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Andrographis paniculata (Burm. F.) Nees. is considered as the herb of the future due to its precious chemical compounds, andrographolide (ANDRO), neoandrographolide (NAG) and 14-deoxyandrographolide (DAG). This study aims to profile the metabolites in young and mature leaf at six different harvest ages using 1HNMR-based metabolomics combined with multivariate data analysis. Principal component analysis (PCA) indicated noticeable and clear discrimination between young and mature leaves. A comparison of the leaves stage indicated that young leaves were separated from mature leaves due to its larger quantity of ANDRO, NAG, DAG, glucose and sucrose. These similar metabolites are also responsible for the PCA separation into five clusters representing the harvest age at 14, 16, 18, 20, 22 weeks of leaves extract. Loading plots revealed that most of the ANDRO and NAG signals were present when the plant reached at the pre-flowering stage or 18 weeks after sowing (WAS). As a conclusion, A. paniculata young leaves at pre-flowering harvest age were found to be richer in ANDRO, NAG and DAG compared to mature leaves while glucose and choline increased with harvest age. Therefore, young leaves of A. paniculata should be harvested at 18 WAS in order to produce superior quality plant extracts for further applications by the herbal, nutraceutical and pharmaceutical industries.

Mast cells (MCs), a type of immune effector cell, have recently become recognized for their ability to cause vascular leakage during dengue virus (DENV) infection. Although MC stabilizers have been reported to attenuate DENV induced infection in animal studies, there are limited in vitro studies on the use of MC stabilizers against DENV induced MC degranulation. 2,4,6-trihydroxy-3-geranyl acetophenone (tHGA) has been reported to be a potential MC stabilizer by inhibiting IgE-mediated MC activation in both cellular and animal models. The present study aims to establish an in vitro model of DENV3-induced RBL-2H3 cells using ketotifen fumarate as a control drug, as well as to determine the effect of tHGA on the release of MC mediators upon DENV infection. Our results demonstrated that the optimal multiplicities of infection (MOI) were 0.4 × 10−2 and 0.8 × 10−2 focus forming units (FFU)/cell. Ketotifen fumarate was proven to attenuate DENV3-induced RBL-2H3 cells degranulation in this in vitro model. In contrast, tHGA was unable to attenuate the release of both β-hexosaminidase and tumor necrosis factor (TNF)-α. Nonetheless, our study has successfully established an in vitro model of DENV3-induced RBL-2H3 cells, which might be useful for the screening of potential MC stabilizers for anti-dengue therapies.

The antioxidant and neuroprotective activity of Glucomoringin isothiocyanate (GMG-ITC) have been reported in in vivo and in vitro models of neurodegenerative diseases. However, its neuroprotective role via mitochondrial-dependent pathway in a noxious environment remains unknown. The main objective of the present study was to unveil the mitochondrial apoptotic genes' profile and prospectively link with neuroprotective activity of GMG-ITC through its ROS scavenging. The results showed that pre-treatment of differentiated SH-SY5Y cells with 1.25 μg/mL purified isolated GMG-ITC, significantly reduced reactive oxygen species (ROS) production level, compared to H

Objective: This research revealed the biochemical outcomes of metabolic dysregulation in serum associated with physiological sickness behavior following lipopolysaccharide (LPS)-induced neuroinflammation in rats, and treatment with Clinacanthus nutans (CN). Verification of 1H NMR analysis of the CN aqueous extract proved the existence of bioactive phytochemical constituents’ in extract. Materials and Methods: Twenty-five rats were subjected to unilateral stereotaxic injection of 10 µL LPS (1 mg/mL), while another ten rats were injected with phosphate-buffered saline (PBS, 10 µL) as control. Then, 29 parameters of rat behavior related to sickness were tracked by a device software (SMART 3.0.1) on days 0 and 14 of CN treatment. The acquired and accumulated data were analyzed using multivariate data analysis with the SIMCA Software package (version 13, Umetrics AB; Umeå, Sweden). The pattern trends of related groups were documented using PCA and OPLS analysis. Results: A similar ameliorated correlation pattern was detected between improvement in physiological sickness behavior and anti-inflammatory biomarkers by the 1H NMR spectra of the sera following treatment with CN (500 and 1000 mg/kg body weight (bw)) and the control drug (dextromethorphan hydrobromide, 5 mg/kg of rats bw) in rats. Here, 21 biomarkers were detected for neuroinflammation. Treatment with the aqueous CN extract resulted in a statistically significant alteration in neuroinflammation metabolite biomarkers, including ethanol, choline, and acetate. Conclusion: This result denotes that the metabolomics approach is a reliable tool to disclose the relationship between central neuroinflammation, and systemic metabolic and physiological disturbances which could be used for future ethno-pharmacological assessments.

The present studies are to evaluate the ability of PB to induce weight loss and urine metabolite profile of Piper betle L. (PB) leaf extracts using metabolomics approach. Dried PB leaves were extracted with ethanol 70% and the studies were performed in different groups of rats fed with high fat (HFD) and normal diet (ND). Then, fed with the PB extract with 100, 300, and 500 mg/kg and two negative control groups given water (WTR). The body weights were monitored and evaluated. Urine was collected and 1H NMR-based metabolomics approach was used to detect the metabolite changes. Results showed that PB-treated group demonstrated inhibition of body weight gain. The trajectory of urine metabolites showed that PB-treated group gave the different distribution from week 12 to 16 compared with the control groups. In 1H NMR metabolomic approach analysis, the urine metabolites gave the best separation in principle component 1 and 3, with 40.0% and 9.56% of the total variation. Shared and unique structures (SUS) plot model showed that higher concentration PB-treated group was characterized by high level of indole-3-acetate, aspartate, methanol, histidine, and creatine, thus caused an increased the metabolic function and maintaining the body weight of the animals treated.

Supplemental material, Supplementary_Material_by_Song,_et_al for Novel 2-Benzoyl-6-(2,3-Dimethoxybenzylidene)-Cyclohexenol Confers Selectivity toward Human MLH1 Defective Cancer Cells through Synthetic Lethality by Dedrick Soon Seng Song, Sze Wei Leong, Kwok Wen Ng, Faridah Abas, Khozirah Shaari, Chee Onn Leong, Felicia Fei-Lei Chung, Chun Wai Mai, Ling Wei Hii, Pei Jean Tan and Vyomesh Patel in SLAS Discovery

Table S1. Maximum percent inhibition of α-glucosidase activity of different EtOH:water (E:W) extracts of C. caudatus. Table S2. Pearson’s correlation coefficient (r) between TPC and α-glucosidase activity of the EtOH:water extract solvent system at different harvesting ages. Table S3. VIP values of the major contributing compounds in the PLS model. Table S4. Maximum percent inhibition of α-glucosidase activity of the C. caudatus E:W (80:20) extract at different harvesting ages. Figure S1. Full spectrum of a representative J-resolved spectrum of an EtOH:water (80:20) C. caudatus extract from δ 0.0 to 8.0. Key to metabolites: 1 – quercetin derivatives; 2 – rutin; 3 - chlorogenic acid; 4 - proline; 5 - fatty acid; 6 - acetic acid; 7- catechin. Figure S2. (A) Full 1H NMR spectra of 6-, 8-, 10-, 12- and 14-week-old samples of C. caudatus. (B) Expanded 1H NMR spectra of the samples in the range of δ 5.5 to 8.5. Samples were dissolved in CH3OH-d4-KH2PO4 buffer in D2O (pH 6.0). Key to metabolites: 1 – quercetin derivatives; 2 - rutin; 3 - chlorogenic acid; 4 - proline; 5 - fatty acid; 6 - acetic acid; 7 – catechin. Figure S3. Positive ion mode MS/MS spectra of peaks 1 to 5. Peak 1, rutin; peak 2, quercetin 3-O-glucoside; peak 3, quercetin 3-O-xyloside; peak 4, quercetin 3-O-arabinofuranoside and peak 5, quercetin 3-O-rhamnoside. Figure S4. Relative quantification of metabolites in 6-, 8-, 10-, 12- and 14-week-old samples of C. caudatus at different harvesting ages. An * indicates significant differences (p

Flacourtia rukam is a species of the Flacourtiaceae. The edible, cherry-like, sweet and sour fruits of the species are not known to be commercially exploited. Phenolic compounds and flavonoids are the main constituents of the fruits, which have also been reported to have good antioxidant properties. However, the available information on the phytochemical and pharmacological aspects of F. rukam indicated it to be very much an underutilized tropical fruit tree. Nevertheless, based on the ethnomedicinal uses of the fruits and other parts of the plant, its economic potential as a health giving food crop should be further investigated and properly evaluated.

Oil palm leaves (OPL) containing flavonoid C-glycosides are abundantly generated as oil palm byproducts. The performances of three macroporous resins with different physical and chemical properties for the enrichment of isoorientin, orientin, vitexin, and isovitexin from acid-hydrolyzed OPL (OPLAH) extract were screened. The XAD7HP resin exhibited the best sorption capacities for the targeted flavonoid C-glycosides and was thus selected for further evaluation. Static adsorption using the XAD7HP resin under optimal conditions (extract adjusted to pH 5, shaken at 298 K for 24 h) gave adsorption kinetics that fit well with a pseudo-second-order kinetic model. The adsorption of isoorientin and orientin was well described by Langmuir isotherms, while vitexin and isovitexin fit well with the Freundlich isotherms. Dynamic sorption trials using the column-packed XAD7HP resin produced 55–60-fold enrichment of isovitexin and between 11 and 25-fold enrichment of isoorientin, vitexin, and orientin using aqueous ethanol. The total flavonoid C-glycoside-enriched fractions (enriched OPLAH) with isoorientin (247.28–284.18 µg/mg), orientin (104.88–136.19 µg/mg), vitexin (1197.61–1726.11 µg/mg), and isovitexin (13.03–14.61 µg/mg) showed excellent antioxidant free radical scavenging activities compared with their crude extracts, with IC50 values of 6.90–70.63 µg/mL and 44.58–200.00 µg/mL, respectively. Hence, this rapid and efficient procedure for the preliminary enrichment of flavonoid C-glycosides by using macroporous resin may have practical value in OPL biomass waste utilization programs to produce high value-added products, particularly in the nutraceuticals, cosmeceuticals, pharmaceuticals, and fine chemicals industries.

Piper betle L. (PB) belongs to the Piperaceae family. The presence of a fairly large quantity of diastase in the betel leaf is deemed to play an important role in starch digestion and calls for the study of weight loss activities and metabolite profile from PB leaf extracts using metabolomics approach to be performed. PB dried leaves were extracted with 70% ethanol and the extracts were subjected to five groups of rats fed with high fat (HF) and standard diet (SD). They were then fed with the extracts in two doses and compared with a negative control group given water only according to the study protocol. The body weights and food intakes were monitored every week. At the end of the study, blood serum of the experimental animal was analysed to determine the biochemical and metabolite changes. PB treated group demonstrated inhibition of body weight gain without showing an effect on the food intake. In serum bioassay, the PB treated group (HF/PB (100mg/kg and 500mg/kg) showed an increased in glucose and cholesterol levels compared to the Standard Diet (SD/WTR) group, a decrease in LDL level and increase in HDL level when compared with High Fat Diet (HF/WTR) group. For metabolite analysis, two separation models were made to determine the metabolite changes via group activities. The best separation of PCA serum in Model 1 and 2 was achieved in principle component 1 and principle component 2. SUS-Plot model showed that HF group was characterized by high-level of glucose, glycine and alanine. Increase in the β-hydroxybutyrate level similar with SD group animals was evident in the HF/PB(500mg/kg) group. This finding suggested that the administration of 500mg/kg PB extracts leads to increase in oxidation process in the body thus maintaining the body weight and without giving an effect on the appetite even though HF was continuously consumed by the animals until the end of the studies and also a reduction in food intake, thus maintaining their body weight although they were continuously consumed HF.

Pharmocological relevance 2,4,6-trihydroxy-3-geranyl acetophenone (tHGA), is a phloroglucinol compound found naturally in Melicope ptelefolia. Melicope ptelefolia has been used traditionally for centuries as natural remedy for wound infections and inflammatory diseases. Aim of the study Endothelial barrier dysfunction is a pathological hallmark of many diseases and can be caused by lipopolysaccharides (LPS) stimulation. Therefore, this study aims to investigate the possible barrier protective effects of tHGA upon LPS-stimulated inflammatory responses in human umbilical vein endothelial cells (HUVECs). Materials and methods HUVECs were pretreated with tHGA prior to LPS stimulation, where inflammatory parameters including permeability, monocyte adhesion and migration, and release of pro-inflammatory mediators were examined. Additionally, the effect of tHGA on F-actin rearrangement and adhesion protein expression of LPS-stimulated HUVECs was evaluated. Results It was found that pretreatment with tHGA inhibited monocyte adhesion and transendothelial migration, reduced endothelial hyperpermeability and secretion of prostaglandin E 2 (PGE 2 ). Additionally, tHGA inhibited cytoskeletal rearrangement and adhesion protein expression on LPS-stimulated HUVECs. Conclusion As the regulation of endothelial barrier dysfunction can be one of the therapeutic strategies to improve the outcome of inflammation, tHGA may be able to preserve vascular barrier integrity of endothelial cells following LPS-stimulated dysfunction, thereby endorsing its potential usefulness in vascular inflammatory diseases.

Herbal medicine has been proven to be an effective therapy offering a variety of benefits, such as moderate reduction in hypoglycemia, in the treatment and prevention of obesity and diabetes. Phyllanthus niruri has been used as a treatment for diabetes mellitus. Herein, the induction of type 2 diabetes in Sprague-Dawley rats was achieved by a low dose of streptozotocin (STZ) (25mg/kgbw). Here, we evaluated the in vivo antidiabetic properties of two concentrations (250 and 500mg/kg bw) of P. niruri via metabolomics approach. The administration of 500mg/kgbw of P. niruri extract caused the metabolic disorders of obese diabetic rats to be improved towards the normal state. The extract also clearly decreased the serum glucose level and improved the lipid profile in obese diabetic rats. The results of this study may contribute towards better understanding the molecular mechanism of this medicinal plant in managing diabetes mellitus.

In the present study, a series of 2-benzoyl-6-benzylidenecyclohexanone analogs have been synthesized and evaluated for their anti-cholinesterase activity. Among the forty-one analogs, four compounds (38, 39, 40 and 41) have been identified as lead compounds due to their highest inhibition on both AChE and BChE activities. Compounds 39 and 40 in particular exhibited highest inhibition on both AChE and BChE with IC50 values of 1.6 μM and 0.6 μM, respectively. Further structure–activity relationship study suggested that presence of a long-chain heterocyclic in one of the rings played a critical role in the dual enzymes’ inhibition. The Lineweaver–Burk plots and docking results suggest that both compounds could simultaneously bind to the PAS and CAS regions of the enzyme. ADMET analysis further confirmed the therapeutic potential of both compounds based upon their high BBB-penetrating. Thus, 2-benzoyl-6-benzylidenecyclohexanone containing long-chain heterocyclic amine analogs represent a new class of cholinesterase inhibitor, which deserve further investigation for their development into therapeutic agents for cognitive diseases such as Alzheimer.

Ipomoea aquatica Forsk is a green leafy vegetable that is a rich source of minerals, proteins, vitamins, amino acids, and secondary metabolites. Different types of I. aquatica cultivars 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 three I. aquatica cultivars 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. The in vitro study revealed that the I. aquatica cultivars 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.

Neptunia oleracea is a plant consumed as vegetable and used as a traditional herb to treat several ailments. This study evaluated metabolite variations among N. oleracea leaf and stem subjected to air drying (AD), freeze drying (FD) and oven drying (OD) using proton nuclear magnetic resonance (1H NMR) based metabolomics. The correlation was also studied for the metabolite content with total phenolic content (TPC), DPPH free radical scavenging and α-glucosidase inhibitory activities. A total of 18 metabolites were identified from N. oleracea extracts, including 10 primary metabolites, 5 flavonoids and 3 phenolic acids using NMR. Ultra-high performance liquid chromatography tandem mass spectrometry analysis (UHPLC-MS/MS) confirmed the presence of the secondary metabolites and revealed the flavonoid derivatives present. All the identified phenolics are first reported from this plant. Multivariate data analysis (MVDA) showed strong correlation between the metabolites with the antioxidant and α-glucosidase inhibitory activities of FD N. oleracea leaves. The compounds suggested to be responsible for the high activity of FD leaves include vitexin-2-O-rhamnoside, catechin, caffeic acid, gallic acid and derivatives of quercetin, kaempferol and myricetin. This study demonstrates that FD N. oleracea leaves are a potential natural source for antioxidant and α-glucosidase inhibitors.

Plants have an extensive diversity of natural phytochemicals that are used as a source of medicine. In this study the methanolic leaf extract of Callicarpa maingayi together with its increasing polarity soluble fractions hexane (Hex), dichloromethane (DCM), ethyl acetate (EtOAc), n-butanol (n-BuOH), and aqueous (Aq) fractions were examined on antioxidant, acetylcholinesterase (AChE), tyrosinase, and α-glucosidase inhibitory activities. Liquid chromatography-diode array detection-electrospray ionization-tandem mass spectrometry (LC-DAD-ESIMS/MS) was used to analyze the most bioactive fractions (EtOAc and n-BuOH). A total of 20 compounds were tentatively identified in these two active fractions. Forsythoside B, acteoside, isoacteoside, isocampneoside II, apigenin 7-O-rutinoside and kaempferol 3-sulfate-7-arabinopyranoside were identified by comparing with the reference standard. Other compounds including cistanoside F, apigenin 6-C-glucoside-8-C-glucoside, β-OH-forsythoside B, campneoside II, rhamnazin 3-O-rutinoside, campneoside I, eukovoside, acacetin diglucoronide, acacetin 7-O-glucuronide, β-OH-poliumoside, poliumoside as well as 3 isomers of 2′-acetylacteoside were tentatively identified based on their UV spectra and MS/MS data. All of the compounds were reported from this species for the first time. The results have indicated the chemical constituents of C. maingayi that contributed to the biological activity and thus, it may have potential application toward the development of nutraceutical and medicinal preparations.

Ethnopharmacological relevance ‘Pegaga’ is a traditional Malay remedy for a wide range of complaints. Among the 'pegaga’, Centella asiatica has been used as a remedy for diabetes mellitus. Thus, we decided to validate this claim by evaluating the in vivo antidiabetic property of C. asiatica (CA) on T2DM rat model using the holistic 1 H NMR-based metabolomics approach. Method In this study, an obese diabetic (mimic of T2DM condition) animal model was developed using Sprague–Dawley rats fed with a high-fat diet and induced into diabetic condition by the treatment of a low dose of streptozotocin (STZ). The effect of C. asiatica extract on the experimental animals was followed based on the changes observed in the urinary and serum metabolites, measured by 1 H NMR of urine and blood samples collected over the test period. Results A long-term treatment of obese diabetic rats with CA extract could reverse the glucose and lipid levels, as well as the tricarboxylic acid cycle and amino acid metabolic disorders, back towards normal states. Biochemical analysis also showed an increase of insulin production in diabetic rats upon treatment of CA extract. Conclusion This study has provided evidence that clearly supported the traditional use of CA as a remedy for diabetes. NMR-based metabolomics was successfully applied to show that CA produced both anti-hyperglycemic and anti-hyperlipidemic effects on a rat model. In addition to increasing the insulin secretion, the CA extract also ameliorates the metabolic pathways affected in the induced diabectic rats. This study further revealed the potential usage of CA extract in managing diabetes mellitus and the results of this work may contribute towards the further understanding of the underlying molecular mechanism of this herbal remedy.

The present study investigated the antidiabetic properties of the extracts and fractions from leaves and stem bark of M. glabra based on dipeptidyl peptidase-4 (DPP-4) and &alpha, amylase inhibitory activity assays. The chloroform extract of the leaves was found to be most active towards inhibition of DPP-4 and &alpha, amylase with IC50 of 169.40 &mu, g/mL and 303.64 &mu, g/mL, respectively. Bioassay-guided fractionation of the leaves&rsquo, chloroform extract revealed fraction 4 (CF4) as the most active fraction (DPP-4 IC50: 128.35 &mu, g/mL, &alpha, amylase IC50: 170.19 &mu, g/mL). LC-MS/MS investigation of CF4 led to the identification of trans-decursidinol (1), swermirin (2), methyl 3,4,5-trimethoxycinnamate (3), renifolin (4), 4&prime, 5,6,7-tetramethoxy-flavone (5), isorhamnetin (6), quercetagetin-3,4&prime, dimethyl ether (7), 5,3&prime, 4&prime, trihydroxy-6,7-dimethoxy-flavone (8), and 2-methoxy-5-acetoxy-fruranogermacr-1(10)-en-6-one (9) as the major components. The computational study suggested that (8) and (7) were the most potent DPP-4 and &alpha, amylase inhibitors based on their lower binding affinities and extensive interactions with critical amino acid residues of the respective enzymes. The binding affinity of (8) with DPP-4 (&minus, 8.1 kcal/mol) was comparable to that of sitagliptin (&minus, 8.6 kcal/mol) while the binding affinity of (7) with &alpha, amylase (&minus, 8.6 kcal/mol) was better than acarbose (&minus, 6.9 kcal/mol). These findings highlight the phytochemical profile and potential antidiabetic compounds from M. glabra that may work as an alternative treatment for diabetes.

Technological advances, coupled with increasing demands by consumers, have led to a drastic increase in plastic production. After serving their purposes, these plastics reach our water bodies as their destination and become ingested by aquatic organisms. This ubiquitous phenomenon has exposed humans to microplastics mostly through the consumption of sea food. This has led the World Health Organization (WHO) to make an urgent call for the assessment of environmental pollution due to microplastics and its effect on human health. This review summarizes studies between 1999 and 2020 in relation to microplastics in aquatic ecosystems and human food products, their potential toxic effects as elicited in animal studies, and policies on their use and disposal. There is a paucity of information on the toxicity mechanisms of microplastics in animal studies, and despite their documented presence in food products, no policy has been in place so far, to monitor and regulates microplastics in commercial foods meant for human consumption. Although there are policies and regulations with respect to plastics, these are only in a few countries and in most instances are not fully implemented due to socioeconomic reasons, so they do not address the problem across the entire life cycle of plastics from production to disposal. More animal research to elucidate pathways and early biomarkers of microplastic toxicity that can easily be detected in humans is needed. This is to create awareness and influence policies that will address this neglected threat to food safety and security.

Arbutus pavarii Pamp is a medicinal plant commonly used by local tribes in East Libya for the treatment of many diseases, such as gastritis, renal infections, cancer and kidney diseases. In this study, the antibacterial activity of the leaf and stem bark extracts of the plant against methicillin-resistant Staphylococcus aureus (MRSA), as well as the metabolite profiles of the bioactive fractions, was investigated. The antibacterial activity was determined by disc diffusion method, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), while the microbial reduction by the bioactive fraction was evaluated using time&ndash, kill test. The bioactive fraction was further subjected to ultrahigh-performance liquid chromatography&ndash, mass spectrometry (UHPLC-ESI-MS/MS) analysis to putatively identify the chemical constituents contained therein. All the extracts and fractions showed different levels of antibacterial activity on the tested MRSA strains. The highest total antibacterial activity, i.e., 4007.6 mL/g, was exhibited by the crude leaf methanolic extract. However, the ethyl acetate fraction of the leaf showed moderate to significant antibacterial activity against MRSA at low MIC (0.08&ndash, 1.25 mg/mL). Metabolite profiling of this fraction using UHPLC-ESI-MS/MS resulted in the putative identification of 28 compounds, which included phenolic acids, flavan-3-ols and flavonols. The results of this study showed that the ethyl acetate fraction of Arbutus pavarii leaf possessed potential antibacterial activity against MRSA and hence can be further explored for pharmaceutical applications as a natural antibacterial agent.

The commercial cultivation of microalgae began in the 1960s and Chlorella was one of the first target organisms. The species has long been considered a potential source of renewable energy, an alternative for phytoremediation, and more recently, as a growth and immune stimulant. However, Chlorella vulgaris, which is one of the most studied microalga, has never been comprehensively profiled chemically. In the present study, comprehensive profiling of the Chlorella vulgaris metabolome grown under normal culture conditions was carried out, employing tandem LC-MS/MS to profile the ethanolic extract and GC-MS for fatty acid analysis. The fatty acid profile of C. vulgaris was shown to be rich in omega-6, -7, -9, and -13 fatty acids, with omega-6 being the highest, representing more than sixty percent (&gt, 60%) of the total fatty acids. This is a clear indication that this species of Chlorella could serve as a good source of nutrition when incorporated in diets. The profile also showed that the main fatty acid composition was that of C16-C18 (&gt, 92%), suggesting that it might be a potential candidate for biodiesel production. LC-MS/MS analysis revealed carotenoid constituents comprising violaxanthin, neoxanthin, lutein, &beta, carotene, vulgaxanthin I, astaxanthin, and antheraxanthin, along with other pigments such as the chlorophylls. In addition to these, amino acids, vitamins, and simple sugars were also profiled, and through mass spectrometry-based molecular networking, 48 phospholipids were putatively identified.

Food sanitizer is important to reduce or eliminate microbial population on the food surface and also to removes dirt. Nowadays, food sanitizer is basically from chemical base. Therefore, this study was to evaluate the toxicity level of S. polyanthum and its stability against different pHs and different temperatures in determining the suitability of leaves extract as natural food sanitizer. Leaves extract was adjusted to pH 3, pH 7, pH 11 conditions and exposed to temperatures at 30°C, 50°C and 80°C for 15 min each. Treated extracts were tested for their antimicrobial properties in term of MICs and MBCs. Toxicity level of S. polyanthum was determined by brine shrimp lethality assay. As a result, antimicrobial activities of S. polyanthum extract was not affected by different pHs and temperatures while toxicity study demonstrated that S. polyanthum, extract was not toxic to Artemia salina with LC50 was 75.85 mg/mL. Therefore, S. polyanthum had the potential to be developed as antimicrobial agent in food sanitizer.

Background: Zanthoxylum rhetsa is a plant used in traditional medicine and is known to possess health benefits such as antibacterial, antidiabetic, and anti-diarrheal activities. Objectives: The objective of this study was to explore and demonstrate the anti-inflammatory activity of various solvent fractions of Z. rhetsa bark. Materials and Methods: The effect of crude methanolic extract and its fractions (hexane, chloroform, ethyl acetate, and butanol) on the levels of pro-inflammatory cytokines (interleukin-1β, tumor necrosis factor-alpha, and interleukin-6) and inflammatory factors was tested via targeting nuclear factor-kappa B (NF-κB) signaling pathways in lipopolysaccharide (LPS)-stimulated mouse RAW 264.7 macrophages. Results: Treatment with all the solvent fractions at various concentrations (50, 100, and 200 μg/ml) suppressed pro-inflammatory cytokine levels in a dose-dependent manner. Among the fractions, the chloroform fractions exhibited a significant inhibition of LPS-induced inflammation. Moreover, these fractions effectively suppressed the expression of various NF-κB signaling targets, including NF-κB, nitric oxide synthase, and cyclooxygenase-2 as well as inhibited the degradation of nuclear factor of kappa light polypeptide gene enhancer in B-cell inhibitor, alpha (inhibitor of kappa B alpha [IκBα]). This anti-inflammatory effect was mediated by the prevention of IκBα degradation, as this protein prevents the translocation of NF-κB from the cytoplasm to the nucleus, initiating the transcription of pro-inflammatory genes. Conclusion: Thus, the Z. rhetsa chloroform fraction may be an effective natural anti-inflammatory agent against inflammation-associated diseases.

Background Although the development of antibiotic and antioxidant manufacturing, the problem of bacterial resistance and food and/or cosmetics oxidation still needs more efforts to design new derivatives which can help to minimize these troubles. Benzimidazo[1,2-c]quinazolines are nitrogen-rich heterocyclic compounds that possess many pharmaceutical properties such as antimicrobial, anticonvulsant, immunoenhancer, and anticancer. Results A comparative study between two methods, (microwave-assisted and conventional heating approaches), was performed to synthesise a new quinazoline derivative from 2-(2-aminophenyl)-1H-benzimidazole and octanal to produce 6-heptyl-5,6-dihydrobenzo[4,5]imidazo[1,2-c]quinazoline (OCT). The compound was characterised using FTIR, 1H and 13C NMR, DIMS, as well as X-ray crystallography. The most significant peak in the 13C NMR spectrum is C-7 at 65.5 ppm which confirms the cyclisation process. Crystal structure analysis revealed that the molecule grows in the monoclinic crystal system P21/n space group and stabilised by an intermolecular hydrogen bond between the N1–H1A…N3 atoms. The crystal packing analysis showed that the molecule adopts zig-zag one dimensional chains. Fluorescence study of OCT revealed that it produces blue light when expose to UV-light and its’ quantum yield equal to 26%. Antioxidant activity, which included DPPH· and ABTS·+ assays was also performed and statistical analysis was achieved via a paired T-test using Minitab 16 software with P

Background Cosmos caudatus is an annual plant known for its medicinal value in treating several health conditions, such as high blood pressure, arthritis, and diabetes mellitus. The α-glucosidase inhibitory activity and total phenolic content of the leaf aqueous ethanolic extracts of the plant at different growth stages (6, 8. 10, 12 and 14 weeks) were determined in an effort to ascertain the best time to harvest the plant for maximum medicinal quality with respect to its glucose-lowering effects. Methods The aqueous ethanolic leaf extracts of C. caudatus were characterized by NMR and LC-MS/MS. The total phenolic content and α-glucosidase inhibitory activity were evaluated by the Folin-Ciocalteu method and α-glucosidase inhibitory assay, respectively. The statistical significance of the results was evaluated using one-way ANOVA with Duncan’s post hoc test, and correlation among the different activities was performed by Pearson’s correlation test. NMR spectroscopy along with multivariate data analysis was used to identify the metabolites correlated with total phenolic content and α-glucosidase inhibitory activity of the C. caudatus leaf extracts. Results It was found that the α-glucosidase inhibitory activity and total phenolic content of the optimized ethanol:water (80:20) leaf extract of the plant increased significantly as the plant matured, reaching a maximum at the 10th week. The IC50 value for α-glucosidase inhibitory activity (39.18 μg mL− 1) at the 10th week showed greater potency than the positive standard, quercetin (110.50 μg mL− 1). Through an 1H NMR-based metabolomics approach, the 10-week-old samples were shown to be correlated with a high total phenolic content and α-glucosidase inhibitory activity. From the partial least squares biplot, rutin and flavonoid glycosides, consisting of quercetin 3-O-arabinofuranoside, quercetin 3-O-rhamnoside, quercetin 3-O-glucoside, and quercetin 3-O-xyloside, were identified as the major bioactive metabolites. The metabolites were identified by NMR spectroscopy (J-resolve, HSQC and HMBC experiments) and further supported by dereplication via LC-MS/MS. Conclusion For high phytomedicinal quality, the 10th week is recommended as the best time to harvest C. caudatus leaves with respect to its glucose lowering potential. Electronic supplementary material The online version of this article (10.1186/s12906-019-2655-9) contains supplementary material, which is available to authorized users.

The present study aims for the first time to provide the in vivo acute toxicological profile of the highest dose of Clinacanthus nutans (Burm. f.) Lindau water leaf extract according to the Organization for economic co-operation and development (OECD) 423 guidelines through conventional toxicity and advanced proton nuclear magnetic resonance (1H-NMR) serum and urinary metabolomics evaluation methods. A single dose of 5000 mg/kg bw of C. nutans water extract was administered to Sprague Dawley rats, and they were observed for 14 days. Conventional toxicity evaluation methods (physical observation, body and organ weight, food and water consumption, hematology, biochemical testing and histopathological analysis) suggested no abnormal toxicity signs. Serum 1H-NMR metabolome revealed no significant metabolic difference between untreated and treated groups. Urinary 1H-NMR analysis, on the other hand, revealed alteration in carbohydrate metabolism, energy metabolism and amino acid metabolism in extract-treated rats after 2 h of extract administration, but the metabolic expression collected after 24 h and at Day 5, Day 10 and Day 15 indicated that the extract-treated rats did not accumulate any toxicity biomarkers. Importantly, the outcomes further suggest that single oral administration of up to 5000 mg/kg bw of C. nutans water leaf extract is safe for consumption.

Background: The official standard for quality control of honey is currently based on physicochemical properties. However, this method is time-consuming, cost intensive, and do not lead to information on the originality of honey. This study aims to classify raw stingless bee honeys by bee species origins as a potential classifier using NMR-LCMS-based metabolomics approach. Methods: Raw stingless bee honeys were analysed and classified by bee species origins using proton nuclear magnetic resonance (1H NMR) spectroscopy and an ultra-high performance liquid chromatography-quadrupole time of flight mass spectrometry (UHPLC-QTOF MS) in combination with chemometrics tools. Results: The honey samples were able to be classified into three different groups based on the bee species origins of Heterotrigona itama, Geniotrigona thoracica, and Tetrigona apicalis. D-Fructofuranose (H. itama honey), β-D-Glucose, D-Xylose, α-D-Glucose (G. thoracica honey), and L-Lactic acid, Acetic acid, L-Alanine (T. apicalis honey) identified via 1H NMR data and the diagnostic ions of UHPLC-QTOF MS were characterized as the discriminant metabolites or putative chemical markers. Conclusion: It could be suggested that the quality of honey in terms of originality and purity can be rapidly determined using classification technique by bee species origins via 1H NMR- and UHPLC-QTOF MS-based metabolomics approach.

The official standard for quality control of honey is currently based on physicochemical properties. However, this method is time-consuming, cost intensive, and does not lead to information on the originality of honey. This study aims to classify raw stingless bee honeys by bee species origins as a potential classifier using the NMR-LCMS-based metabolomics approach. Raw stingless bee honeys were analysed and classified by bee species origins using proton nuclear magnetic resonance (1H-NMR) spectroscopy and an ultra-high performance liquid chromatography-quadrupole time of flight mass spectrometry (UHPLC-QTOF MS) in combination with chemometrics tools. The honey samples were able to be classified into three different groups based on the bee species origins of Heterotrigona itama, Geniotrigona thoracica, and Tetrigona apicalis. d-Fructofuranose (H. itama honey), &beta, d-Glucose, d-Xylose, &alpha, d-Glucose (G. thoracica honey), and l-Lactic acid, Acetic acid, l-Alanine (T. apicalis honey) ident d-Fructofuranose identified via 1H-NMR data and the diagnostic ions of UHPLC-QTOF MS were characterized as the discriminant metabolites or putative chemical markers. It could be suggested that the quality of honey in terms of originality and purity can be rapidly determined using the classification technique by bee species origins via the 1H-NMR- and UHPLC-QTOF MS-based metabolomics approach.

A group of 126 endophytic fungi was isolated from dragon fruit plants in Malaysia. Dual culture and disc diffusion test revealed that seven strains could suppress the growth of Colletotrichum gloeosporioides. Of all the potential strains, strain T3.3 exhibited the best antagonistic activity by producing an inhibition zone of 12±1.0 mm in dual culture test. Disc diffusion test using crude extract produced by this fungus could inhibit the growth of C.gloeosporioides by 33.33±2.89%. The diffused non volatile metabolite produced by this strain suppressed the growth of the pathogen by 97% after 7 days of incubation. Based on morphological characteristics and ITS region sequence, strain T3.3 was identified as Penicillium oxalicum. The carbon utilization profile in Biolog FF Microplate analysis revealed that P.oxalicum T3.3 is a versatile microorganism. P.oxalicum T3.3 was found to produce β-glucanase and chitinase with activity of 3.38 U/mL and 1.19 U/mL respectively. In addition, the ethylacetate extract of P.oxalicum T3.3 could suppress the growth of C.gloeosporioides. Scanning electron microscopy study showed that the release of extracellular antifungal metabolites from the endophytic P.oxalicum T3.3 had caused abnormal hyphal growth of C.gloeosporioides. Treatment with the crude extract from this fungus on dragon fruit could control anthracnose disease in this fruit in vivo. Thus, endophytic Penicillium oxalicum T3.3 is considered as a potential biological control agent of anthracnose disease in dragon fruit.

Clinacanthus nutans (Burm.f.) Lindau (Acanthaceae), commonly known as Sabah snake grass, is a vegetable and a well-known herb that is considered an alternative medicine for insect bites, skin rashes, herpes infection, inflammation, and cancer and for health benefits. Current review aims to provide a well-tabulated repository of the phytochemical screening, identification and quantification, and the pharmacological information of C. nutans according to the experimental design and the plant preparation methods which make it outstanding compared to existing reviews. This review has documented valuable data obtained from all accessible library databases and electronic searches. For the first time we analyzed the presence of flavonoids, triterpenoids, steroids, phytosterols, and glycosides in C. nutans based on the results from phytochemical screening which are then further confirmed by conventional phytochemical isolation methods and advanced spectroscopic techniques. Phytochemical quantification further illustrated that C. nutans is a good source of phenolics and flavonoids. Pharmacological studies on C. nutans revealed that its polar extract could be a promising anti-inflammation, antiviral, anticancer, immune and neuromodulating, and plasmid DNA protective agent; that its semipolar extract could be a promising antiviral, anticancer, and wound healing agent; and that its nonpolar extract could be an excellent anticancer agent.

Additional file 1. Additional tables.

Clinacanthus nutans is a medicinal herb and a traditional remedy for herpes viral infection, cancer, and diabetes mellitus. Despite its popular use, there is limited information on the chemical constituents and their relationship with the bioactivities of the herb. The choice of drying and extraction methods will greatly influence the metabolite profile and the bioactivities of an herbal extract. In order to maximize retention of the original chemical profile of the herb and quality assurance, optimization of processing methods is needed. Using nuclear magnetic resonance (NMR) based metabolomics approach, we have carried out a discriminative analysis of the metabolite profiles of the leaves and stems of the herb when different drying (air, oven, and freeze) and extraction (soaking and sonication) methods were used and correlated the metabolite profiles with the total phenolic content (TPC), antioxidant and α-glucosidase inhibitory activities. Identification of primary and secondary metabolites was performed using 1D- and 2D-NMR techniques as well as ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS). Results showed that the leaf extracts, which were richer in phenolic compounds and terpenoids, showed significantly higher bioactivities compared to the stem extracts. Several newly reported compounds in the herb, identified using tandem mass spectrometry, included gendarucin A, a gendarucin A isomer, 3,3-di-O-methylellagic acid, ascorbic acid, and two isomeric oxoprolinates. From the NMR metabolomics analysis, the PLS biplot model indicated that the presence of some phenolics compounds, terpenoids, and sulfur-containing glucosides in the oven and air dried leaf extracts are the main components responsible for the antioxidant and α-glucosidase inhibitory activities. This study has provided additional information on the chemistry and biology of the herb that may be useful in future development phytomedicinal preparations of C. nutans.

Several studies have suggested that plants are potential sources of bioactive compounds. Phyllanthus is a plant genus that has been used in traditional medicine due to its phytomedicinal metabolites content. The variation between two Phyllanthus species (P. niruri and P. urinaria) was studied using proton nuclear magnetic resonance (1H NMR) combined with multivariate data analysis (MVDA). The total phenolic content (TPC), DPPH radical scavenging activity and α-glucosidase inhibitory activity of the Phyllanthus species were also evaluated and correlated with their phytochemical constituents at different growth stages (8, 10 and 12 weeks) using partial least square regression (PLS). Principal component analysis (PCA) and PLS indicated separation between the two species based on the identified metabolites and the screened bioactivities. A comparison of the two species indicated that P. urinaria was separated from P. niruri due to its larger quantity of fatty and amino acids, choline, phyllanthin and sucrose. However, P. niruri contained higher quantities of hypophyllanthin and phenolic compounds. The loading column plot, which was used to compare the P. niruri at different growth stages, indicated that the eight-week-old plant contained a higher amount of fatty acids, amino acids (leucine and alanine), phyllanthin and choline. The dominant substances in the P. niruri at 10 weeks of growth by PC1 were identified as hypophyllanthin, malic acid, sucrose, and identified phenolics. The 12 week sample was differentiated by its higher sugars contents as well as malic acid and leucine. The harvested samples of both Phyllanthus species at ten weeks of age exhibited significant bioactivities with the highest content and number of metabolites.

Cannabinoids are terpenophenolic compounds consisting of an aromatic polyketide and derived from the geranyl diphosphate C10 terpenoid unit. They are the active constituents in Cannabis sativa and have been utilized in a number of cannabis-based medicines. Biotransformation of cannabinoids is an important field of xenobiochemistry and toxicology and the study of the metabolism of these compounds can lead to the discovery of new compounds, unknown metabolites with unique structures and new therapeutic entities. Different fungi, bacteria, plants and animal cells have been used for the regio- and stereoselective transformation of cannabinoids. All of the above mentioned organisms have distinct enzymes which catalyze the conversion of a specific cannabinoid at different positions and thus provide a variety of derivatives. All organisms are able to transform the alkyl side chain where as mammalians are unique in the formation of the carboxy derivatives. This review article assesses the current knowledge on the biotransformation of tetrahydrocannabinol and with particular focus on ∆9-THC.

Ipomoea aquatica Forsk is a green leafy vegetable that is a rich source of secondary metabolites, vitamins and amino acids with many health benefits. Proton Nuclear Magnetic Resonance (1H NMR) spectroscopy was combined with multivariate data analyses to distinguish Ipomoea aquatica samples at different developmental stages. A principal component analysis (PCA) of Ipomoea leaves provides clusters based on the different developmental stages by combining principal components PC1 and PC2 with a total variance of 65.1%. A loading plot analysis revealed that the initial stages (weeks 3 and 4) showed comparatively low contents of phenolic and organic acids, such as citric and maleic acid; the latter stages (weeks 5 and 6) exhibited higher glucose and phenolic compound contents. The sugar, phenolic compound and organic acid contents increased based on the developmental stage of the Ipomoea aquatica. The latent structures were projected using a partial least squares (PLS) model to predict the biological activity of the Ipomoea extracts based on their 1H NMR spectra. The results of this study indicate that 1H NMR-based metabolomics data could be used to discriminate the different developmental stages and to predict the biological activities of Ipomoea leaves. The present study may be applied when monitoring the quality of fruits and vegetables.

Article history: Received on: 25/12/2014 Revised on: 10/01/2015 Accepted on: 22/01/2015 Available online: 27/02/2015 The aims of this study was to isolate compounds from the leaves of methanol extract of Garcinia cowa and to evaluated their cytotoxic activity against breast (MCF-7) and lung (H-460) cell lines. The dichloromethane fraction was separated by successive silica gel column chromatography to give three compounds. Based on spectroscopic comparison with those of the literature these compounds were elucidated as methyl 2,4,6trihydroxy-3-(3-methylbut-2-enyl)benzoate (1), garcinisidone-A (2) and methyl 4,6dihydroxy-2-(4-methoxy-5(3-methylbut-2-enyl)-3,6-dioxocylohexa-1,4-dienyloxy)-3-(3-methylbut-2-enyl)benzoate (3). Compound 1, 2 and 3 had IC50 value of 21.0 ± 10.2 µM, 21.2 ±8.4 µM and 17.2 ± 6.2µM against MCF-7, while only compound (2) was found to be in active against H-460 with IC50 value of 18.1 ± 6.7 µM. Conclusion: The results indicate that G. cowa leaves could be important sources of natural cytotoxic compounds and only compound (2) had activity against H-460 cell lines.

A series of forty-five 1,5-diphenylpenta-2,4-dien-1-one analogues were synthesized and evaluated for their nitric oxide (NO) inhibition activity in IFN-γ/LPS-activated RAW 264.7 cells. Compounds 3h, 7a, 7d and 7e exhibited comparable or significantly higher activity than the standard, curcumin (IC50 = 14.69 ± 0.24 μM). Compound 7d, a 5-methylthiophenyl-bearing analogue, displayed the most promising NO-inhibitory activity with an IC50 value of 10.24 ± 0.62 μM. The 2D and 3D QSAR analyses performed revealed that a para-hydroxyl group on ring B and an α,β-unsaturated ketone moiety on the linker are crucial for a remarkable anti-inflammatory activity. Based on ADMET and TOPKAT analyses, compounds 3h, 7a and 7d are predicted to be nonmutagenic and to exhibit high blood–brain barrier (BBB) penetration, which indicates that they are potentially effective drug candidates for treating central nervous system (CNS) related disorders.

C. nutans (Burm. f.) Lindau is a shrub that is commonly found in South East Asia, particularly in Malaysia, Thailand and Indonesia. This plant has diverse and potential medicinal uses in traditional herbal medicine for treating many diseases which include skin rashes. Skin rash is one of the symptoms that primarily occur in people with allergies. Till date, there has been no scientific evidence to support this traditional use of C. nutans as well as its potential of possessing anti-allergic properties. IgE and mast cells play an important role in the pathogenesis of allergy. Upon cross-linking of allergen to IgE antibodies bound to FcεRI on mast cells, pro-inflammatory mediators will be released as a result of degranulation. The release of pro-inflammatory mediators by mast cells activated by allergen in the skin causes large, itchy, red swellings of the skin. This study aimed to identify the most active C. nutans extract in inhibiting in vitro IgE-mast cell degranulation. Rat basophilic leukaemic cells (RBL-2H3) were pre-sensitised with IgE, pre-treated with C. nutans ethanolic (100%, 70% and 50%) or aqueous extracts and challenged with dinitrophenyl-bovine serum albumin (DNP-BSA) accordingly to analyse the release of β-hexosaminidase, histamine, interleukin (IL)-4 and tumour necrosis factor (TNF)-α. It was found that C. nutans aqueous extract was the most active extract compared to the ethanolic extracts as it was able to inhibit the release of β-hexosaminidase and histamine at concentrations of 5 mg/ml and above.

Epithelial-mesenchymal transition (EMT) is currently recognized as the main cellular event that contributes to airway remodeling. Eosinophils can induce EMT in airway epithelial cells via increased transforming growth factor (TGF)-β production. We assessed the effect of synthetic 2,4,6-trihydroxy-3-geranyl acetophenone (tHGA) upon eosinophil-induced EMT in a cellular model. The human eosinophil cell line EoL-1 was used to induce EMT in BEAS-2B human bronchial epithelial cells. The induction of EMT was dose-dependently suppressed following tHGA treatment in which the epithelial morphology and E-cadherin expression were not altered. Protein and mRNA expression of vimentin, collagen I and fibronectin in eosinophil-induced epithelial cells were also significantly suppressed by tHGA treatment. Following pathway analysis, we showed that tHGA suppressed eosinophil-induced activator protein-1-mediated TGF-β production by targeting c-Jun N-terminal kinase and phosphoinositide 3-kinase signaling pathways. These findings corroborated previous findings on the ability of tHGA to inhibit experimental murine airway remodeling.

BATMAN and BAYESIL are software tools, which can provide a solution for automated metabolite quantifications based on the proton nuclear magnetic resonance (1H-NMR) spectral data of bio-fluids. However, their specific application for the quantitative 1H-NMR based metabolomics of urine has not been investigated. The aim of this study is to evaluate the performance of BATMAN and BAYESIL in the quantitative metabolite analysis of urine based on its 1H-NMR spectra. BATMAN and BAYESIL were used for automated metabolite quantification based on the 1H-NMR spectra of the urine from the lean, obese and obese-diabetic rat groups. PLS-DA model was used to discriminate the three different groups based on the results from the quantifications. BATMAN was found to be superior to BAYESIL in identifying and quantifying the metabolites in the urine samples, owing to its flexibility that allows users to define and adjust the relevant signals of the pure standard metabolites in the database in order to fit the signals in the samples, a necessary step since variations and peak shift are natural in most 1H-NMR spectra. The results of BATMAN also agreed well with that of the manual deconvolution method, which indicated the higher accuracy in metabolite quantification, despite the need of pre-processing and longer processing time than BAYESIL. However, in the case where the problems in baseline correction and peak shift of 1H-NMR spectra are absent, the use of BAYESIL is more advantageous. Application of quantitative 1H-NMR based metabolomics of the urine showed that PLS-DA model derived from BATMAN could satisfactorily discriminate the lean, obese, and obese-diabetic rat groups. Both BATMAN and BAYESIL are useful for the quantitative automation of urine metabolites based on its 1H-NMR spectra. The results from BATMAN method is superior to BAYESIL but require expertise in spectroscopy and longer computer time. Both methods help in simplifying the interpretation of metabolite status in the VIP analysis.

Phyllanthus niruri is an important medicinal plant. To standardize the extract and guarantee its maximum benefit, processing methods optimization ought to be amenable and beneficial. Herein, three dried P. niruri samples, air (AD), freeze (FD) and oven (OD), extracted with various ethanol to water ratios (0%, 50%, 70%, 80% and 100%) were evaluated for their metabolite changes using proton nuclear magnetic resonance (1H-NMR)-based metabolomics approach. The amino acids analysis showed that FD P. niruri exhibited higher content of most amino acids compared to the other dried samples. Based on principal component analysis (PCA), the FD P. niruri extracted with 80% ethanol contained higher amounts of hypophyllanthin and phenolic compounds based on the loading plot. The partial least-square (PLS) results showed that the phytochemicals, including hypophyllanthin, catechin, epicatechin, rutin, quercetin and chlorogenic, caffeic, malic and gallic acids were correlated with antioxidant and α-glucosidase inhibitory activities, which were higher in the FD material extracted with 80% ethanol. This report optimized the effect of drying and ethanol ratios and these findings demonstrate that NMR-based metabolomics was an applicable approach. The FD P. niruri extracted with 80% ethanol can be used as afunctional food ingredient for nutraceutical or in medicinal preparation.

Diabetes mellitus (DM) is a chronic disease that can affect metabolism of glucose and other metabolites. In this study, the normal- and obese-diabetic rats were compared to understand the diabetes disorders of type 1 and 2 diabetes mellitus. This was done by evaluating their urine metabolites using proton nuclear magnetic resonance (1H NMR)-based metabolomics and comparing with controls at different time points, considering the induction periods of obesity and diabetes. The biochemical parameters of the serum were also investigated. The obese-diabetic model was developed by feeding the rats a high-fat diet and inducing diabetic conditions with a low dose of streptozotocin (STZ) (25 mg/kg bw). However, the normal rats were induced by a high dose of STZ (55 mg/kg bw). A partial least squares discriminant analysis (PLS-DA) model showed the biomarkers of both DM types compared to control. The synthesis and degradation of ketone bodies, tricarboxylic (TCA) cycles, and amino acid pathways were the ones most involved in the variation with the highest impact. The diabetic groups also exhibited a noticeable increase in the plasma glucose level and lipid profile disorders compared to the control. There was also an increase in the plasma cholesterol and low-density lipoprotein (LDL) levels and a decline in the high-density lipoprotein (HDL) of diabetic rats. The normal-diabetic rats exhibited the highest effect of all parameters compared to the obese-diabetic rats in the advancement of the DM period. This finding can build a platform to understand the metabolic and biochemical complications of both types of DM and can generate ideas for finding targeted drugs.

The antiphoto aging property of Zanthoxylum rhetsa obtained from Pangkor Island, Malaysia, was evaluated. Solvent fractions of different polarity obtained from the methanolic extract of the bark material were initially tested for anticollagenase and antielastase activities. The ethyl acetate fraction showed bioactivity against the protease enzymes. Hence, it was subjected to further purification via column chromatography, to yield a major constituent, hesperidin. Subsequently, the ethyl acetate fraction and hesperidin were tested for their effects against UVB-induced cytotoxicity and expressions of inflammatory cytokines (IL-6, IL-1β, and TNF-α), NF-κB, and MMPs (MMP1, 3, and 9) in human dermal fibroblasts (HDF). Both fraction and pure compound prevented UVB-induced cytotoxicity in HDF cells, in a dose dependent manner. Moreover, the ethyl acetate fraction inhibited the increase of pro-inflammatory cytokines induced by UVB to a level similar to the control (without UV treatment). Additionally, the fraction significantly inhibited the expressions of NF-κB, MMP 1, MMP 3, and MMP 9 in HDF cells treated with UVB. Similar effects were observed with hesperidin. The results obtained suggested that the ethyl acetate fraction of Z. rhetsa and its bioactive constituent, hesperidin, have the potential to be used as active ingredients in sunscreen and antiphoto aging formulations.

VIP score and variable regions of the assigned metabolites. Figure S2. Body weight of rats in day 0, day 7 and day 14 of treatment. Data are expressed as mean ± S.D.; n = 5 rats per group. N = normal rat, D = diabetic rat, DG = diabetic rat treated with glibenclamide, D-E = diabetic rat treated with OS ethanol extract, D-A = diabetic rat treated with OS water extract. *p ≤ 0.05 versus normal control in same day, **p ≤ 0.05 versus day 0 and other day in same group. (PDF 243 kb)