ß-sitosterol isolated from the leaves of Trema orientalis (Cannabaceae) promotes viability and proliferation of BF-2 cells.

Trema orientalis is a pioneer species in the cannabis family (Cannabaceae) that is widely distributed in Thai community forests and forest edges. The mature leaves are predominantly used as an anti-parasite treatment and feed for local freshwater fish, inspiring investigation of their phytochemical composition and bioactivity. The purpose of this work was to investigate the bioactive compounds in T. orientalis leaf extract and their cytotoxicity in the BF-2 fish cell line (ATCC CCL-91). Flash column chromatography was used to produce 25 mL fractions with a mixture solvent system comprised of hexane, diethyl ether, methanol, and acetone. All fractions were profiled with HPLC-DAD (mobile phase methanol:aqueous buffer, 60:40 v/v) and UV detection (wavelengths 256 and 365 nm). After drying, a yellowish powder was isolated from lipophilic leaf extract with a yield of 280 µg/g dry weight. Structure elucidation by nuclear magnetic resonance (NMR) indicated it to consist of pure ß-sitosterol. The lipophilic extract and pure compound were evaluated for cytotoxicity using BF-2 cells. MTT assays showed both leaf extract and pure compound at 1 µg/mL to increase cell viability after 24 h treatment. The respective half maximal inhibitory concentration (IC50) values of leaf extract and ß-sitosterol were 7,027.13 and 86.42 µg/ml, indicating a lack of toxicity in the BF-2 cell line. Hence, T. orientalis can serve as a source of non-toxic natural lipophilic compounds that can be useful as bioactive ingredients in supplement feed development.

Profiling of historical rag papers by their non-cellulosic polysaccharide composition.

Hemicellulose and pectin are noteworthy components of historical European rag papers, and have not been studied in detail so far. Rag papers were made from used textiles, and fiber-based utilities, such as ropes and bags. These had been prepared until the mid-19th century from plant-based fibers. Their polysaccharide composition could relate to their condition and history. This information can be expected to hold importance for the preservation and conservation of historical objects. We investigated a collection of rag papers of different age for their composition of non-cellulosic polysaccharides, and compared the findings with modern rag papers and wood pulps. Furthermore, a non-destructive determination of the hemicellulose and pectin content by near-infrared spectroscopy was developed. Historical rag papers had a lower hemicellulose/pectin content than pulps; the fractions of rhamnose, galactose, and arabinose were higher, while xylose was lower. In modern rag papers, xylose tended to be at the higher end of the range, which suggests a degradation of hemicelluloses/pectin over time or a change in raw materials and manufacturing. Rag papers also showed higher crystallinity than wood pulp papers. These findings provide insights into rag paper characteristics and offer potential classification methods.

Antimicrobial, antioxidant, and sun protection potential of the isolated compounds from Spermacoce princeae (K. Schum).

BACKGROUND: Spermacoce princeae (K. Schum) has been used in the treatment of bacterial skin infections in Uganda. Pharmacological studies revealed that extracts of S. princeae exhibited antibacterial, antioxidant, and sun protection potential. This study aimed at isolating and identifying pure compounds from the extracts based on comprehensive analytical characterization by multiple analytical techniques. METHODS: The plant samples were extracted by sequential maceration using n-hexane, ethyl acetate, methanol, and distilled water. The compounds were isolated using a combination of chromatographic techniques and their structures were elucidated by multiple spectroscopic techniques. The antibacterial and antifungal activity determination of the isolated compounds was carried out using an agar well diffusion and potato dextrose assay against Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Candida albicans, and Aspergillus flavus while the antioxidant activity was screened with the 2,2-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging assay. The sun protection factor was determined using a Shimadzu Ultra Violet-visible (UV-VIS) double beam spectrophotometer between 290 to 320 nm. RESULTS: Eleven compounds; quercetin (1), kaempferol-3-O-rutinoside (2), rutin (3, 12), myo-inositol (4), asperulosidic acid (5), hexadecanoic acid (6), ß-sitosterol (7), stigmasterol (8), campesterol (9), ursolic acid (10), and ß-sitosterol glucoside (11) were identified in the S. princeae extracts. Compound 2 had good antifungal activity against C. albicans (zone of inhibition, 23.0 ± 0.1 mm). Compound 10 showed antibacterial and antifungal activity against S. aureus, P. aeruginosa, C. albicans, and A. flavus. Compound 2 had a good percentage radical scavenging effect (IC50 = 64.81 µg/ml) and a good sun protection factor (SPF = 26.83). CONCLUSION: This study reports the first-time isolation and identification of compounds 1 to 11 from S. princeae, which contribute to its antimicrobial, antioxidant, and sun protection potential.

Silviridoside: A New Triterpene Glycoside from Silene viridiflora with Promising Antioxidant and Enzyme Inhibitory Potential.

A new triterpene glycoside, silviridoside, was isolated from the aerial parts of Silene viridiflora (Caryophyllaceae) using different chromatographic techniques. The structure of silviridoside was comprehensively elucidated as 3-O-ß-D-galacturonopyranosyl-quillaic acid 28-O-ß-D-glucopyranosyl-(1→2)-[α-L-rhamnopyranosyl-(1→3)]-ß-D-fucopyranosyl ester by one- and two-dimensional nuclear magnetic resonance (NMR) spectroscopy and high-resolution mass spectrometry (HR-MS). Silviridoside showed promising antioxidant activity in different antioxidant assays such as 2,2-diphenyl-1-picrylhydrazyl (DPPH) (2.32 mg TE/g), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) (1.24 mg TE/g), cupric-reducing antioxidant capacity (CUPRAC) (9.59 mg TE/g), ferric-reducing antioxidant power (FRAP) (5.13 mg TE/g), phosphomolybdenum (PHD) (0.28 mmol TE/g), and metal-chelating (MCA) (6.62 mg EDTA/g) assays. It exhibited a good inhibitory potential on acetylcholinesterase (AChE) (2.52 mg GALAE/g), butyrylcholinesterase (BChE) (7.16 mg GALAE/g), α-amylase (0.19 mmol ACAE/g), α-glucosidase (1.21 mmol ACAE/g), and tyrosinase (38.83 mg KAE/g). An in silico evaluation of the pharmacodynamic, pharmacokinetic, and toxicity properties of silviridoside showed that the new compound exhibited reasonable pharmacodynamic and pharmacokinetic properties without any mutagenic effect, but slight toxicity. Thus, it could be concluded that silviridoside could act as a promising lead drug for pharmaceutical and nutraceutical developments to combat oxidative stress and various disorders, but a future optimization is necessary.

Cannabinoids from inflorescences fractions of Trema orientalis (L.) Blume (Cannabaceae) against human pathogenic bacteria.

BACKGROUND: Cannabinoids; tetrahydrocannabinol (THC), cannabidiol (CBD) and cannabinol (CBN), might show antibacterial activity. Trema orientalis is a species in the Cannabaceae that is closely related to Cannabis through plastome phylogenetic evidence. This species is widely distributed throughout tropical Asia and is used as traditional medicine, particularly for the treatment of infectious diseases. However, no studies on the antibacterial activity of cannabinoid-containing inflorescences extracts are available. Thus, the aim of this study was to determine cannabinoid content and antibacterial activity of inflorescences fractions from T. orientalis native to Thailand. METHODS: We hypothesized that inflorescences from T. orientalis might display cannabinoids similar to Cannabis because of their close taxonomic relationship. We extracted the mature inflorescences and infructescence of T. orientalis in three disparate populations from different Thailand floristic regions. Extractions were subsequently partitioned into hydrophilic and lipophilic fractions using distilled water and chloroform. The lipophilic extracts were further fractionated by the column chromatography with gradient elution and analyzed by gas chromatography-mass spectrometry (GC-MS). Characterized cannabinoids were used in bioassays with multidrug-resistance bacteria. RESULTS: Lipophilic extracts and fractions of inflorescences from all Thailand floristic regions consistently displayed cannabinoids (THC, CBD and CBN) in various quantities. These extracts exhibited inhibitory activity for Staphylococcus aureus, Pseudomonas aeruginosa, and Acinetobacter baumannii strains with minimum inhibitory concentration values varying from 31.25 to 125 µg/mL. CONCLUSION: Our study is the first to report cannabinoid detection in extracts from inflorescences of T. orientalis, a species in the Cannabaceae. These extracts and their fractions containing cannabinoids showed pronounced antibacterial activity. The use of analytic methods also demonstrated reproducible cannabinoid extraction.

Yanangdaengin, a dihydrochalcone glucoside galloyl ester as active antioxidative agent from leaves of Lysiphyllum strychnifolium (syn. Bauhinia strychnifolia).

Objective: To isolate and identify the major bioactive components from the leaves of Lysiphyllum strychnifolium, an indigenous herb used in traditional Thai medicine for detoxification, longevity, and some other health related issues. Methods: Comparative HPLC analyses of the crude extracts from three provenances were carried out for an overview of characteristic compound profiles. Isolation of the major compounds was undertaken with chromatographic methods. Chemical structures were elucidated by NMR spectroscopic techniques and mass spectrometry. DPPH scavenging assay was carried out to determine the free radical scavenging activity of isolated compounds. Results: Yanangdaengin (3), a dihydrochalcone glucoside galloyl ester, has been isolated together with its corresponding dihydrochalcone glucoside trilobatin (2) as major compounds from the leaves of L. strychnifolium. Additionally, gallic acid (1) was co-chromatographically identified. Free radical scavenging activity of isolated compounds were determined. Compound 3 exhibited higher free radical scavenging activities in comparison to Trolox and quercetin. Conclusion: The isolated compounds could be used as chemical markers for quality assessment. The present work could promote the quality control and herbal medicinal product development of this plant.

Chemovariation and antibacterial activity of extracts and isolated compounds from species of Ixora and Greenea (Ixoroideae, Rubiaceae).

BACKGROUND: A large number of secondary metabolites can be obtained from plants used for traditional medicine in two related genera (Ixora and Greenea) in the subfamily Ixoroideae (Rubiaceae), but there are only a few detailed studies on their bioactivities. Therefore, the main goals of this study were to determine the antibacterial activities of lipophilic extracts from plants of some Ixora and Greenea species native to Thailand, and to isolate some pure compounds from those extracts. Moreover, we compared the occurrence of compounds in different plant parts of samples from different habitats to better understand their variation. METHODS: A total of 56 lipophilic extracts were obtained from the leaves, stem bark, and root bark of eight Ixora and two Greenea species collected at various locations in Thailand. Isolated compounds were identified using nuclear magnetic resonance. Antimicrobial activities were evaluated against four Gram-positive and nine Gram-negative human pathogenic bacterial strains. RESULTS: Extracts from I. javanica, I. nigricans, I. brunonis, and G. montana, along with isolated scopoletin, exhibited antibacterial activities against Gram-positive methicillin-resistant Staphylococcus aureus ATCC 43300, with minimum inhibitory concentration values ranging from 64 to 256 µg/mL. The occurrence of scopoletin, isofraxidin, and geniposidic acid in lipophilic extracts showed some variation among different plant parts and species. CONCLUSIONS: Lipophilic extracts of Ixora and Greenea species have the potential to be developed as anti-Gram-positive agents, in particular to counter infections of methicillin-resistant S. aureus strains. The chemical profiles showed differences between floristic regions but similarity within the same plant parts.

Flavone glucosides from Artemisia juncea.

A new flavone glucoside, 4',5-dihydroxy-3',5',6-trimethoxyflavone-7-O-ß-D-glucoside was obtained from aerial parts of Artemisia juncea, together with the known flavone eupatilin (5,7-dihydroxy-3',4',6-trimethoxyflavone). The compounds were comprehensively analytically characterized by IR, UV, NMR and HR-MS, and their chemical structures ascertained. The EtOAc fraction of A. juncea showed the strongest DPPH radical scavenging ability as well as reducing power (in CUPRAC and FRAP assays) and phosphomolybdenum activity. This fraction also exhibited the strongest inhibitory effects on tyrosinase. Additionally, the best antidiabetic effects were observed for eupatilin and the CHCl3 fraction.

Interaction of ascaridole, carvacrol, and caryophyllene oxide from essential oil of Chenopodium ambrosioides L. with mitochondria in Leishmania and other eukaryotes.

The antileishmanial activity of the essential oil (EO) from Chenopodium ambrosioides L. has been demonstrated in vitro and in animal models, attributed to the major components of the EO. This study focused on the effects of the three major EO compounds carvacrol, caryophyllene oxide (Caryo), and the antileishmanial endoperoxide ascaridole (Asc) on mitochondrial functions in Leishmania tarentolae promastigotes (LtP). EO and Caryo were able to partially inhibit the leishmanial electron transport chain, whereas other components failed to demonstrate a direct immediate effect. Caryo demonstrated inhibition of complex III activity in LtP and in isolated complex III from other species. The formation of superoxide radicals was studied in Leishmania by electron spin resonance spectroscopy in the presence of iron chelators wherein selected compounds failed to trigger a significant immediate additional superoxide production in LtP. However, upon prolonged incubation of Leishmania with Asc and especially in the absence of iron chelators (allowing the activation of Asc), an increased superoxide radical production and significant impairment of mitochondrial coupling in Leishmania was observed. Prolonged incubation with all EO components resulted in thiol depletion. Taken together, the major components of EO mediate their leishmanicidal activity via different mitochondrial targets and time profiles. Further studies are required to elucidate possible synergistic effects of carvacrol and Asc and the influence of minor compounds.

Quality assessment and antiplasmodial activity of West African Cochlospermum species.

The present study focuses on development of phytochemical methods for quality assessment of two West-African Cochlospermum species (Cochlospermum planchonii and Cochlospermum tinctorium) traditionally used for malaria treatment in Burkina Faso. Antimalarial activity of preparations from dried rhizomes (decoction) was tested against the chloroquine-sensitive Plasmodium strain 3D7 using the histidine-rich protein II (HRP2) drug susceptibility assay and compared with extract preparations using organic solvents of different polarity. Two main apocarotenoids were isolated from rhizomes of C. planchonii and unambiguously identified as dihydrocochloxanthine and cochloxanthine by spectroscopic methods. Comparative HPLC analyses of thirty-nine (39) samples from markets and from collections in natural habitats of both species showed a high variability in the accumulation of cochloxanthines and related carotenoids which were proven to be characteristic for rhizomes of both species and generally absent in leaves. Furthermore, content of total phenolics and antioxidant activities (DPPH and FRAP) as well as haemolytic activity of various extracts was tested. The HPLC method presented here was validated and provides a good separation of both compounds including 10 minor carotenoids. Extracts from both species and pure cochloxanthine offered pronounced antioxidant activities and weak haemolytic activity while, in contrast, dihydrocochloxanthine had a strong haemolytic effect at the highest concentration analysed. However, cochloxanthine as well as dihydrocochloxanthine showed erythroprotective effects against the haemolytic activity of the reference saponin. Moderate antiplasmodial activity between 16 and 63 µg/ml were observed with all tested extracts, and lower IC50 values were obtained with pure dihydrocochloxanthine (IC50=6.9 µg/ml), cochloxanthine (IC50=6.8 µg/ml), the DCM fraction (IC50=2.4 µg/ml) and the ethyl acetate fraction (IC50=11.5µg/ml) derived from a methanolic extract of C. planchonii. This study shows a major variability of carotenoid content and antiplasmodial activity of both C. planchonii and C. tinctorium. The high haemolytic activity of dihydrocochloxanthine (at 100 µg/ml) should be considered as a selection criterion for choosing species phenotypes for treatment.