Ultrasound Treated Samples Research Articles

Effect of ultrasound-assisted pH-shifting treatment on the physicochemical properties of melon seed protein

Melon seeds have received considerable attention in recent years because of their high protein content, but they have not yet been fully used. The modification of melon seed protein (MSP) using ultrasound-assisted pH-shifting treatment was investigated in this study by analyzing structural characteristics and functional properties. The particle size, free sulfhydryl content, surface hydrophobicity, solubility, secondary structure, water-holding capacity, oil-holding capacity, emulsification activity index, and emulsification stability index of MSP were determined. MSP treated with ultrasound-assisted, pH-shifting had a smaller particle size, lower free sulfhydryl content, higher surface hydrophobicity, and solubility increased from 43.67 % to 89.12 %. The secondary structure of MSP was affected by ultrasonic treatment, manifesting as an α-helix increase and β-helix, β-turn, and random coil content decrease, which may be the reason why the protein structure became more compact after treatment. The water and oil holding capacities of MSP increased from 2.74 g/g and 3.14 g/g in untreated samples to 3.19 g/g and 3.97 g/g for ultrasound-treated samples, and further increased to 3.97 g/g and 5.02 g/g for ultrasound-assisted, pH-shifting treatment at pH 9.0, respectively. The emulsification activity index of MSP was 21.11 m2/g before treatment and reached a maximum of 32.34 m2/g after ultrasound-assisted, pH-shifting treatment at pH 9.0. The emulsification stability of MSP was maximized by ultrasonic treatment at pH 7.0. Ultrasound-assisted, pH-shifting treatment can effectively improve the functional properties of MSP by modifying the protein structure, which improves the potential application of melon seed protein in the food industry.

Modification of pea protein isolates by high-intensity ultrasonication: Functional, structural and nutritional properties

The current study aims to modify the functional, physical, structural and nutritional characteristics of pea protein isolate. High-intensity ultrasound treatment was used at 20 kHz frequency and 25 % amplitude for 10 (US10), 20 (US20), or 30 (US30) min. Results indicated that ultrasound application enhanced the protein solubility and zeta potential. When compared to control samples, the foaming capacity (FC) and stability (FS) as well as emulsion activity (EA) and stability (ES) were also increased from 157.5, 42.03, 46.25 and 53.75 % up to 182.5, 81.57, 72.50 and 67.50 %, respectively. Besides, particle size was found to be lower for ultrasound treated samples (92.9–131.1 nm) in comparison to that of untreated commercial pea protein isolate (161.9 nm). Moreover, while the bioaccessibility of pea protein in untreated sample was calculated as 28.90 %, ultrasonication increased the retention of pea protein up to 49.36 %. It can be concluded from the results that the ultrasonication process can be used as an advantageous, green and non-thermal tool for obtaining protein isolates with improved techno-functional properties and nutritional quality. Therefore, this treatment might improve the characteristics, and thus increase the utilization of plant-based proteins, especially pea protein, in various food systems.

Effect of High-Power Ultrasound Treatment on Bioactive Compound Content and Chromatic Characteristics of Red Wines

The objective of this work was to investigate, at a laboratory scale, the effect of ultrasound treatment (frequency 20 kHz) on the extraction of total phenolic compounds and monomeric anthocyanins from crushed Merlot grapes in order to reduce maceration time. Antioxidant capacity expressed as ferric reducing antioxidant power (FRAP) and chromatic properties were also investigated. Microvinification was performed on samples both sonicated with a probe-type ultrasonic instrument and untreated (control) and the maceration operation was carried out with sequential extraction (after 3, 5 and 7 days) to evaluate the effect of two factors such as amplitude (A), 70 and 90% and treatment time (t), 3, 4 and 5 minutes on the extraction kinetics of bioactive compounds and chromatic characteristics of the samples. The results showed that throughout the maceration process, the sonicated samples exhibited higher levels of bioactive compounds and chromatic characteristics compared to the control, with the values obtained correlating with maceration time, US treatment time and amplitude. Improvement in all investigated characteristics is evident for ultrasound treated samples compared to untreated ones. Our study provided useful data concerning the impact of ultrasound on the bioactive properties and sensory attributes of red wine, thus offering a theoretical basis for the implementation of this technique in winemaking.

Effect of carbonyl-amino condensation, non-covalent cross-linking and conformational changes induced by ultrasound-assisted Maillard reaction on lysinoalanine formation in silkworm pupa protein

The inhibition of cross-linked lysinoalanine (LAL) formation in silkworm pupa protein isolates (SPPI) by Maillard reaction (using varying xylose concentration) and ultrasound treatment was studied. Results showed that sonicated SPPI was effectively grafted with high concentration of xylose (5 %), resulting in the lowest LAL content, which was 48.75 % and 30.64 % lower than the control and ultrasound-treated samples, respectively. Chemical bond analysis showed that the combined treatment destroyed the ionic bonds, intrachain (g-g-t), and interchain (g-g-g) disulfide bonds, but stimulated the polymerization of hydrogen and hydrophobic bonds between SPPI and xylose, and as well enhanced the net negative charge between SPPI/Xylose complexes. The particles of the complexes were more loose, dispersed and rough, and had a stronger hydrophilic microenvironment, accompanied by alterations in microscopic, secondary and tertiary structures. Ultrasound treatment induced the breakdown of the oxidative cross-linking in SPPI, and promoted the sulfhydryl group-dehydroalanine binding and the carbonyl-amino condensation of the protein and xylose, and thus inhibited the formation of cross-linked LAL. Furthermore, the physicochemical and structural parameters were highly interrelated with cross-linked LAL content (|r| > 0.9). The outcomes provided a novel avenue and theoretical basis for minimizing LAL formation in SPPI and improving the nutrition and safety of SPPI.

The Role of Ultrasound Cavitation Combined with Alcalase Enzyme in Improving the Structural and Antioxidant Attributes of Sulfated Polysaccharide from Yellowfin (Thunnus albacares) Skin

ABSTRACT Alcalase enzyme (1 mg) and ultrasound (400 W) at 0–20 min were applied to obtain sulfated polysaccharides from yellowfin skin. Applying 15 min ultrasound resulted in a 0.12% increase in the rate of recovery yield. The intact structure of polysaccharides was determined by Fourier transform infrared spectroscopy, X-ray diffractometry, and scanning electron microscope. Ultrasound improved the thermal behavior of sulfated polysaccharides by affecting their microstructure, as analyzed by DSC. The color characterization (L*, a*, and b*) of ultrasound-treated samples was found to be superior compared to the control. Applying 15 min led to 16% and 9% enhancing DPPH and ABTS radical scavenging activities along with improving ferric-reducing antioxidant power of the polysaccharide.

The alterations in granule, shell, blocklets, and molecular structure of pea starch induced by ultrasound

Understanding the alterations to starch multi-scale structure induced by ultrasound treatment can help in determining the effective application of ultrasound in functional-starch preparation. This study aimed to comprehensively characterize and understand the morphological, shell, lamellae, and molecular structures of pea starch granules treated by ultrasound under different temperatures. Scanning electron microscopy and X-ray diffraction analyses showed that UT (ultrasound treatment) did not change C-type of crystalline, but caused a pitted surface and endowed a looser structure and higher enzyme susceptibility as the temperature increased above 35 °C for pea starch granules. Fourier transform infrared spectroscopy and small-angle X-ray scattering analyses revealed that UT reduced the short-range ordering and increased the thickness of semi-crystalline and amorphous lamellae by inducing starch chain depolymerization, which was manifested by molecule weight and chain length distribution analysis. The sample ultrasound-treated at 45 °C had the higher proportion of B2 chains compared with the other ultrasound-treated samples because the higher ultrasonic temperature altered the disruption sites of starch chains.

Ultrasound probe assisted extraction of Paramignya trimera (Oliver) Guillaum root

Xao tam phan (XTP), a Paramignya genus of the Citrus family, is a renowned medicinal plant in Southeast Asian countries, including Vietnam, and is associated with treating cancer, hepatitis, diabetes, and nose infections. Extraction is a key operation in the preparation of XTP extracts rich in phytochemical compounds. Recently, the application of ultrasound has resulted in a large number of successes in the extraction of medicinal plants for improvement in yield and quality of extracts. Until present, ultrasonic-assisted extraction has not been favored for XTP. In this work, the effect of sonication treatment on collecting the extract of total phenolics and saponin from the XTP root was investigated, then the experimental data were analyzed and optimized by Response Surface Methodology (RSM). The results indicated that the optimal sonication power, temperature, and time were 200 W, 58°C, and 10 min, respectively. Under those conditions, the total phenolic and saponin contents in the XTP extract were maximized and validated to be 20.70 ± 0.77 mg GAE/g dried matter and 326.30 ± 2.03 mg EE/g dried matter, respectively. For comparison, a higher extraction yield of ultrasound-treated samples, in terms of total phenolics (~ 27%) and total saponin (~ 76%), was found as compared to the untreated ones. In conclusion, the results reveal that the application of ultrasound on xao tam phan root significantly enhanced the phytochemical contents during extraction.

Application of ultrasounds to improve oak aging of white wines

Increasing the efficiency of the release for certain compounds during the maturation of some wines in the presence of oak chips is an important topic for oenology research. This study followed the influence of the use of ultrasound waves with a frequency of 35 kHz on the physicochemical and sensory characteristics of some white wines from Iasi, Romania. Oak chips (1 g/L and 2 g/L, fresh, light, and medium toasted) were added to the Fetească regală and Sauvignon blanc stabilized wines. The samples were kept at a constant temperature of 15oC for 10 and 15 days, respectively. For comparison, 15 mins. ultrasound-treated samples were used. It has been observed that this technique in addition to reducing the aging time, significantly influences the organoleptic characteristics of the treated wines. New ways to continue research to clarify, optimize and streamline the methods of artificial aging of wines have also been identified.

Extracted pepsin of trout waste and ultrasound-promoted method for green recovery of fish collagen

Collagen type I is a prominent protein in animal tissues. This structural protein can be commercially derived from fish processing waste without any health risk. The current study aimed to investigate the efficacy of rainbow trout pepsin and the ultrasound-promoted method on collagen recovery. Pepsin extract was purified from rainbow trout stomach in order to use in collagen extraction from wasted yellowfin skin through ultrasound technique in five durations (0, 5, 10, 15, and 20 min). The 15 min ultrasound treated pepsin soluble collagen indicated the highest recovery yield (23.8%) as well as the highest imino acid content (18.2%). The structure of type I collagen was observed in the SDS-PAGE pattern of collagens and no structural change occurred during the extraction process. The presence of the helical structure in collagen samples was confirmed by UV and FTIR spectra. The ultrasound cavitation advanced the viscosity of collagens and resulted in excellent solubility in acidic environments and lower salt concentrations. Ultrasound-treated samples suggested superior water holding capacity along with emulsifying, and foam attributes. The improved gel strength of collagens was parallel with increasing the time of ultrasound up to 15 min. Free radicals scavenging ability and ferric reducing power of collagens were positively stimulated by increasing the time of the ultrasound up to 15 min. Therefore, this study contributes to the development of green technology and the promotion of by-product utilization for collagen recovery as a potential practical protein in biomaterial, wound dressing, drug delivery, food, and cosmetics products.

Effect of ultrasonic pretreatment on flavor characteristics of brewer's yeast‐peanut meal hydrolysate/xylose Maillard reaction products

In this study, we investigated the effect of ultrasonic pretreatment on the flavor characteristics of Maillard reaction products (MRPs) because ultrasound enhances the Maillard reaction (MR). Brewer's yeast-peanut meal hydrolysate was subjected to an ultrasonic treatment, followed by a thermal treatment to induce the MR. Then, several properties of the MRPs were investigated. Compared to the untreated samples, the ultrasound-treated samples presented looser microstructures, which were conducive to the combination of xylose with amino acids or peptides in MR. Consequently, more intermediates were produced and a higher browning degree of the MRPs was observed, compared to those obtained by thermal MR. These findings were supported by the results of MRP analysis through fluorescence and FT-IR spectroscopies. The volatile compounds of samples treated by exposure to 200 W ultrasound endowed the MRPs with prominent flavor characteristics, as confirmed by the scores recorded using Electronic-tongue. Novelty impact statement The flavor characteristics of the MRPs from the brewer's yeast-peanut meal hydrolysate have been reported for the first time in our study. Our findings demonstrate that ultrasonic pretreatment can improve the flavor characteristics of the MRPs, thereby supporting their use in the production of condiments. However, the benefits of the ultrasonic treatment do not always increase with the applied power. This study exploits an agricultural by-product as a raw material, fostering new ways to reduce waste and support a sustainable circular economy.

The effect of high-power ultrasound pretreatment on drying efficiency and bioactive compounds of chokeberry (Aronia melanocarpa L.).

Chokeberry (aronia) fruit is a rich source of bioactive compounds with various health benefits. However, it has a limited shelf life and a firm (tight) structure that makes it difficult for moisture transfer during drying. Therefore, the aim of the study is to investigate the effects of high-power ultrasound pretreatment (550 W, 30 min) and air drying temperature (65, 70 and 75 °C) on the drying rate and bioactive compounds of chokeberry fruit. Accordingly, chokeberries were dried directly or after ultrasound pretreatment, and the retention rate of bioactive properties, i.e., total phenolic, flavonoid and monomeric anthocyanin contents along with antioxidant capacity values, were examined. The required time for drying as well as the activation energy values were significantly lower in ultrasound treated samples. Logarithmic and Midilli models exhibited the best fit for drying kinetics of samples. The pretreatment led to significantly lower moisture content and aw values. Compared to fresh sample, the highest retention rate for some bioactive properties was found in ultrasound pretreated sample dried at 75 °C. The ultrasonication also reduced the electricity consumption (kWh) in all temperature levels of drying operations. As a conclusion, the ultrasound pretreatment prior to drying had dual role by facilitating drying kinetics and by providing higher bioactive contents.

Novel application of ultrasound and microwave-assisted methods for aqueous extraction of coconut oil and proteins.

Alternative methods for wet extraction of coconut oil and protein assisted by ultrasound or microwave were developed and compared. Coconut milk was prepared by milling the pulp (5:1 water to coconut pulp ratio), further destabilised at pH 4 and centrifuged to obtain the cream and cream protein fractions (control process). Microwave-assisted treatment applied in milk (1min, 3 pulses of 20s; 2.5GHz; 4.31kW/kg by pulse) generated a significant increase in cream obtained, and in the coconut oil extraction yield (~ 20%) compared to its control. The ultrasound-assisted treatment (2.5min; 24kHz; 0.573kW/kg, 6.85W/cm2) also improved oil extraction (10-16%). Moreover, a higher protein yield was achieved in ultrasound treated samples when compared to their control (49.6-86.1%). Large particles of 11m , probably aggregates of particles, and smaller particles of 3.6m , were detected in coconut milk, which were reduced by ultrasound effect. Alternative treatments caused a greater liberation of total phenols in coconut cream. Coconut proteins in water (0.1%) showed high negative electrokinetic potential. The surface pressure of coconut proteins at the air/water interface was not modified by assisted treatments.

Effects of Microwaves, Ultrasonication, and Thermosonication on the Secondary Structure and Digestibility of Bovine Milk Protein.

Cow’s milk is considered an excellent protein source. However, the digestibility of milk proteins needs to be improved. This study aimed to evaluate the relationship between the functional properties of milk proteins and their structure upon microwave, ultrasound, and thermosonication treatments. The protein content, digestibility, and secondary-structure changes of milk proteins were determined. The results demonstrated that almost 35% of the proteins in the untreated samples had a α-helix structure and approximately 29% a β-sheet and turns structure. Regarding the untreated samples, the three treatments increased the α-helices and correspondingly decreased the β-sheets and turns. Moreover, the highest milk protein digestibility was observed for the ultrasound-treated samples (90.20–94.41%), followed by the microwave-treated samples (72.56–93.4%), whereas thermosonication resulted in a lower digestibility (68.76–78.81%). The milk protein content was reduced as the microwave processing time and the temperature increased. The final milk protein available in the sample was lower when microwave processing was conducted at 75 °C and 90 °C compared to 60 °C, whereas the ultrasound treatment significantly improved the protein content, and no particular trend was observed for the thermosonicated samples. Thus, ultrasound processing shows a potential application in improving the protein quality of cow’s milk.

Effect of ultrasonication on the metabolome and transcriptome profile changes in the fermentation of Ganoderma lucidum

Development of an efficient liquid fermentation method is helpful for food and pharmaceutical applications. This study investigated the effect of ultrasonication on the liquid fermentation of Ganoderma lucidum, a popular edible and medical fungi. Significant changes at both metabolic and transcriptional levels in mycelia were induced by ultrasound treatment. Compared with the control, 857 differential metabolites were identified (578 up- and 279 down-regulated metabolites), with more metabolites biosynthesis after sonication; 569 differentially expressed genes (DEGs) (267 up- and 302 down-) and 932 DEGs (378 up- and 554 down-) were identified in ultrasound-treated samples with recovery time of 0.5 and 3 h, respectively. Furthermore, 334 DEGs were continuously induced within the recovery time of 3 h, indicating the lasting influence of sonication on mycelia. The DEGs and differential metabolites were mainly involved in pathways of carbohydrate, energy metabolism, amino acids, terpenoids biosynthesis and metabolism and membrane transport, suggesting that ultrasound induced multifaceted effects on primary and secondary metabolism. Ultrasonication enhanced the triterpenoids production of G. lucidum (34.96 %) by up-regulating the expression of terpenoids synthase genes. This study shows that the application of ultrasound in liquid fermentation of G. lucidum is an efficient approach to produce more metabolites.

The Use of Ultrasound for Preventing Honey Crystallization.

The aim of this study was to evaluate the effect of ultrasound treatment for preventing honey crystallization on the physicochemical and microbiological properties of unifloral honey and polyfloral honey. Honey samples without any treatment were used as a control group for comparison. The effect of applying ultrasound treatment was evaluated by studying over time the tendency of crystallization, the rheological properties of honey and chemical and microbiological properties. The parameters analyzed for the two groups of samples (treated and untreated with ultrasound), which did not vary or had small variations during the research were water content, acidity, water activity, glucose, fructose, sucrose, glucose/water ratio, glucose/fructose ratio. The crystallization process was installed in the control samples from the first month of the study, and much later in the treated samples. The color of the untreated samples varied considerably, and the color of the treated ones remained stable or slightly varied. For the control samples, the smallest variation in hydroxymethylfurfural (HMF) concentration was in raspberry honey (5%), and the most significant variation was in honeydew honey (30%). For the treated samples, the largest variation of this parameter was found in tillia honey (127%), and the smallest variation was in rapeseed honey (26%). The microbiological quality was higher for the treated samples. In the ultrasound-treated samples of acacia honey, honeydew honey and grassland honey, yeasts, molds or standard plate counts (SPCs) were undetectable. For control samples, SPC values were <10–50 cfu/g. Ultrasound-treated samples maintained their SPC parameter levels or were thus reduced (<10–20 cfu/g). Yeasts and molds were undetectable or had value between <10 and 10 cfu/g. The yeasts and the molds ranged in the control samples between <10 and 40 cfu/g.

Effects of preliminary treatment by ultrasonic and convective air drying on the properties and oil absorption of potato chips

The initial water content was closely related to the oil absorption and properties of fried food. The effects of convective air drying (D) and ultrasound combined convective air drying (UD) pretreatment on the properties and oil absorption of potato chips have been investigated. The oil contents were 48.48 ± 1.42% and 39.78 ± 3.08% for control samples (without D and UD pretreatment) and ultrasound treated samples (without D pretreatment). When the mass loss of samples was reached the proportion of quality to without drying samples quality 80%, 50%, and 20%, the oil contents of D pretreated samples decreased by 12.67%, 28.24% and 62.07%, respectively, and the oil contents of UD pretreated samples decreased by 7.42%, 24.10% and 51.76% (compared to the ultrasound pretreated samples ), respectively. By applying ultrasound before frying, more cracks and pores were exhibited of fried potato chips. After drying process, potato chips exhibited less disruption of cell structure and less deformation of cell irregular. The hardness of the D and UD pretreated potato chips increased with the extension of drying. The FTIR analysis stated the formation of amylose-lipid complexes. This research could contribute to providing evidence for the development and application of the pretreatment strategies.

Application of ultrasonication at different microbial growth stages during apple juice fermentation by Lactobacillus plantarum: Investigation on the metabolic response

In this work, low-intensity ultrasonication (58.3 and 93.6 W/L) was performed at lag, logarithmic and stationary growth phases of Lactobacillus plantarum in apple juice fermentation, separately. Microbial responses to sonication, including microbial growth, profiles of organic acids profile, amino acids, phenolics, and antioxidant capacity, were examined. The results revealed that obvious responses were made by Lactobacillus plantarum to ultrasonication at lag and logarithmic phases, whereas sonication at stationary phase had a negligible impact. Sonication at lag and logarithmic phases promoted microbial growth and intensified biotransformation of malic acid to lactic acid. For example, after sonication at lag phase for 0.5 h, microbial count and lactic acid content in the ultrasound-treated samples at 58.3 W/L reached 7.91 ± 0.01 Log CFU/mL and 133.70 ± 7.39 mg/L, which were significantly higher than that in the non-sonicated samples. However, the ultrasonic effect on microbial growth and metabolism of organic acids attenuated with fermentation. Moreover, ultrasonication at lag and logarithmic phases had complex influences on the metabolism of apple phenolics such as chlorogenic acid, caffeic acid, procyanidin B2, catechin and gallic acid. Ultrasound could positively affect the hydrolysis of chlorogenic acid to caffeic acid, the transformation of procyanidin B2 and decarboxylation of gallic acid. The metabolism of organic acids and free amino acids in the sonicated samples was statistically correlated with phenolic metabolism, implying that ultrasound may benefit phenolic derivation by improving the microbial metabolism of organic acids and amino acids.

Development of NIR-HSI and chemometrics process analytical technology for drying of beef jerky

Beef jerky samples immersed in brine or water, both with and without ultrasound treatment were dried at 60 °C for 30, 60, 90, 120, 150, 180, 210 and 240 min. Drying behaviour was evaluated using ten drying kinetic models and hyperspectral imaging. All samples reached water activity values (Aw) < 0.85 during the first 90 min of drying. Moisture content (MC) of beef jerky was predicted using near infrared hyperspectral imaging and chemometrics. Partial least squares regression, band selection and spectral pre-treatments were applied to develop MC prediction models using beef jerky spectra. Most of the MC prediction models developed in this work had RPD values >4, indicating their suitability for process control applications. The best performing MC prediction models for the non-ultrasound and ultrasound treated samples were developed using the ensemble Monte Carlo variable selection (EMCVS) on second derivative of log(1/R) spectra and EMCVS-selectivity ratio on linear detrended log (1/R) spectra, respectively. This study demonstrated the potential of NIR-HSI and chemometrics as a PAT tool for drying of beef jerky.

Free radical detection in water after processing by means of high voltage electrical discharges and high power ultrasound

Nonthermal processing technologies have been researched in lab scale for more than 15 years. In this study, the influence of sonification and high voltage electrical discharge—plasma treatment of water on the formation and stabilization of hydroxyl radicals has been investigated by electron paramagnetic resonance (EPR) spectroscopy and spin-trapping method. As analysis tool, an agglomerative hierarchical clustering was used in grouping samples recorded by EPR and near infrared spectroscopy (NIR) spectroscopy. Also, the multivariate analysis was performed using the principal component analysis (PCA) and Principal Regression models (PCR). Results for ultrasound-treated samples showed that number of radicals is increasing with treatment time and increase in amplitude. High-voltage electrical discharge treatment at 120 Hz resulted in the formation of the largest number of hydroxyl radicals. Developed PCR models are highly effective in prediction of NIR spectra, regardless the sample treatment what is an additional step forward in green technology. Practical applications Usage of electron paramagnetic resonance measurement can be used in laboratory and scale-up treatments by nonthermal processing. It is of outmost importance to monitor the production of free radicals, from starting of processing, in order to avoid overtreatment of foods and to avoid possible oxidation and unwanted radical chain reactions. Monitoring of processing can be used to optimize nonthermal processing and to avoid production of unwanted oxidative products and compounds.

Retention and stability of bioactive compounds in functional peach beverage using pasteurization, microwave and ultrasound technologies.

The peach functional beverages pasteurized for 10min at 90°C, microwaved for 1.5min at 850W of power and sonicated for 90min at 20kHz of frequency were selected to keep in storage for up to 30days in refrigerator to examine the changes happened to their physicochemical characteristics and functional components. It was observed that the pH and the cloud values of all processed juice samples reduces with the storage time, whereas, the total soluble solids almost remain consistent particularly in microwave and ultrasound treated samples. While storage period causes the decrement in total phenolic content (TPC) and total flavonoid content of treated beverage samples, but ultrasound processing showed greater retention of TPC value up to 5.7% more than other techniques during storage. The similar trend was observed for antioxidant activity where the ultrasound treatment showed improved free radicals (2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) scavenging activities except ferric ion reducing antioxidant power after 30days of storage.