Identification Of Secondary Metabolites Research Articles

Phytochemical composition and antitumor activity of a new arctic lichen Anamylopsora pakistanica

For the first-time, chemical composition and in vitro antitumor activity was investigated of a newly described lichen Anamylopsora pakistanica Usman & Khalid from the second highest plateau of the world (Deosai Plains, Pakistan). HPLC–UV method was used for identification of secondary metabolites and the acetone extract had higher values of TPC (41.90 mg GA/g) and TFC (75.37 mg RE/g) as compared to methanol extract. As chemical constituents 5,7-dihydroxy-6-methylphthalide, haematommic acid and alectorialic acid, were identified as major compounds. Atranol, alectorialin, gyrophoric acid and usnic acid were detected as minor substances. Acetone and methanol extracts induced a dose-dependent and time-dependent decrease in the viability of three types of tumour cells HeLa, HCT116 and MDA-MB-231. This lichen extract can induce S phase arrest in HeLa as compared to the untreated cells. Extract of this unique lichen, A. pakistanica, can be used safely as a significant source of biologically active compounds.

Identification of Secondary Metabolites from Actinomycetes Isolated from the Hilly Region of Nepal

Actinomycetes are a unique class of Gram-positive bacteria with a fungus-like filamentous mycelium. They are often called ‘antibiotics factories’ because they can produce many potent secondary metabolites. At present, multidrug-resistant (MDR) pathogens are the main problem for humans and animals and to overcome this problem, more potent antibiotics are urgently required. Fortunately, Nepal has great geographical and climatic diversity serving as a home for various actinomycetes. This study identified several actinomycetes using morphological and biochemical assays from seven soil samples collected from different elevations in Nepal. The primary objective of this study was to screen the potent metabolites-producing actinomycetes. For this purpose, antimicrobial assays of seven isolates were carried out against Escherichia coli ATCC 25922, Salmonella typhi ATCC 14028, Shigella sonnei ATCC 25931, and Staphylococcus aureus ATCC 43300. The sample BT36 demonstrated the highest zone of inhibition against the tested microorganisms among others. Similarly, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of all isolates were carried out. Among them, the MIC of samples BT36, PT7, and AB1 were found to be 1.952 mg/mL, 0.012 mg/mL, and 0.029 mg/mL, respectively. Ethyl acetate extracts of resulting potent isolates PT7 and AB1 were subjected to mass spectrometric analysis. A total of 22 bioactive compounds were annotated from liquid chromatography-high resolution mass spectroscopy (LC-HRMS), including epopromycin A, myxopyronin B, gilvocarcin HE, and okilactomycin A. It is concluded that this study will be useful in setting up a strategy to isolate useful secondary metabolites from soil microbes from the hilly region of Nepal.

In vitro antileishmanial activities of hydro-methanolic crude extracts and solvent fractions of Clematis simensis fresen leaf, and Euphorbia abyssinica latex.

As a result of increasing drug resistance, crossover resistance development, prolonged therapy, and the absence of different agents with innovative methods for implementation, the efficacy of recent antileishmanial medications is severely declining. So, it is vital to look for other medications from botanical remedies that have antileishmanial activity. The latex of Euphorbia abyssinica (E abyssinica) and the leaves of Clematis simensis fresen (C simensis) were macerated in methanol (80%). In vitro antileishmanial activity of the preparation was tried on promastigotes of Leishmania aethiopica (L aethiopica) and Leishmania donovani (L donovani) using resazurin assay, and fluorescence intensity was measured. One percent of dimethyl sulfoxide (DMSO) and media as negative control and amphotericin B as positive control were used. Additionally, hemolytic & phytochemical tests of the preparation were done. The mean and standard errors of each extract were evaluated and interpreted for statistical significance using one-way analysis of variance. From sigmoidal dose-response curves of % inhibition, half maximal inhibitory concentration (IC50) values were determined by GraphPad Prism and Microsoft Excel; outcomes were presented as mean ± standard error of mean of triplicate trials. P < .05 was statistical significance. The phytochemical screening of C simensis and E abyssinica confirmed the existence of steroids, phenols, tannins, saponins, alkaloids, terpenoids, flavonoids and glycosides. C simensis possesses antileishmanial activity with IC50 outcomes of 46.12 ± 0.03 and 8.18 ± 0.10 µg/mL on the promastigotes of L aethiopica and L donovani, respectively. However, E abyssinica showed stronger activity with IC50 outcomes of 16.07 ± 0.05 µg/mL and 4.82 ± 0.07 µg/mL on L aethiopica and L donovani, respectively. C simensis and E abyssinica have a less hemolytic effect on human red blood cells at low concentrations. The outcomes from this investigation demonstrated that the preparation of C simensis and E abyssinica indicated significant antileishmanial activity. Therefore, further in vivo assessment of antileishmanial, cytotoxicity activity and quantitative identification of secondary metabolites are highly recommended.

Evaluation of Antifungal Activity of Endophytic Bacillus spp. and Identification of Secondary Metabolites Produced Against the Phytopathogenic Fungi.

Endophytic bacteria serve as a rich source of diverse antimicrobial compounds. Recently, there has been a growing interest in utilizing endophytic Bacillus spp. as biological agents against phytogenic fungi, owing to their potential to produce a wide range of antimicrobial substances. The objective of this research was to investigate the protective abilities of 15 endophytic Bacillus spp. isolated from previous study from wheat plant, against the phytopathogenic fungi, Fusarium graminearum and Macrophomina phaseolina. A dual culture plate assay was conducted as a preliminary analysis, revealing that 7 out of 15 endophytic Bacillus spp. demonstrated inhibition against one or both of the phytopathogenic fungi used in this study. All seven endophytes were further assessed for the presence of diffusible antifungal metabolites. The cultures were grown in potato dextrose broth for 120h, and the cell-free supernatant was extracted and analyzed using the cup plate method. The methanolic extract yielded similar results to the dual culture plate analysis, except for WL2-15. Additionally, deformities in the mycelial structure were examined under the light microscope upon exposure to methanolic extract. Furthermore, the analysis and identification of metabolites were carried out via gas chromatography-mass spectrometry of methanolic extract from selected seven endophytic Bacillus spp. The chromatogram revealed the presence of some major peaks such as tridecanoic acid, methyl ester, hydroperoxide, 1-methylbutyl, 9-octadecenamide, (z)-, hexane-1,3,4-triol, 3,5-dimethyl- tetradecanoic acid. To the best of our knowledge, this is the first report of these biocontrol agents in endophytic Bacillus spp. Interestingly, volatile organic compound production was also seen in all the isolates against the phytopathogenic fungi.

Comparative Analysis of Phytochemical Profiles and Selected Biological Activities of Various Morphological Parts of Ligustrum vulgare.

Ligustrum vulgare (LV), widely cultivated in Europe and often used in hedges, has been historically recognized in folk medicine for its potential health benefits. This study focused on exploring the untargeted identification of secondary metabolites in ethanol extracts (70% v/v) from different morphological parts (young shoots, leaves, flowers and fruits) of LV at various stages of plant development, using ultra-high-performance liquid chromatography with high-resolution mass spectrometry (UHPLC-HRMS). Additionally, the selected biological activities (antioxidant activity, cyclooxygenase-2 inhibition (COX-2), α-amylase inhibition and cytotoxicity) of the tested extracts were determined. Untargeted metabolomics showed that LV extracts were a rich source of phenylethanoid compounds, flavonoids, iridoids and their derivatives. The flowers of LV had the highest content of oleuropein (33.43 ± 2.48 mg/g d.w.). The lowest antioxidant activity was obtained for ripe and post-seasonal fruits, while in the case of other samples, the activity was at a similar level. All tested extracts showed α-amylase and COX-2 inhibitory activity. In addition, LV extracts showed strong antiproliferative properties in colorectal (HT29) and liver (HepG2) cancer cell lines. The obtained results show the difference in the content of bioactive compounds in various morphological parts of Ligustrum vulgare. These differences may influence the multifaceted medicinal potential of this plant.

Antioxidant and anticholinesterase properties of Echinometra mathaei and Ophiocoma erinaceus venoms from the Persian Gulf.

Introduction: The Persian Gulf is home to a diverse range of marine life, including various species of fish, crustaceans, mollusks, and echinoderms. This study investigates the potential therapeutic properties of venoms from echinoderms in the Persian Gulf, specifically their ability to inhibit cholinesterases (Acetylcholinesterase and butyrylcholinesterase) and act as antioxidants. Methods: Four venoms from two echinoderm species, including the spine, gonad, and coelomic fluids of sea urchins, as well as brittle star venoms, were analyzed using various methods, including LD50 determination, protein analysis, antioxidant assays, GC-MS for secondary metabolite identification, and molecular docking simulations. Results and discussion: The study's results revealed the LD50 of the samples as follows: 2.231 ± 0.09, 1.03 ± 0.05, 1.12 ± 0.13, and 6.04 ± 0.13mg/mL, respectively. Additionally, the protein levels were 44.037 ± 0.002, 74.223 ± 0.025, 469.97 ± 0.02, and 104.407 ± 0.025μg/mL, respectively. SDS-PAGE and total protein studies indicated that at least part of the venom was proteinaceous. Furthermore, the study found that the brittle star samples exhibited significantly higher antioxidant activity compared to other samples, including the standard ascorbic acid, at all tested concentrations. GC-MS analysis identified 12, 23, 21, and 25 compounds in the samples, respectively. These compounds had distinct chemical and bioactive structures, including alkaloids, terpenes, and steroids. Conclusion: These venoms displayed strong cholinesterase inhibitory and antioxidant activities, likely attributed to their protein content and the presence of alkaloids, terpenes, and steroids. Notably, the alkaloid compound C 7 was identified as a promising candidate for further research in Alzheimer's disease therapy. In conclusion, echinoderms in the Persian Gulf may hold significant potential for discovering novel therapeutic agents.

Alexandrium spp.: From Toxicity to Potential Biotechnological Benefits.

Many dinoflagellates of the genus Alexandrium are well known for being responsible for harmful algal blooms (HABs), producing potent toxins that cause damages to other marine organisms, aquaculture, fishery, tourism, as well as induce human intoxications and even death after consumption of contaminated shellfish or fish. In this review, we summarize potential bioprospecting associated to the genus Alexandrium, including which Alexandrium spp. produce metabolites with anticancer, antimicrobial, antiviral, as well as anti-Alzheimer applications. When available, we report their mechanisms of action and targets. We also discuss recent progress on the identification of secondary metabolites with biological properties favorable to human health and aquaculture. Altogether, this information highlights the importance of studying which culturing conditions induce the activation of enzymatic pathways responsible for the synthesis of bioactive metabolites. It also suggests considering and comparing clones collected in different locations for toxin monitoring and marine bioprospecting. This review can be of interest not only for the scientific community, but also for the entire population and industries.

In-vitro and in-silico anti-HSV-1 activity of a marine steroid from the jellyfish Cassiopea andromeda venom

In this study, we investigated the potential in vitro anti-HSV-1 activities of the Cassiopea andromeda jellyfish tentacle extract (TE) and its fractions, as well as computational work on the thymidine kinase (TK) inhibitory activity of the identified secondary metabolites. The LD50, secondary metabolite identification, preparative and analytical chromatography, and in silico TK assessment were performed using the Spearman-Karber, GC-MS, silica gel column chromatography, RP-HPLC, LC-MS, and docking methods, respectively. The antiviral activity of TE and the two purified compounds Ca2 and Ca7 against HSV-1 in Vero cells was evaluated by MTT and RT-PCR assays. The LD50 (IV, mouse) values of TE, Ca2, and Ca7 were 104.0 ± 4, 5120 ± 14, and 197.0 ± 7 (μg/kg), respectively. They exhibited extremely effective antiviral activity against HSV-1. The CC50 and MNTD of TE, Ca2, and Ca7 were (125, 62.5), (25, 12.5), and (50, 3.125) μg/ml, respectively. GC-MS analysis of the tentacle extract revealed seven structurally distinct chemical compositions. Four of the seven compounds had a steroid structure. According to the docking results, all compounds showed binding affinity to the active sites of both thymidine kinase chains. Among them, the steroid compound Pregn-5-ene-3,11-dione, 17,20:20,21 bis [methylenebis(oxy)]-, cyclic 3-(1,2-ethane diyl acetal) (Ca2) exhibited the highest affinity for both enzyme chains, surpassing that of standard acyclovir. In silico data confirmed the experimental results. We conclude that the oxosteroid Ca2 may act as a potent agent against HSV-1.

Salazinic Acid and Norlobaridone from the Lichen Hypotrachyna cirrhata: Antioxidant Activity, α-Glucosidase Inhibitory and Molecular Docking Studies

The present study was intended for the identification of secondary metabolites in acetone extract of the lichen Hypotrachyna cirrhata using UPLC-ESI-QToF-MS/MS and the detection of bioactive compounds. This study led to the identification of 22 metabolites based on their MS/MS spectra, accurate molecular masses, molecular formula from a comparison of the literature database (DNP), and fragmentation patterns. In addition, potent antioxidant and α-glucosidase inhibitory potentials of acetone extract of H. cirrhata motivated us to isolate 10 metabolites, which were characterized as salazinic acid (11), norlobaridone (12), atranorin (13), lecanoric acid (14), lichesterinic acid (15), protolichesterinic acid (16), methyl hematommate (17), iso-rhizonic acid (18), atranol (19), and methylatratate (20) based on their spectral data. All these isolates were assessed for their free radicals scavenging, radical-induced DNA damage, and intestinal α-glucosidase inhibitory activities. The results indicated that norlobaridone (12), lecanoric acid (14), methyl hematommate (17), and atranol (19) showed potent antioxidant activity, while depsidones (salazinic acid (11), norlobaridone (12)) and a monophenolic compound (iso-rhizonic acid, (18)) displayed significant intestinal α-glucosidase inhibitory activities (p &lt; 0.001), which is comparable to standard acarbose. These results were further correlated with molecular docking studies, which indicated that the alkyl chain of norlobaridione (12) is hooked into the finger-like cavity of the allosteric pocket; moreover, it also established Van der Waals interactions with hydrophobic residues of the allosteric pocket. Thus, the potency of norlobaridone to inhibit α-glucosidase enzyme might be associated with its allosteric binding. Also, MM-GBSA (Molecular Mechanics-Generalized Born Surface Area) binding free energies of salazinic acid (11) and norlobaridone (12) were superior to acarbose and may have contributed to their high activity compared to acarbose.

Untargeted metabolomic analyses support the main phylogenetic groups of the common plant-associated Alternaria fungi isolated from grapevine (Vitis vinifera)

Alternaria, a cosmopolitan fungal genus is a dominant member of the grapevine (Vitis vinifera) microbiome. Several Alternaria species are known to produce a variety of secondary metabolites, which are particularly relevant to plant protection and food safety in field crops. According to previous findings, the majority of Alternaria species inhabiting grapevine belong to Alternaria sect. Alternaria. However, the phylogenetic diversity and secondary metabolite production of the distinct Alternaria species has remained unclear. In this study, our aim was to examine the genetic and metabolic diversity of endophytic Alternaria isolates associated with the above-ground tissues of the grapevine. Altogether, 270 Alternaria isolates were collected from asymptomatic leaves and grape clusters of different grapevine varieties in the Eger wine region of Hungary. After analyses of the nuclear ribosomal DNA internal transcribed spacer (ITS) and RNA polymerase second largest subunit (rpb2) sequences, 170 isolates were chosen for further analyses. Sequences of the Alternaria major allergen gene (Alt a 1), endopolygalacturonase (endoPG), OPA10-2, and KOG1058 were also included in the phylogenetic analyses. Identification of secondary metabolites and metabolite profiling of the isolates were performed using high-performance liquid chromatography (HPLC)–high-resolution tandem mass spectrometry (HR-MS/MS). The multilocus phylogeny results revealed two distinct groups in grapevine, namely A. alternata and the A. arborescens species complex (AASC). Eight main metabolites were identified in all collected Alternaria isolates, regardless of their affiliation to the species and lineages. Multivariate analyses of untargeted metabolites found no clear separations; however, a partial least squares-discriminant analysis model was able to successfully discriminate between the metabolic datasets from isolates belonging to the AASC and A. alternata. By conducting univariate analysis based on the discriminant ability of the metabolites, we also identified several features exhibiting large and significant variation between A. alternata and the AASC. The separation of these groups may suggest functional differences, which may also play a role in the functioning of the plant microbiome.

Evidence of antagonism in vitro and in vivo of extracts from Banisteriopsis laevifolia (A. Juss) B. Gates against the rice blast fungus

Rice blast is a common disease caused by Magnaporthe oryzae which affects rice production worldwide, including Brazil. To reduce the use of harmful pesticides, sustainable control options are necessary for managing rice blast. One possible option is to use plant extracts because they produce secondary metabolites capable of inhibiting the pathogen and controlling the disease. This study aimed to evaluate the potential of ethanolic extracts of leaves and flowers from Banisteriopsis laevifollia, a native Brazilian plant, to antagonise M. oryzae. In vitro antifungal activity assays were carried out, in which mycelium growth reduction, conidia germination, and appressorium formation of M. oryzae were evaluated. In addition, in vivo assay for suppression of leaf blast severity by extracts was done. We also explored the identification of secondary metabolites from ethanolic extracts of leaves and flowers by HPLC–HRMS. Results showed that both leaf and flower extracts inhibited mycelial growth in 21.72% and 30.49%, respectively, due to the inhibition of ergosterol production. Additionally, both extracts inhibited melanin production in the mycelium of M. oryzae by inhibiting tyrosine production. Both leaf and flower extracts significantly inhibited conidia germination and appressorium formation. Furthermore, rice leaves treated with the extracts suppressed by 18.17% and 18.97% leaf blast severity, with the 1.00 mg/mL concentration of flower extract and 0.75 mg/mL concentration of leaf extract, respectively, showing the most significant reduction of leaf blast. The extracts contain phenolic compounds, including 2,3,7,8-tetrahydroxy-chromeno-(5,4,3)-chromene-5,10-dione, 3-(4-hydroxyphenyl)-2-propenoic acid, 2-(3,4-dihydroxyphenyl)-3,4-dihydro-1(2H)-benzopyran-3,5,7-triol, 3,4′,5,7-tetrahydroxyflavone, 3′,4′,5,7-tetrahydroxyflavone, 4′,5,7-trihydroxyflavanone, 3,3′,4′,5,7-pentahydroxyflavone, 3,3′,4′,5,7-pentahydroxyflavone -3-O-glucoside, and 3,3′,4′,5,7-pentahydroxyflavone-3-O-rutinoside, which are likely responsible for the antifungal activity of the extracts. Overall, this study demonstrates the potential of plant extracts as a sustainable alternative for the management of rice blast.

Qualitative Identification of Secondary Metabolites and Determination of the Toxicity of Extracts Obtained from the Flower of Kalanchoe Pinnata

Kalanchoe pinnata, also known as air leaf or life leaf, is a plant used in traditional medicine in different world regions. In Mexico, it is included in the Atlas de la Medicina Tradicional Mexicana with a wide variety of applications, such as antimicrobial, anti-inflammatory, anticancer, and antihistamine, among others. However, neither a secondary metabolite profile of the flower has been reported nor information on its possible toxicity. The latter is the purpose of this work. A phytochemical profile of extracts with solvents of different polarity (aqueous, methanol, ethanol, and ether) was carried out. In this profile, the structural compounds could be qualitatively determined by chemical reactions, and some changes in coloring or precipitation were observed. The acute toxicity test of the extracts was performed with an aquatic organism, Artemia sp, and a terrestrial organism Eisenia foetida, as well as the evaluation of the antioxidant capacity of the extracts in the organism of Eisenia foetida. The ABTS radical method and TROLOX were applied as synthetic antioxidants for the evaluation of the inhibition percentage. Most important secondary metabolites were qualitatively identified in the extracts of K. pinnata flowers. Mainly in the alcoholic extracts (methanol and ethanol) tannins, alkaloids, and flavonols were found. As mentioned above, they are reported to have toxicological effects. The toxicity and antioxidant activity tests confirm the preliminary results obtained in the identification of secondary metabolites. It is therefore concluded that the flower of Kalanchoe pinnata contains secondary metabolites that may be of great therapeutic interest.

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.