Butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), and tert-butylhydroquinone (TBHQ) are synthetic antioxidants used in the food industry. Herein, we describe the development of a novel graphite nanocomposite-based electrochemical sensor for the multiplex detection and measurement of BHA, BHT, and TBHQ levels in complex food samples using a linear sweep voltammetry technique. Moreover, our newly established analytical method exhibited good sensitivity, limit of detection, limit of quantitation, and selectivity. The accuracy and reliability of analytical results were challenged by method validation and comparison with the results of the liquid chromatography method, where a linear correlation of more than 0.99 was achieved. The addition of sodium dodecyl sulfate as supporting additive further enhanced the LSV response (anodic peak current, Ipa) of BHA and BHT by 2- and 20-times, respectively.
The present study aims to assess the antioxidant activities (AOA) and total phenolic content (TPC) of water extracts of selected edible wild mushrooms: Pleurotus porrigens, Schizophyllum commune, Hygrocybe conica, and Lentinus ciliatus. The AOA were evaluated against DPPH radical and ABTS radical cation scavenging ability, ferric-reducing antioxidant power (FRAP) and beta-carotene-linoleate bleaching (beta-CB) assays, and the Folin-Ciocalteu method for TPC. BHA was used as reference. P. porrigens showed significantly higher (p < 0.05) DPPH* scavenging ability (90.78 +/- 0.30%) and FRAP (6.37 +/- 0.22 mM FE/100g), while Sch. commune showed significantly higher (p < 0.05) ABTS*+ inhibition activity (94.96 +/- 0.70%) and beta-CB inhibition activity (94.18 +/- 0.17%), respectively. TPC was found in a descending order of P. poriggens > L. ciliatus = Pleurotus ostreatus (cultivated) > H. conica = Sch. commune. Positive correlation was observed between the AOA and TPC. When compared to BHA (2 mM), P. porrigens showed significantly higher (p < 0.05) DPPH* scavenging ability and reducing power, while Sch. commune showed comparable DPPH* scavenging ability and ABTS*+ inhibition activity. All the mushrooms have better ABTS*+ inhibition activity than BHA (1 mM). The beta-CB inhibition activity of BHA was significantly higher than those of edible wild mushrooms. The water extracts of edible wild mushrooms showed potent antioxidant activities compared to BHA to a certain extent.
An HPLC method with ultraviolet-visible spectrophotometry detection has been optimized and validated for the simultaneous determination of phenolic compounds, such as butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) as antioxidants, and octyl methyl cinnamate (OMC) as UVB-filter in several personal care products. The dynamic range was between 1 to 250 mg/L with relative standard deviation less than 0.25% (n = 4). Limits of detection for BHA, BHT, and OMC were 0.196, 0.170, and 0.478 mg/L, respectively. While limits of quantification for BHA, BHT, and OMC were 0.593, 0.515, and 1.448 mg/L, respectively. The recovery for BHA, BHT, and OMC was ranged from 92.1-105.9%, 83.2-108.9%, and 87.3-103.7%, respectively. The concentration ranges of BHA, BHT, and OMC in 12 commercial personal care samples were 0.13-4.85, 0.16-2.30, and 0.12-65.5 mg/g, respectively. The concentrations of phenolic compounds in these personal care samples were below than maximum allowable concentration in personal care formulation, that is, 0.0004-10 mg/g, 0.002-5 mg/g, and up to 100 mg/g for BHA, BHT, and OMC, respectively.
Introduction: Hygrocybe conica (HC), a wild mushroom commonly consumed by the indigenous people (Orang Asli) in Peninsular Malaysia, was assessed for its antioxidant content. Methods: The HC mushroom was extracted using distilled water and the crude extract partitioned using different solvents and open column chromatography to evaluate its potential antioxidant properties. The mushroom extract was partitioned using liquid-liquid extraction into the hexane (Fl), chloroform (F2), butanol (F3) and formic acid (F4) fractions. Based on solvent polarity, the water extract of the mushroom was fractionated into non-polar (FI), semi-polar (Fii), and polar fractions (Fiii) using open column chromato graphy. Antioxidant capacities were determined using DPPH, ABTS, and ferric reducing antioxidant power (FRAP) assays while Folin-Ciocalteu reagent assay was used to determine total phenolic content (TPC). Results: The HC extract had the highest TPC and DPPH scavenging capacity compared to its extract fractions. TE values (ABTS assay) of F2 and F4 were not significantly higher than the HC extract. Among the extract fractions of different polarities, Fiii had the highest antioxidant capacities (DPPH and FRAP) compared to FI and Fii while FRAP values of these fractions were not significantly lower than the FRAP value of HC extract. The HC extract had significantly lower antioxidant capacity than antioxidant standards (ascorbic acid and BHA). Tannie acid as the main bioactive component in HC mushroom was detected using HPLC method. The presence of phenolics in HC extract was also confirmed using TLC. Conclusion: Due to the presence of potent phenolic components, the mycelia of HC could be consumed for potential antioxidative benefits.
This study was undertaken to evaluate the potential of fruit waste materials as source of natural antioxidant. The fruit peels including mango, guava and papaya peel were used in this study. The total phenolic content (TPC) was determined by Folin-Ciocalteu assay while antioxidant activities were determined by using ferric reducing antioxidant power (FRAP), 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging, ferric thiocynate (FTC) and thiobarbituric acid (TBA) assays. These antioxidant activities were compared to synthetic antioxidants, BHA/BHT combination and ascorbic acid. The results demonstrated that TPC ranged from 3.23 to 15.84 g GAE/100 g extract. Mango peels extract exhibited highest TPC compared to guava peel and papaya peel extract. In the FRAP assay, mango peel extract at 200 ppm, guava peel extract at 400 ppm and papaya peel extract at 1200 ppm, exhibited reducing power comparable to the permissible amount of BHA/BHT at 200 ppm. At concentration of 250 μg/ml, the DPPH radical scavenging activity of extracts and standards decreased significantly in the order of mango peel extract > guava peel extract > BHA/BHT > ascorbic acid > papaya peel extract. For the FTC assay, the antioxidant activity of mango peel extract was significantly higher than ascorbic acid, guava peel and papaya peel extract but lower than BHA/BHT while in the TBA assay, percentage inhibition of BHA/BHT and ascorbic acid were found to be higher than fruit peel extracts. The quantitative analysis for flavonoids showed the presence of catechin, epicatechin and kaempferol in the peel extracts.
Efficacy of some Malaysian herbal aqueous extracts, BHA/BHT (synthetic antioxidants) and ascorbic acid in retarding oxidative rancidity was tested with cakes. The development of lipid oxidation products during 15 days at room temperature was evaluated by means of Peroxide Value (PV) and Thiobarbituric acidreactive substances (TBARS) value. The six formulations consists of control sample (cake without addition of antioxidant) (F1), cake added with curry leaves extract (F2), cake incorporated with kesum leaves extract (F3), cake added with tenggek burung leaves extract (F4), cake incorporated with ascorbic acid (F5) and cake added with BHA/BHT (F6). Formulation with the incorporation of tenggek burung leaves extract showed powerful oxidative stability effect compared to the formulations with othe plant extracts and control sample. However, cakes with BHA/BHT showed the strongest oxidative stability throughout the storage period. Therefore, it is suggested that tenggek burung leaves extract can be added into the food system for effectiveness as antioxidant to prolong the shelf life of the product.
This study was conducted on selected local herbs such as ulam raja (Cosmos caudatus), kesum (Polygonum minus), selom (Oenanthe javanica), pegaga (Centella asiatica) and curry leaves (Murraya koenigii) to investigate their antioxidative activities. The water extracts of the herbs were analysed for total phenolic content, reducing antioxidant power, ferric thiocyanate (FTC) and the thiobarbituric acid (TBA) test was also accried out. Polygonum minus showed the highest total phenolic content and reducing power among the herbs. Increasing the concentration of the extracts resulted in increased Fe3+ reducing antioxidant power for all the herbs. FTC and TBA tests on the extracts during seven days of storage showed that all the herbs extracts had the ability to reduce oxidation compared to the control (P < 0.05). From the FTC analysis, Murraya koenigii leaves was best in reducing the oxidation rate (67.67%) compared to the other herbs studied. Analysis of TBA showed that Centella asiatica extract had the highest antioxidant effect. However, both TBA and FTC analysis for these two herbs showed no significant difference (P >0.05) from Polygonum minus and butylated hydroxyanisole (BHT) a synthetic antioxidant. Correlation analysis showed positive correlations between amount of total phenolic content and reducing power (r = 0.75) and antioxidative activities (r = 0.58) in linoleic acid emulsion system. This shows that antioxidative activities of these Malaysian herbal plants especially Polygonum minus may be a potential source of natural antioxidants with similar characteristics to the synthetic antioxidant, BHT.
The efficacy of pandan leaf extract (PLE) addition on the oxidative degradation of sunflower oil (SFO) during microwave heating was studied. 80% of methanol extract showed better antioxidant action than the 100% methanol or ethanol extract and the total phenolic contents, DPPH radical scavenging activity and linoleic acid system of PLE were found to be 1845.50 mg GAE/100 g, 60.62-89.87% and 82.21%, respectively. 80% of methanolic extracts at different concentrations (0.1, 0.2 and 0.4 wt. %) were added to SFO. The antioxidant treated and control oil samples were subjected to microwave heating and were analyzed at regular intervals for the extent of oxidative changes following the measurements of peroxide value, p-anisidine value, TOTOX, free fatty acid, specific extinction, iodine value, viscosity, polar compounds and fatty acid composition. The PLE were found to be quite effective towards suppressing the primary and secondary oxidation products in the tested oil. The order of effectiveness (p<0.05) was BHA > 0.4% PLE > 0.2% PLE > 0.1% PLE > control. The present results suggested that antioxidant extract from pandan leaf might be used to protect vegetable oils from oxidation.
With the concern of adverse effects of lipid oxidation on food deterioration and human health, the antioxidant activities of kenaf seed extracts (KSE), roselle seed extracts (RSE) and roselle extracts (RE) were evaluated by comparing its oxidative stability in refined, bleached & deodorised (RBD) sunflower oils with that of in synthetic antioxidant, BHA. Established methods such as peroxide values (PV), p-anisidine values (AV), TOTOX values, free fatty acids (FFA), iodine values (IV), total phenolic contents (TPC), conjugated dienes (CD) and conjugated triene (CT) were employed to assess the extent of oil deterioration. During 24 days storage, consensus was accomplished based on the results assessed by PV, TOTOX, CD, CT, IV and TPC at which the antioxidant activities of KSE, RSE and RE were better than BHA. Surprisingly, the results obtained by AV and FFA assays showed the reversed. Among the extracts, RSE exhibited the best antioxidant activities. These suggest that KSE, RSE and RE may be used as potential source of natural antioxidants in the application of food industry to prevent lipid oxidation.
The aim of this study is to determine the antioxidant capacity of underutilized fruits in Malaysia namely Milk apple (Syzygium malaccense), Malay apple (Syzygium malaccense (L.) Merr. and Perry), and Water apple (Syzygium aqueum). Synthetic antioxidants (BHA and BHT) commonly used in the food industries may not be as safe as it was presumed earlier. As BHA and BHT may be carcinogenic, it is important to look for new sources of natural antioxidants from fruits and vegetables. Freeze dried samples extracted with acetone and water were measured by ferric 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and Ferric Reducing Antioxidant Power (FRAP) assays. Acetone extract (50%) showed higher values for both DPPH and FRAP assays compared with water extract. Milk apple has the highest DPPH value of 95.26% inhibition of DPPH. Milk apple also showed the highest FRAP value with 8722.22 µM of Fe (II) per gram of freeze dried sample. There was a significant difference (P < 0.05) in the types of extraction used. Antioxidant capacities of the samples are in the following order: Milk apple > Malay apple > Water apple. Proximate compositions and mineral contents of the samples were determined too. The samples can be used as a source of natural antioxidants.
Cinnamon deodorised aqueous extract (CinDAE) was prepared and evaluated for its total phenolic (315.3 ± 35.4 mg GAE/g) and flavonoid (99.3 ± 9.6 mg RE/g) contents. Stabilizing efficiency of CinDAE, for chicken meatballs, was measured against oxidative deterioration as function of storage time under chilled conditions. For this purpose, oxidative stability [2-thiobarbituric acid reactive substances (TBARS); peroxide value (PV)], colour and sensory acceptability were measured in the control meatballs (C), and those stabilized with 200 ppm of: CinDAE (T1), ascorbic acid (T2), BHA/BHT (50/50; w/w) (T3). In comparison to "C", induction period (IP) and redness (a* value) of the stabilized samples (T1, T2 and T3) were increased, while PV and TBARS were decreased throughout storage (8 ± 1 °C) significantly (p
Curcuminoids have been used for the management of burns and wound healing in traditional Chinese medicine practices but the wide application of curcuminoids as a healing agent for wounds has always been a known problem due to their poor solubility, bioavailability, colour staining properties, as well as due to their intense photosensitivity and the need for further formulation approaches to maximise their various properties in order for them to considerably contribute towards the wound healing process. In the present study, a complex coacervation microencapsulation was used to encapsulate curcuminoids using gelatin B and chitosan. This study also focused on studying and confirming the potential of curcuminoids in a microencapsulated form as a wound healing agent. The potential of curcuminoids for wound management was evaluated using an in vitro human keratinocyte cell (HaCaT) model and the in vivo heater-inflicted burn wound model, providing evidence that the antioxidant activities of both forms of curcuminoids, encapsulated or not, are higher than those of butylated hydroxyanisole and butylated hydroxytoluene in trolox equivalent antioxidant capacity (TEAC) and (2,2-diphenyl-1-picryl-hydrazyl-hydrate) (DPPH) studies. However, curcuminoids did not have much impact towards cell migration and proliferation in comparison with the negative control in the in vitro HaCaT study. The micoencapsulation formulation was shown to significantly influence wound healing in terms of increasing the wound contraction rate, hydroxyproline synthesis, and greater epithelialisation, which in turn provides strong justification for the incorporation of the microencapsulated formulation of curcuminoids as a topical treatment for burns and wound healing management as it has the potential to act as a crucial wound healing agent in healthcare settings.
Natural preservatives having the great antioxidant and antimicrobial activity have been utilized in the food industry for many years. In the present study, the effect of of two brands of commercial Assam green tea infusion (represented by A and B) and 0.02% BHA/BHT on microbial growth, anti-lipid oxidation and color change were investigated in cooked beef. The green tea concentration has influenced to the results. It was found that A and B at the concentration of 250 mg/mL significantly reduced the population of Staphylococcus aureus, Listeria monocytogenes, Salmonella typhimurium and E. coli in the cooked beef to an undetectable level within 2 days of storage at 4oC. A and B also exhibited higher anti-lipid oxidation activity compared to 0.02% BHA/BHT, and control. Assam green tea infusions in cooked beef significantly increased ∆ L*
value and decreased ∆ a* and ∆ b* value (p ≤ 0.05). These indicate that Assam green tea infusion might be a potential candidate as a natural preservative for beef and other types of food.
Glutathione S-transferase isoenzymes (GSTs) catalyze the conjugation reaction between glutathione and electrophilic
compounds. GSTs are involved in the detoxification of toxic and carcinogenic compounds, thus protecting the body from
toxic injuries. Tocotrienols are part of the vitamin E family and is believed to possess potent antioxidant activity. The
objective of this study was to determine the effect of increasing doses of tocotrienol rich fraction (TRF) supplementation
on liver GSTs gene and protein expression. A total of 30 male ICR white mice were divided into five groups (n=6 for each
group) and given treatment for 14 days through oral supplementation. Groups were divided as follows: - three groups
administered with TRF at doses of 200, 500 and 1000 mg/kg, respectively, a positive control group administered with 100
mg/kg butylated hydroxyanisole (BHA) and a control group administered with only the vehicle (corn oil). At day 15, the
mice were sacrificed and their livers isolated. Total RNA was extracted from the liver and quantitative real-time polymerase
chain reaction (qPCR) assays were performed to analyze GSTs gene expression. Total liver protein was also extracted
and the protein expression of GSTs was determined by Western blotting. The results showed that TRF oral supplementation
caused a significant dose-dependent increase in liver GST isoenzymes gene and protein expression, compared to controls.
In conclusion, TRF oral supplementation for 14 days resulted in increased gene and protein expression of GST isoenzymes
in mice liver dose-dependently, with the highest expression seen in mice treated with 1000 mg/kg TRF.
The role of diets in causing cancers necessitates the ongoing search for natural antimutagens of promising anticancer therapeutics. This study determined the potential anticancer efficacy of the leaf extract of Myristica fragrans (Houtt.). Methanol leaf extract of M. fragrans (Houtt.) alone was screened for mutagenicity in the bacterial reverse mutation (Ames) test, using the Salmonella typhimurium TA100 strain, the Allium cepa, and the mouse in vivo bone marrow micronucleus tests. The antimutagenicity of this extract against benzo[a]pyrene- and cyclophosphamide-induced mutations was evaluated. An antioxidant test on the extract was performed with 2,2-diphenyl-1-picrylhydrazyl, using butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) as the standards, whereas its phytochemicals were elucidated by following the gas chromatography/mass spectrometry protocol. In S. typhimurium (TA100), the mutagenicity ratio at 200,500 and 1,000 µg/well was >2. Cell division in the A. cepa root tips and mouse bone marrow was significantly (P ≤ 0.05) inhibited at 2,000 and 4,000 mg/kg, whereas the observed chromosomal aberrations and micronucleated polychromatic erythrocytes were non-dose-related and were insignificantly (P ≥ 0.05) different from the negative control. Inhibition of benzo[a]pyrene- and cyclophosphamide-induced mutagenicity by this extract was above 40%. Half-maximal inhibitory concentration of the extract in the antioxidant test was lower than that of BHA and BHT. Phytochemical compounds, possessing antioxidant activity, may be responsible for the observed effects, suggesting a strong antimutagenic activity of the MeOH leaf extract of M. fragrans, a necessary characteristic of a promising anticancer agent.