhibition of cell proliferation (GI50) of 341.5 nM, being 12- and 9-fold less than GI50 concentrations for normal lymphocytes and Vero cells, respectively. MLT-401 caused nuclear fragmentation and DNA laddering as seen by electrophoresis. Jurkat cells showed a time-dependent accumulation of sub G0/G1 cells after MLT-401 treatment. Mitochondrial membrane-bound Na+/K+ ATPase, Ca2+ ATPase, and Mg2+ ATPase activities were inhibited by MLT-401 in a dose-dependent manner. Conclusion MLT-401 possesses significant antiproliferative activity and scavenges free radicals released through mitochondrial membrane damage in a Jurkat cell line model of cancer cells. Further investigation of MLT-401 as a chemotherapeutic anticancer agent and development of other arylidene indanone analogues are warranted. A detailed elucidation of mechanistic pathways is required for further development.

Antioxidant Properties of Stingless Bee Honey and Its Effect on the Viability of Lymphoblastoid Cell Line

Research on the medical benefit of stingless bee honey (kelulut honey) is rather new although it has been used as traditional food and additive for ages. The primary objective of our study was to evaluate the antioxidant properties of kelulut honey and its effect on lymphoblastoid cell line. We analysed the antioxidant properties of kelulut honey by ferric reducing antioxidant potential assay, total phenolic and flavonoid contents using UV spectrophotometry. The total phenolic content, total flavonoid content and ferric reducing antioxidant potential of Malaysian kelulut honey produced by Trigona spp. were found to be 844.45 mg RE/kg honey, 78.29 mg RE/kg honey and 1132.66 mM FE/kg honey, respectively. Our findings showed a strong correlation between total phenolics and flavanoids contents with its antioxidant potential at R2 = 0.920 and R2 = 0.951, respectively. The effect of honey on cell viability of lymphoblastoid cell line (LCL) was also investigated. The cells were cultured in RPMI-1640 medium supplemented with 0 - 500 μg/mL of kelulut honey for 24 hours. Cell viability was quantitated using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, MTS assay showed that honey supplementation boosted the viability of LCL up to 164.64% (p< 0.01). The significant increase in cell viability might be modulated by the antioxidant properties of kelulut honey.

A comparison of the performance of the cupric reducing antioxidant potential assay and the ferric reducing antioxidant power assay for the analysis of antioxidants using reaction flow chromatography

The cupric reducing antioxidant capacity (CUPRAC) assay is an established procedure used to measure antioxidant content. Recently, the CUPRAC assay was adapted for use in post column derivatisation coupled with high performance liquid chromatography (HPLC-PCD). In prior works reported in the literature, CUPRAC HPLC-PCD assays used large post column mixing coils leading to a deterioration in the observed separation efficiency. In this work we used the CUPRAC PCD assay with reaction flow (RF) chromatography. No reaction loop was required, thus preserving the efficiency of the separation. We subsequently compared the performance of the CUPRAC assay to that of an antioxidant assay that we have previously converted to RF chromatography, namely the ferric reducing antioxidant potential (FRAP) assay, both in RF mode. The relative response factor with respect to trolox was measured for both RF assays via the separation of a standard mixture of 16 compounds. Sample analysis of coffee and green tea via both RF assays was used to compare their ability to screen for bioactive candidates in complex natural products. In general, the CUPRAC assay is recommended as the RF PCD methodology of choice as it provided a higher level of performance compared to the FRAP assay, demonstrating less baseline noise interference, greater sensitivity, wider linear dynamic range and better assay precision.

Antioxidant Activity of Musa Paradisiaca (Banana) Soft Pith and Its Cytotoxicity Against Oral Squamous Carcinoma Cell Lines

Musa paradisiaca also known as banana possess great medicinal value such as haematopoietic, immunomodulatory, antifungal, anti-diabetic and antioxidant effects. Antioxidant and antiproliferative activities between two different species, banana soft pith (BSP) Awak and Abu were conducted in this study. Briefly, BSP were collected (Baling, Kedah) and extracted by 70% ethanol forming crude extract. The crude extracts were then fractionated by n-hexane, ethyl acetate, butanol and water. All BSP extracts and fractions were subjected to total phenolic content (TPC), total flavonoid content (TFC), 2,2-diphenyl-1-picrylhydrazil (DPPH) free radical scavenging assay, ferric reducing antioxidant potential (FRAP) assay and cytotoxic assay screening (MTT). Fraction ethyl acetate from BSP Abu showed the highest cytotoxic activity towards HSC-4 cell lines (26.95 ± 1.80 µg/mL) and the highest TPC (12.25 ±0.39 mg GAE/g) and FRAP assay (191.03 ± 0.83 mg FeSO4/g) values as compared to other fractions among BSP Abu. However, TPC (13.93 ± 0.28 mg GAE/g) and TFC (441.59 ± 19.31 mg QE/g) values of BSP Awak were found to be higher as compared to BSP Abu.

Antioxidant Activity and Bioactive Compounds of Babassu (Orbignya phalerata) Virgin Oil Obtained by Different Methods of Extraction

Background:The investigation of new sources of raw materials and the knowledge of the composition of the food is fundamental for the evaluation of their potential and the availability of nutrients for the consumer population.Objective:This work aimed to deepen the knowledge about the crude oil of babassu fruit obtained by two different methods of extraction, cold pressing and extraction by cooking the fruit almond.Method:Total phenolic compounds contents and antioxidant activity were determined by ferric reducing antioxidant potential assay and 2,2-diphenyl-1-picrylhydrazyl radical scavenging capacity assay. By liquid chromatography, the content of different bioactive compounds was determined. Data was submitted to Analysis of Variance (ANOVA) and compared by f test (p &lt;0.05).Results:The results showed that for most of the bioactive compounds there was no difference between the two types of babassu oil. For those compounds where the oils differed, the virgin oil had about three times the content of the extra-virgin oil. In addition, the antioxidant activity was higher for the oil extracted by cooking of the babassu mass, ranging from approximately 2.5 times higher up to 19.2 times higher than the antioxidant activity of the babassu oil extracted by pressing.Conclusion:The process of extraction by cooking the almond mass can incorporate a larger number of bioactive components and improve the antioxidant activity of the virgin babassu oil. However, the extraction method does not influence the content of tocopherols of distinct types of babassu oil.

Effect of compost and NPK fertilizer on improving biochemical and antioxidant properties of Moringa oleifera

Organic and inorganic fertilizers play a significant role in improving the nutritional quality of Moringa plant (Moringa oleifera). A field experiment of six treatments (control; 5 t/ha Compost; 120 kg/ha NPK; 50 + 50%/ha (NPK + Compost); 10 + 50%/ha (NPK + Compost); and 50 + 10%/ha (NPK + Compost) with three replications was conducted in a randomized complete block design in order to evaluate the effect of the application of compost and NPK fertilizer in various combinations on the biochemical and antioxidant potential of the Moringa plant. The results showed a significant influence of the compost and NPK fertilizer in various combination on the biochemical and antioxidant properties of the plant. Higher amount of carbohydrate, phenolic and flavonoid contents (406.24, 45.23 and 1.69 mg g−1 dry wt., respectively) were observed in the plant treated with 50 + 50%/ha (NPK + Compost). Protein content (27.91 mg g−1 dry wt.) was improved by the application of 120 kg/ha NPK fertilizer. The antioxidant properties such as, DPPH, ABTS and nitric oxide were analyzed as percentage scavenging and half maximal effective concentration (EC50) whereas ferric-reducing antioxidant potential (FRAP) was determined in terms of absorbance at 700 nm and EC0.50. The half maximal effective concentrations in combined application of NPK and compost (50 + 50%/ha (NPK + Compost)) were the lowest among the fertilizer treatments, resulting in EC50 value of 27.61, 17.18, 104.24 μg mL−1 for DPPH, ABTS and nitric oxide assays, respectively whereas in case of the FRAP assay the EC0.50 value as found out to be 292.35 μg mL−1.