Gelatin is widely used in food and pharmaceutical products. However, the addition of gelatin especially in food products becomes a controversial issue among Muslims due to its animal origin. Thus, the present study was aimed to detect and differentiate the origin of gelatin added in processed foods using a combination method of sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and Principal Component Analysis (PCA). Porcine gelatin had exhibited 11 prominent polypeptides compared to bovine gelatin with 2 prominent polypeptides. Polypeptides of both gelatin sources at molecular weight ranged from 53 to 220 kDa can be used to differentiate between porcine and bovine gelatins using PCA. The efficiency in extracting gelatin from processed foods by different solutions was also evaluated. Extraction of gelatin in processed foods by cold acetone and deionised water had exhibited a similar polypeptide patterns, suggesting both solutions are suitable. The study indicated that approach of a simple gelatin extraction combined with SDS-PAGE and PCA, may provide robust information for gelatin species differentiation of processed foods.
The effect of solvent type in antioxidant compounds extraction from banana tissues was studied. The solvent system used was pure methanol, ethanol, acetone and their aqueous solution at 50% and 70% concentrations. Comparison among three common cultivars of banana in Malaysia (Berangan, Mas and Raja) had been done and their antioxidant activities were determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging system, ferric reducing ability in plasma (FRAP)
assays and total phenolic content (TPC) assays. Acetone 70% had the strongest antioxidant compounds extraction power as compared to other solvent. All banana samples were found to be low in primary antioxidant but powerful secondary antioxidant source of fruit. The ascending order of banana cultivars in term of their antioxidant activities in all antioxidant assays carried out were Berangan < Mas < Raja. FRAP-TPC assays were highly correlated (R2>0.70) than FRAP-DPPH
and TPC-DPPH assays due to the same mechanism that occurred in the reaction of FRAP and TPC assays.
The galls of Quercus infectoria are commonly used in Malay traditional medicine to treat wound infections after childbirth. In India, they are employed traditionally as dental applications such as that in treatment of toothache and gingivitis. The aim of the present study was to evaluate the antibacterial activity of galls of Quercus infectoria Olivier against oral bacteria which are known to cause dental caries and periodontitis. Methanol and acetone extracts were screened against two Gram-positive bacteria (Streptococcus mutans ATCC 25175 and Streptococcus salivarius ATCC 13419) and two Gram-negative bacteria (Porphyromonas gingivalis ATCC 33277 and Fusobacterium nucleatum ATCC 25586). The screening test of antibacterial activity was performed using agar-well diffusion method. Subsequently, minimum inhibitory concentration (MIC) was determined by using twofold serial microdilution method at a concentration ranging between 0.01 mg/mL and 5 mg/mL. Minimum bactericidal concentration (MBC) was obtained by subculturing microtiter wells which showed no changes in colour of the indicator after incubation. Both extracts showed inhibition zones which did not differ significantly (P < 0.05) against each tested bacteria. Among all tested bacteria, S. salivarius was the most susceptible. The MIC ranges for methanol and acetone extracts were the same, between 0.16 and 0.63 mg/mL. The MBC value, for methanol and acetone extracts, was in the ranges 0.31-1.25 mg/mL and 0.31-2.50 mg/mL, respectively. Both extracts of Q. infectoria galls exhibited similar antibacterial activity against oral pathogens. Thus, the galls may be considered as effective phytotherapeutic agents for the prevention of oral pathogens.
Henna plant (Lawsonia inermis) is an Indian medicinal plant used in traditional medicine for the treatment of various diseases, besides its popularity as a natural dye to colour hand and hair. Research in the recent past has accumulated enormous evidence revealing henna plant to be an excellent source of antioxidants such as total phenolics. In this study, the extraction of total phenolics from henna stems was evaluated using the Folin-Ciocalteu assay. A set of single factor experiments was carried out for identifying the optimum condition of each independent variable affecting total phenolic content (TPC) extraction efficiency of henna stems, namely the solvent type, solvent concentration (v/v, %), extraction time (min) and extraction temperature (oC). Generally, high extraction yield was obtained using aqueous acetone (about 40%) as solvent and the extraction yield could further be increased using a prolonged time of 270 min and a higher incubation temperature of 55°C. Under these optimized conditions, the experimental maximum yield of TPC of 5554.15 ± 73.04 mg GAE/100 g DW was obtained.
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.
Palm kernel cake (PKC) was used for biobutanol production by Clostridium saccharoperbutylacetonicum N1-4 in acetone-butanol-ethanol (ABE) fermentation. PKC was subjected to acid hydrolysis pretreatment and hydrolysates released were detoxified by XAD-4 resin. The effect of pH, temperature and inoculum size on butanol production was evaluated using an empirical model. Twenty ABE fermentations were run according to an experimental design. Experimental results revealed that XAD-4 resin removed 50% furfural and 77.42% hydroxymethyl furfural. The analysis of the empirical model showed that linear effect of inoculums size with quadratic effect of pH and inoculum size influenced butanol production at 99% probability level (P<0.01). The optimum conditions for butanol production were pH 6.28, temperature of 28°C and inoculum size of 15.9%. ABE fermentation was carried out under optimum conditions which 0.1g/L butanol was obtained. Butanol production was enhanced by diluting PKC hydrolysate up to 70% in which 3.59g/L butanol was produced.
Bioactive compounds from Quercus Infectoria (manjakani) were extracted with six different types of solvents: 100% methanol, ethanol, acetone, aqueous and 70% methanol and ethanol. High Performance liquid chromatography (HPLC) was used to identify and quantify the active compounds, namely gallic acid and tannic acid. Total phenolics content were determined by Folin-Ciocalteu while antioxidant and antibacterial activity were tested using DPPH free radicals scavenging and disc diffusion assay. The result revealed that aqueous extract contained the highest concentration of bioactive compounds compared to other types of solvents which are 51.14 mg/g sample and 1332.88 mg/g sample of gallic acid and tannic acid respectively.. The highest level of phenolic compound was found in 100% acetone extract (121 mg GAE/g). The results demonstrated that aqueous extract gives the highest antioxidant activity approximately 94.55% while acetone extract gives the largest inhibition zone for disc diffusion assay which is 19.00 mm respectively. The results revealed rich sources of gallic acid and tannic acid in Q. infectoria which might provide a novel source of these natural antioxidant and antibacterial activity.
Protease was extracted from two maturity stages of noni fruits (Morinda citrifolia L.), unripe (stage 1) and ripe (stage 5). The crude extract was partially purified by acetone precipitation method followed by dialysis, gel filtration chromatography and freeze drying. Protein concentrations, proteolytic activity, molecular weight distribution, pH stability, temperature stability and storage efficiency of the resulting protease were evaluated. The unripe and ripe noni fruit contains 0.65 and 0.35% protein, respectively. Molecular weight of the proteases from both stages ranged approximately between 3 to 28 kDa based on the SDS-PAGE results. The optimum activity were at pH 7s and 6, temperatures of 40 and 50°C, respectively for proteases obtained from the unripe and ripe fruit. Analysis from the freeze dried protease indicated that protease from ripe noni fruits had higher protein concentration and specific activity compared to those from unripe fruit. However, it is more sensitive to pH and temperature and less stable during storage as it shows lower proteolytic activity compared to protease from unripe fruit. Based on its high proteolytic activity reaching up to 70.31 U/mg and storage stability (30% lost of activity), noni fruit could be an alternative source of plant protease.
This study aimed to optimise potential extraction conditions using response surface methodology (RSM) for yielding maximum levels of total phenolic content (TPC) and 2,2-diphenyl-1-picrylhydrazyl radical (DPPH•) scavenging capacity of henna (Lawsonia inermis) stems. The ranges for selected independent variables, namely acetone concentration (20−90%, v/v), extraction time (10−90 min), and extraction temperature (25−45°C) were identified by screening tests. Optimum conditions obtained for extraction of TPC were 47.0% acetone, extraction time of 47.6 min and extraction temperature of 37.3oC. The result also showed that 75.8% acetone, extraction time of 26.2 min and extraction temperature of 41oC yielded the highest DPPH• scavenging capacity. The optimized extraction conditions have resulted in TPC and DPPH• scavenging capacity of 5232.4 mg GAE/100 g DW and 6085.7 mg TE/100 g DW, respectively which similar to the predicted values. Therefore, RSM has successfully optimized the extraction conditions for TPC and radical scavenging capacity of henna stems.
Novel, fast, selective, eco-friendly and reproducible solid-phase membrane tip extraction and gas chromatography with mass spectrometry methods were developed and validated for the analysis of triazine herbicides (atrazine and secbumeton) in stream and lake waters. The retention times of atrazine and secbumeton were 7.48 and 8.51 min. The solid-phase membrane tip extraction was carried out in semiautomated dynamic mode on multiwall carbon nanotubes enclosed in a cone-shaped polypropylene membrane cartridge. Acetone and methanol were found as the best preconditioning and desorption solvents, respectively. The extraction and desorption times for these herbicides were 15.0 and 10.0 min, respectively. The percentage recoveries of atrazine and secbumeton were 88.0 and 99.0%. The linearity range was 0.50-80.0 μg/L (r(2) > 0.994), with detection limits (<0.47 μg/L, S/N = 3) and good reproducibility (<8.0%). The ease of operation, eco-friendly nature, and low cost of solid-phase membrane tip extraction made these methods novel. The Solid-phase membrane tip extraction method was optimized by considering the effect of extraction time, desorbing solvents and time.
Oil palm (Elaeis guineensis) is an important economic crop cultivated for its nutritional palm oil. A significant amount of effort has been undertaken to understand oil palm growth and physiology at the molecular level, particularly in genomics and transcriptomics. Recently, proteomics studies have begun to garner interest. However, this effort is impeded by technical challenges. Plant sample preparation for proteomics analysis is plagued with technical challenges due to the presence of polysaccharides, secondary metabolites and other interfering compounds. Although protein extraction methods for plant tissues exist, none work universally on all sample types. Therefore, this study aims to compare and optimize different protein extraction protocols for use with two-dimensional gel electrophoresis of young and mature leaves from the oil palm. Four protein extraction methods were evaluated: phenol-guanidine isothiocyanate, trichloroacetic acid-acetone precipitation, sucrose and trichloroacetic acid-acetone-phenol. Of these four protocols, the trichloroacetic acid-acetone-phenol method was found to give the highest resolution and most reproducible gel. The results from this study can be used in sample preparations of oil palm tissue for proteomics work.
In the title complex salt, [Au2{(C6H5)2PCH2P(C6H5)2}]Cl2·(CH3)2C=O·H2O, the dication forms an eight-membered {-PCPAu}2 ring with a transannular aurophilic inter-action [Au⋯Au = 2.9743 (2) Å]. The ring approximates a flattened boat conformation, with the two methyl-ene C atoms lying ca 0.58-0.59 Å above the least-squares plane defined by the Au2P4 atoms (r.m.s. deviation = 0.0849 Å). One Cl(-) anion functions as a weak bridge between the Au(I) atoms [Au⋯Cl = 2.9492 (13) and 2.9776 (12) Å]. The second Cl(-) anion forms two (water)O-H⋯Cl hydrogen bonds about a centre of inversion, forming a centrosymmetric eight-membered {⋯HOH⋯Cl}2 supra-molecular square. Globally, the dications and loosely associated Cl(-) anions assemble into layers lying parallel to the ac plane, being connected by C-H⋯Cl,π(phen-yl) inter-actions. The supra-molecular squares and solvent acetone mol-ecules are sandwiched in the inter-layer region, being connected to the layers on either side by C-H⋯Cl,O(acetone) inter-actions.
Acacia species are multipurpose trees, widely used in the traditional systems of medicine to treat various ailments. The major objective of the present study was to determine the gene expression of enzymatic antioxidants by acetone extract from the stem bark of three Acacia species (Acacia dealbata, Acacia ferruginea and Acacia leucophloea) in hydrogen peroxide (H2O2)-induced human hepatoma (HepG2) cells. The expression of antioxidant enzymes such as superoxide dismutase containing copper-zinc (CuZnSOD)/manganese (MnSOD), catalase (CAT) and glutathione peroxidase (GPx) in HepG2 cells was evaluated by real-time PCR. The results of antioxidant enzyme expression in real-time PCR study revealed that the H2O2 (200 μM) challenged HepG2 cells reduced the expression of enzymes such as SOD, GPx and CAT. However, the cells pre-treated with acetone extracts of all the three Acacia species significantly (P > 0.05) up-regulated the expression of antioxidant enzymes in a concentration dependent manner (25, 50 and 75 μg/mL). In conclusion, the findings of our study demonstrated that the acetone extract of Acacia species effectively inhibited H2O2 mediated oxidative stress and may be useful as a therapeutic agent in preventing oxidative stress mediated diseases.
Acetone soluble oil palm empty fruit bunch cellulose acetate (OPEFB-CA) of DS 2.52 has been successfully synthesized in a one-step heterogeneous acetylation of OPEFB cellulose without necessitating the hydrolysis stage. This has only been made possible by the mathematical modeling of the acetylation process by manipulating the variables of reaction time and acetic anhydride/cellulose ratio (RR). The obtained model was verified by experimental data with an error of less than 2.5%. NMR analysis showed that the distribution of the acetyl moiety among the three OH groups of cellulose indicates a preference at the C6 position, followed by C3 and C2. XRD revealed that OPEFB-CA is highly amorphous with a degree of crystallinity estimated to be ca. 6.41% as determined from DSC. The OPEFB-CA films exhibited good mechanical properties being their tensile strength and Young's modulus higher than those of the commercial CA.
Improvement in the butanol production selectivity or enhanced butanol:acetone ratio (B:A) is desirable in acetone-butanol-ethanol (ABE) fermentation by Clostridium strains. In this study, artificial electron carriers were added to the fermentation medium of a new isolate of Clostridium acetobutylicum YM1 in order to improve the butanol yield and B:A ratio. The results revealed that medium supplementation with electron carriers changed the metabolism flux of electron and carbon in ABE fermentation by YM1. A decrease in acetone production, which subsequently improved the B:A ratio, was observed. Further improvement in the butanol production and B:A ratios were obtained when the fermentation medium was supplemented with butyric acid. The maximum butanol production (18.20 ± 1.38 g/L) was gained when a combination of methyl red and butyric acid was added. Although the addition of benzyl viologen (0.1 mM) and butyric acid resulted in high a B:A ratio of 16:1 (800% increment compared with the conventional 2:1 ratio), the addition of benzyl viologen to the culture after 4 h resulted in the production of 18.05 g/L butanol. Manipulating the metabolic flux to butanol through the addition of electron carriers could become an alternative strategy to achieve higher butanol productivity and improve the B:A ratio.
Antioxidants in seaweeds have attracted increasing interest for its role in protecting human health. Therefore, the aim of this study was to assess the Total phenolic content (TPC) values and antioxidant activities in red seaweeds Kappaphycus alvarezii and Kappaphycus striatum of different solvent extracts. Total phenolic content (TPC) and antioxidant activities (DPPH scavenging assay and Trolox equivalent antioxidant capacity assay, TEAC) for both K. alvarezii and K. striatum extracts were determined using different solvents at different concentrations (ethanol: 50%, 70%, 100%; acetone: 50%, 70%, 100%; methanol: 50%, 70%, 100%). The TPC value was measured using the Folin-Ciocalteu’s method. The antioxidant activities were measured by 2, 2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay and Trolox Equivalent Antioxidant Capacity (TEAC) assay. The highest TPC value of K. alvarezii antioxidant extract was obtained by 50% ethanol extracts while for K. striatum obtained by 50% methanol extract. The highest percentage of DPPH free radical inhibition for K. alvarezii was shown by 50% acetone extract while K. striatum was shown using 50% methanol extract. The highest TEAC value for K. alvarezii was shown by 50% acetone while K. striatum extract was shown by 50% ethanol extract. The TPC values and antioxidant activities of all solvent extracts of K. striatum were significantly higher (p< 0.05) than K. alvarezii antioxidant extracts. The TPC values showed strong correlation (r = 0.797) with TEAC values for K. alvarezii antioxidant extract (p< 0.01). The TEAC values also showed strong correlation (r = 0.735) with percentage of DPPH free radical inhibition for K. alvarezii (p< 0.01). The TPC value, DPPH free radical scavenging assay and TEAC assay for K. striatum extracts showed strong correlation (r> 0.8) with each other (p< 0.01). In summary, K. striatum showed better antioxidant activity and higher TPC value than K. alvarezii.
Proteases were extracted from starfruit at maturity Index 2 (unripe, light green) and Index 7 (very ripe, orange) and partially purified using acetone and 40% ammonium sulfate precipitations. Higher yield and proteolytic activity were observed for proteases purified using acetone than 40% ammonium sulfate. As for maturity index, yield and protein concentration of proteases from Index 2 were higher than those from Index 7. SDS-PAGE result showed intense bands for acetone proteases while a distinct band at 50 kDa was observed in all the proteases. Enzyme activity decreased during the seven days storage at 4°C with minimum relative activity of 70% achieved for acetone proteases at day seven. This study suggested that acetone precipitation is more effective method for purifying starfruit protease based on the yield and proteolytic activity compared to using 40% ammonium sulphate precipitation. In order to obtain higher protein concentration and proteolytic activity, starfruit at the unripe stage, Index 2 is a better raw material than Index 7 to be used for protease production.
Effects of different types of solvent on the antioxidant and antibacterial activity of Quercus infectoria extract have not been well documented. Therefore, extraction process was conducted using conventional Soxhlet extraction with six different types of solvent (100% methanol, ethanol, acetone, water and 70% methanol, and ethanol). High performance liquid chromatography was implemented to identify gallic acid and tannic acid in the extracts. Water extracts contained the highest concentration of both gallic acid and tannic acid compared to other types of solvent; 51.14 mg/g sample and 1332.88 mg/g sample of gallic acid and tannic acid. Meanwhile, antioxidant and antibacterial activity were tested using DPPH free radicals scavenging and disc diffusion assay. Results demonstrated that water extracts gave the highest antioxidant activity (approximately 94.55%), while acetone extract gave the largest inhibition zone for disc diffusion assay (19.00mm respectively). The results also revealed rich sources of gallic acid and tannic acid in Q. infectoria which might provide a novel source of these natural antioxidant and antibacterial activity.
It is crucial to determine several protein-related parameters at the initial stages of proteomic analysis of any biological samples. In this study, crude protein content, total soluble protein, total phenolic content and the SDS-PAGE profile of fifteen varieties of seaweed from Semporna, Sabah, Malaysia were analysed. The crude protein, total soluble protein and total phenolic content of all seaweed samples were in the range of 3.99 to 13.18 % of dry weight, 0.52 to 1.45 mg/mL in acetone dried powder samples and 8.59 to 48.98 mg PGE/g dry weight, respectively. In general, the differences (crude protein, total soluble protein and total phenolic content) among all fifteen varieties of seaweeds were significant (p< 0.05). There was also a strong positive correlation between crude protein and total soluble protein concentration (Pearson’s Correlation Coefficient (r)=0.923; p=0.01) in these fifteen varieties of seaweed. A distinctive protein pattern was observed in the SDS-PAGE gels between three different seaweed classes of green, red and brown colours. All of these results are important in sample preparations (extractions) before furthering proteomic analysis in order to identify and characterize seaweed proteomes.
We investigated the effect of different solvents (ethyl acetate, methanol, acetone, and chloroform) on the extraction of phytoconstituents from Lantana camara leaves and their antioxidant and antibacterial activities. Further, GC-MS analysis was carried out to identify the bioactive chemical constituents occurring in the active extract. The results revealed the presence of various phytocompounds in the extracts. The methanol solvent recovered higher extractable compounds (14.4% of yield) and contained the highest phenolic (92.8 mg GAE/g) and flavonoid (26.5 mg RE/g) content. DPPH radical scavenging assay showed the IC50 value of 165, 200, 245, and 440 μg/mL for methanol, ethyl acetate, acetone, and chloroform extracts, respectively. The hydroxyl scavenging activity test showed the IC50 value of 110, 240, 300, and 510 μg/mL for methanol, ethyl acetate, acetone, and chloroform extracts, respectively. Gram negative bacterial pathogens (E. coli and K. pneumoniae) were more susceptible to all extracts compared to Gram positive bacteria (M. luteus, B. subtilis, and S. aureus). Methanol extract had the highest inhibition activity against all the tested microbes. Moreover, methanolic extract of L. camara contained 32 bioactive components as revealed by GC-MS study. The identified major compounds included hexadecanoic acid (5.197%), phytol (4.528%), caryophyllene oxide (4.605%), and 9,12,15-octadecatrienoic acid, methyl ester, (Z,Z,Z)- (3.751%).