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Govindarajan Karthivashan,1 Mas Jaffri Masarudin,2 Aminu Umar Kura,1 Faridah Abas,3,4 Sharida Fakurazi1,5 1Laboratory of Vaccines and Immunotherapeutics, Institute of Bioscience, 2Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, 3Department of Food Science, Faculty of Food Science and Technology, 4Laboratory of Natural Products, Institute of Bioscience, 5Department of Human Anatomy, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia Abstract: This study involves adaptation of bulk or sequential technique to load multiple flavonoids in a single phytosome, which can be termed as “flavonosome”. Three widely established and therapeutically valuable flavonoids, such as quercetin (Q), kaempferol (K), and apigenin (A), were quantified in the ethyl acetate fraction of Moringa oleifera leaves extract and were commercially obtained and incorporated in a single flavonosome (QKA–phosphatidylcholine) through four different methods of synthesis – bulk (M1) and serialized (M2) co-sonication and bulk (M3) and sequential (M4) co-loading. The study also established an optimal formulation method based on screening the synthesized flavonosomes with respect to their size, charge, polydispersity index, morphology, drug–carrier interaction, antioxidant potential through in vitro 1,1-diphenyl-2-picrylhydrazyl kinetics, and cytotoxicity evaluation against human hepatoma cell line (HepaRG). Furthermore, entrapment and loading efficiency of flavonoids in the optimal flavonosome have been identified. Among the four synthesis methods, sequential loading technique has been optimized as the best method for the synthesis of QKA–phosphatidylcholine flavonosome, which revealed an average diameter of 375.93±33.61 nm, with a zeta potential of -39.07±3.55 mV, and the entrapment efficiency was >98% for all the flavonoids, whereas the drug-loading capacity of Q, K, and A was 31.63%±0.17%, 34.51%±2.07%, and 31.79%±0.01%, respectively. The in vitro 1,1-diphenyl-2-picrylhydrazyl kinetics of the flavonoids indirectly depicts the release kinetic behavior of the flavonoids from the carrier. The QKA-loaded flavonosome had no indication of toxicity toward human hepatoma cell line as shown by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide result, wherein even at the higher concentration of 200 µg/mL, the flavonosomes exert >85% of cell viability. These results suggest that sequential loading technique may be a promising nanodrug delivery system for loading multiflavonoids in a single entity with sustained activity as an antioxidant, hepatoprotective, and hepatosupplement candidate. Keywords: quercetin, kaempferol, apigenin, phytosome, antioxidant, HepaRG cell line

Abubakar Amali Muhammad,1 Palanisamy Arulselvan,1 Cheah Pike See,2 Farida Abas,3 Sharida Fakurazi1,2 1Laboratory of Vaccine and Immunotherapeutics, Institute of Bioscience, 2Unit of Anatomy, Department of Human Anatomy, Faculty of Medicine and Health Sciences, 3Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia, Serdang, Malaysia Abstract: Diabetic foot ulcer is a serious complication of diabetes, which affects a significant percentage (15%) of diabetics and up to 15%–24% of those affected may require amputation. Therefore, the economic burden of diabetic foot ulcers is enormous and is associated with high cost of treatment and prolongs hospitalization. The present study was conducted to evaluate antibacterial and in vivo wound healing activities of an aqueous fraction of Moringa oleifera on a diabetic condition. Antibacterial activity testing was carried out using agar well and tube dilution techniques. The in vivo study was conducted using six groups of animals that comprise of one normal and diabetic control group each, three treatment groups of 0.5%, 1%, and 2% w/w aqueous fraction, and a positive control group (1% w/w silver sulfadiazine). Rats were induced with diabetes using a combination of streptozotocin 65 and 150 mg/kg nicotinamide daily for 2 days, and excision wounds were created and treated with various doses (0.5%, 1%, and 2% w/w aqueous fraction) daily for 21 days. Biophysical, histological, and biochemical parameters were investigated. The results of the study revealed that aqueous fraction possessed antibacterial activity through inhibition of growth of Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli organisms. The topical application of aqueous fraction revealed enhancement of wound healing under sustained hyperglycemic condition for the duration of the experiment. This enhancement was achieved through decreased wound size, improved wound contraction, and tissue regeneration, as well as downregulation of inflammatory mediators, such as tumor necrosis factor-α, interleukin-1β, interleukin-6, inducible nitric oxide synthase, and cyclooxygenase-2, and upregulation of an angiogenic marker vascular endothelial growth factor in wound tissue treated with various doses of aqueous fraction of M. oleifera. The findings suggest that aqueous fraction of M. oleifera containing Vicenin-2 active compound may accelerate wound healing in hyperglycemic condition. Keywords: aqueous fraction, Vicenin-2, wound healing, inflammatory mediators

Bacterial growth in food sources after harvesting and during food manufacturing has emerged as a major agricultural concern. Currently, several chemical preservatives have been added to foods to prevent or inhibit bacterial growth. These phenomena, however, become a concern when consumers begin to demand the safety of the food they consume, thus resulting in a search for natural plant extracts as an alternative. The antimicrobial activity of Cosmos caudatus leaf extract against six food pathogens was thus investigated in the present work. The disc-diffusion test, the minimum inhibitory concentration (MIC), and the minimum bactericidal concentration (MBC) tests were used to determine the antibacterial potential. Time-kill kinetics were used to determine their killing endpoints at various incubation times. Thereafter, Bacillus cereus was selected as a representative bacterium to be viewed under the light sheet electron microscope (LSEM) to observe the morphological changes. For the application as a sanitiser, different concentrations of C. caudatus extract were applied to chicken meat and oyster mushroom to determine bacterial reduction and sensory acceptability. The toxicity test using brine shrimp lethality assay was also performed to determine the toxicity level of C. caudatus extract. Results showed that C. caudatus extract contain five major peaks of flavonoids. In the antibacterial test, all tested pathogens were inhibited, with B. cereus being the most susceptible, with a significant reduction time of more than 3 log10 at 3.125 mg/mL. Under LSEM, the population of B. cereus was reduced, and the shape was wrinkled. In the application, C. caudatus extract at 0.05% for 5 min treatment time was chosen as the best formulation, whereas the brine shrimp tests showed that C. caudatus extract was safe for human consumption (LC50: 3.54 mg/mL). In conclusion, C. caudatus extract could have the potential to be further explored as a natural food sanitiser. [ABSTRACT FROM AUTHOR]

Hyperlipidaemia is one of the essential public health risk factors that can cause other metabolic diseases such as cardiovascular diseases and diabetes. Dieting and healthy lifestyle have been among the primary approaches. However, medication is required to regulate the lipid profile in some instances. Therefore, there has been an increase in interest in using or integrating herbal medicine with modern medicine in treating hyperlipidaemia. Nonetheless, preparing standardised herbal extract or products has been one of the major challenges in the herbal industry. Standardising herbal extract or product (single plant-based or mixture of multiple herbs) is needed to ensure the quality, safety, and efficacy of the herbal maintained from batch to batch before it is released to the market. The present review thus evaluates several herbal plants with anti-hyperlipidaemic activities, quality control using chemical markers, and metabolomics application in herbal plants. [ABSTRACT FROM AUTHOR]

The use of chemical preservatives in food products to inhibit the growth of microorganisms is widely used nowadays. However, their use has become a concern due to several negative side effects, and when consumers question the safety of the foods they eat. Therefore, the present work was conducted to investigate the potential of plant natural sanitiser from Cosmos caudatus Kunth extract to reduce the natural microflora present in raw beef and shrimp meat samples. The present work aimed to investigate the reduction of natural microflora (B. cereus, E. coli, Pseudomonas spp., S. aureus, and L. monocytogenes) in raw beef and shrimp meat samples following sanitisation with different concentrations of C. caudatus extract (0.05, 0.50, and 5.00%) at different soaking times (5, 10, and 15 min). The sanitised samples were further evaluated with sensory acceptability (colour, odour, texture, and overall acceptability) to determine their acceptance level after treatment. Based on the results, the microflora in beef and shrimp meat samples were reduced significantly (p < 0.05) started from 0.05% at varied soaking times. The decrease in bacterial populations was proportional to the increase in extract concentrations and soaking times. In sensory acceptability, all cooked samples achieved acceptance level by the panellists at 0.05% after 10 min of soaking time. Food samples treated with 0.05% of C. caudatus extract and 10 min of soaking time showed the best combination in terms of bacterial reduction and the level of acceptance by the panellists. Hence, it can be concluded that C. caudatus extract has a high potential as a natural-based food sanitiser that can prevent bacterial contamination while maintaining the sensory acceptability of the foods. [ABSTRACT FROM AUTHOR]

Problem statement: Plant antioxidants are natural plant products that inhibit the adverse effects of the Reactive Oxygen Species (ROS) produced in plants. Thus, antioxidants enable plants to survive. Depending on this principle, human beings started using some plants' parts to cure from diseases many centuries ago, even before the discovery of microbes or disclosing the etiology of many human diseases. Shed light on the importance of diet and natural products that have proved of great health impact on human traditionally and scientifically, disclosing the central role of natural antioxidants in this matter. Moreover, this review tried to give an overview on the research has so far done in Asian countries particularly in the field of natural antioxidants during the period 2000-2008. Therefore, the aim of this review is to present a comprehensive index for the antioxidant property of herbs and plants that have been traditionally used as well as experimentally proved beneficial in these countries. Approach: We had compared Asian and other nation's diet in the popularity of the traditional medicine and the type of daily meal in respect to their antioxidants contents and valuable health promoting effects. Many literatures had been collected using online search engines and the main library of University Putra Malaysia. The gathered data had been arranged alphabetically in a table depending on the plant's scientific name, common name, the used part(s) and the family name. Results: The research had shown that there were seventy-six plants with proved antioxidant activity which had been well studied by Asian scientists in the chosen period of time. Conclusion: The antioxidant components of the natural products constitute the major source of human health promotion and maintenance. The nature is still the perfect source for health promotion and for the supplementation of safe drugs. Great attention is mandatory from all the scientists around the world to explore many underestimated plants with highly effective antioxidant activity. Key words: Dietary supplements, herbs, medicinal food, natural products, spice, INTRODUCTION Since antiquity, man has used plants to treat common infectious diseases and some of these traditional medicines are still included as part of the habitual treatment of various maladies [...]

The present work aimed to evaluate the potential of Carica papaya seed extract (CPSE) as an antibacterial agent against Salmonella Enteritidis, Bacillus cereus, Vibrio vulnificus, and Proteus mirabilis. The bioautography of the CPSE on thin-layer chromatography (TLC) plates was performed, followed by fractionation of the CPSE by column chromatography using hexane:ethyl acetate (1:1) eluent. Minimum Inhibitory Concentration (MIC), toxicity, composition, and stability of the crude, fractions, and sub-fractions of the CPSE were evaluated. The bioautographic fractions of the CPSE at MIC of 5.63 mg/mL had shown that hexane:ethyl acetate (1:1) fraction and its sub-fraction 3C (Rf = 0.94 ± 0.03) demonstrated the equivalent MIC value (5.63 mg/mL) with the crude CPSE. However, the hexane:ethyl acetate (1:1) fraction and sub-fraction 3C had higher toxicity (LC50 = 1.797 ± 0.305 and 0.332 ± 0.059 mg/mL, respectively) than the crude CPSE (LC50 = 5.505 ± 0.718 mg/mL). Thus, only the crude CPSE was subjected to stability study. The dominant cis-vaccenic acid in sub-fraction 3C demonstrated the lowest MIC against B. cereus (1.41 mg/mL), P. mirabilis (1.41 mg/mL), and S. Enteritidis (0.70 mg/mL) in its pure form. Hence, these results signified the potency of the cis-vaccenic acid as an antibacterial compound from the CPSE. The stability study of the crude CPSE solution showed that at MIC of 5.63 mg/mL, the crude CPSE solution acted as a potent antibacterial agent in acidic condition (pH 4), water activity (Aw) < 0.950, and temperature < 40°C. [ABSTRACT FROM AUTHOR]

Mangifera odorata, or known as "kuini" in Malay, is considered an underutilised climacteric tropical fruit. Recent studies had suggested that the by-products of M. odorata may serve as functional food ingredients based on their nutritional values and biological activities. Thus, the present work was aimed to determine the nutritional composition, antidiabetic properties through α-glucosidase and α-amylase inhibition assays, and phytochemical profiling of M. odorata pulp using ultra-high-performance liquid chromatography-electrospray ionisation orbitrap tandem mass spectrometry (UHPLC-ESI-Orbitrap-MS/MS). Proximate analyses found that M. odorata pulp contained high amounts of soluble fibre and moisture. The main sugar in the pulp was identified as sucrose, while ß-carotene, potassium, calcium, iron, and zinc were the major vitamins and minerals. The extraction of the sample in water, acetone, methanol, and ethanol were performed for the determination of antidiabetic assay. The results showed that 60% of ethanolic extract had the highest α-amylase inhibitory activity among the four extraction solvents. The findings revealed high levels of mangiferin in the active extract, which may be responsible for the α-amylase inhibition activity, thus contributing to lower post-prandial blood glucose. The present work provides evidence on the nutritional benefits of M. odorata pulp, as well as the potential use of its ethanolic extract as an α-amylase inhibitor. [ABSTRACT FROM AUTHOR]

The present work aimed to investigate the phytochemical profile and biological activities of Sudanese baobab (Adansonia digitata L.) fruit pulp extract. Baobab fruit pulp serves as food, and has been used in traditional medicine in Africa for the treatment of several diseases, and believed to possess many biological activities. The sample of baobab fruit was collected from Blue Nile State in Sudan. The fruit pulp was obtained from baobab pods, sieved, extracted with methanol (80%), and analysed for its antioxidant activity, and alpha-glucosidase and nitric oxide (NO) inhibitory activities. The phytochemical constituents of the pulp were determined by LC-MS and ¹H-NMR techniques. The results for antioxidant activity evaluated by DPPH and FRAP for methanolic extract were 232.70 mg Ascorbic Acid Equivalent Antioxidant Capacity (AEAC)/g and 222.28 mmol/g of Fe2+, respectively. Baobab pulp extract showed greater capability in inhibiting the generation of NO from the stimulated RAW264.7 cells at 98.45% inhibition and IC50 of 36.55 µg/mL; and α-glucosidase enzyme activity at 97.94% inhibition and IC50 of 58.59 µg/mL. The LC-MS/MS analysis of the baobab extract showed that 52 compounds were found in the baobab pulp, including 19 flavonoids, ten phenolic acids, four lipids (glycero-3-phosphocholine derivative, 1,2-didodecanoyl- sn-glycero-3-phospho-(1'-sn-glycerol), 1-pentadecanoyl-2-(13Z,16Z-docosadienoyl)-glycero-3-phosphate, and glycero-3-hosphocholine derivative), sugars, organic acid, iridoids, adenosine, scopoletin, and taraxerone. The NMR detected 21 metabolites from baobab pulp extract including carbohydrates, organic acids, vitamin, amino acids, phenolic compounds, alkaloids, and fatty acids. The present work is the first of its kind in comprehensively analysing the Sudanese baobab fruit pulp, combined with qualitative characterisation of chemical components using NMR, LC-MS, α-glucosidase, and NO inhibitors. Baobab fruit pulp contains several beneficial phytochemicals and biological activities, and it has potential positive effect on general human's health. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Ipomoea aquaticaForsk is a green leafy vegetable that is a rich source of minerals, proteins, vitamins, amino acids, and secondary metabolites. Different types ofI. aquaticacultivars are grown for consumption but little is known about the metabolites variation. Proton nuclear magnetic resonance (1H NMR) spectroscopy combined with multivariate data analysis was applied for metabolic profiling of threeI. aquaticacultivars including “broad leaf (K-25)”, “bamboo leaf (K-88)”, and “special pointed leaf (K-11)”. The orthogonal partial least squares discriminant analysis (OPLS-DA) indicated a clear separation among cultivars. The relative levels of various compounds, such as amino acids, organic acids, sugars, and phenolic compounds were specific to each cultivar. The K-11 cultivar was different from the other cultivars due to a high phenolic content. The content of sugars and some amino acids was higher in K-88 and K-25 possessed a higher content of organic acids. Thein vitrostudy revealed that theI. aquaticacultivars exhibited potent antioxidant and α-glucosidase activities. The results of this study indicate that the K-11 cultivar was the most active due to the abundance of epicatechin, 4,5-dicaffeoylquinic, protocatechuic acid, and rutin. [ABSTRACT FROM PUBLISHER]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]