Soo Yee Lee

Paddy plants tend to accumulate heavy metals from both natural and anthropogenic sources, and this poses adverse risks to human health. The objective of this study was to investigate heavy metal contamination in paddy plants in Kelantan, Malaysia, and its health risk assessment. The bioaccumulation of heavy metals was studied by means of enrichment (EF) and translocation factors (TF). The health risk assessment was performed based on USEPA guidelines. The EF for heavy metals in the studied areas was in the descending order of Cu > As > Cr > Cd > Pb. Meanwhile, Cr and Pb exhibited higher TF values from stem to grain compared with the others. The combined hazard index (HI) resulting from five heavy metals exceeded the acceptable limit (HI >1). The lifetime cancer risk, in both adult and children, was beyond the acceptable limit (10⁻⁴) and mainly resulted from exposure. The total cancer risk (CRt) due to simultaneous exposures to multiple carcinogenic elements also exceeded 10⁻⁴. In conclusion, intake of heavy metal through rice ingestion is likely to cause both non-carcinogenic and carcinogenic health risks. Further research is required to investigate the extent of heavy metal contamination in agricultural soils and, moreover, to establish human exposure as a result of rice consumption.

Exposure to polycyclic aromatic hydrocarbons (PAHs) through diet is gaining concern due to the risk it poses to human health. This study evaluated the bioaccessibility of PAHs contained in charcoal-grilled beef and chicken in different segments of the gastrointestinal tract (GIT) with regard to the degree of doneness and fat content of the meats. The levels of 15 PAHs in the grilled meat samples and bioaccessible fractions were determined using high-performance liquid chromatography (HPLC) equipped with PAH column, and UV and fluorescence detectors. Total PAHs were found in beef (30.73 ng/g) and chicken (70.93 ng/g) before its digestion, and different PAHs’ bioaccessibility were observed in the different segments of GIT, with the highest in the stomach followed by the small intestine, despite the relatively higher bioaccessibility of individual PAHs in grilled beef as compared to those in grilled chicken. Additionally, the PAHs’ bioaccessibility increased with the increase in the degree of doneness. Positive linear correlation was observed for the PAHs’ bioaccessibility and the fat contents of grilled meat. Overall, this study highlights the influence of meat doneness (cooking time) and fat contents on the bioaccessibility and bioaccumulation of PAHs.

Rice is one of the major crops as well as the staple food in Malaysia. However, historical mining activity has raised a concern regarding heavy metal contamination in paddy plants, especially in Perak, a state with major tin mining during the late nineteenth century. Therefore, the objective of this study is to investigate the heavy metals (As, Cd, Pb, Cu, Cr) contamination in paddy soils and paddy plants in three districts in Perak. The content of heavy metals was determined using ICP-MS, while the absorption and transferability of heavy metals in the paddy plants were investigated through enrichment (EF) and translocation (TF) factors. Principal component analysis (PCA) was employed to recognize the pattern of heavy metal contaminations in different sampling areas. Health risk assessment was performed through calculation of various indices. The quantification results showed that root contained highest concentration of the studied heavy metals, with As exhibiting the highest concentration. The EF results revealed the accumulation of As, Cu, and Cr in the rice grains while PCA showed the different compositional pattern in the different sampling areas. The health risk assessment disclosed both noncarcinogenic and carcinogenic risks in the local adults and children. Overall, findings from this study show that heavy metal contamination poses potential health risks to the residents and control measure is required.

Research background. Curcuma species (Zingiberaceae) are well known medicinal herbs in India and Southeast Asia. Despite various findings reporting their beneficial biological activities, very little information has been recorded on the Curcuma caesia. Thus, this study aims to determine the phenolic content, antioxidant and α-glucosidase inhibitory activity of both rhizome and leaves of C. caesia. Experimental approach. Rhizome and leaves of C. caesia were dried with oven (OD) and freeze (FD)-drying methods, and extracted with different Φ(ethanol,water)=100:0, 80:20, 50:50 and 0:100. The bioactivities of C. caesia extracts were evaluated using in vitro tests; total phenolic content (TPC), antioxidant (DPPH and FRAP) and α-glucosidase inhibitory activity. Proton nuclear magnetic resonance (1H NMR)-based metabolomics approach was employed to differentiate the most active extracts based on their metabolite profiles and correlation with bioactivities. Results and conclusions. The FD rhizome extracted with Φ(ethanol,water)=100:0 was observed to have potent TPC expressed as gallic acid equivalents, FRAP expressed as Trolox equivalents and α-glucosidase inhibitory activity with values of (45.4±2.1) mg/g extract, (147.7±8.3) mg/g extract and (265.5±38.6) μg/mL (IC50), respectively. Meanwhile, for DPPH scavenging activity, the Φ(ethanol,water)=80:20 and 100:0 extracts of FD rhizome showed the highest activity with no significant difference between them. Hence, the FD rhizome extracts were selected for further metabolomics analysis. Principal component analysis (PCA) showed clear discrimination among the different extracts. Partial least square (PLS) analysis showed positive correlations of the metabolites, including xanthorrhizol derivative, 1-hydroxy-1,7-bis(4-hydroxy-3-methoxyphenyl)-(6E)-6-heptene-3,4-dione, valine, luteolin, zedoardiol, β-turmerone, selina-4(15),7(11)-dien-8-one, zedoalactone B and germacrone, with the antioxidant and α-glucosidase inhibition activities, whereas curdione and 1-(4-hydroxy-3,5-dimethoxyphenyl)-7-(4-hydroxy-3-methoxyphenyl)-(lE,6E)-1,6-heptadiene3,4-dione were correlated with α-glucosidase inhibitory activity. Novelty and scientific contribution. C. caesia rhizome and leaf extracts contained phenolic compounds and had varies antioxidant and α-glucosidase inhibitory capacities. These findings strongly suggest that the rhizomes of C. caesia are an invaluable natural source of active ingredients for applications in pharmaceutical and food industries.

Oil palm (Elaeis guineensis Jacq.) leaf (OPL) is abundantly generated from oil palm plantations as biomass that is rich in bioactive metabolites, primarily flavonoids. Six natural deep eutectic solvents (NaDES) were synthesized using a direct heating technique from different combinations of choline chloride with 1,2-propanediol (PD), 1,4-butanediol (BD), glycerol (GLY), glucose (GLU), maltose (MAL) and lactic acid (LA). The synthesized NaDES were subjected to physicochemical and biological evaluations comprising physical appearance, density, water activity, viscosity, polarity, thermal behaviors, spectroscopic analysis, cytotoxicity, radical scavenging activities and solubility tests. Compared to aqueous methanol, the synthesized NaDES, which appeared as a slightly to moderately viscous transparent liquid, showed favorable physicochemical properties as extraction solvents with a low cytotoxicity profile on cultured fibroblast cells. Further, the NaDES obtained from the choline chloride:lactic acid (LA) combination showed high free radical scavenging characteristics. Hydrogen bonding interactions were shown to play a significant role in the formation of the NaDES. Further, ultra-high-performance liquid chromatography ultraviolet/photodiode array (UHPLC-UV/PDA) analysis revealed that the NaDES from the choline chloride:glycerol (GLY) combination had comparable efficiencies with aqueous methanol regarding extracting flavonoids (luteolin and apigenin derivatives) from OPLs. The results of the present study suggested that the tailor-made NaDES were not only easy-to-use, stable and safe solvents but also suitable for extracting bioactive phytochemical compounds. The study highlighted their potential as an alternative green technology for applications in oil palm biomass utilization programs.

The differences in pungency of “sirih” imply the probable occurrence of several variants of Piper betle L. in Malaysia. However, the metabolite profiles underlying the pungency of the different variants remain a subject of further research. The differences in metabolite profiles of selected Malaysian P. betle variants were thus investigated; specifically, the leaf aqueous methanolic extracts and essential oils were analyzed via ¹H-NMR and GC-MS metabolomics, respectively. Principal component analysis (PCA) of the ¹H-NMR spectral data showed quantitative differences in the metabolite profiles of “sirih melayu” and “sirih india” and revealed an ambiguous group of samples with low acetic acid content, which was identified as Piper rubro-venosum hort. ex Rodigas based on DNA sequences of the internal transcribed spacer 2 (ITS2) region. The finding was supported by PCA of two GC-MS datasets of P. betle samples obtained from several states in Peninsular Malaysia, which displayed clustering of the samples into “sirih melayu” and “sirih india” groups. Higher abundance of chavicol acetate was consistently found to be characteristic of “sirih melayu”. The present research has provided preliminary evidence supporting the notion of occurrence of two P. betle variants in Malaysia based on chemical profiles, which may be related to the different genders of P. betle.

Huge quantities of oil palm (Elaeis guineensis Jacq.) leaves (OPL) are generated as agricultural biomass from oil palm plantations. OPL are known to contain significant amounts of flavonoids. For maximal exploitation of these valuable antioxidant compounds, an innovative and sustainable extraction method employing natural deep eutectic solvents (NaDES) combined with ultrasonic assisted extraction was developed. Various NaDES composed of choline chloride as the hydrogen bond donor (HBD) and 1,2 propanediol (PD), 1,4 butanediol (BD), glycerol (GLY), glucose (GLU), maltose (MAL), and lactic acid (LA) as the hydrogen bond acceptor (HBA) were synthesized. The influence of these compositions, the methods of their synthesis, molar ratios, and water contents on their capacity to extract flavonoids from OPL was evaluated. Based on the results, it was found that methods which incorporate a heating step produced NaDES with the best capacity to extract OPL flavonoids. These thermal methods combined with molar ratios of 1:3 or 1:4 and water contents of 17 to 50% were found to be the optimal conditions for preparing NaDES, specifically when applied to the PD, BD, and GLY NaDES. Subsequently, UHPLC-UV/PDA-MS/MS analysis revealed NaDES extracts recovered by macroporous adsorption resin XAD7HP were able to optimally extract at least twelve luteolin and apigenin derivatives in OPL NaDES extracts prepared from glycerol and 1,4-butanediol demonstrated better and comparable efficiency as aqueous methanol in extracting flavonoids from OPL. The in vitro studies of antioxidant and wound healing properties supported these findings by exhibiting good free radical scavenging, cell proliferation, and migration activities. Additionally, the NaDES extracts also showed non-cytotoxicity effects at 1000 µg/mL and below on 3T3 fibroblast cells. Results of the study showed that NaDES could be a promising eco-friendly green solvent to extract bioactive OPL flavonoids that have great potential for applications as wound healing agents.

Terminalia catappa and Terminalia subspathulata are two species of the Combretaceae family of medium to large forest trees. The fruits of T. catappa are known for the edible nuts commonly known as tropical almonds due to their similarity in taste with almonds of commerce. Therefore, the chemical profiles of the fruits of the two Terminalia species were examined to ascertain their potential value for food or health uses. Gas chromatography-mass spectrometry (GCMS) and ultrahigh-pressure liquid chromatography-electrospray ionisation tandem mass spectrometry (UHPLC-ESI-MS/MS) techniques were employed to profile the extracts to ensure good coverage of the classes of metabolites of the fruit extracts. The GCMS results revealed that T. catappa nuts were rich in palmitic acid (33.2%), linoleoyl chloride (29.1%), and oxacyclohexadecan-2-one commonly known as pentadecanolide (16.2%). In comparison, the major constituents of T. subspathulata fruits were palmitic acid (18.1%) and its methyl ester, methyl palmitate (9.3%). Furthermore, a total of 38 compounds were putatively identified in the 70% aqueous methanolic extracts of both species via UHPLC-ESI-MS/MS analysis, comprising three organic acids, sixteen hydrolysable tannins, ten phenolic acids, eight flavonoids, and a diarylheptanoid. The GCMS- and liquid chromatography-mass spectrometry-(LCMS-) LCMS-based metabolite profiles obtained in the present study have revealed the diversity of chemical constituents in the T. catappa nuts and T. subspathulata fruits, potentially valorised as functional foods nutraceutical ingredients for plant-based health products.

Introduction: Stevia is known for its sweet taste, attributed to the presence of steviol glycosides. Although reports on the dynamic changes of steviol glycosides during development of stevia are available, the data are mainly focused on stevioside and rebaudioside A. Information concerning the comprehensive metabolite profile of stevia in relation to different developmental stages is still lacking.Objective: This study investigated the metabolite changes along the developmental stages of a local stevia cultivar.Methodology: Stevia leaves were harvested at 4 different developmental stages (early vegetative, late vegetative, budding, and flowering). Samples were then subjected to LC–MS metabolomics analysis to determine the metabolite variations.Results: A total of 55 metabolites, comprising phenolic acids, flavonoids, and terpenoids were identified by MS/MS analysis of the stevia leaf extracts, revealing a metabolite profile which was comparatively similar with those of cultivars grown in other countries. PLS‐DA differentiated the early vegetative stage stevia leaf samples from those of the later stages by higher content of phenolic acids. The leaf metabolomes of the later 3 stages (late vegetative, budding, and flowering) were collectively richer in flavonoids. Meanwhile, the content of steviol glycosides is highest during the late vegetative and budding stages.Conclusion: The present study provided, for the first time, a general overview of the metabolite variations with regard to the different developmental stages of stevia. The information may facilitate decision making of suitable harvesting times for higher yields of steviol glycosides or a more balanced metabolite profile in terms of pharmacologically useful metabolites.

The aim of the present study was to evaluate the variations in the phytochemical compositions and the antimicrobial activities of Syzygium grande and Oenanthe javanica leaf extracts against Bacillus sp. The antibacterial activity of the methanol, 70% methanol, ethyl acetate, and hexane extracts of both plant species were examined in vitro, against various Bacillus species. In addition, the 1H Nuclear Magnetic Resonance (1H NMR) spectroscopy-based metabolomics was employed to gain insights into the correlations between the chemical constituents of the bioactive extract to the biological activity. The hexane extract of S. grande leaves and methanolic extract of O. javanica leaves showed more potent anti-Bacillus activity, among the test extracts. Principal component analysis (PCA) successfully differentiated the extracts on the basis of their metabolite profiles. The results of the Partial Least Square (PLS) analysis showed that the non-polar compounds of the hexane extract of S. grande (alanine, betulin, β-sitosterol, β-caryophyllene, acetic acid and 3-hydroxybutyric acid) were strongly correlated to its anti-Bacillus activity. On the other hand, choline, ellagic acid, and gallic acid, the metabolites present in the methanolic extract of O. javanica, were strongly correlated to its anti-Bacillus activity. On this basis, it was therefore concluded that these conmpounds could be the potential bioactive constituents, contributing to the anti-Bacillus activity of the individual plant species. Further in-depth investigations into the potential utilization of the two plants for useful applications in managing and control of Bacillus sp., will help pave the way towards their valorization.