The objective of this study was to investigate the effect of nutmeg (Myristica fragrans Houtt.) extract at different concentrations on chemical characteristics of raw beef under frozen storage. Nutmeg extracts at concentrations of 0.25%, 0.65%, 1.25%, 2.50% and 5.00% (g/ml) were used to treat raw beef (2.5 × 2.5 × 1.0 cm; 4 ± 0.5 g) with dilution method. Treated samples were then individually packed in overwrapped trays and stored for 3 weeks at -18 ± 1oC. The effects of the extract on the chemical characteristics such as lipid oxidation, colour, pH, moisture, fat, and protein content of raw beef were evaluated at 0, 4, 7, 10, 14 and 21 days of storage. Lipid oxidation was evaluated based on thiobarbituric acid-reactive substance (TBARS) content. Colour of beef was observed by spectrophotometer in colorimetic parameters CIELabs. Values of pH were measured using pH meter. Moisture, fat and protein content were determined using method by Analysis Association of Official Analytical Chemists (AOAC). The result showed that extract at concentration of 1.25% inhibited TBARS value meaning that extract of 1.25% or more was able to maintain the oxidative stability of beef at -18oC. A 1.25% of extract was also able to maintain the redness (a*) of treated beef compared to untreated during frozen storage. The pH values of all samples beef decreased starting from 10th day of storage. Untreated samples (0.00%) showed the lowest pH values compared to other treated samples at the end day of storage. There was no significant different in term of protein content in all treated or untreated samples. However, fat and moisture content were significantly affected by the concentration of nutmeg extract. Treated beef was able to retain its moisture with only loss of moisture ranging from 0.2% – 2.00% while untreated samples had 5.00% loss of moisture. The fat content of untreated samples (0.00%) showed a reduction of 0.2% of fat content at the end of storage compared to all treated sample with only loss of 0.1% - 0.05%. Overall, nutmeg extract can be used to maintain the chemical characteristics of raw beef during storage for 3 weeks.
Edible medicinal plants are often used in the treatment of various ailments and spice in traditional food preparation. In this study, 45 of tropical edible medicinal plants extracts from Indonesia, Malaysia, and Thailand were screened for their antimicrobial activity against five standard microorganisms for food preservative namely Aspergillus niger, Candida albicans, Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus. The methanol extracts of Piper nigrum L. seed, Piper cubeba L. seed, and the root of Ligusticum acutilobum Siebold and Zucc. showed antimicrobial activity against five species of standard microorganisms. Among them, P. cubeba L. extract demonstrated the most susceptible against all tested microorganisms. Minimal inhibitory concentration (MIC) and minimal bactericidal or fungicidal concentration (MBC or MFC) were performed by the broth microdilution techniques as described by the Clinical and Laboratory Standard Institute. MIC values of P. cubeba L. extract to A. niger, C. albicans, E. coli, P. aeruginosa and S. aureus were 12.8, 1.6, 3.2, 6.4, and 1.6 mg/ml, respectively. P. cubeba extract killed A. niger, C. albicans, E. coli, P. aeruginosa and S. aureus with MBC values of 25.6, 3.2, 6.4, 12.8, and 3.2 mg/ml, respectively. The potent antimicrobial activity of P. cubeba L. extract may support its use for natural food preservative.
Persicaria hydropiper, locally known as kesum, is an herb belongs to the family Polygonaceae. It has been used widely in many countries as food flavoring and possesses a wide range of medicinal values. The total phenolic content and xanthine oxidase inhibitory activity of the methanolic extract of P. hydropiper and fractions were determined spectrophotometrically. The butanol fraction was found to contain high phenolic content and was able to inhibit xanthine oxidase activity. Online profiling using liquid chromatography coupled with electrospray ionisation spectrometry (LC-ESIMS/MS) has revealed ten constituents in this active fraction. The major components were flavonoid derivatives and flavonoid sulphates, which were confirmed by comparison with an authentic standards as well as their MS/MS fragmentation patterns and UV spectra.
Neptunia oleracea Lour. is a tropical plant cultivated in Southeast Asia. It is consumed as vegetable and traditional herb for the treatment of several disorders. The objective of the present work was to isolate the phenolic compounds from N. oleracea, followed by their bioactivity evaluation and quantitative analysis. The ethyl acetate (EtOAc) and methanol (MeOH) fractions of N. oleracea were subjected to various chromatographic techniques to isolate the phenolic compounds. The isolated phenolic compounds were characterised by several spectroscopic methods, including mass spectrometry (MS) and nuclear magnetic resonance (NMR). Then, these compounds were subjected to DPPH free radical scavenging as α-glucosidase inhibitory assays for the evaluation of their activities. Their contents in the fractions were analysed via high performance liquid chromatography (HPLC) quantitative analysis. Five phenolic compounds including quercetin-3-O-β-D-xylopyranoside (1), quercetin-3-O-α-Larabinopyranoside (2), quercetin-3-O-α-L-rhamnoside (3), methylgallate (4) and rutin (5) were isolated from N. oleracea for the first time. Evaluation on the DPPH free radical scavenging and α-glucosidase inhibitory activities of these compounds showed that methylgallate (4) was the most potent antioxidant and α-glucosidase inhibitors among them, with IC50 values of 17.25 and 50.76 μM, respectively. The HPLC quantitative analysis revealed the high content of the quercetin derivatives (compounds 1, 2 and 3) in the EtOAc fraction (ranging from 125.68 to 157.55 μg/mg) and methylgallate (4) in the MeOH fraction (75.25 μg/mg). Comparison of the bioactivities of the isolated phenolic compounds with the fractions indicated their significant contribution for the DPPH free radical scavenging of N. oleracea; while they might be working synergistically for the α-glucosidase inhibitory activity. The results of the present work could help to validate the contribution of phenolic compounds for the studied bioactivities of N. oleracea.
Algal have attracted attention from biomedical scientists as they are a valuable natural
source of secondary metabolites that exhibit antioxidant activities. In this study, singlefactor
experiments were conducted to investigate the best extraction conditions (ethanol
concentration, solid-to-solvent ratio, extraction temperature and extraction time) in extracting
antioxidant compounds and capacities from four species of seaweeds (Sargassum polycystum,
Eucheuma denticulatum , Kappaphycus alvarezzi variance Buaya and Kappaphycus alvarezzi
variance Giant) from Sabah. Total phenolic content (TPC) and total flavonoid content (TFC)
assays were used to determine the phenolic and flavonoid concentrations, respectively, while
2,2-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and 2,2-diphenyl-1-picylhydrazyl
(DPPH) radical scavenging capacity assays were used to evaluate the antioxidant capacities of
all seaweed extracts. Results showed that extraction parameters had significant effect (p < 0.05)
on the antioxidant compounds and antioxidant capacities of seaweed. Sargassum polycystum
portrayed the most antioxidant compounds (37.41 ± 0.01 mg GAE/g DW and 4.54 ± 0.02 mg
CE/g DW) and capacities (2.00 ± 0.01 μmol TEAC/g DW and 0.84 ± 0.01 μmol TEAC/g DW)
amongst four species of seaweed.
The study was aimed to determine the antioxidant and α-glucosidase inhibition activities of
the stem and leaf of five different traditional medicinal plants. The studied plants exhibited
varied antioxidant and α-glucosidase inhibition activities. The antioxidant activities of the
plants were determined through their free radical scavenging capabilities using DPPH assay.
The most potent antioxidant activity was demonstrated by Neptunia oleracea with an IC50 of
35.45 and 29.72 μg/mL for leaf and stem, respectively. For α-glucosidase inhibition activity,
Neptunia oleracea exhibited potential α-glucosidase inhibition activity with IC50 value of
19.09 and 19.74 μg/mL for leaf and stem, respectively. The highest total phenolic content
(TPC) was also marked in Neptunia oleracea leaf and stem with value of 40.88 and 21.21 mg
GAE/g dry weight, respectively. The results also showed that Strobilanthes crispus collected
from two different locations possessed different levels of phenolic content, antioxidant and
α-glucosidase inhibition activities. The study revealed that phenolic compounds could be the
main contributors to the antioxidant and α-glucosidase inhibition activities with R values of 78.9
and 67.4%, respectively. In addition, antioxidant and α-glucosidase were positively correlated
(R = 81.9%). Neptunia oleracea could be suggested as a potential natural source of antioxidant
and antidiabetic compounds that can be used for the prevention or treatment of diabetes.
The main purpose of this study was to evaluate the antioxidant and α-glucosidase inhibitory
activities of Phyllanthus acidus. The P. acidus fruits were dried using three different methods,
namely oven (OD), air (AD) and freeze (FD) dryings and extracted with ethanol at different
ratios (50 and 100%). The proximate analysis and total phenolic content (TPC) as well as
free radical scavenging and α-glucosidase inhibitory activities were determined. The proximate
analysis of P. acidus fruit indicated that all the dried samples contained potential nutrient
contents. The highest TPC value, α-glucosidase inhibitory and antioxidant activities were
observed for 50% ethanolic extract from OD method with TPC value of 28.39 mg GAE/g dried
extract, IC50 value of 12.394 μg/mL and 64.17% inhibition, respectively. The study revealed
that phenolic compounds could be the main contributors to the antioxidant and α-glucosidase
inhibitory activities based on the Pearson correlation coefficients with R values of 95.0 and
73.8%, respectively. The study could provide scientific evidence for some folk uses in the
treatment of diseases related to the production of reactive oxygen species and oxidative stress.
The aim of the present work was to compare and choose the best method to extract incurred
pesticide residues from green tea. Accelerated solvent extraction (ASE) with in-cell cleanup
and the quick, easy, cheap, effective rugged and safe (QuEChERS) methods were tested on
green tea samples with incurred beta-endosulfan pesticide. The extracts were analyzed by
GC-MS/MS and the recovery and the precision of both methods were compared. The average
recovery using ASE with the in-cell cleanup method was in the range of 89 to 92% which is
better than that obtained using a QuEChERS method. Both the ASE with in-cell cleanup and
the QuEChERS methods provided good precision with RSDs in the range of 12 to 15% and
17 to 18%, respectively. This finding indicates that the ASE method with the in-cell cleanup is
more suitable for the accurate determination of pesticides incurred in tea.
Malaysia is a surplus poultry producing country with well-established commercial slaughtering and processing plants. Immense quantity of heads, feet, viscera, blood and feathers are usually discarded and not optimally utilized. Chicken heads are rich in protein, and could be a potential source of gelatin. The aim of the present work was therefore to find a simpler, faster, cheaper and greener gelatin extraction technology as compared to current available methods of gelatin extraction from poultry heads. A comparison of three different gelatin extraction methods with alkaline-acid pretreatment (E1), single acid pretreatment (E2) and single alkaline pretreatment (E3) were studied to extract gelatin from chicken heads. E1 and E2 produced gelatins of Type A, while E3 produced gelatin of Type B. High bloom gelatin (>300 g) with
Clinacanthus nutans (Acanthaceae) is a local plant consumed as tisane in Indonesia and ‘ulam’ in Malaysia. This plant has been claimed for its ability to prevent many diseases including diabetes. However, the scientific proof on this claim is still lacking. Therefore, the present work study was designed to evaluate the antidiabetic potential and antioxidant capacity of C. nutans leaves extracts using in vitro bioassay tests. The 80% methanolic crude extract of this plant was further partitioned using different polarity solvents namely hexane, hexane:ethyl acetate (1:1, v/v), ethyl acetate, ethyl acetate:methanol (1:1, v/v), and methanol. All the sub-fractions were analysed for antioxidant effect via 2, 2-diphenyl-2-picrylhydrazil (DPPH) scavenging activity, ferric reducing power (FRAP) and xanthine oxidase (XO) assays followed by antidiabetic evaluation via α-glucosidase and dipeptidyl peptidase-IV (DPP-IV) inhibitory assays and glucose uptake experiment. The ethyl acetate fraction showed a good antioxidant potential while the hexane fraction exhibited high α-glucosidase and DPP-IV enzyme inhibition. The hexane fraction also improved glucose uptake in a dose-dependent manner. The present work thus provides an informative data on the potential of C. nutans to be developed as a functional food in preventing diabetes.
The use of chitosan as a delivery carrier has attracted much attention in recent years. In this study, chitosan nanoparticles (CS-NP) and chitosan-ΦKAZ14 bacteriophage-loaded nanoparticles (C-ΦKAZ14 NP) were prepared by a simple coercavation method and characterized. The objective was to achieve an effective protection of bacteriophage from gastric acids and enzymes in the chicken gastrointestinal tract. The average particle sizes for CS-NP and C-ΦKAZ14 NP were 188 ± 7.4 and 176 ± 3.2 nm, respectively. The zeta potentials for CS-NP and C-ΦKAZ14 NP were 50 and 60 mV, respectively. Differential scanning calorimetry (DSC) of C-ΦKAZ14 NP gave an onset temperature of -17.17 °C with a peak at 17.32 °C and final end set of 17.41 °C, while blank chitosan NP had an onset of -20.00 °C with a peak at -19.78 °C and final end set at -20.47. FT-IR spectroscopy data of both CS-NP and C-ΦKAZ14 NP were the same. Chitosan nanoparticles showed considerable protection of ΦKAZ14 bacteriophage against degradation by enzymes as evidenced in gel electrophoresis, whereby ΦKAZ14 bacteriophage encapsulated in chitosan nanoparticles were protected whereas the naked ΦKAZ14 bacteriophage were degraded. C-ΦKAZ14 NP was non-toxic as shown by a chorioallantoic membrane (CAM) toxicity assay. It was concluded that chitosan nanoparticles could be a potent carrier of ΦKAZ14 bacteriophage for oral therapy against colibacillosis in poultry.
Disease inflicted by avian pathogenic Escherichia coli (APEC) causes economic losses and burden to the poultry industry worldwide. In this study, the efficacy of chitosan nanoparticles loaded ΦKAZ14 (C-ΦKAZ14 NPs) as an oral biological therapy for Colibacillosis was evaluated. C-ΦKAZ14 NPs containing 10(7) PFU/ml of ΦKAZ14 (Myoviridae; T4-like coliphage) bacteriophage were used to treat experimentally APEC-infected COBB 500 broiler chicks. C-ΦKAZ14 NPs and ΦKAZ14 bacteriophage were administered orally in a single dose. The clinical symptoms, mortality, and pathology in the infected birds were recorded and compared with those of control birds that did not receive C-ΦKAZ14 NPs or naked ΦKAZ14 bacteriophage. The results showed that C-ΦKAZ14 NP intervention decreased mortality from 58.33 to 16.7% with an increase in the protection rate from 42.00 to 83.33%. The bacterial colonization of the intestines of infected birds was significantly higher in the untreated control than in the C-ΦKAZ14 NP-treated group (2.30×10(9) ± 0.02 and 0.79×10(3) ± 0.10 CFU/mL, respectively) (P ≤ 0.05). Similarly, a significant difference in the fecal shedding of Escherichia coli was observed on d 7 post challenge between the untreated control and the C-ΦKAZ14 NP-treated group (2.35×10(9) ± 0.05 and 1.58×10(3) ± 0.06 CFU/mL, respectively) (P ≤ 0.05). Similar trends were observed from d 14 until d 21 when the experiment was terminated. Treatment with C-ΦKAZ14 NPs improved the body weights of the infected chicks. A difference in body weight on d 7 post challenge was observed between the untreated control and the C-ΦKAZ14 NP-treated group (140 ± 20 g and 160 ± 20 g, respectively). The increase was significant (P ≤ 0.05) on d 21 between the 2 groups (240 ± 30 g and 600 ± 80 g, respectively). Consequently, the clinical signs and symptoms were ameliorated upon treatment with C-ΦKAZ14 NPs compared with infected untreated birds. In all, based on the results, it can be concluded that the encapsulation of bacteriophage could enhance bacteriophage therapy and is a valuable approach for controlling APEC infections in poultry.
Momordica charantia, also known as bitter melon or ‘peria katak’ in Malaysia, is a member of the family Cucurbitaceae. Bitter melon is an excellent source of vitamins and minerals that made it extensively nutritious. Moreover, the seed, fruit and leave of the plant contain bioactive compounds with a wide range of biological activities that have been used in traditional medicines in the treatment of several diseases, including inflammation, infections, obesity and diabetes. The aim of this study was to evaluate changes in urinary metabolite profile of the normal, streptozotocin-induced type 1 diabetes and M. charantia treated diabetic rats using proton nuclear magnetic resonance (1H-NMR) -based metabolomics profiling. Study had been carried out by inducing diabetes in the rats through injection of streptozotocin, which exhibited type 1 diabetes. M. charantia extract (100 and 200 mg/kg body weight) was administrated to the streptozotocin-induced diabetic rats for one week. Blood glucose level after administration was measured to examine hypoglycemic effect of the extract. The results obtained indicated that M. charantia was effective in lowering blood glucose level of the diabetic rats. The loading plot of Partial Least Square (PLS) component 1 showed that diabetic rats had increased levels of lactate and glucose in urine whereas normal and the extract treated diabetic rats had higher levels of succinate, creatine, creatinine, urea and phenylacetylglycine in urine. While the loading plot of PLS component 2 showed a higher levels of succinate, citrate, creatine, creatinine, sugars, and hippurate in urine of normal rat compared to the extract treated diabetic rat. Administration of M. charantia extract was found to be able to regulate the altered metabolic processes. Thus, it could be potentially used to treat the diabetic patients.
Optimization process is an important aspect in the natural product extractions. Herein, an alternative approach is proposed for the optimization in extraction, namely, the Generalized Likelihood Uncertainty Estimation (GLUE). The approach combines the Latin hypercube sampling, the feasible range of independent variables, the Monte Carlo simulation, and the threshold criteria of response variables. The GLUE method is tested in three different techniques including the ultrasound, the microwave, and the supercritical CO2 assisted extractions utilizing the data from previously published reports. The study found that this method can: provide more information on the combined effects of the independent variables on the response variables in the dotty plots; deal with unlimited number of independent and response variables; consider combined multiple threshold criteria, which is subjective depending on the target of the investigation for response variables; and provide a range of values with their distribution for the optimization.
Curcumin has poor in vivo absorption and bioavailability, highlighting a need for new curcumin analogues with better characteristics in these aspects. The aim of this study is to determine the anti-cancer properties of four selected curcumin analogues, on the cytotoxicity, proliferative and apoptotic effects on androgen-independent human prostate cancer cells (PC-3 and DU 145). Initial cytotoxicity screening showed MS17 has the highest cell inhibitory effect, with EC50 values of 4.4 ± 0.3 and 4.1 ± 0.8 µM, followed by MS13 (7.5 ± 0.1 and 7.4 ± 2.6 µM), MS49 (14.5 ± 1.2 and 12.3 ± 2.3 µM) and MS40E (28.0 ± 7.8 and 30.3 ± 1.9 µM) for PC-3 and DU 145 cells, respectively. Time-dependent analysis also revealed that MS13 and MS17 displayed a greater anti-proliferative effect than the other compounds. MS17 was chosen based on the high selectivity index value for further analysis on the morphological and biochemical hallmarks of apoptosis. Fluorescence microscopy analysis revealed apoptotic changes in both treated prostate cancer cells. Relative caspase-3 activity increased significantly at 48 h in PC-3 and 12 h in DU 145 cells. Highest enrichment of free nucleosomes was noted at 48 h after treatment with MS17. In conclusion, MS17 demonstrated anti-proliferative effect and induces apoptosis in a time and dose-dependent manner suggesting its potential for development as an anti-cancer agent for androgen-independent prostate cancer.
Blue-green alga (Spirulina platensis) is a well renowned nutri-supplement due to its high nutritional and medicinal properties. The aim of this study was to examine the wound healing efficiency of Spirulina platensis at various solvent extracts using in vitro scratch assay on human dermal fibroblast cells (HDF). Various gradient solvent extracts (50 μg/ml of methanolic, ethanolic and aqueous extracts) from Spirulina platensis were treated on HDF cells to acquire its wound healing properties through scratch assay and in this investigation we have used allantoin, as a positive control to compare efficacy among the phytoextracts. Interestingly, aqueous extract were found to stimulate proliferation and migration of HDF cells at given concentrations and enhanced closure rate of wound area within 24 hours after treatment. Methanolic and ethanolic extracts have shown proliferative effect, however these extracts did not aid in the migration and closure of wound area when compared to aqueous extract. Based on phytochemical profile of the plant extracts analyzed by LC-MS/MS, it was shown that compounds supposedly involved in accelerating wound healing are cinnamic acid, narigenin, kaempferol, temsirolimus, phosphatidylserine isomeric derivatives and sulphoquinovosyl diacylglycerol. Our findings concluded that blue-green algae may pose potential biomedical application to treat various chronic wounds especially in diabetes mellitus patients.
Photodynamic therapy (PDT) is an alternative treatment for cancer that involves administration of a photosensitive drug or photosensitizer that localizes at the tumor tissue followed by in situ excitation at an appropriate wavelength of light. Tumour tissues are then killed by cytotoxic reactive oxygen species generated by the photosensitizer. Targeted excitation and photokilling of affected tissues is achieved through focal light irradiation, thereby minimizing systemic side effects to the normal healthy tissues. Currently, there are only a small number of photosensitizers that are in the clinic and many of these share the same structural core based on cyclic tetrapyrroles. This paper describes how metabolic tools are utilized to prioritize natural extracts to search for structurally new photosensitizers from Malaysian biodiversity. As proof of concept, we analyzed 278 photocytotoxic extracts using a hyphenated technique of liquid chromatography-mass spectrometry coupled with principal component analysis (LC-MS-PCA) and prioritized 27 extracts that potentially contained new photosensitizers for chemical dereplication using an in-house UPLC-PDA-MS-Photocytotoxic assay platform. This led to the identification of 2 new photosensitizers with cyclic tetrapyrrolic structures, thereby demonstrating the feasibility of the metabolic approach.
Canarium odontophyllum, also known as CO, is a highly nutritious fruit. Defatted parts of CO fruit are potent sources of nutraceutical. This study aimed to determine oxidative stress and lipid peroxidation effects of defatted CO pericarp and peel extracts using in vitro bioassays. Cell cytotoxic effect of the CO pericarp and peel extracts were also evaluated using HUVEC and Chang liver cell lines. The crude extracts of defatted CO peel and pericarp showed cytoprotective effects in t-BHP and 40% methanol-induced cell death. The crude extracts also showed no toxic effect to Chang liver cell line. Using CD36 ELISA, NAD(+) and LDL inhibition assays, inhibition of oxidative stress were found higher in the crude extract of defatted CO peel compared to the pericarp extract. Hemoglobin and LDL oxidation assays revealed both crude extracts had significantly reduced lipid peroxidation as compared to control. TBARS values among defatted CO pericarp, peel, and cyanidin-3-glucoside showed no significant differences for hemoglobin and LDL oxidation assays. The protective effects of defatted CO parts, especially its peel is related to the presence of high anthocyanin that potentially offers as a pharmaceutical ingredient for cardioprotection.
Plants that help in slowing down the digestion of triacylglycerols (TAGs) in the pancreas and small intestine of humans play an important role in the reduction of obesity. On the other hand, there may be plants or plant parts that stimulate intestinal lipolytic activity, thus contributing to greater TAG assimilation. The aim of this study was to evaluate the aqueous methanolic extracts of ninety eight (98) medicinal, herbal and aquatic plant materials from Malaysia for their effect on porcine pancreatic lipase (PPL) activity and to identify the structure of an anti-lipase compound from one of the sources. The degree of inhibition was also quantified as relative to orlistat activity against PPL (orlistat equivalents). Results revealed that while 19.4% of the extracts were found to have anti-lipase activity ≥80%, 12% were actually found to promote PPL activity. Twenty two percent (22.4%) exhibited moderate inhibition (41%-80%) and 2% were neutral toward PPL activity. The ripe fruit of Averrhoa carambola and the leaves of Archidendron jiringa (Jack) I.C Nielsen L. (jering), Cynometra cauliflora (nam-nam) and Aleurites moluccana (L.) Willd (candle nut/buah keras) had the highest (100%) anti-lipase activity and are equivalent to 0.11 µg orlistat/mL. Plants that stimulated lipase activity included Pimpinella anisum L. (aniseed/jintan manis), activating the enzyme by 186.5%. Kaempferol 3-O-rhamnoside was isolated from the ethyl acetate fraction of C. cauliflora leaves and found to be an active lipase inhibitor. The structure was elucidated using 1H-NMR, 13C-NMR and 2D-NMR analyses.