Identification Of Secondary Metabolites Research Articles

Identification of secondary metabolites containing a diketopiperazine core in extracts from myxobacterial strains with growth inhibition activity against a range of prey species.

Myxobacteria produce a variety of bioactive secondary metabolites, and with a wealth of under-researched species they hold vast potential for undiscovered compounds. With the ever-increasing need for new antibiotics, the development of novel therapeutics is vitally important. Therefore, this study aimed to extract and elucidate antimicrobial metabolites from the following myxobacteria: Myxococcus xanthus CA010 and AB022; Corallococcus exiguus DSM 14696T; Myxococcus stipitatus DSM 14675T; and Corallococcus aberystwythensis AB050AT. Metabolite mixtures were extracted in acetone from XAD-16 resin incubated in liquid cultures and analysed using GC-MS. Bioactivity was identified using a growth inhibition assay against a panel of clinically relevant prey species including Gram-positive and Gram-negative bacteria and a fungus. Growth of Klebsiella pneumoniae and Enterococcus faecalis was most affected by the metabolite mixtures and the mixtures from AB022 and AB050AT were effective against the most prey. GC-MS analysis revealed metabolites with roles in the synthesis and degradation of amino acids and fatty acids, but also identified compounds A and B with a diketopiperazine (DKP) core. With previously confirmed bioactivity of compound A, it is suggested that these DKP compounds are contributing to the antimicrobial activity observed. Furthermore, many compounds could not be identified and so these unknowns present further potential for novel bioactive compounds.

Antioxidant Activity, Cytotoxicity, and Identification of Secondary Metabolites of Kigelia africana from Waterpark Platinum Riau

Antioxidants can prevent reactive oxygen-associated diseases, which trigger carcinogenesis, cardiovascular disease, and premature aging. India and Africa have mostly practiced sausage trees (Kigelia africana) as traditional medicine, whereas Indonesia is still limited. The research aims to determine the phytochemistry and bioactivity of the n-hexane fractions from crude methanol extract of the leaves and bark of the sausage tree as antioxidants and their toxicity. The sausage tree is derived from Waterpark Platinum Riau. The sausage trees were macerated with methanol and fractionated by n-hexane and ethyl acetate. All samples were tested for their antioxidant to 1,1-diphenyl-2-picrylhydrazyl (DPPH) and cytotoxicity to Artemia salina Leach larvae. Phytochemical results of methanol crude extract, n-hexane, and ethyl acetate fractions of leaves and sausage bark showed the presence of all secondary metabolites except alkaloids in the n-hexane fraction. Antioxidant and cytotoxic activity of stem and leaf bark ethyl acetate fraction was stronger than crude methanol extract and n-hexane fraction. Based on LC-MS/MS data, the secondary metabolite components that have contributed strongly antioxidant activity of this study are flavonoid compounds such as kaempferol and the derivatives, lignans (cubebin), and steroids (pregnant).

Bioprospecting of Endophytic Fungi from Phyllanthus acidus Linn. Leaf, Identification of Secondary Metabolite: Quercetin

Bioprospecting of Endophytic Fungi from Phyllanthus acidus Linn. Leaf, Identification of Secondary Metabolite: Quercetin

Exploring eco-sensitive strategies for effective powdery mildew management in grapevines

Grapevine powdery mildew is one of the most important plant diseases widely affecting crops in many countries. The main aim of the present study was to use Ampelomyces quisqualis isolates to suppress the powdery mildew of grapes under field conditions. The findings revealed a strong correlation between the mycoparasite, Ampelomyces quisqualis and the grape powdery mildew pathogens, suggesting its potential for effective control. The results revealed that a single spray of A. quisqualis @ 2.0 per cent (MDU1) succeeded by two sprays of Trichoderma asperellum @ 2.0 per cent (Tasp,7) were found to be effective against Erysiphe necator. In addition, liquid formulation of A. quisqualis isolate MDU1 (2% w/v) as a foliar spray at 2% containing 2 × 106 spores ml-1 the during initial occurrence of the disease up to three sprayings at 15 days interval was found to be best. Thus, a liquid formulation of A. quisqualis isolate MDU1 proved effective in managing grapes powdery mildew disease under field conditions. The rDNA ITS region of A. quisqualis isolates was analyzed at the molecular level, and the resulting sequences were subjected to GC-MS analysis. The secondary metabolite identification using GC-MS revealed the presence of antimicrobial compounds, including squalene with the highest peak of 4.643 percent, octadecanoic acid with 3.862 percent, tetradecanoic acid with 3.600 percent, and 9,12-octadecadienoic acid (Z,Z) with 1.451 percent. These bioactive compounds revealed by GC-MS analysis in crude extracts of A. quisqualis had a stronger antifungal activity against E. necator.

Identification of Secondary Metabolites from the Mangrove-Endophyte Lasiodiplodia iranensis F0619 by UPLC-ESI-MS/MS.

Lasiodiplodia is a widely distributed fungal genus, frequently found in tropical and subtropical regions where it can cause disease in important crops. It represents a promising source of active secondary metabolites with uses in chemical, pharmaceutical, and agrochemical processes. In this study, the strain Lasiodiplodia iranensis F0619 was isolated from the mangrove Avicennia ger-minans, collected from Sarigua National Park in the Republic of Panama. Fractions of crude extract were analyzed by UPLC-ESI-MS/MS, and five compounds, previously reported from Lasiodiplodia genus were identified, including 11,12-didehydro-7-iso-jasmonic acid (1), 4,5-didehydro-7-iso-jasmonic acid (2), cyclo-(L-Leu-L-Pro) (3), jasmonate-threonine (4), and abscisic acid (5). We describe and analyze their MS/MS fragmentation patterns to confirm the compounds 'chemical structures.

High-speed countercurrent chromatography with offline detection by electrospray mass spectrometry and nuclear magnetic resonance detection as a tool to resolve complex mixtures: A practical approach using Coffea arabica leaf extract.

Many secondary metabolites isolated from plants have been described in the literature owing to their important biological properties and possible pharmacological applications. However, the identification of compounds present in complex plant extracts has remained a great scientific challenge, is often laborious, and requires a long research time with high financial cost. The aim of this study was to develop a method that allows the identification of secondary metabolites in plant extracts with a high degree of confidence in a short period of time. In this study, an ethanolic extract of Coffea arabica leaves was used to validate the proposed method. Countercurrent chromatography was chosen as the initial step for extraction fractionation using gradient elution. Resulting fractions presented a variation of compounds concentrations, allowing for statistical total correlation spectroscopy (STOCSY) calculations between liquid chromatography coupled with high-resolution tandem mass spectrometry (LC-HRMS/MS) and NMR across fractions. The proposed method allowed the identification of 57 compounds. Of the annotated compounds, 20 were previously described in the literature for the species and 37 were reported for the first time. Among the inedited compounds, we identified flavonoids, alkaloids, phenolic acids, coumarins, and terpenes. The proposed method presents itself as a valid alternative for the study of complex extracts in an effective, fast, and reliable way that can be reproduced in the study of other extracts.

Primary and secondary metabolites identification in Hamelia patens leaves

The aim of the present study was to investigate the presence of phytochemicals in leaves extracts (aqueous, methanol, ethanol) of Hamelia patens, the selected medicinal plant. Ethanolic and methanolic (organic) extract showed good number of compounds compared to aqueous extracts. Phyto-chemical analysis of organic extracts of Hamelia patens has shown the biological compounds like protein and amino acids, carbohydrates, cardiac glycosides, tannin, phenol, alkaloids, steroids, terpenoids, flavonoides and saponin. Quinones, phlobatannins, coumarins, emodins and fatty acids were absent in both the aqueous and organic extracts.

Phytochemical analysis, antioxidant activity and identification of bioactive constituents from three wild medicinally important underutilized plants of Ladakh, India using GCMS and FTIR based metabolomics approach

Underutilized wild medicinal plants have been used in various traditional system of medicine including Chinese, Turkish, African and Brazilian in the treatment of pain, purgative, constipation, tonic, hemostatic, headaches, arthritis, backache, rheumatism, and eczema. The aim of the present study is to quantify and identify the bioactive constituents of three ethnobotanically important wild plants of cold desert Ladakh including Rheum spiciforme, Rumex patientia, and Sonchus oleraceus. Using spectrophotometry, total phenol, flavonoid, phytosterol and antioxidant activity was analyzed. Rheum spiciforme has highest phenol content (77.2 mg/g), Rumex patientia was highest in phytosterol (133.3 mg/g) and flavonoids (88.7 mg/g). Sonchus oleraceus showed highest antioxidant activity by DPPH (IC50 = 39.51 μg/mL) and FRAP assay (98.93 mg/g). GCMS and FTIR metabolomics approach is used for the identification of secondary metabolites for further use in pharmaceutical preparations. FTIR analysis of the plant extract shows the presence of functional groups such as carboxylic acids, aromatics, alkanes, alcohols, phenols, aliphatic amines, alkenes and amine groups. GCMS analysis of methanolic extract revealed 29 different bioactive compounds with high content of 1-Naphthalenol, methylcarbamate (13.22%), Cyclopentadecane (12.43%) in Rheum spiciforme, 22 peak with high content of Benzene, 1,1′-sulfonylbis[4-chloro- in Rumex patientia (31.18%) and 13 compounds identified with the highest % area of compound 2,2-Dimethoxybutane in Sonchus oleraceus (19.33%). The functional groups and identified compounds have various health benefits including antioxidant, anti-inflammatory, immunomodulatory and antifungal activity which could be used for production of food supplements and novel drugs.

Antioxidant Activity of n-hexane and Etil Acetate Fractions of Bangkal (Nauclea subdita (Korth.) Steud.) Leaves

Bangkal (Nauclea subdita (Korth.) Steud.) is a tropical plant belonging to the Rubiaceae family, commonly found in South Kalimantan. This plant is one of the plants that has efficacy as a medicinal plant. This study aimed to quantitatively identify secondary metabolites and antioxidant activity in the n-hexane and ethyl acetate fractions of N. subdita leaves. The method of identification of secondary metabolites using the test tube. Antioxidant activity using the DPPH method based on IC50 value. The results of identifying secondary metabolites in the n-hexane fraction of N. subdita leaves contain alkaloids, flavonoids, steroids, and phenolic compounds, while the ethyl acetate fraction of N. subdita leaves contain alkaloids, flavonoids, steroids, tannins, saponins, and phenolics. The results of the antioxidant activity test of the n-hexane fraction and the ethyl acetate fraction of the leaves of N. subdita showed IC50 values ​​of 229.61178±3.65919 and 54.54296±0.02236 ppm, respectively. Based on the IC50 value, the n-hexane fraction of N. subdita leaves had weak antioxidant activity, and the ethyl acetate fraction of N. subdita leaves had strong antioxidant activity.

Diversity of Fungal Communities on Diseased and Healthy Cinnamomum burmannii Fruits and Antibacterial Activity of Secondary Metabolites.

The composition and structure of fungal communities on healthy and diseased fruits of Cinnamomum burmannii (Nees and Nees) Blume were characterized, with evaluation of the antibacterial activity of secondary metabolites from culturable fungi following the first identification of secondary metabolites in the fungus Medicopsis romeroi (Esf-14; GenBank accession number OK242756). These results are significant for understanding the functional variation in bioactivity in fungal communities and developing a broader range of bioactive resources. High-throughput sequencing results indicated that the fungal community in diseased fruit differed from that in healthy fruit at the phylum, class, order, or genus level, with significant differences in the species and relative abundance of the dominant flora. A total of 49 (healthy fruit) and 122 (diseased fruit) artificially cultivable endophytic fungi were isolated, and 41 different strains (11 from healthy fruit and 30 from diseased fruit) were successfully identified by morphological and molecular biological analyses, which were classified into 8 groups and 23 genera by phylogenetic tree analysis, with Pleosporales, Glomerellales, and Hypocreales being the dominant groups at the order level and Colletotrichum being the dominant group at the genus level. The results of the antibacterial assay demonstrated that the secondary metabolites of all strains had different degrees of antibacterial activity, while the secondary metabolites of endophytic fungi from diseased fruit were generally stronger than those of fungi from healthy fruit, with the active secondary metabolites dominated by small and moderately polar compounds. Combined analysis of fungal communities, phylogenetic tree analysis, and bioactivity analysis of culturable strains revealed strong antibacterial activity of both upregulated and downregulated flora in diseased fruit. Five compounds, including two new (5,6-dimethoxy-[1',1:4,1″-terphenyl]-2-ol [compound 1] and 5-(methoxycarbonyl)-2-methylbenzo[d][1,3]dioxole-2-carboxylic acid [compound 2]) and three known compounds (3,7-dihydroxy-1,9-dimethyldibenzofuran [compound 3], methyl 3-hydroxybenzoate [compound 4], and uracil [compound 5]), were isolated and identified for the first time from the endophytic fungus Medicopsis romeroi. In general, the diversity of fungal communities on diseased fruit was lower than that on healthy fruits, while the antibacterial activity of artificially cultured endophytic fungi on diseased fruits was generally stronger than that on healthy fruits, suggesting excellent promise for the development of secondary metabolites from active strains on diseased fruit as antibacterial agents. IMPORTANCE Powdery fruit disease is a notorious disease of Cinnamomum burmannii that causes severe loss in fruit production. Studies on the function of endophytic fungal communities in healthy plant tissues are not new, while little is known about the functional changes of fungal communities in disease-causing plant tissues. Our results demonstrate that fungal communities in diseased fruits differ from those in healthy fruits at the level of phylum, class, order, or genus, with significant differences in the species and relative abundance of dominant groups. Endophytic fungi in diseased fruits appeared to produce secondary metabolites with stronger antibacterial properties, although the community diversity was not as varied as that in healthy fruits. In addition, secondary metabolites of the Medicopsis romeroi strain from diseased fruits were identified for the first time. These results have important implications for understanding the functional variation of bioactivity in fungal communities and for developing a broader resource of bioactivity.

Chemical Identification of Secondary Metabolites from Rhizospheric Actinomycetes Using LC-MS Analysis: In Silico Antifungal Evaluation and Growth-Promoting Effects.

The rhizosphere is a rich source of actinomycetes which can produce several potential biologically active secondary metabolites. The principal goal for this research is to extract, purify, and characterize the bioactive secondary metabolites produced by three different strains of actinomycetes isolated from the rhizosphere of rosemary, black locust, and olive. The plant growth-promoting effect (PGPE) of the studied strains of actinomycetes on Ocimum basilicum L. (basil) and the disease-control effect on necrotic stem lesions of "black leg" caused by Fusarium tabacinum on basil were evaluated in silico. The cell-free culture filtrates from the studied actinomycetes isolates were evaluated in vitro for their antimicrobial activity against some common phytopathogens. The secondary metabolites obtained from the cell-free culture filtrates have been chemically characterized using high-resolution electrospray ionization of liquid-chromatography/mass-spectrometric detection (ESI-(HR)Orbitrap-MS). Results of the in silico trial showed that all studied isolates demonstrated PGPE on basil seedlings, improved some eco-physiological characteristics, and reduced the disease incidence of F. tabacinum. The extracted metabolites from the studied actinomycetes demonstrated antimicrobial activity in a Petri-plates assay. The chemical analysis revealed the presence of 20 different components. This research emphasizes how valuable the examined isolates are for producing bioactive compounds, indicating their putative antimicrobial activity and their potential employment as fungal biocontrol agents. In particular, the obtained results revealed the possibility of green synthesis of some important secondary metabolites, such as N-Acetyl-l-histidinol, Rhizocticin A, and Eponemycin, from actinomycetes. The bioactive metabolites may be successively used to develop novel bio-formulations for both crop protection and/or PGPE.

Antibacterial Activity, TLC-Bioautography Analysis, and Determination of Bioactive Components in Ethyl Acetate Extract of Robusta Coffee Leaf (Coffea canephora L.) From Aceh, Indonesia

Infectious diseases affect many people in the world, including Indonesia. One of the causes of infection is bacteria, including scherichia coli and Staphylococcus aureus. Robusta coffee (Coffea canephora L.) is one of the plants that is widely encountered on Indonesian plantations. The leaves are usually discarded after being trimmed to maximize the growth of coffee beans. The results of several studies show that robusta coffee leaves demonstrate antibacterial and antioxidant activity due to the presence of flavonoids, alkaloids, tannins, saponins, polyphenols, and chlorogenic acids. The aim of this research to investigate antibacterial activities, total flavonoid content and inspect the phytochemicals of the bioactive compounds as antibacterial with TLC Bioautography in the ethyl acetate extract of robusta coffee leaves. The identification of secondary metabolites was conducted by using the TLC-Bioautography method against Escherichia coli and Staphylococcus aureus. The determination of total flavonoid levels was carried out by using Spectrophotometry UV-Vis. Phytochemical testing showed that the extract contained alkaloids, steroids, phenolics, tannins, and flavonoids. The ethyl acetate extract inhibits the growth of Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 25922 (P=0.00) with 100% extract as the most potent. The results of TLC-Bioautography showed the presence of polyphenolic compounds (Rf 0.36) which was confirmed by a clear zone in the disc diffusion test. The total flavonoid content in robusta coffee leaves was 57.255±0.0348 mg QE/g extract. Ethyl acetate extract from coffee leaves (Coffea canephora L). inhibits the growth of S. aureus and E. coli due to the presence of polyphenolic compounds

Karakterisasi Fraksi Aktif Antioksidan dari Biji Gandaria (Bouea macrophylla)

Gandaria (Bouea macrophylla) plants contain antioxidant compounds that can reduce free radicals. Free radicals are one of the causes of various degenerative diseases. The purpose of this study was to determine the fraction that had the highest antioxidant activity, and to determine the secondary metabolites contained in the most active fraction of B. macrophylla seeds based on LCMS/MS analysis. Maceration of B. macrophylla seeds with ethanol solvent. Ethanol maserate was concentrated and fractionated by liquid-liquid extraction method using n-hexane, ethyl acetate and ethanol as solvents. Antioxidant activity was measured by the free radical scavenger method with 1,1-diphenyl-2-picrylhidrazil as a source of radicals and characterization of the secondary metabolites contained in the most active fraction using a Liquid Chromatography-Mass Spectrometer. B. macrophylla seed ethanol extract has a % yield of 31.75%. The ethyl acetate fraction of B. macrophylla seeds has the highest mass compared to other fractions and has strong antioxidant activity with an IC50 value of 2.95 ppm. The ethyl acetate fraction of B. macrophylla seeds contains tannins, phenolic, flavonoid dan alkaloid compounds. Based on the results of the identification of secondary metabolites by LC-MS/MS, the ethyl acetate fractions of B. macrophylla seeds contained phenylalalnine compounds.

Metabolomics Analysis Reveals the Effect of Fermentation to Secondary Metabolites of Chenopodium album L. Based on UHPLC-QQQ-MS

Chenopodium album L. (CAL) is an excellent vegetable crop that is rich in nutrients and possesses potential pharmaceutical value. However, the research on the secondary metabolites and the processing utilization of CAL has been rarely reported. In this study, the polyphenol content, microstructure and secondary metabolite composition of aerial parts of CAL (AC), including stems, leaves, inflorescence and grain, before and after fermentation were investigated. The results showed that the polyphenol content of fermented AC (FAC) was significantly higher than that of AC (increased by 38.62%). The AC had a compact surface, while FAC had a loose and cracked surface with large holes. A total of 545 secondary metabolites, including 89 alkaloids, 179 flavonoids, 25 lignans and coumarins, 163 phenolic acids, 35 terpenoids, 9 quinones, 6 tannins and 39 others, were identified in the AC and FAC by UHPLC-QQQ-MS metabolomics. Differential metabolites analysis reviewed 285 differential metabolites (117 upregulated and 168 downregulated) between AC and FAC. The decrease in parts of toxic alkaloids accompanied with the increase in some biologically active substances with small molecules, such as quercetin, kaempferol, p-coumaric acid and protocatechuic acid, indicated that fermentation is beneficial to enhance the bioavailability of AC. This study provides a reference value for the identification of secondary metabolites from AC and the application of fermentation in the deepness development of AC.

Metabolite Profiling and Bioactivities of Leaves, Stems, and Flowers of Rumex usambarensis (Dammer) Dammer, a Traditional African Medicinal Plant.

The comprehensive identification of secondary metabolites represents a fundamental step for the assessment of bioactivities and pharmacological properties of traditional herbal drugs. Rumex usambarensis (Dammer) Dammer has been described as a multipurpose remedy in different African traditional pharmacopoeias, but its phytochemical profile has not been properly investigated. Herein we report a high throughput metabolomic screening, based on ultra-high performance liquid chromatography-travelling wave ion mobility spectrometry quadrupole time-of-flight (UHPLC-TWINS-QTOF), which was performed for the first time on different R. usambarensis plant parts. By applying high-resolution mass spectrometry-based metabolomics and chemometric analysis, a complete discrimination of different aerial parts was obtained, with the annotation of 153 significant metabolites in leaves, stems, and flowers, suggesting an easy authentication and discrimination route. Phytochemical data were correlated to antimicrobial and antioxidant properties. Flavonoids, benzopyranes, chromones, and xanthones derivatives, along with a richer phytocomplex, might be responsible for the stronger bioactivities obtained from flowers.

Antioxidant Activity of Usnic Acid Compound from Methanol Extract of Lichen Usnea sp.

Lichen Usnea sp. is one of the sources of natural bioactive compounds which are currently being developed as medicinal ingredients. The purpose of this study was the isolation and identification of secondary metabolites from methanol extract, toxicity test and antioxidant activity of Usnea sp. Lichen was isolated by maceration using methanol solvent, then separated by liquid-liquid partition and separation using vacuum chromatography. Based on the results of the study, NMR-1D spectral data and FTIR spectrum is the presence of functional groups showed the presence of F15 compound is usnic acid consisting of 18 carbons with 3 carbons from the C=O carbonyl group. The results of the toxicity test showed that all of them were active against Artemia salina L. shrimp larvae with LC50 values of 0.820 µg/mL (Usnea sp.), 1.030 µg/mL (n-hexane), 1.056 µg/mL (ethyl acetate), and 1.236 µg/mL (methanol extract). The results of the antioxidant activity test showed that the inhibitory activity of usnic acid isolate was very active with an IC50 value of 11.696 µg/mL. Meanwhile, methanol extracts and ethyl acetate showed antioxidant activity with IC50 values of 18.098 and 26.917 µg/mL, respectively.

Metabolites Profiling of Penicillium citrinum Recovered from Endophytic of Ramie (Boehmeria nivea) as a Potential Biocontrol Against Pathogenic Fungi

Endophytes are a potent source of bioactive compounds that mimic plant-based metabolites. Fungi Fusarium spp. and Clonostachys rosea have been identified as pathogenic microorganisms in ramie (Boehmeria nivea). The antifungal test was carried out using the diffusion method and the MIC50 and Minimum Fungicidal Concentration (MFC) values using the tested microbial pathogens were Fusarium solani isolate 3248941, F. solani isolate Colpat-359, F. oxysporum isolate N-61-2, dan Clonostachys rosea strain B3042. Identification of secondary metabolites of the extract was carried out using GC-MS. The chromatogram of GC-MS analysis of this ethyl acetate extract (EA) showed seven dominant chemical compounds with various biological activities. Ethyl asetate extract of P. citrinum showed inhibition zone ranged from 19.10–22.07 mm with strong-very strong category at 200 mg ml-1 concentration against the tested microbial pathogens. Results revealed that the significant MIC values were observed against F. solani isolate 3248941 and Clonostachys rosea strain B3042 by less than 6.3 mg ml-1 and against F. solani isolate Colpat-359 and F. oxysporum isolate N-61-2 with 12.5 mg ml-1 and 25 mg ml-1 respectively. Minimum Fungicidal concentrations (MFC) for EA of P. citrinum were also reported against F. solani isolate 3248941 and C. rosea strain B3042, by 12.5 mg ml-1, whilst Fusarium solani isolate Colpat-359 by 25 mg ml-1 and Fusarium oxysporum isolate N-61-2 ,by 50 mg ml-1 respectively. Results of this study showed that P. citrinum, endophytic fungi of ramie, could be a promising source of compounds for antifungal agents.

Soil samples from sporotrichosis transmission belt area: Searching for fungal species and their antagonistic activity against Sporothrix brasiliensis.

Since 1998, the state of Rio de Janeiro, Brazil, has become a public health problem regarding sporotrichosis, a disease caused by Sporothrix spp. involving contact with infected cats. Efforts to isolate these species from environmental sources are not always successful. In our study, soil from residences situated in cities of Rio de Janeiro where cats with sporotrichosis live was collected and cultured an attempt to isolate Sporothrix spp. but it was not successful. However, other saprophytic fungal species were isolated from soil and identified and among them Purpureocillium lilacinum was the most frequent. From there, we decided to study the in vitro interaction of this species with S. brasiliensis, the principal agent that causes sporotrichosis in this state. The results showed that ten isolates of P. lilacinum inhibited the radial mycelial growth of S. brasiliensis with different percentage of inhibition. The interaction between them revealed the pattern described as overgrowth by antagonist. In conclusion, our data suggest that fungal species with very fast growth and capable of producing metabolites could hinder the growth of Sporothrix spp., it also opens the way for the identification of secondary metabolites with biological activity that could be tested against pathogenic fungi.

HPLC-HRESI-MSn Characterization of Polyphenolic Compounds in the Stem Bark of Chlorophora regia A. Chev (Moraceae)

Isolation and identification of secondary metabolites from medicinal plants could be tedious and time consuming. Therefore, any technique that could be used to confirm the identity of medicinal plant constituents without isolating them will save time and resources. Chlorophora similar to many genera in the Moraceae family have been demonstrated to be rich sources of polyphenolic compounds with important biological activities. The current study was designed to employ HRESI-MSn analyses to qualitatively examine isolated polyphenolic compounds from the stem bark of Chlorophora regia. Based on the HRESI-MSn data obtained, the fragmentation patterns of the compounds under study will be proposed and could be used in their identification in a matrix. Five polyphenolic compounds were successfully isolated and purified using various chromatographic techniques including column chromatography, thin-layer chromatography (TLC) and preparative HPLC. The structures of the isolated compounds were elucidated by in-depth analyses of their 1D and 2D NMR and mass spectroscopic data. HRESI-MS/MS was further used to characterize the isolated compounds. Five polyphenolic compounds including three Diels-Alder type adducts: sanggenon C, kuwanol E and chalcomoracin; two stilbene derivatives: chlorophorin and isochlorophorin were isolated from the stem bark. The tandem MS fragmentation patterns of the compounds in positive mode, were successfully proposed. The fragments obtained and proposed fragmentation patterns of the isolated compounds could be employed qualitatively in the identification of the studied polyphenolic compounds in a matrix.

Inhibition Activity Alpha-Glucosidase Enzyme And Toxicity From Ethanol Extract Of Caulerpa Seaweed

Caulerpa is a green macroalgae found on Lemukutan Island, West Kalimantan. Caulerpa contains bioactive compounds that have potential to inhibit the alpha-glucosidase enzyme for people with diabetes mellitus. Inhibition of the alpha-glucosidase enzyme will have an impact on delaying glucose absorption. In addition, it is necessary to carry out a toxicity test to detect toxic effects and assess safety in the use of compounds. Therefore, the purpose of this study was to determine the inhibitory activity of the alpha-glucosidase enzyme and the toxicity of the ethanolic extract of Caulerpa seaweed in Lemukutan Island waters. Identification of secondary metabolites was determined from phytochemical tests. Inhibitory activity of the alpha-glucosidase enzyme determined by using enzymatic reactions, while the toxicity by method of BSLT. The showed that the percentage yield of Caulerpa ethanol extract was 3.66%. The content of bioactive compounds in the ethanolic extract of Caulerpa contains positive alkaloids, flavonoids, saponins, tannins, steroids, and phenolics. The inhibitory of the alpha-glucosidase enzyme had the highest percentage of inhibition with a concentration of 300 ppm indicating a percent inhibition of 16.85%. Toxicity obtained LC50 value of 598.19 ppm which is classified as low toxic.