• Antioxidant, Anticancer and Antimicrobial, Effects of Rubia cordifolia Aqueous Root Extract

      Barlow, R.; Barnes, D. A.; Campbell, Anna M.; Nigam, Poonam S.; Owusu-Apenten, Richard K.; University of Chester, University of Ulster (Sciencedomain international, 2015-11-10)
      Aims: To evaluate the total antioxidant capacity (TAC) of Rubia cordifolia root extracts, to test anticancer activity against MDA-MB-231 breast cancer cell lines, and to evaluate antimicrobial activity of the same extract versus six Gram-positive and negative bacteria. Study Design: In vitro. Place of Study and Duration: School of Biomedical Sciences, Ulster University, July 2014-Sept 2015. Methodology: TAC was tested using ABTS, DPPH, FRAP and Folin assays and values were expressed as mg-gallic acid equivalents per 100 g (GAE/100 g) of sample. Anticancer properties were examined against MDA-MB-231 breast cancer cell lines using Sulforhodamine B assay. Antimicrobial activity was examined using a disk diffusion assay with three Gram-positive (Staphylococcus epidermidis, Staphylococcus aureus and Bacillus cereus) and three Gram-negative (Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi) bacteria. Results: TAC of dry extracts of Rubia cordifolia ranged from 523±43 to 4513±208 (mg GAE mg/100 g) depending on the method of analysis, ABTS> FRAP> Folin > DPPH methods. R. cordifolia dry extract showed cytotoxicity against MDA-MB-231 with IC50 = 44 µg/ml or 5.1µM GAE. No antimicrobial activity was observed against the three Gram-positive, or three Gram-negative bacterial species using the water extract or R. cordifolia. Conclusion: R. cordifolia aqueous extract possess high total antioxidant capacity but values depend on the method of analysis. R. cordifolia extract inhibits MDA-MB-231 breast cancer cells proliferation but nil anti-bacterial activity was observed for three Gram-positive and three Gram-negative bacterial strains tested.
    • Determination of Iron (III) Reducing Antioxidant Capacity for Manuka Honey and Comparison with ABTS and Other Methods

      Yusof, Hasif I. M.; Owusu-Apenten, Richard K.; Nigam, Poonam S.; Universiti Kebangsaan Malaysia Medical Centre; Ulster University; University of Chester (SCIENCEDOMAIN International, 2018-06-11)
      Aims: Applying multiple assays with trolox as the sole reference compound is a recent AOAC proposal to improve the reliability of total antioxidant capacity determinations. The aim of this study was to evaluate, iron (III) reducing antioxidant capacity (iRAC) for Manuka honey samples and comparisons with ABTS and other well-known assays. Study Design: In-vitro, laboratory-based study. Place and Duration of Study: School of Biomedical Sciences, Faculty of Life and Health Sciences, Ulster University, Cromore Road, Coleraine, BT52 1SA, UK; September 2015-May 2016. Methodology: Manuka honey rated Unique Manuka Factor (UMF) 5+, 10+, 15+, 18+ and a nonrated (NR) sample were analysed using five assays for total antioxidant capacity namely, iRAC, ABTS, DPPH, FRAP, and Folin assays. Values for total antioxidant capacity were normalized as Trolox Equivalent Antioxidant capacity (TEAC) for comparison within and between assays. Results: The TAC were correlated for all methods (R2 = 0.83-0.99) and also correlated with the total phenols content. Actual TEAC value for a given honey ranged by 21-70-fold depending on the assay method with the following general order of increase; DPPH < FRAP (pH 3.6) < iRAC (pH 7.0) <ABTS (pH7) < Folin (pH ~11). The trends in TAC values are discussed alongside of TEAC values for 50 food items and some challenges for comparing different antioxidant methods are highlighted. Conclusion: Total antioxidant capacity of Manuka honey changes in a regular manner probably affected by assay pH. The findings are important for attempts to standardize antioxidant methods as currently applied to foods, beverages and dietary supplements. Further research is recommended to examine the effect of normalizing antioxidant methods for solvent composition and pH.
    • Effect of pH on the Radical Quenching Capacity of Tea Infusions Using the ABTS•+ Assay

      Chan, Yuk Man; Cheng, Nga Kwan; Nigam, Poonam S.; Owusu-Apenten, Richard K.; University of Chester, University of Ulster (Sciencedomain International, 2016-06-22)
      Aims: The aims of this study were to assess the impact of pH on the free radical quenching activity of tea infusions using a modified 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid (ABTS) assay and three antioxidant compounds as reference. Study Design: In-vitro method. Place and Duration of Study: Faculty of Life and Health Science, School of Biomedical Sciences, Ulster University, UK. From Sept 2014 and May 2016. Methodology: Free radical quenching capacity of tea (Earl grey, black tea, Ceylon tea, & green tea) infusions were investigated using persulfate activated ABTS with acetate buffer (pH 4.5) or phosphate buffer saline (pH 7.0) as solvent. Tests were performed using 96-well microplates, 20 µl of sample and 280 µl of ABTS reagent, and calibrated using ascorbic acid, trolox or gallic acid as reference antioxidants. Results: Gallic acid free radical quenching was pH dependent and unsuitable as reference. The free radical quenching capacity of trolox and ascorbic acid was not significantly different at pH 4.5 and pH 7.0. The radical quenching capacity of tea infusions expressed as Trolox Equivalent Antioxidant Capacity (TEAC) or Ascorbic Acid Equivalent Antioxidant Capacity (AAEAC) was greater by 50-300% at pH 7 compared to pH 4.5. Conclusion: The modified ABTS assay is suitable for examining the influence of pH on free radical quenching ability of tea samples. Gallic acid was not a suitable reference compound. The radical quenching capacity of tea infusions increases with rising pH.