Ferric Ion Reducing Power Assay Research Articles

IN VITRO ANTIOXIDANT AND ANTICANCER ACTIVITY OF MACRANGA PELTATA LEAF EXTRACTS ON LUNG CANCER CELL LINES

Objective: To determine the phytochemical constituents, antioxidant and anticancer activities of Macranga peltata leaf extracts on A549 and P 51 human lung cancer cell lines.
 Methods: Leaf sample was subjected to soxhlet extraction method and phytochemical screening was done using different biochemical tests. The antioxidant activity was tested using 2,2–diphenyl-1-picrylhydrazyl, ferric ion reducing power assay and 2,2–azino bis 3–ethyl benzothiazoline–b–sulfonic acid assay. In vitro anticancer activity on A549 and P 51 human lung cancer cell lines was evaluated by (3-(4,5-dimethyl thiazole-2yl)-2,5–diphenyl tetrazolium bromide) MTT assay.
 Results: Phytochemical screening confirmed the presence of phytoconstituents like tannins, flavanoids, alkaloids, saponins, glycosides and reducing sugar. The antioxidant assays exhibited significant inhibiting activity. MTT cell proliferation assay of methanolic extract of plant drug produced a cytotoxic effect on lung cancer cell lines A549 and P 51 with an IC50 value of 197.10 µg/ml, while it had no cytotoxic effect on normal cell lines.
 Conclusion: It has been reported the effect of flavanoid characteristics in reducing the cancer. M. peltata contains a wide variety of secondary metabolites that hold strong antioxidant capacity. Their leaf have found to be potential source of antioxidants and anticancer activity.

Elite Zingiber extracts as potential free radical scavengers and radioprotectors

ABSTRACT Fresh rhizome extracts of four elite Zingiber species, viz., Zingiber kangleipakense, Zingiber montanum, Zingiber officinale, and Zingiber zerumbet, prepared in 60% aqueous ethanol/60% aqueous methanol solutions were used for assessing their comparative free radical scavenging and radioprotective properties. Various methods, such as ferric ion reducing power assay, DPPH ion scavenging assay, nitric oxide scavenging assay, metal chelating assay, thiyl/sulfur free radical reactivity assay, and hydroxyl ion scavenging assay, were employed for radical scavenging capabilities. Bio-active principles, such as total phenolic, flavonoid, and ascorbic acid contents, were also assessed. Z. officinale showed the highest TPC, TFC, and ascorbic acid content of 650 mg GAE/100 g, 889 mg QE/100 g, and 3.19 mg/100 g, respectively, as well as the greatest efficacy in ferric ion reducing and DPPH scavenging ability having lowest IC50 value of 0.22 mg/mL. Z. montanum showed the lowest IC50 value of 0.91 mg/mL in NO scavenging assay and also exhibited the highest curcumin protection in thiyl free radical scavenging assay. Z. kangleipakense showed the lowest IC50 value of 0.24 mg/mL and 0.25 mg/mL in metal ion chelating assay and hydroxyl radical assay, respectively. 60% aqueous ethanol extracts of Z. officinale and Z. kangleipakense were used for radioprotective assays both for in vitro and in vivo using plasmid pBR322 DNA and bone marrow cells of albino rats, respectively, based on their significant free radical scavenging properties. At the same concentration of 0.25 mg/mL, Z. officinale and Z. kangleipakense showed 71.50% and 56.54% protection, respectively, of DNA from exposure to 6 Gy gamma radiation. Also, Z. officinale and Z. kangleipakense exhibited 51.16% and 38.96% protection from radiation-induced chromosomal damages in rats, respectively, when exposed to 2 Gy. Reduction in the DNA open circular form as well as the chromosomal aberrations in treatments with plant extracts compared to the controls confirmed the radioprotective potentials of the extracts. The results showed that the extracts possessed potential antioxidant compounds having capacities to scavenge radiation-induced free radical species. The accessibility of the extracts in radiation countermeasures can provide immense nutraceutical potentials.

Evaluation of in vitro antioxidant and Antityrosinase activities of Ixora coccinea Linn. Roots

Ixora coccinea Linn. Commonly known as ‘vetchi’ is a flowering shrub of the Rubiaceae family used in the treatment of various ailments in the Ayurvedic system of medicines. This research aimed to study the qualitative phytochemistry and to determine the antioxidant and antityrosinase activities of Ixora coccinea roots. Phytochemical analysis indicated the presence of various plant bioactive metabolites in Ixora coccinea roots. Different solvent extracts were tested for their in vitro antioxidant potential using a 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay and ferric ion-reducing power assay. The total phenolic content (TPC) of the extracts was determined by spectrophotometric methods. Methanolic and aqueous extracts of the roots were evaluated for antityrosinase activity by inhibitory potency of mushroom tyrosinase. The methanolic extracts of Ixora coccinea roots possessed the highest DPPH radical scavenging activity with an IC50 value of 26.88μg/ml compared with aqueous and ethyl acetate extracts. The estimation of total phenolic content revealed that methanolic extract has the maximum phenolic content and exhibited the highest ferric reducing power compared with the other two extracts. The methanolic extract of Ixora coccinea roots showed a comparable antioxidant activity with that of standard ascorbic acid and also exhibited potent tyrosinase inhibition with an IC50value of 120.03μg/ml using Kojic acid as a reference standard. The current study showed that the methanolic extract of Ixora coccinea roots have superior skin whitening and antioxidant activities and could be used as a potential source of natural ingredient for formulating cosmetic products.

Integrating of Aqueous Enzymolysis with Ethanolic Ultrasonication to Ameliorate Cordycepin Content from Vietnamese Cordyceps militaris and Biological Analysis of Extracts

AbstractIn this study, the employment of aqueous enzymolysis using alcalase with ethanolic ultrasonication extraction method was investigated. The cordycepin content was analyzed by using reversed‐phase high‐performance liquid chromatography with a diode array detector. The result showed that the highest cordycepin content of 0.4365±0.0309 mg/g could be obtained under extraction conditions including total enzyme volume of 240 μL, incubation time of 50 min, incubation temperature of 55 °C, and ultrasonic power of 840 W. The comparative studies of antioxidant activities of the extract were determined in terms of the DPPH free radical scavenging assay, ferric ion reducing power assay, and H2O2 scavenging assay. Additionally, the MTT assay was conducted to evaluate the anticancer activities of the extract, illustrating the cytotoxic property against Hep‐G2 cancer cells with the half‐maximal inhibitory concentration (IC50) value of 12.20±0.03 μg/mL compared to that of 0.36±0.03 μg/mL for ellipticine, MCF‐7 cancer cells with the IC50 value of 11.60±0.09 μg/mL compared to that of 0.51±0.04 μg/mL for ellipticine, and Lu cancer cells with the IC50 value of 34.50±0.25 μg/mL compared to that of 0.32±0.03 μg/mL for ellipticine.

Rapid Green Synthesis of Biogenic Silver Nanoparticles Using Cinnamomum tamala Leaf Extract and its Potential Antimicrobial Application Against Clinically Isolated Multidrug-Resistant Bacterial Strains.

Cinnamomum tamala is Indian bay leaves also known as Tej patta commonly used in the preparation of delicious food for its sweet aroma and tremendous medicinal values. In this study, the significant concentration-dependent free radical scavenging and antioxidant efficacy of the aqueous extracts of bay leaves has been determined using DPPH (2, 2-diphenyl-l-picrylhydrazyl) radical scavenging, ferric ion-reducing power assay, and hydrogen peroxide radical scavenging assay. The leaf extract has also been utilized in the rapid synthesis of silver nanoparticles (AgNPs) under mild conditions (30min reaction time at 70°C) without the addition of extra stabilizing or capping agents. Mostly spherical shaped particles were formed with diameter ranging from 10 to 12nm as evident by HRTEM imaging. The silver nanoparticles were also characterized using FTIR, XRD, and UV-visible spectroscopic techniques. The antibacterial effect of the synthesized AgNPs was studied against three clinically isolated multidrug-resistant bacterial strains (Escherichia coli (EC-1), Klebsiella pneumonia (KP-1), and Staphylococcus aureus (SA-1)). The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of AgNPs against EC-1 were 12.5 and 15μg/mL and in SA-1 were 10 and 50μg/mL, and in the case of KP-1, both values were 12.5μg/mL. It was also noted that 8h treatment duration using AgNPs was sufficient to eliminate all types of bacterial growth as evidenced by time-dependent killing kinetic assays. The biocompatibilities of AgNPs were also tested against human health RBCs, and it was observed that it did not show any significant toxicity up to 50μg/mL concentration.

Neuroprotective and anti-inflammatory activities of Aegiceras corniculatum (L.) Blanco

Objectives: The objective of the study is to evaluate the antioxidant, anti-inflammatory, and neuroprotective potential of Aegiceras corniculatum (L.) Blanco. Materials and Methods: Sequential Soxhlet extraction of titled plant material was carried out. Phytochemical analysis was performed using standard procedures. The total phenolic and flavonoid content was estimated along with in vitro antioxidant property of the extracts which was assessed by ferric ion-reducing power assay, phosphomolybdenum assay, 2,2-diphenyl-2-picrylhydrazyl assay, and hydrogen peroxide scavenging assay. Further, the extracts were assessed for anti-inflammatory property by in vitro protein denaturation assay, whereas neuroprotective activity was performed by pretreatment 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Results: Qualitative phytochemical analysis revealed the presence of alkaloids, phenols, flavonoids, tannins, anthraquinones, glycosides, lignins, and sterols. Quantitative phytochemical analysis revealed that the aqueous extract possessed high phenolic content and methanol extract possessed high flavonoid content when compared to other extracts. All the extracts showed antioxidant activity revealed through different assays. Among all the extracts, methanol extract showed significant anti-inflammatory activity with 85.32 ± 1.63 percentage inhibition of protein degradation. Methanol extract at the concentration of 5 μg showed comparatively significant neuroprotection against trimethyltin (TMT)-induced toxicity with 70.02% SK-S-NH cell survival against 56.30% cell survival TMT-intoxicated group. Conclusion: The leaf extracts of A. corniculatum possess antioxidant, anti-inflammatory, and neuroprotective properties.

Ionic Liquid⁻Ultrasound-Based Extraction of Biflavonoids from Selaginella helvetica and Investigation of Their Antioxidant Activity.

As a new and green solvent, ionic liquids (ILs) have received more attention during the green extraction and separation process for natural medicines. In this paper, IL-ultrasound-assisted extraction (IL-UAE) of total biflavonoids (TBFs) from Selaginella helvetica was firstly developed, and different ILs were employed and compared. Based on single-factor experiment, solid–liquid ratio (1:10–1:14 g/mL), IL concentration (0.6–1.0 mmol/mL), and extract temperature (40–60 °C) were further explored, according to response surface methodology (RSM), with TBF yields as the index. Moreover, antioxidant activity of TBF extract was analyzed by four methods, i.e., 2,2-di(4-tert-octylphenyl)-1-picrylhydrazyl (DPPH) and 2,2′-azinobis-(3-ethylbenzth-iazoline-6-sulphonate (ABTS) free radical scavenging assay, ferric ion reducing power assay, and chelation of ferrous ions assay. The results indicated that [C6mim]PF6 had a high selectivity and efficiency. Moreover, important parameters for the extraction process were investigated and optimized. Through parameter optimization (0.8 mmol/L, 250 W, 40 min, 1:12.7 g/mL, and 47 °C), a yield of 18.69 mg/g biflavonoids was obtained from the extract of S. helvetica. Compared with ethanol-UAE, heat-reflux extraction, Soxhlet extraction, and percolation extraction, IL-UAE could not only obtain higher yield in a shorter time, but also reduce the solvent consumption. In addition, TBF extract showed potential antioxidant activity based on the above four antioxidant methods. In short, IL-UAE was first employed to develop a novel and green extraction method for TBF content, and this experiment provides valuable references for further utilization of S. helvetica.

Evaluation of Antioxidant Activities of Alpinia galanga (L.) Willd

Present research was designed to evaluate the free radical scavenging capacities and antioxidant activities of rhizome extracts of Alpinia galanga prepared in different solvent systems (60% aqueous methanol, 60% aqueous ethanol and distilled water) using different in vitro chemical assays. Antioxidant components such as total phenolic content (TPC), total flavonoid content (TFC) and ascorbic acid contents of the ginger species were screened. Antioxidant assays employed included sulphur free radical reactivity assay, ferric ion reducing power assay, DPPH free radical scavenging capacity assay, hydroxyl radical scavenging assay, nitric oxide scavenging activity assay and hydrogen peroxide scavenging assay. The obtained data reveal that the plant extracts contained significant amount of the observed antioxidant components and also exhibited significant free radical scavenging capacities. Methanol (60%) extract exhibited highest antioxidant activity than other solvents. The polyphenolic constituents of the plant extracts appear to be largely responsible for the radical scavenging capacity. The plant extracts act as promising source of antioxidants, and may be useful for development of nutraceuticals and pharmaceutical drugs.

English

The study was conducted to evaluate the preservative potentials of essential oils (EOs) of Piper guineense, Xylopica aethiopica and Tetrapleura tetraptera in mixed fruit juice and determine their antioxidant capacity. The preservative effect at varying concentrations was evaluated using S. cerevisiae, S. aureus and B. cereus as test isolates. The antioxidant properties were determined using 1,1- diphenyl-2-picrylhydrazyl (DPPH) and ferric ion reducing power assay (FRPA) methods. Concentrations (10 and 5%) of P. guineense and X. aethiopica EOs reduced the number of S. cerevisiae by one log cycle on the 10th day of storage, while 3.3% reduced it by one log cycle after 14 days. The 10% concentration produced a log cycle reduction in the growth of S. aureus and B. cereus on the 12th day. However, 5% concentration reduced the growth of B. cereus by one log cycle on the 14th day. T. tetraptera had the largest amounts of vitamin A (544.41 iu/100 mL), phenol (1.81%) and flavonoids (0.45%). P. guineense had the highest value for vitamin E (37.03 iu/100 mL). X. aethiopica EO produced the highest scavenging activity (46.04%), while P. guineense EO produced the strongest reducing power activity (38.64%). The EOs can act as natural preservative in mixed fruit juice. Their inclusion in such products can improve the products thereby serving as functional foods/beverages, and thus can reduce the risks of cancer formation and chronic diseases in humans. Key words: Mixed fruit juice, preservation, microbial isolates, essential oils, antioxidant ability.

Development of Phaleria macrocarpa (Scheff.) Boerl Fruits Using Response Surface Methodology Focused on Phenolics, Flavonoids and Antioxidant Properties.

In this study, the optimal conditions for the extraction of antioxidants from the Buah Mahkota Dewa fruit (Phaleria macrocarpa) was determined by using Response Surface Methodology (RSM). The optimisation was applied using a Central Composite Design (CCD) to investigate the effect of three independent variables, namely extraction temperature (°C), extraction time (minutes) and extraction solvent to-feed ratio (% v/v) on four responses: free radical scavenging activity (DPPH), ferric ion reducing power assay (FRAP), total phenolic content (TPC) and total flavonoid content (TFC). The optimal conditions for the antioxidants extraction were found to be 64 °C extraction temperature, 66 min extraction time and 75% v/v solvent to-feed ratio giving the highest percentage yields of DPPH, FRAP, TPC and TFC of 86.85%, 7.47%, 292.86 mg/g and 3.22 mg/g, respectively. Moreover, the data were subjected to Response Surface Methodology (RSM) and the results showed that the polynomial equations for all models were significant, did not show lack of fit, and presented adjusted determination coefficients (R2) above 99%, proving that the yield of phenolic, flavonoid and antioxidants activities obtained experimentally were close to the predicted values and the suitability of the model employed in RSM to optimise the extraction conditions. Hence, in this study, the fruit from P. macrocarpa could be considered to have strong antioxidant ability and can be used in various cosmeceutical or medicinal applications.

Total phenolic, flavonoid contents, in-vitro antioxidant activities and hepatoprotective effect of aqueous leaf extract of Atalantia ceylanica.

BackgroundDecoction prepared from leaves of Atalantia ceylanica is used in traditional medicine in Sri Lanka for the treatment of various liver ailments since ancient times. Lyophilized powder of the water extract of A. ceylanica leaves was investigated for its phytochemical constituents, antioxidant and hepatoprotective activity in-vitro.MethodsThe total phenolic and flavonoid contents were determined using Folin Ciocalteu method and aluminium chloride colorimetric assay respectively. The antioxidant activities of the decoction were investigated using 1,1-Diphenyl-2-picrylhydrazyl (DPPH), hydroxyl radical, nitric oxide scavenging assays and ferric ion reducing power assay. Hepatotoxicity was induced on porcine liver slices with ethanol to study hepatoprotective activity. Porcine liver slices were incubated at 37°C with different concentrations of the water extract of A. ceylanica in the presence of ethanol for 2 hours. The hepatoprotective effects were quantified by the leakage of alanine transaminase (ALT), aspartate transaminase (AST) and lactate dehydrogenase (LDH) to the medium. Thiobarbituric acid reactive substances (TBARS) assay was performed to examine the anti-lipid peroxidation activity caused by the plant extract.ResultsThe mean ± SD (n =9) for the levels of total phenolics and flavonoids were 4.87 ± 0.89 w/w% of gallic acid equivalents and 16.48 ± 0.63 w/w% of (-)-Epigallocatechin gallate equivalents respectively. The decoction demonstrated high antioxidant activity. The mean ± SD values of EC50 were 131.2 ± 36.1, 48.4 ± 12.1, 263.5 ± 28.3 and 87.70 ± 6.06 μg/ml for DPPH, hydroxyl radical, nitric oxide scavenging assays and ferric ion reducing power assay respectively.A significant decrease (p <0.05) was observed in ALT, AST and LDH release from porcine liver slices treated with A. ceylanica extract at a concentration of 2 mg/ml in the presence of ethanol (5 M) compared to that of ethanol (5 M) treated slices. Furthermore, a reduction in lipid peroxidation was also observed in liver slices treated with the leaf extract of A. ceylanica (2 mg/ml) compared to that of ethanol induced liver toxicity (p <0.05).ConclusionsThe results suggest that aqueous extract of A. ceylanica exerts hepatoprotective activity against ethanol induced liver toxicity of porcine liver slices which can be attributed to the antioxidant properties possessed by the plant material.Electronic supplementary materialThe online version of this article (doi:10.1186/1472-6882-14-395) contains supplementary material, which is available to authorized users.

Anti‐Microbial Activity and Composition of Manuka and Portobello Honey

Recently renewed interest in the therapeutic properties of honey has led to the search for new antimicrobial honeys. This study was undertaken to assess the antimicrobial activity and composition of a locally produced Portobello honey (PBH) on three bacteria known to infect wounds. Manuka honey (MH) was used for comparative purposes. Broth culture and agar disc diffusion assays were used to investigate the antimicrobial properties of honey. The honeys were tested at four concentrations: 75%, 50%, 10% and 1% (v/v) and compared with an untreated control. The composition of honey was determined by measuring: polyphenol content by Folin Ciocalteau method, antioxidant capacity by ferric ion reducing power assay, hydrogen peroxide (H2 O2 ) by catalase test, pH and sugar content by pH strips and refractometer, respectively. Both honeys at 75% and 50% inhibited the majority of the three bacteria tested. 10% PBH exhibited antimicrobial activity to the lesser extent than 10% MH. The difference was very significant (p ≤ 0.001). Both honeys were acidic with pH 4, and both produced H2 O2 . The sugar content of PBH was higher than MH, but the difference was not significant. The MH had significantly higher levels of the polyphenols and antioxidant activity than PBH.