We evaluated the phenolic content and antioxidant capacities of pod and seed extracts (in methanol, ethanol, and water) of an underutilized legume, Clitoria fairchildiana (Howard). The antioxidant capacity of the extracts was determined using the ferric reducing antioxidant potential assay, and the free radical-scavenging capacity was evaluated using 2,2-diphenyl-1-picrylhydrazyl radical-scavenging and ABTS assays. In addition, the total flavonoids, flavonols, and tannin contents were also determined. Overall, the methanol extracts of the pod contained high concentration of phenolics and showed high antioxidant capacities compared to seed extracts. In addition, a positive correlation was found between total phenol and tannin versus antioxidant capacity. Results of the present study indicate pods and seeds of C. fairchildiana to possess rich amount of natural antioxidants, and can be further explored for their possible use as a natural additive in food or in pharmaceutical industries.
The effect of room temperature ionic liquid (RTIL) on the formation of the fluorescence ternary complex oxalate-sodium morin-5-sulfonate (NaMSA)-Aluminium(III) has been studied. In weakly acidic medium and in the presence of RTIL, 1-Butyl-3-methylimidazolium hexafluorophosphate (BMIM-PF6), total complex formation is achieved as compared with the formation of the binary complex of NaMSA-Aluminium(III). The fluorescence characteristics of the system allowed the establishment of a very sensitive method for the spectrofluorimetric determination of oxalate ion. The ternary complex formed its highest fluorescence signal at 513 nm and excitation at 420 nm. In these conditions, the method produces a detection limit of 0.57 ng mL(-1). The procedure has been satisfactorily applied to the determination of oxalate ion in a vegetal tissue (spinach leaves).
The interest to sulfonated methyl esters of fatty acids (SME) has been growing during the last decade, because these surfactants are considered as an environmentally friendly and renewable alternative of the linear alkyl-benzene sulfonates (LAS). Here, we present a quantitative study on the properties of aqueous SME solutions, and especially on their surface tension isotherms, critical micelle concentration (CMC) and its dependence on the concentration of added NaCl. It is demonstrated that the CMC of an ionic surfactant determined by electrical conductivity is insensitive to the presence of a small nonionic admixture, so that the CMC values determined by conductivity represent the CMC of the pure surfactant. Using SME as an example, we have demonstrated the application of a new and powerful method for determining the physicochemical parameters of the pure ionic surfactant by theoretical data analysis ("computer purification") if the used surfactant sample contains nonionic admixtures, which are present as a rule. This method involves fits of the experimental data for surface tension and conductivity by a physicochemical model based on a system of mass-balance, chemical-equilibrium and electric-double-layer equations, which allows us to determine the adsorption and micellization parameters of C12-, C14-, C16- and C18-SME, as well the fraction of nonionic admixtures (if any). Having determined these parameters, we can further predict the interfacial and micellization properties of the surfactant solutions, such as surface tension, adsorption, degree of counterion binding, and surface electric potential at every surfactant, salt and co-surfactant concentrations.
UV-curable hyperbranched urethane acrylate (HBPUA) from oleic acid of palm oil has been synthesized through a medium aided by p-toluene sulfonic acid as a catalyst. This mixture was then used as the core (HBP) and reacted with palm oil oleic acid to form the hyperbranched polyol (HBP-1). HBPUA was prepared by reacting HBP-1 resin with diisocyanate and hydroxyl-containing acrylate monomer with the presence of 0.1-2 wt% dibutyltin dilaurate as a catalyst. The reaction was confirmed by several analytical data i.e. hydroxyl value (OHV), Fourier Transform infrared (FTIR) spectroscopy, gel permeation chromatography (GPC) and nuclear magnetic resonance (NMR) spectroscopy analyses. The HBPUA was easily curable when subjected to ultraviolet (UV) radiation.
Pleurotus porrigens is a well-known edible, wild mushroom enjoyed as a delicacy by aborigines in Sabah and as source of income for the aborigines who collect and sell them at tamu (local market). This study aimed to evaluate the antioxidant activity in vitro and identify potent antioxidative components of aqueous extracts of P. porrigens. The antioxidant activities were evaluated using DPPH radical scavenging ability, ABTS radical cation inhibition activity, ferric reducing/antioxidant power, and total phenolic content. Activity-guided purifications based on DPPH radical scavenging ability resulted in 5 subfractions (SF). The highest DPPH radical scavenging ability was found in SF-III and SF-IV, but all were lower than butylated hydroxyanisole (BHA) and α-tocopherol. Analysis with high-performance liquid chromatography-diode array detectors found presence of ascorbic acid and (+)-catechin in SFs of P. porrigens, as well as some unidentified components that may have contributed to the radical scavenging ability. In conclusion, aqueous extract of P. porrigens possesses promising antioxidant activities, although they are lesser in their partially purified SFs. Nonetheless, P. porrigens could be promoted as an antioxidant-rich food as part of a normal diet that provides antioxidative benefit.
A central composite design of response surface methodology (RSM) was employed to optimize the extraction time (X1: 266.4-393.6 min) and temperature (X2: 42.9-57.1°C) of Pleurotus ostreatus aqueous extract with high antioxidant activities, namely DPPH radical-scavenging activity, ABTS radical cation inhibition, and ferric reducing/antioxidant power, as well as total phenolic content (TPC). Results showed that the data were adequately fitted into four second-order polynomial models developed by RSM. The extraction time and temperature were found to have significant quadratic effects on antioxidant activities and TPC. The optimal extraction time and temperature were 282.3 min and 42.9°C (DPPH), 393.6 min and 42.9°C (ABTS), 340.4 min and 49.8°C (FRAP), and 347.6 min, 49.7°C (TPC), with corresponding yields of 53.32%, 73.20%, 37.14 mM Fe2+ equivalents/100 g, and 826.33 mg gallic acid equivalents/100 g, respectively. These experimental data were close to their predicted values. The establishment of such a model provides a good experimental basis for employing RSM to optimize the extraction time and temperature for high antioxidant activities from P. ostreatus.
A simple and rapid gas chromatography (GC) method with flame ionization detector was developed for detection of isopropyl para-toluenesulphonate (IPTS) in palm-based isopropyl palmitate (IPP) and isopropyl myristate (IPM). The method involved spiking the IPP/IPM samples with IPTS and directly injecting the spiked samples into GC without undergoing clean-up steps. The calibration curves for IPTS showed good linearity with coefficient correlation of 0.9999 for six-point calibration from 0.5 to 50 μg mL(-1) and 0.9996 for six-point calibration from 0.5 to 200 μg mL(-1) . IPTS recoveries from IPP were 98.6-103.5% with relative standard deviation (RSD) of 0.40-2.80%, whereas recoveries from IPM were 97.0-107.2% with RSD of 0.42-4.21%. The identity of IPTS recovered from the isopropyl esters was confirmed by a GC-mass spectrometer detector. The method was successfully applied to the analyses of IPTS in commercial samples. It was found that there were IPTS in the range of 34.8-1303.0 μg g(-1) in the palm-based esters for some of the samples analysed.
Syzygium aqueum, a species in the Myrtaceae family, commonly called the water jambu is native to Malaysia and Indonesia. It is well documented as a medicinal plant, and various parts of the tree have been used in traditional medicine, for instance as an antibiotic. In this study, we show S. aqueum leaf extracts to have a significant composition of phenolic compounds, protective activity against free radicals as well as low pro-oxidant capability. Its ethanolic extract, in particular, is characterized by its excellent radical scavenging activity of EC(50) of 133 μg mL(-1) 1,1-diphenyl-2-picryl-hydrazyl (DPPH), 65 μg mL(-1) 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) and 71 μg mL(-1) (Galvinoxyl), low pro-oxidant capabilities and a phenolic content of 585-670 mg GAE g(-1) extract. The extract also displayed other activities, deeming it an ideal cosmetic ingredient. A substantial tyrosinase inhibition activity with an IC(50) of about 60 μg mL(-1) was observed. In addition, the extract was also found to have anti-cellulite activity tested for its ability to cause 98% activation of lipolysis of adipocytes (fat cells) at a concentration of 25 μg mL(-1). In addition, the extract was not cytotoxic to Vero cell lines up to a concentration of 600 μg mL(-1). Although various parts of this plant have been used in traditional medicine, this is the first time it has been shown to have cosmeceutical properties. Therefore, the use of this extract, alone or in combination with other active principles, is of interest to the cosmetic industry.
Health hazards associated with usage of plastic films for food preservation demands for development of active films from non-toxic and antioxidant rich bio-sources. The reported work highlights the development, characterization and application studies of chitosan films enhanced for their antioxidant activity by mango leaf extract (MLE) incorporation. Effect of MLE variation (1-5%) on the morphology, optical nature, water exposure and mechanical characteristics of the chitosan-MLE composite films was studied. Increase in the MLE concentration resulted in films with increased thickness and decreased moisture content. Contact angle, water solubility and vapor permeability analysis demonstrated the reduced hydrophilicity and water vapor penetrability of the films due to MLE inclusion. MLE films possessed better tensile strength (maximum of 23.06 ± 0.19 MPa) with reduced elongation ratio than the pure chitosan film (18.14 ± 0.72 MPa). Antioxidants assessment in terms of total phenolic content, DPPH radical scavenging, ferric reducing power and ABTS radical scavenging showed improved antioxidant activity with the incremental amounts of MLE in the chitosan films. Microscopic studies revealed the smooth, compact and dense nature of the MLE-chitosan films favouring low oxygen transport rates. Application studies to cashew nuts preservation for 28 days storage indicated 56% higher oxidation resistance for the 5% MLE film than a commercial polyamide/polyethylene film. Results highlight the potential and promising nature of MLE impregnated chitosan films as suitable alternative for active packaging films for food preservation.
Enzyme immobilization has been known to be one of the methods to improve the stability and reusability of enzyme. In this study, a strategy to optimize laccase immobilization on polyethylene terephthalate grafted with maleic anhydride electrospun nanofiber mat (PET-g-MAH ENM) was developed. The development involves the screening and optimization processes of the crucial factors that influence the immobilization yield such as enzyme concentration, pH values, covalent bonding (CV) time, CV temperature, crosslinking (CL) time, CL temperature and glutaraldehyde concentration using two-level factorial design and Box-Behnken design (BBD), respectively. It was found that laccase concentration, pH values and glutaraldehyde concentration play important role in enhancing the immobilization yield of laccase on PET-g-MAH ENM in the screening process. Subsequently, the optimization result showed at 0.28 mg/ml laccase concentration, pH 3 and 0.45% (v/v) glutaraldehyde concentrations gave the highest immobilization yield at 87.64% which was 81.2% increment from the immobilization yield before optimization. Under the optimum condition, the immobilized laccase was able to oxidize 2, 2-azino-bis 3-ethylbenzothiazoline-6- sulfonic acid (ABTS) in a broad range of pH (pH 3-6) and temperature (20- 70 °C). Meanwhile, the kinetic parameters for Km and Vmax were 1.331 mM and 0.041 mM/min, respectively. It was concluded that the optimization of immobilized laccase on PET-g-MAH ENM enhance the performance of this biocatalyst.
Vigna sinensis also known as long-podded cowpea or Chinese long bean (Family:Fabaceae) is most widely grown in Southeast Asia. They are a good source of protein, vitamin A, vitamin C, iron, phosphorus, and potassium. The antioxidant potential of crude methanol extract, chloroform, and ethyl acetate soluble fractions of Vigna sinensiswas screened for in- vitro antioxidant activity using total phenolic content, ferric reducing power, 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) assay, ferric thiocyanate (FTC) and thiobarbituric acid (TBA) tests. It was found that ethyl acetate fraction have maximum amount of polyphenolics compounds (2.69 mg/g GAE in concentration 0.5 mg/mL); more effective than methanol and chloroform extract.This fraction also exhibited fairly good antioxidant activity with in both TBA (17.39% mg/g GAE) and FTC (12.65% mg/g GAE) methods.
This study investigated the effects of different percentages of ethanol (0 - 100%), extraction times (1 - 5 h) and temperatures (25 - 60°C) on total phenolic content (TPC) and antioxidant activity (AA) of sapodilla pulp and peel. TPC was determined by Folin-Ciocalteu reagent method, while AA was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, 2,2-azino-bis-(3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS) radical scavenging assay, ferric reducing antioxidant power (FRAP) assay and β-carotene bleaching (BCB) assay. Based on the optimal extraction conditions used, sapodilla pulp extract had TPC of 3.89 mg GAE/g, 63.20% of DPPH scavenging activity, 4.30% of ABTS scavenging activity, 19.17% of BCB activity, and FRAP value of 15.24 mg TE/g; while its peel extract had TPC of 9.23 mg GAE/g, 92.95% of DPPH scavenging activity, 5.36% of ABTS scavenging activity, 8.14% of BCB activity, and 27.85 mg TE/g (FRAP value). Using the optimal extraction conditions for sapodilla pulp (40% ethanol as extraction solvent that extracted at 60°C for 4 h) and sapodilla peel (80% ethanol and 2 h extraction time at 40°C), highest antioxidants can be extracted from the pulp and peel.
The objective of this study was to evaluate the effects of solid-to-solvent ratio (1:5. 1:10, 1:15 and 1:20) on the extraction of phenolic compounds (TPC and TFC) and antioxidant capacity (ABTS and DPPH radical scavenging capacity) of P. niruri. Solid-to-solvent ratio showed a significant effect for both phenolic compounds (TPC and TFC) and antioxidant capacity (ABTS and DPPH radical scavenging capacity) with 1:20 was the condition for extracting the highest of phenolic compounds (TPC and TFC) with a value of 5788.7 mg GAE/100 g DW and 1906.5 mg CE/100 g DW, respectively and exhibited high antioxidant capacities (ABTS and DPPH radical scavenging capacities) with a value of 0.820 mM and 1.598 mM, respectively among the four levels studied. TPC was positively and significantly correlated with ABTS and DPPH (r=0.999 and r=0.999) under the effects of solid-to-solvent ratio as compared to TFC, positively and strongly correlated (r=0.865 and r=0.868) with ABTS and DPPH.
Ultrasound-assisted extraction (UAE) was applied for the extraction of bioactive valuable compounds from winter melon (Benincasa hispida) seeds. Effects of amplitude (25-75%), temperature (40-60°C) and sonication time (20-60 min) on crude extraction yield (CEY) and radical scavenging activities (RSA, % inhibition of DPPH˙ and ABTS˙+ free radicals) of extracts were determined using complete randomised design (CRD). The results showed that the CEY and RSA of extracts significantly affected by independent variables. The maximum value of CEY (97.14±0.36 mgg-1), scavenging of DPPH˙ radicals (32.12 ± 0.38%) and scavenging of ABTS˙+ radicals (40.52±0.73%) were obtained at the combined treatment conditions of 75%, 55°C and 40 min. The UAE results obtained were compared with those achieved by using conventional Soxhlet extraction (CSE) method. It was found UAE allowed extraction at lower temperature and the extracts obtained posses higher quality compare with CSE. UAE is a promising environment friendly technique for the extraction of bioactive compounds from winter melon (Benincasa hispida) seeds.
Clinacanthus nutans (Burm. F.) Lindau or locally known in Sabah, Malaysia as ‘Sabah Snake Grass’ has been ethnobotanically used to treat various diseases in Asian countries. This study was conducted to determine the total phenolics content (TPC), flavonoids content (TFC) and antioxidant activity of herbal teas developed from C. nutans leaves with different drying techniques (microwave-oven dried and freeze dried) and infusion time (1, 2, 5, 10, 15 and 20 min). Ferric reducing/antioxidant power (FRAP) assay, 2,2’-azino-bis(3-ethylbenzothiazoline- 6-sulphonic acid (ABTS) and 2, 2-diphenyl-1-pycryl-hydrazyl (DPPH) free radical scavenging assays were used to investigate the antioxidant capacity. The highest TPC of herbal tea was observed in 20 min infusion of unfermented microwave-oven dried leaves (177.80 ± 19.10 mg TAE/L), while the highest TFC was observed in 10 min infusion of fermented microwave-oven dried leaves (22.13 ± 1.53 mg CE/L). Short infusion times from 5 min to 15 min were able to extract high amount of phenolics compounds. Unfermented tea contained higher TPC content (P < 0.05) as compared to fermented tea, while, TFC showed no significant difference between both types. Freeze dried infusion shows no significant difference (P > 0.05) as compared to microwave-oven dried for TPC, TFC and antioxidant capacity. Moderate and low correlation was observed between TPC and FRAP values (r = 0.507) and between TFC and ABTS values (r = 0.256). Preparation of C. nutans herbal tea as potential natural antioxidant source can be used as a basic reference for future research on the dietary intake of these herbal teas.
Although Malaysia is one of the important pineapple fruit producing and exporting country, the production of MD-2 pineapple fruit only started in 2009. Pineapple fruit has been harvested at different ripening stages for different markets. The information on Malaysian grown MD-2 pineapple fruit quality is lacking. Therefore this work was carried out with the aim to determine physicochemical quality, antioxidant compounds and activity of MD-2 pineapple fruit at five ripening stages. Ripening stage affected physicochemical quality of MD-2 pineapple fruit. Soluble solids concentration of MD-2 pineapple fruit increased from 15.41 to 18.02% SSC when fruit ripened from stage 1 to 4 and no significant difference was found in fruit between stage 4 and 5. The ascorbic acid content decreased while total carotenoids content increased as ripening stage advanced. The total phenolic content of both 80% methanol and water extraction solvents increased significantly as fruit ripened from stage 1 to 3 and reduced as fruit ripened to stage 5. The antioxidant activity of MD-2 pineapple fruit as assayed using DPPH, FRAP and ABTS showed similar trend as total phenolic content. These results suggest that ripening stage affect MD-2 pineapple fruit quality and nutritional values.
The effects of ethanol concentration (0-100%, v/v), extraction time (60-300 min) and extraction temperature (25-65°C) on the extraction of phenolic antioxidants from Andrographis paniculata was evaluated using single-factor experiments. The following complementary assays were used to screen the antioxidant properties of the crude extracts: total phenolic content (TPC), total flavonoid content (TFC), condensed tannin content (CTC), 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical-scavenging capacity and 2,2’-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging capacity. The extraction conditions chosen had significant effects (p < 0.05) on the extraction of phenolic compounds and antioxidant capacity. The optimal conditions were 60% ethanol for 60 min at 65oC for phenolic compounds and at 25oC for antioxidant capacity. Strong negative significant (p < 0.05) correlations were observed between the phenolic compounds (TPC, TFC and CTC) and antioxidant capacity comprising ABTS (-0.924, -0.909, -0.887, respectively) and DPPH radical-scavenging capacities (-0.992, -0.938, -0.928, respectively) were determined under the influence of extraction temperature.
Papaya (Carica papaya L. cv. Hongkong) is an economically important fruit crop grown in Malaysia. During its ripening stages, (C. papaya L.) exhibits different physicochemical properties, antioxidant capacities, and sensory quality results. The objective of this study was to elucidate in detail the antioxidant capacity of C. papaya as determined by total phenol content (TPC), total flavonoid content (TFC), ferric reducing antioxidant power (FRAP),2,2-diphenyl-1-picrylhydrazyl (DPPH) and scavenging systemand (ABTS). The study also aimed to study physicochemical changes of papaya fruits based on measured pH, titratable acidity (TA), total soluble solids (TSS), moisture and fruit color at five different stages of ripening. The fruits were harvested at five different, stages RS1, RS2, RS3, RS4, and RS5 corresponding to 12, 14, 16, 18, and 20 weeks after anthesis, respectively. Significant differences were found at different stages of ripening. The pH of the fruit decreased significantly (P < 0.05), whereas TA, moisture, and TSS increased significantly (P < 0.05) during the ripening process. The redness (a*) and yellowness (b*) values of fruit color both increased significantly (P < 0.05), whereas
lightness (L*) varied. The total phenol content TPC, TFC, FRAP, DPPH and ABTS values increased significantly (P < 0.05) with the ripening process. Sensory evaluation based on the color, sweetness, sourness, flavor, and overall acceptance for the last three maturity stages was also performed. RS5 had a better score than RS3 or RS4. The results showed the important role of the ripening stage in increasing the antioxidant content of papaya fruits.
Four types of soft dates (SD), three types of semi-dried dates (SDD) and one type of dried dates (DD) were used in this study. The antioxidant activities were assessed using TEAC method (ABTS assay) and the ferric reducing/antioxidant power method (FRAP assay), while total phenolic content (TPC) and total flavonoid content (TFC) were measured using Folin-Ciocalteau and aluminum chloride colorimetric methods. Multivariate analysis of variance (MANOVA), discriminant analysis (DA) and principal component analysis (PCA) were used to analyze the data. MANOVA showed a strong significant difference between the eight types of dates. DA identified the relative contribution of each parameter in distinguishing the dates. DA also identified two functions responsible for discriminating the dates and showed the difference between different types of dates. The first function distinguished DD from other types of dates, whilst the second function discriminated SD and SDD, affording 100% correct assignation. PCA identified only one component responsible for explaining 98.85% of the total variance in antioxidant data. It is suggested that the TEAC method and the quantitative determination of TPC and TFC was suitable for differentiation of dates and quality control.
In this study, the effect of different solvent including ethanol, n-hexane and ethyl acetate on antioxidant
activity and total phenolic content (TPC) of winter melon (Benincasa hispida) seeds extract was investigated using conventional Soxhlet extraction (CSE). DPPH and ABTS scavenging activity and TPC results indicated that the seed extracts obtained using ethanol possessed the highest antioxidant activity and followed by ethyl acetate and n-hexane. By considering obtained results, it was clear that there was a high positive correlation between TPC and antioxidant activity. Linoleic acid forms a significant percentage of unsaturated fatty acids of the seed extract (60.6%). It is well known that essential fatty acids including linoleic acid and linolenic acid which are detected in extracts play important roles in preventing many disease and abnormal differentiation problems. B. hispida seeds are potential source of natural antioxidant compounds to replace synthetic antioxidants.