Nur Khaleeda Zulaikha Zolkeflee

Ethnopharmacological relevance: The roots of Heterostemma piperifolium (Apocynaceae) have been documented in tribal ethnomedicine of Orissa state, India, in preparations used to treat menorrhagia and fever. In the same tribal community, the congeneric H. tanjorense was used for neuropsychological disturbances, a traditional category encompassing acute mental agitation and behavioral dysregulation. Members of the Apocynaceae family, including closely related Heterostemma species, are known sources of purine-type alkaloids structurally homologous to established neuromodulators such as caffeine and theobromine, compounds recognized for their roles in mood regulation and stress response.Aim of the study: To evaluate the neurobehavioral and brain metabolomic effects of alkaloid-enriched fractions of H. piperifolium in an adult zebrafish acute stress model.Method: Alkaloid-enriched fractions were prepared from leaves of H. piperifolium via Sephadex LH-20 chromatography and chemically profiled by UHPLC-MS/MS. Adult zebrafish subjected to standardized acute stress were treated by immersion with the fraction at three concentrations (25, 250, and 1250 ng/mL) for five consecutive days and evaluated across swim, novel tank, and light-dark behavioral paradigms. Brain metabolomics was performed using untargeted LC-MS/MS with pathway enrichment analysis.Results: Nine putative compounds were identified, predominantly purine alkaloids including heteromines G and H. The fraction attenuated stress-induced behavioral disturbances across all three paradigms, with 250 ng/mL producing the most consistent recovery. Untargeted brain metabolomics identified 23 differential metabolites, with hierarchical clustering revealing dose-dependent realignment of stress-disrupted profiles toward fluoxetine-like signatures. Key metabolic alterations spanned the tryptophan–kynurenine pathway, energy metabolism, and phospholipid turnover.Conclusions: H. piperifolium alkaloid-enriched fractions exert anxiolytic-relevant neurobehavioral effects and multi-pathway metabolic modulation in a zebrafish acute stress model, supporting their potential as plant-derived neuroactive compounds warranting further investigation.

Anti-diabetic compounds from natural sources are now being preferred to prevent or treat diabetes due to adverse effects of synthetic drugs. The decoction of Muntingia calabura leaves was traditionally consumed for diabetes treatment. However, there has not been any published data currently available on the processing effects on this plant’s biological activity and phytochemical profile. Therefore, this study aims to evaluate the effect of three drying methods (freeze-drying (FD), air-drying (AD), and oven-drying (OD)) and ethanol:water ratios (0, 50, and 100%) on in vitro anti-diabetic activities of M. calabura leaves. In addition, an ultrahigh-performance-liquid chromatography–electrospray ionization tandem mass spectrometry (UHPLC-ESI-MS/MS) method was used to characterize the metabolites in the active extract. The FD M. calabura leaves, extracted with 50% ethanol, is the most active extract that exhibits a high α-glucosidase and α-amylase inhibitory activities with IC₅₀ values of 0.46 ± 0.05 and 26.39 ± 3.93 µg/mL, respectively. Sixty-one compounds were tentatively identified by using UHPLC-ESI-MS/MS from the most active extract. Quantitative analysis, by using UHPLC, revealed that geniposide, daidzein, quercitrin, 6-hydroxyflavanone, kaempferol, and formononetin were predominant compounds identified from the active extract. The results have laid down preliminary steps toward developing M. calabura leaves extract as a potential source of bioactive compounds for diabetic treatment.

The 70% ethanolic extracts from eight neglected fruits; Muntingia calabura, Leucaena leucocephala, Spondias dulcis, Syzygium jambos, Mangifera caesia, Ardisia elliptica, Cynometra cauliflora and Ficus auriculata were evaluated for their 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging, α-glucosidase inhibitory activities as well as total phenolic content. The results of this study revealed that M. caesia fruit extract demonstrated the most potent radical scavenging activity. Among the fruits examined for α-glucosidase inhibitory activity, M. calabura and F. auriculata exhibited strong activity with no significant difference. The Pearson correlation indicated that the activities of M. caesia and F. auriculata contributed by phenolic compounds. A total of 65 metabolites were tentatively identified by using ultra-high-performance liquid chromatography tandem mass spectrometry (UHLPC-MS/MS). These findings suggested that the possible application of M. caesia and F. auriculata as a functional food with antioxidant and α-glucosidase inhibitory properties.

Plants and plant-based products have been used for a long time for medicinal purposes. This study aimed to determine the antioxidant and anti-α-glucosidase activities of eight selected underutilized plants in Malaysia: Leucaena leucocephala, Muntingia calabura, Spondias dulcis, Annona squamosa, Ardisia elliptica, Cynometra cauliflora, Ficus auriculata, and Averrhoa bilimbi. This study showed that the 70% ethanolic extract of all plants exhibited total phenolic content (TPC) ranging from 51 to 344 mg gallic acid equivalent (GAE)/g dry weight. A. elliptica showed strong 2,2-diphenyl-1-picrylhydrazyl (DPPH) and nitric oxide (NO) scavenging activities, with half maximal inhibitory concentration (IC₅₀) values of 2.17 and 49.43 μg/mL, respectively. Most of the tested plant extracts showed higher inhibition of α-glucosidase enzyme activity than the standard, quercetin, particularly A. elliptica, F. auriculata, and M. calabura extracts with IC₅₀ values of 0.29, 0.36, and 0.51 μg/mL, respectively. A total of 62 metabolites including flavonoids, triterpenoids, benzoquinones, and fatty acids were tentatively identified in the most active plant, i.e., A. elliptica leaf extract, by using ultra-high-performance liquid chromatography (UHPLC)–electrospray ionization (ESI) Orbitrap MS. This study suggests a potential natural source of antioxidant and α-glucosidase inhibitors from A. elliptica.

Pineapple (Ananas comosus) waste is a promising source of metabolites for therapeutics, functional foods, and cosmeceutical applications. This study strives to characterize the complete metabolite profiles of a variety of MD2 pineapple waste extracts. Metabolomics strategies were utilized to identify bioactive metabolites of this variety prepared with different solvent ratios. Each pineapple waste extract was first screened for total phenolic content, 2,2-diphenyl-1-picrylhydrazyl free radical scavenging, nitric oxide scavenging, and α-glucosidase inhibitory activities. The highest TPC was found in all samples of the peel, crown, and core extracted using a 50% ethanol ratio, even though the results were fairly significant than those obtained for other ethanol ratios. Additionally, crown extracted with a 100% ethanol ratio demonstrated the highest potency in DPPH and NO scavenging activity, with IC₅₀ values of 296.31 and 338.52 µg/mL, respectively. Peel extracted with 100% ethanol exhibited the highest α-glucosidase inhibitory activity with an IC₅₀ value of 92.95 µg/mL. Then, the extracts were analyzed and the data from ¹H NMR were processed using multivariate data analysis. A partial least squares and correlogram plot suggested that 3-methylglutaric acid, threonine, valine, and α-linolenic acid were the main contributors to the antioxidant activities, whereas epicatechin was responsible for the α-glucosidase inhibitory activity. Relative quantification further supported that 100% crown extract was among the extracts that possessed the most abundant potential metabolites. The present study demonstrated that the crown and peel parts of MD2 pineapple extracted with 100% ethanol are potentially natural sources of antioxidants and α-glucosidase inhibitors, respectively.