The main objective of the present study was to investigate the effect of solvent type and ratio as well as the extraction techniques (i.e. supercritical fluid extraction (SFE) and conventional solvent extraction) on betacyanins and antioxidant activity of the peel and fresh extract from the red pitaya (Hylocereus polyrhizus). The peel and flesh extracts obtained by SFE at 25MPa pressure and 10% EtOH/water (v/v) mixture as a co-solvent contained 24.58 and 91.27mg/100ml total betacyanin, respectively; while the most desirable solvent extraction process resulted in a relatively higher total betacyanin in the peel and flesh extracts (28.44 and 120.28mg/100ml, respectively). The major betacyanins identified in the pitaya peel and flesh extracts were betanin, isobetanin, phyllocactin, butyrylbetanin, isophyllocactin and iso-butyrylbetanin. The flesh extract had the stronger antioxidant activity than the peel extract when the higher proportion of ethanol to water (E/W) was applied for the extraction.
The primary objective of this study was to evaluate the physicochemical and rheological properties of butter produced by Lactobacillus helveticus fermented cream. The incorporation of putative probiotic - the L. helveticus, to ferment cream prior to butter production was anticipated to alter the nutritional composition of butter. Changes in crude macronutrients and the resultant modification relating to textural properties of butter induced upon metabolic activities of L. helveticus in cream were focused in this research. Fermented butter (LH-butter) was produced by churning the cream that was fermented by lactobacilli at 37 °C for 24 h. Physicochemical analysis, proximate analysis and rheology properties of LH-butter were compared with butter produced using unfermented cream (control). LH-butter showed a significantly (P<0.05) higher fat content and acid value; lower moisture and ash; and was softer than the control. Cream fermentation modified nutritional and textural properties of butter in which LH-butter contained higher health beneficial unsaturated fatty acids than the control and thus rendered the product softer. Its enrichment with probiotics could thus further enhance its functional property.
The purpose of this study was to identify porcine-specific peptide markers from thermally processed meat that could differentiate pork from beef, chevon and chicken meat. In the initial stage, markers from tryptic digested protein of chilled, boiled and autoclaved pork were identified using LC-QTOF-MS. An MRM method was then established for verification. A thorough investigation of LC-QTOF-MS data showed that only seven porcine-specific peptides were consistently detected. Among these peptides, two were derived from lactate dehydrogenase, one from creatine kinase, and four from serum albumin protein. However, MRM could only detect four peptides (EVTEFAK, LVVITAGAR, FVIER and TVLGNFAAFVQK) that were consistently present in pork samples. In conclusion, meat species determination through a tandem mass spectrometry platform shows high potential in providing scientifically valid and reliable results even at peptide level. Besides, the specificity and selectivity offered by the proteomics approach also provide a robust platform for Halal authentication.
Little is known about the bioavailability of heavy metal contamination and its health risks after rice ingestion. This study aimed to determine bioavailability of heavy metal (As, Cd, Cu, Cr, Co, Al, Fe, Zn and Pb) concentrations in cooked rice and human Health Risk Assessment (HRA). The results found Zn was the highest (4.3±0.1 mg/kg), whereas As showed the lowest (0.015±0.001 mg/kg) bioavailability of heavy metal concentration in 22 varieties of cooked rice. For single heavy metal exposure, no potential of non carcinogenic health risks was found, while carcinogenic health risks were found only for As. Combined heavy metal exposures found that total Hazard Quotient (HQtotal) values for adult were higher than the acceptable range (HQTotal<1), whereas total Lifetime Cancer Risk (LCRTotal) values were higher than the acceptable range (LCRTotal values >1×10(-4)) for both adult and children. This study is done to understand that the inclusion of bioavailability heavy metal into HRA produces a more realistic estimation of human heavy metal exposure.
In this paper, we investigated the antibacterial mechanism and potential therapeutic targets of three antibacterial medicinal plants. Upon treatment with the plant extracts, bacterial proteins were extracted and resolved using denaturing gel electrophoresis. Differentially-expressed bacterial proteins were excised from the gels and subjected to sequence analysis by MALDI TOF-TOF mass spectrometry. From our study, seven differentially expressed bacterial proteins (triacylglycerol lipase, N-acetylmuramoyl-L-alanine amidase, flagellin, outer membrane protein A, stringent starvation protein A, 30S ribosomal protein s1 and 60 kDa chaperonin) were identified. Additionally, scanning electron microscope study indicated morphological damages induced on bacterial cell surfaces. To the best of our knowledge, this represents the first time these bacterial proteins are being reported, following treatments with the antibacterial plant extracts. Further studies in this direction could lead to the detailed understanding of their inhibition mechanism and discovery of target-specific antibacterial agents.
This study aimed to isolate a potent antiglucosidase and antioxidant fraction from Stenochlaena palustris. Extraction was performed with hexane, chloroform, ethyl acetate, methanol, and water. Antiglucosidase, 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging and ferric reducing antioxidant power (FRAP) assays found methanol extract (ME) to be the most active. Water fraction (WF) of ME was a stronger α-glucosidase inhibitor (EC50 2.9 μg/mL) than quercetin, with weak antiamylase activity. WF was a competitive α-glucosidase inhibitor. DPPH scavenging activity of WF (EC50 7.7 μg/mL) was weaker than quercetin. WF (EC50 364 μg/mL) was a stronger hydrogen peroxide scavenger than gallic acid (EC50 838 μg/mL) and was equally strong as quercetin in scavenging superoxide. WF possessed moderate copper chelating activity. WF was enriched in total phenolics (TP) and hydroxycinnamic acids (THC). TP correlated with antioxidant activity (R(2) > 0.76). Only THC correlated with antiglucosidase activity (R(2) = 0.86). Overall, WF demonstrated concurrent, potent antiglucosidase and antioxidant activities.
This research was conducted to investigate the effects of different grinding techniques (dry, semi-wet and wet) of milled rice grains on the damaged starch and particle size distribution of flour produced from a new variety, MR263, specifically related to the pasting and thermal profiles. The results indicated that grinding techniques significantly (p<0.05) affected starch damage content and particle size distribution of rice flour. Wet grinding process yields flour with lowest percentage of starch damage (7.37%) and finest average particle size (8.52μm). Pasting and gelatinization temperature was found in the range of 84.45-89.63°C and 59.86-75.31°C, respectively. Dry ground flour attained the lowest pasting and gelatinization temperature as shown by the thermal and pasting profiles. Correlation analysis revealed that percentage of damaged starch granules had a significant, negative relationship with pasting temperature while average particle size distribution had a significant, strong negative relationship with gelatinization temperature.
Antioxidant and α-amylase inhibitor peptides were successfully extracted from Pinto bean protein isolate (PBPI) using Protamex. A factorial design experiment was conducted and the effects of extraction time, pH and temperature were studied. pH 7.5, extraction time of 1h, S/E ratio of 10 (w/w) and temperature of 50 °C gave the highest antioxidant activities (i.e., ABTS scavenging activity (53.3%) and FRAP value (3.71 mM)), whereas pH 6.5 with the same extraction time, S/E ratio and temperature, gave the highest α-amylase inhibitory activity (57.5%). It was then fractioned using membrane ultrafiltration with molecular weight cutoffs of 100, 50, 30, 10 and 3 kDa. Peptide fraction <3 kDa, which exhibited the highest antioxidant activities (i.e., ABTS (42.2%) and FRAP (0.81 mM)) and α-amylase inhibitory activity (62.1%), was then subjected to LCMS and MS/MS analyses. Six sequences were identified for antioxidant peptides, whereas seven peptides for α-amylase inhibitor.
Fragaria chiloensis fruit has a short postharvest life mainly due to its rapid softening. In order to improve its postharvest life, preharvest applications of methyl jasmonate (MeJA) and chitosan were evaluated during postharvest storage at room temperature. The quality and chemical parameters, and protection against decay were evaluated at 0, 24, 48 and 72 h of storage from fruits of two subsequent picks (termed as first harvest and second harvest). In general, fruits treated with MeJA and chitosan maintained higher levels of fruit firmness, anthocyanin, and showed significant delays in decay incidence compared to control fruit. MeJA-treated fruits exhibited a greater lignin content and SSC/TA ratio, and delayed decay incidences. Instead, chitosan-treated fruits presented higher antioxidant capacity and total phenol content. In short, both the elicitors were able to increase the shelf life of fruits as evidenced by the increased levels of lignin and anthocyanin, especially of the second harvest.
Various 6-methoxytetrahydro-β-carboline derivatives, namely BEN (6-methoxy-1-phenyl-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole), ANI (6-methoxy-1-(4-methoxyphenyl)-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole), ACE (6-methoxy-1-methyl-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole) and VAN (2-methoxy-4-(6-methoxy-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indol-1-l)phenol), were prepared via the Maillard reaction using food flavours and 5-methoxytryptamine in aqueous medium and were investigated for their in vitro antioxidant and cytotoxicity properties. These derivatives were found to exhibit moderate antioxidant properties, based on a combination of DPPH, ABTS and FRAP assays. The results suggested that the Maillard reaction could be used to generate β-carboline antioxidants. It was beneficial that VAN showed the highest antioxidant activity but the least cytotoxic activities on non-tumourous cell lines of NIH/3T3, CCD18-Co and B98-5 using MTT assay. ACE, ANI and BEN showed mild toxicity at effective antioxidative concentrations derived from DPPH and ABTS assays. Furthermore, they are safer compared to 5-fluorouracil, cisplatin and betulinic acid on NIH/3T3, CCD18-Co and B98-5 cells. In conclusion, the antioxidant and cytotoxicity properties of 6-methoxytetrahydro-β-carbolines were demonstrated for the first time.
The large amount of waste produced by the food industries causes serious environmental problems and also results in economic losses if not utilized effectively. Different research reports have revealed that food industry by-products can be good sources of potentially valuable bioactive compounds. As such, the mango juice industry uses only the edible portions of the mangoes, and a considerable amount of peels and seeds are discarded as industrial waste. These mango by-products come from the tropical or subtropical fruit processing industries. Mango by-products, especially seeds and peels, are considered to be cheap sources of valuable food and nutraceutical ingredients. The main uses of natural food ingredients derived from mango by-products are presented and discussed, and the mainstream sectors of application for these by-products, such as in the food, pharmaceutical, nutraceutical and cosmetic industries, are highlighted.
5-Hydroxymethylfurfural (HMF) is formed during heat treatment of carbohydrate-containing foods, especially in a deep-fat frying process. This study aimed to investigate the effect of amino acids on the formation and reduction of HMF from glucose, fructose and sucrose at frying temperature in model systems containing binary mixtures of an amino acid and a sugar in equal concentrations (0.3M). The results revealed that the formation of HMF from sugars accelerated in the presence of acidic amino acids (i.e. glutamic and aspartic acids). Conversely, the presence of basic amino acids (i.e. lysine, arginine and histidine) led to reduced concentrations of HMF to non-detectable levels in model systems. The results showed that both pH and heating time significantly affected the formation of HMF from fructose in the presence of glutamic acid. In this regard, a higher amount of HMF was formed at lower pH.
Tamarillo (Solanum betaceum Cav.) is an underutilised fruit in Malaysia. The fruit, however, contains good proportions of soluble fibre, protein, starch, anthocyanins and carotenoids. Amongst the fruits, only tamarillo mesocarp contains both polar (anthocyanins) and non-polar (carotenoids) pigments. The ability to retain both polar and non-polar pigments in the mesocarp could be related to the unique properties of its hydrocolloids. To understand the pigment-hydrocolloid interaction in the fruit, information on the physicochemical characteristics of the hydrocolloids is required. Therefore, hydrocolloids from the anthocyanin-rich seed mucilage fraction of the tamarillo and its carotenoid-rich pulp fraction were extracted and characterised. Water and 1% citric acid were used to extract the seed mucilage hydrocolloid while 72% ethanol and 20mM HEPES buffer were used for pulp hydrocolloid extraction. Seed mucilage hydrocolloid was primarily composed of arabinogalactan protein-associated pectin whereas pulp hydrocolloid was composed of hemicellulosic polysaccharides with some naturally interacting proteins and neutral polysaccharides.
In this study, the methods for extraction and purification of miraculin from Synsepalum dulcificum were investigated. For extraction, the effect of different extraction buffers (phosphate buffer saline, Tris-HCl and NaCl) on the extraction efficiency of total protein was evaluated. Immobilized metal ion affinity chromatography (IMAC) with nickel-NTA was used for the purification of the extracted protein, where the influence of binding buffer pH, crude extract pH and imidazole concentration in elution buffer upon the purification performance was explored. The total amount of protein extracted from miracle fruit was found to be 4 times higher using 0.5M NaCl as compared to Tris-HCl and phosphate buffer saline. On the other hand, the use of Tris-HCl as binding buffer gave higher purification performance than sodium phosphate and citrate-phosphate buffers in IMAC system. The optimum purification condition of miraculin using IMAC was achieved with crude extract at pH 7, Tris-HCl binding buffer at pH 7 and the use of 300 mM imidazole as elution buffer, which gave the overall yield of 80.3% and purity of 97.5%. IMAC with nickel-NTA was successfully used as a single step process for the purification of miraculin from crude extract of S. dulcificum.
Jackfruit is a sweet tropical fruit with very pleasant aroma, and the ripe seeds are edible. In this study, jackfruit seed proteins were isolated and subjected to trypsin digestion. The resultant protein hydrolysate was then subjected to antioxidant assay-guided purification, using centrifugal filtration, C18 reverse-phase and strong cation exchange (SCX) fractionations. The purified SCX fraction was further analyzed by de novo peptide sequencing, and two peptide sequences were identified and synthesized. Peptide JFS-2 (VGPWQK) was detected with antioxidant potential, with EC50 value comparable to that of commercial GSH antioxidant peptide. Additionally, the identified peptides were tested with protein protection potential, in an albumin protein denaturation inhibitory assay. Concurrently, we also investigated the pH, temperature, and gastrointestinal-digestion stability profiles for the identified peptide. With further research efforts, the identified peptides could potentially be developed into preservative agent for protein-rich food systems or as health-promoting diet supplements.
Palm-pressed mesocarp oil has been found to contain plenty of naturally occurring valuable phytonutrients. The application and study of the oil are limited, therefore, quality assessment of refined red palm-pressed mesocarp olein (PPMO) is deemed necessary to provide data in widening the applications as a niche products or raw material for the nutraceutical industry. Results showed that refined PPMO has comparable physicochemical properties and oxidative stability with commercial cooking oil, palm olein (PO). The food safety parameters and contaminants (PAH, 3-MCPD ester, 2-MCPD ester, glycidyl ester and trace metals) analyses proven that refined PPMO is safe to be consumed. Besides, refined PPMO contains remarkably greater concentrations of phytonutrients including carotenoids, phytosterols, squalene and vitamin E than PO, postulating its protective health benefits. The overall quality assessment of refined PPMO showed that it is suitable for human consumption and it is a good source for food applications and dietary nutritional supplements.
Lysozyme from crude chicken egg white (CEW) feedstock was successfully purified using a stirred fluidized bed adsorption system ion exchange chromatography where STREAMLINE SP and SP-XL high density adsorbents were selected as the adsorption carrier. The thermodynamic and kinetic studies were carried out to understand the characteristics of lysozyme adsorption by adsorbents under various conditions, including adsorption pH, temperature, lysozyme concentration and salt concentrations. Results showed that SP and SP-XL adsorbents achieved optimum lysozyme adsorption at pH 9 with capacity of ~139.77 and ~251.26 mg/mL, respectively. The optimal conditions obtained from batch studies were directly employed to operate in SFBA process. For SP-XL adsorbent, the recovery yield and purification factor of lysozyme were 93.78% and ~40 folds, respectively. For SP adsorbent, lysozyme can be eluted ~100% with purification factor of ~26 folds. These two adsorbents are highly suitable for use in direct recovery of lysozyme from crude CEW.
The redfleshed pulp discarded from pink guava puree industry is a rich source of lycopene and pectin. In this study, we developed a facile extraction process employing water as the primary extraction medium to isolate the lycopene and pectin from pink guava decanter. When the decanter was suspended in water, the complexation of lycopene and pectin formed the cloudy solution, where the colloidal complexes were recovered through centrifugation. The presence of lycopene and pectin in the complex was confirmed by the spectroscopic, microscopic and chromatographic analyses. The lycopene fractionated from the complexes had a purity level of 99% and was in all-trans configuration. The colloidal complexes yielding the highest concentration of lycopene was obtained at pH 7, 1% (w/v) solid loading and 25 °C. The experimental data of time-course extraction of lycopene-pectin complex were best fitted with two-site kinetic model, hinting the fast- and slow-release phases in the extraction process.
Solution-enhanced dispersion by supercritical carbon dioxide (SEDS) and spray drying (SD) were used to microencapsulate red palm oil (RPO) to prolong the functionality of carotenes and vitamin E. The protective effects provided by SEDS and SD were evaluated in terms of the oxidative stability (65 °C for 35 days), fatty acid compositions, color change and degradation kinetics of carotenes and vitamin E (25 °C, 45 °C, 65 °C, and 85 °C for up to 198 days). SEDS microcapsules (SEDS-M) were the most oxidatively stable (total oxidation (Totox): 26.5), followed by SD microcapsules (SD-M) (34.9) and RPO (56.7). Degradation of carotenes and vitamin E fitted well a first-order kinetic model (average absolute relative deviation = 2-16%). SEDS-M offered better protection to vitamin E (Ea = 36 kJ/mol), whereas SD-M provided better protection for α + β carotene (Ea = 29 kJ/mol). Overall, encapsulation protected RPO during storage, with SEDS-microencapsulated RPO performing better than SD-microencapsulated RPO.
In this article, an easy and quick method based on microwave assisted acid digestion technique prior to quantification using inductively coupled plasma mass spectrometry for the analysis of heavy metals in cocoa beans, cocoa powder and chocolate was established and validated for arsenic (As), cadmium (Cd), lead (Pb), and antimony (Sb). Limit of quantification for all elements were product dependent and varies from 7.84 to 194.52 µg/kg. The recoveries of the heavy metals at 250 and 1000 µg/kg spiking levels were ranged between 96.27-108.75%, 90.43-101.97% and 89.72-106.26% for cocoa beans, cocoa powder, and chocolate, respectively. Relative standard deviation values obtained were all below 20% and the expanded uncertainty measurements for the elements were less than 25%. The analysis of real samples found that the concentration level is far from the national alarming level except for cadmium in cocoa beans.