Secondary Metabolite Profiles Research Articles

Multivariate Data Analysis Guided Identification of Anticancer Constituents of Spina Gleditsiae for Human Hepatoma Cells

Spina Gleditsia has been traditionally used as a lead ingredient of many formulas for cancer treatment in Traditional Chinese Medicine. This work aimed to reveal the phytochemical foundation of Spina Gleditsiae and underpin a fundamental understanding of its anticancer activities. The growth inhibition activities were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay of 7 solvent extracts of Spina Gleditsia in the human liver cancer cell line HepG2. The activities were then mapped onto the secondary metabolite profile of crude extracts by principal components analysis (PCA) of High Performance Liquid Chromatography-Ultraviolet (HPLC-UV) data. The top 5 PCA components of the map discriminated extract activities, mainly based on the differential content of 3 compounds. The identified compounds were then tested individually using cytotoxic assay on cancer cells in vitro. Principal components analysis constructed secondary metabolite mapping quickly identified functional compounds from the crude herbal extracts. (2 R,3 R)-7,3′,4′-Trihydroxy-5-methoxy-flavanonol, (2 R,3 R)-5,7,3′-trihydroxy-4′-methoxy-flavanonol, and isovitexin inhibited the growth of human liver cancer cell line growth with activities of 38.6 ± 1.9, 40.7 ± 0.9, and 41.5 ± 3.1 μg/mL, respectively, comparable with that of cisplatin (IC50: 41.6 ± 2.9 μg/mL). (2 R,3 R)-7,3′,4′-Trihydroxy-5-methoxy-flavanonol, (2 R,3 R)-5,7,3′-trihydroxy-4′-methoxy-flavanonol, and isovitexin are, for the first time, identified as the anticancer constituents of Spina Gleditsia. It thus strongly underpins a fundamental understanding of the anticancer activities of Spina Gleditsia and supports its ongoing clinical uses in China.

Non‐targeted metabolomics approach to assess the brown marine macroalga Dictyota dichotoma as a functional food using liquid chromatography with mass spectrometry

AbstractThe study elucidates presence of 16 potential putative metabolites in Dictyota dichotoma through non‐targeted secondary metabolite profile analysis using electrospray ionization negative mode of liquid chromatography with time‐of‐flight mass spectrometry. Two fatty acyl glycosides viz; 5‐megastigmen‐7‐yne‐3,9‐diol 9‐glucoside (biomarker for food consumption) and (3S,7E,9S)‐9‐hydroxy‐4,7‐megastigmadien‐3‐one 9‐glucoside (nutrient, emulsifier, surfactant), an o‐glycosyl compound 5‐megastigmen‐7‐yne‐3,9‐diol 3‐glucoside (biomarker for food consumption), all with m/z value 370.19 were detected. An acyclic monoterpenoid and diterpenoid viz; isodigeranyl and digeranyl (both with m/z 274.26; food biomarkers) were detected. Additionally, (3′x,5′a,9′x,10′b)‐O‐(3‐hydroxy‐6‐oxo‐7‐drimen‐11‐yl) umbelliferone (m/z 396.19; nutrient, emulsifier, surfactant) and kamolonol (m/z 398.2; antibacterial, anti‐tumor) were observed. Two angular pyranocoumarins: mammea C/AB cyclo D (m/z 398.2; food consumption biomarker) and 5‐hydroxy‐8, 8‐dimethyl‐6‐(3‐methyl‐1‐oxobutyl)‐4‐pentyl‐2H,8H‐benzo[1,2‐b:3,4‐b']dipyran‐2‐one (m/z 398.2; fruit constituent) were present. Two terpene lactones ‐ galbanic acid (m/z 398.2; antimicrobial, anticoagulant, antiviral, anti‐inflammatory, anti‐cancer) and asacoumarin A (m/z 398.2; antiprotozoal, antiglycation activity) were detected. Armillaribin (m/z 398.2; food biomarker) along with glycinoeclepin B (m/z 558.28; nutrient, emulsifier, surfactant) was noted. Two flavonoids; (S)‐(E)‐2′‐(3,6‐dimethyl‐2‐heptenyl)‐3′,4′,7‐trihydroxyflavanone and (S)‐(E)‐8‐(3,6‐dimethyl‐2‐heptenyl)‐4′,5,7‐trihydroxyflavanone (m/z 396.19) and a drug triclofos (m/z 227.89; sedative) were detected. This study highlights the metabolic makeup of D. dichotoma collected from Kachchh coast thereby, accentuating the probability of its inclusion as a promising ingredient in functional foods from algal origin.

Epigenetic modulation of secondary metabolite profiles in Aspergillus calidoustus and Aspergillus westerdijkiae through histone deacetylase (HDAC) inhibition by vorinostat.

The effects of epigenetic modulation on secondary metabolite biosynthesis were investigated with five Aspergillus species cultured in the presence of either the DNA methyltransferase inhibitor 5-azacitidine or the histone deacetylase inhibitor vorinostat. With Aspergillus calidoustus and Aspergillus westerdijkiae, fermentation in the presence of vorinostat (100 μM) induced significant changes in secondary metabolite profile with examples of both induction and repression. We identified putative biosynthetic gene clusters for emericellamide in A. calidoustus and ochratoxin in A. westerdijkiae. A substantial induction in production levels was observed for two secondary metabolites: the diketopiperazine alkaloid phenylahistin in A. calidoustus and the polyketide penicillic acid in A. westerdijkiae, indicating the potential of epigenetic regulation for the activation of silent fungal biosynthetic pathways.

Variation in secondary metabolites in a unique set of tomato accessions collected in Turkey

In this study, 50 tomato landraces grown in Turkey were investigated in terms of their secondary metabolite profiles. Each accession was planted in 2016 and 2017 in 3 replicates in an open field. In this study, color, pH and brix of the fruit samples were measured and an unbiased LCMS-based metabolomics approach was applied. Based on Principal Components Analysis (PCA) and Hierarchical Cluster Analysis (HCA) of the relative abundance levels of >250 metabolites, it could be concluded that fruit size was the most influential to the biochemical composition, rather than the geographical origin of accessions. Results indicated substantial biodiversity in various metabolites generally regarded as key to fruit quality aspects, including sugars; phenolic compounds like phenylpropanoids and flavonoids; alkaloids and glycosides of flavour-related volatile compounds. The phytochemical data provides insight into which Turkish accessions might be most promising as starting materials for the tomato processing and breeding industries.

Chemical profiling of secondary metabolites from Himatanthus drasticus (Mart.) Plumel latex with inhibitory action against the enzymes α-amylase and α-glucosidase: In vitro and in silico assays

Ethnopharmacological relevanceHimatanthus drasticus is an important medicinal plant whose latex is traditionally used in Northeast Brazil to treat various diseases, including diabetes. The use of α-amylase and α-glucosidase inhibitors can be an effective strategy to modulate levels of postprandial hyperglycemia via control of starch metabolism. Aims of the studyThis study aimed to verify if H. drasticus latex has inhibitory activity against enzymes linked to type 2 diabetes, besides chemically characterizing the metabolites responsible for such activities. In addition, in silico analysis was performed to support the traditional claim of possible antidiabetic activity of this latex. Materials and methodsLatex from H. drasticus stems was sequentially partitioned with n-hexane (FHDH), CHCl3 (FHDC) and EtOH (FHDHA). Wash extraction of the FHDHA fraction was performed to obtain the other extract fractions. The FHDHA was submitted to chromatography in a SPE C18 cartridge using gradient elution with MeOH/H2O to produce five fractions: FHDHA1, FHDHA2, FHDHA3, FHDHA4 and FHDHA5. The FHDHA1 was subjected to semi-preparative reverse phase HPLC. Lineweaver-Burk plots were used to investigate the kinetic parameters of α-amylase and α-glucosidase inhibitory activity. The interactions between plumieride and porcine pancreatic α-amylase and α-glucosidase were analyzed through an in silico molecular docking study. ResultsPhytochemical identification of compounds present in the FHDHA fraction of H. drasticus latex was possible by 1H, 13C NMR analysis and mass spectrometry, and the results were compared with the literature. The identified compounds were α-ethyl glucoside, protocatechuic acid, 3-O-caffeoylquinic acid, 15-demethylplumieride acid, 5-O-caffeoylquinic acid, caffeic acid, vanillic acid, plumieride, and catechin. The inhibition results of the fractions tested against α-amylase and α-glucosidase showed inhibitory activities dependent on the increase of fractions and compound concentrations. The IC50 results obtained from FHDHA, FHDHA1 and plumieride fractions against α-amylase were 36.46, 72.61, 33.87 μg/mL respectively. The IC50 of plumieride was the closest to that of acarbose (22.52 μg/mL), a result similar to that obtained for α-glucosidase. The type of inhibition was competitive for both enzymes. ConclusionsThere was strong inhibition of α-amylase and α-glucosidase by FHDHA, FHDHA1 and plumieride, suggesting that these enzymes slow glucose absorption.

Effect of cut on secondary metabolite profile in hydroponically-grown Rumex acetosa L. seedlings: a metabolomic approach

Seedlings of Rumex acetosa L. (sorrel) were grown in floating system and two consecutive cuts took place: 15 (C1) and 30 (C2) days after sowing. An untargeted metabolomics approach was utilised to fingerprint phenolics and other health-related compounds in sorrel leaves, as well as to unveil differences between the two cuts. The untargeted approach allowed to putatively identify 458 metabolites considering both the cuts. Three new terpenoids and two new phenylpropanoid glycosides (dihydrosyringin and dihydroconiferin) with antifungal and anti-inflammatory activity were annotated. Overall, leaves from C2 had lower level of secondary metabolites (44 were down-accumulated), especially sesquiterpenes and stilbenes. Conversely, anthocyanins showed a relevant increase in C2 than in C1 leaves. The dataset suggests that sorrel leaves represent a good source of nutraceutical compounds and unveils the pivotal effect of pre-harvest factor in secondary metabolite profile.

Identification of Rosellinia species as producers of cyclodepsipeptide PF1022 A and resurrection of the genus Dematophora as inferred from polythetic taxonomy.

Rosellinia (Xylariaceae) is a large, cosmopolitan genus comprising over 130 species that have been defined based mainly on the morphology of their sexual morphs. The genus comprises both lignicolous and saprotrophic species that are frequently isolated as endophytes from healthy host plants, and important plant pathogens. In order to evaluate the utility of molecular phylogeny and secondary metabolite profiling to achieve a better basis for their classification, a set of strains was selected for a multi-locus phylogeny inferred from a combination of the sequences of the internal transcribed spacer region (ITS), the large subunit (LSU) of the nuclear rDNA, beta-tubulin (TUB2) and the second largest subunit of the RNA polymerase II (RPB2). Concurrently, various strains were surveyed for production of secondary metabolites. Metabolite profiling relied on methods with high performance liquid chromatography with diode array and mass spectrometric detection (HPLC-DAD/MS) as well as preparative isolation of the major components after re-fermentation followed by structure elucidation using nuclear magnetic resonance (NMR) spectroscopy and high resolution mass spectrometry (HR-MS). Two new and nine known isopimarane diterpenoids were identified during our mycochemical studies of two selected Dematophora strains and the metabolites were tested for biological activity. In addition, the nematicidal cyclodepsipeptide PF1022 A was purified and identified from a culture of Rosellinia corticium, which is the first time that this endophyte-derived drug precursor has been identified unambiguously from an ascospore-derived isolate of a Rosellinia species. While the results of this first HPLC profiling were largely inconclusive regarding the utility of secondary metabolites as genus-specific chemotaxonomic markers, the phylogeny clearly showed that species featuring a dematophora-like asexual morph were included in a well-defined clade, for which the genus Dematophora is resurrected. Dematophora now comprises all previously known important plant pathogens in the genus such as D. arcuata, D. bunodes, D. necatrix and D. pepo, while Rosellinia s. str. comprises those species that are known to have a geniculosporium-like or nodulisporium-like asexual morph, or where the asexual morph remains unknown. The extensive morphological studies of L.E. Petrini served as a basis to transfer several further species from Rosellinia to Dematophora, based on the morphology of their asexual morphs. However, most species of Rosellinia and allies still need to be recollected in fresh state, cultured, and studied for their morphology and their phylogenetic affinities before the infrageneric relationships can be clarified.

Endophytic Fungi of Native Salvia abrotanoides Plants Reveal High Taxonomic Diversity and Unique Profiles of Secondary Metabolites.

Endophytic fungi are often embedded in their host’s metabolic networks, which can result in alterations of metabolite production and higher amounts of active compounds in medicinal plants. This study reports the occurrence, diversity, and secondary metabolite profiles of endophytic fungi isolated from Salvia abrotanoides plants obtained from three geographically distinct sites in Iran. A total of 56 endophytic fungi were isolated from roots and leaves of S. abrotanoides; site-specificity and root-dominated colonization was found to be a general characteristic of the endophytes. Based on molecular identification, the endophytic fungi were classified into 15 genera. Mycelial extracts of these isolates were subjected to high-resolution mass spectrometry analyses and revealed a broad spectrum of secondary metabolites. Our results demonstrated that Penicillium canescens, P. murcianum, Paraphoma radicina, and Coniolariella hispanica are producers of cryptotanshinone, which is a main bioactive compound of S. abrotanoides. Moreover, it was shown that it can be produced independent of the host plant. The effect of exogenous gibberellin on S. abrotanoides and endophytic fungi was shown to have a positive effect on increasing the cryptotanshinone production in the plant as well as in endophytic fungi cultivated under axenic conditions. Our findings provide further evidence that endophytic fungi play an important role in the production plant bioactive metabolites. Moreover, they provide an exploitable basis to increase cryptotanshinone production in S. abrotanoides.

Comprehensive Analysis of the Alternaria Mycobolome Using Mass Spectrometry Based Metabolomics.

Alternaria fungi are widely distributed plant pathogens infecting grains and vegetables and causing major harvest losses in the field and during postharvest storage. Besides, consumers are endangered by the formation of toxic secondary metabolites. Some of these secondary metabolites are chemically characterized as mycotoxins, but the majority of the Alternaria mycobolome still remains unknown. Fourier-transform ion cyclotron resonance mass spectrometry (FTICR-MS) and LC-MS/MS are combined for the non-targeted and targeted analysis of the metabolome of three A.alternata isolates and one A.solani isolate. Due to the ultra-high resolution of FTICR-MS, unique molecular formulae are assigned to measured m/z signals. The molecular formulae are matched to entries of the databases Antibase and Kyoto Encyclopedia of Genes and Genomes. The non-targeted analysis of the fungal extracts reveals variations in the secondary metabolite profile of A.alternata and A.solani. Differences in the biosynthesis of dibenzo-α-pyrones, perylene quinones, tentoxin, and tenuazonic acid of the A.alternata and A.solani isolates are determined applying targeted LC-MS/MS. FTICR-MS analyses reveal clear differences in the metabolic profile of the A.solani and the A.alternata isolates.

Phytochemical Screening of Callus and Cell Suspensions Cultures of Thevetia peruviana

Abstract Thevetia peruviana is an ornamental shrub grown-up in many tropical region of the world. This plant produces secondary metabolites with biological properties of interest for the pharmaceutical industry. The objective was to determine the secondary metabolites profile of callus and cell suspension cultures of T. peruviana and compare them with those from explant (fruit pulp). Extracts in 50% aqueous ethanol and ethyl acetate were prepared. The phytochemical analysis was performed using standard chemical tests and thin layer chromatography. In addition, total phenolic and flavonoids compounds (TPC and TFC), total cardiac glycosides (TCG) and total antioxidant activity (TAA) was determined during the cell suspension growth. Phenolic chemical profile was also analyzed by high performance liquid chromatography (HPLC). Common metabolites (alkaloids, amino acids, antioxidants, cardiac glycosides, leucoanthocyanidins, flavonoids, phenols, sugars and triterpenes) were detected in all samples. The maximum production of extracellular TCG, TPC, TFC and TAA in cells suspensions were at 6-12 days; in contrast, intracellular content was relatively constant during the exponential grown phase (0 to 12-days). HPLC analysis detected one compound with retention time at 11.6 min; this compound was tentatively identified as dihydroquercetin, a flavonoid with anti-cancer properties. These results provide evidence on the utility of the in vitro cell cultures of T. peruviana for valuable pharmaceutical compounds production.

Characterization of Secondary Metabolites Profile of Flavonoid from Salam Leaves (Eugenia polyantha) Using TLC and UVSpectrophotometry

Characterization of Secondary Metabolites Profile of Flavonoid from Salam Leaves (Eugenia polyantha) Using TLC and UVSpectrophotometry

Effect of Gender, Rearing, and Cooking on the Metabolomic Profile of Porcine Muscles.

To clarify the relationship between the fiber type composition and meat quality, we performed metabolomic analysis using porcine longissimus dorsi (LD) muscles. In the LD of pigs raised outdoors, the expression of myosin heavy chain (MyHC)1 (slow-twitch fiber marker protein) was significantly increased compared with that of MyHC1 in pigs raised in an indoor pen, suggesting that rearing outdoors could be considered as an exercise treatment. These LD samples were subjected to metabolomic analysis for examining the profile of most primary and secondary metabolites. We found that the sex of the animal and exercise stimulation had a strong influence on the metabolomic profile in the porcine skeletal muscles, and this difference in the metabolomic profile is likely in part due to the changes in the muscle fiber type. We also examined the effects of cooking (70 °C for 1 h). The effect of exercise on the metabolomic profile was also maintained in the cooked muscle tissues. Cooking treatment resulted in an increase in some of the metabolite levels while decreasing in some other metabolite levels. Thus, our study could indicate the effect of the sex of the animal, exercise stimulus, and cooking on the metabolomic profile of pork meat.

Harvest time–related changes in biomass, phenolics and antioxidant potential in Knautia sarajevensis shoot cultures after elicitation with salicylic acid and yeast

Elicitation is one of the effective methods for enhancement of metabolite production in plants. In this study we evaluated for the first time how elicitors affect biomass and phenolic production and antioxidant potential in relation to type (salicylic acid and yeast extract) and concentration of the elicitor and harvesting time in Knautia sarajevensis. Biomass production was estimated according to the control plants cultivated in liquid MS medium without eliciting agents. Total phenol, flavonoid and flavanol contents were estimated along with antioxidant activities at different harvesting times (0, 3, 7 and 15 d post-elicitation). The differences of phenolic metabolites were analysed by HPLC-UV for all treatments at all harvesting times. The best eliciting effect was recorded after 7 d by using 100 mM salicylic acid. Secondary metabolite profile analysis showed a significant increase in the content of rosmarinic acid, as well as chlorogenic and salicylic acids. The DPPH radical scavenging activity was decreased after elicitation. Increase in peroxidase activity was recorded especially after elicitation with 100 mM salicylic acid 3 d post-elicitation.

Pigmentosins from Gibellula sp. as antibiofilm agents and a new glycosylated asperfuran from Cordyceps javanica.

In the course of our exploration of the Thai invertebrate-pathogenic fungi for biologically active metabolites, pigmentosin A (1) and a new bis(naphtho-α-pyrone) derivative, pigmentosin B (2), were isolated from the spider-associated fungus Gibellula sp. Furthermore, a new glycosylated asperfuran 3, together with one new (6) and two known (4 and 5) cyclodepsipeptides, was isolated from Cordyceps javanica. The pigmentosins 1 and 2 showed to be active against biofilm formation of Staphylococcus aureus DSM1104. The lack of toxicity toward the studied microorganism and cell lines of pigmentosin B (2), as well as the antimicrobial effect of pigmentosin A (1), made them good candidates for further development for use in combination therapy of infections involving biofilm-forming S. aureus. The structure elucidation and determination of the absolute configuration were accomplished using a combination of spectroscopy, including 1D and 2D NMR, HRMS, Mosher ester analysis, and comparison of calculated/experimental ECD spectra. A chemotaxonomic investigation of the secondary metabolite profiles using analytical HPLC coupled with diode array detection and mass spectrometry (HPLC–DAD–MS) revealed that the production of pigmentosin B (2) was apparently specific for Gibellula sp., while the glycoasperfuran 3 was specific for C. javanica.

Utilisation of Rhododendron luteum Sweet bioactive compounds as valuable source of enzymes inhibitors, antioxidant, and anticancer agents

Ethnobotanical evidences report the use of Rhododendron luteum Sweet (Ericaceae) in traditional medicinal systems. However, R. luteum has been associated to the occurrence of ‘mad honey’ poisoning. In the present study, the ethyl acetate, methanol, and water extracts of R. luteum were investigated for their in vitro antioxidant, enzyme inhibition, and cytotoxic properties. The cytotoxicity of R. luteum extracts on A549 lung cancer cell line was evaluated using MTT cell viability assay. Besides, HPLC-ESI-MSn approach was employed to elucidate the secondary metabolite profiles of R. luteum in order to establish any structure-activity relationship. Methanol and water extracts of R. luteum possessed highest radical scavenging and reducing properties while the ethyl acetate extract showed highest metal chelating properties. In terms of enzyme inhibition, the methanol and ethyl acetate extracts of R. luteum, possessing epigallocatechin, were active inhibitors of cholinesterase enzymes, α-glucosidase, and tyrosinase. Water extract caused growth inhibition of A549 cells with 207.2 μg/ml IC50 value. Though R. luteum has received little scientific attention due to the occurrence of grayanotoxins in the plant, however, data presented in this work shows promising biological activity of R. luteum and highlighted its role as a potential source of antioxidant and key enzyme inhibitors.

The dynamic responses of plant physiology and metabolism during environmental stress progression.

At adverse environmental conditions, plants produce various kinds of primary and secondary metabolites to protect themselves. Both primary and secondary metabolites play a significant role during the heat, drought, salinity, genotoxic and cold conditions. A multigene response is activated during the progression of these stresses in the plants which stimulate changes in various signaling molecules, amino acids, proteins, primary and secondary metabolites. Plant metabolism is perturbed because of either the inhibition of metabolic enzymes, shortage of substrates, excess demand for specific compounds or a combination of these factors. In this review, we aim to present how plants synthesize different kinds of natural products during the perception of various abiotic stresses. We also discuss how time-scale variable stresses influence secondary metabolite profiles, could be used as a stress marker in plants. This article has the potential to get the attention of researchers working in the area of quantitative trait locus mapping using metabolites as well as metabolomics genome-wide association.

Hydroponically Grown Sanguisorba minor Scop.: Effects of Cut and Storage on Fresh-Cut Produce.

Wild edible plants have been used in cooking since ancient times. Recently, their value has improved as a result of the scientific evidence for their nutraceutical properties. Sanguisorba minor Scop. (salad burnet) plants were hydroponically grown and two consecutive cuts took place at 15 (C1) and 30 (C2) days after sowing. An untargeted metabolomics approach was utilized to fingerprint phenolics and other health-related compounds in this species; this approach revealed the different effects of the two cuts on the plant. S. minor showed a different and complex secondary metabolite profile, which was influenced by the cut. In fact, flavonoids increased in leaves obtained from C2, especially flavones. However, other secondary metabolites were downregulated in leaves from C2 compared to those detected in leaves from C1, as evidenced by the combination of the variable important in projections (VIP score > 1.3) and the fold-change (FC > 2). The storage of S. minor leaves for 15 days as fresh-cut products did not induce significant changes in the phenolic content and antioxidant capacity, which indicates that the nutraceutical value was maintained. The only difference evidenced during storage was that leaves obtained from C2 showed a lower constitutive content of nutraceutical compounds than leaves obtained from C1; except for chlorophylls and carotenoids. In conclusion, the cut was the main influence on the modulation of secondary metabolites in leaves, and the effects were independent of storage.

Effect-directed profiling and identification of bioactive metabolites from field, in vitro-grown and acclimatized Musa spp. accessions using high-performance thin-layer chromatography-mass spectrometry

Bananas and plantains (Musa spp.) are used as nutritious foods, and at the same time, are a source of phytoconstituents for the pharmaceutical industry. As biological activities of especially the pulp and peel of Musa spp. have been documented, this study investigated the variation in the secondary metabolite profiles of the leaves from field, in vitro-grown and acclimatized accessions. The genetic fidelity of the diverse accessions was assessed using diversity array technology sequencing. It showed that the in vitro-grown accessions were true-to-type with the field samples. The antioxidant and anticholinesterase activities of the samples from different culture systems (field and in vitro) were evaluated by UV-spectrophotometry and compared to high-performance thin-layer chromatography-effect-directed analysis (HPTLC-EDA). The latter was applied for the first time for effect-directed profiling of the polar and medium polar sample components via different biochemical and biological assays. Compound zones showed acetyl-/butylrylcholinesterase inhibition (zones 1–4), α-/β-glucosidase inhibition (zones 1 and 2) as well as antioxidative (zones 1–3) and antimicrobial (zone 4) activities. Structures were preliminary assigned by HPTLC-HRMS. The HPTLC was effective for bioactivity-guided characterization of the bioactive constituents in Musa spp. accessions. Accumulation of useful metabolites, especially compounds with antioxidant and anticholinesterase properties, was higher in samples from in vitro system. This validated the use of plant tissue culturing as an alternative method for large scale production of plant material and supply of bioactive constituents.

Metabolomic study of Virola surinamensis (Rol.) Warb raw seed butter from different Amazonian regions

Introduction: Virola surinamensis, known in the Brazilian Amazonian region as ucuuba, produces seeds with high lipid content. The butter extracted from ucuuba seeds has a high moisturize potential, skin repairing properties and soft texture, which is interesting for cosmetic application. Methods: Virola surinamensis seeds from three different Amazonian regions were collected and studied: Para’s Northeast (Para/Brazil), Marajo Island (Para/Brazil) and RESEX Medio Jurua (Amazon as/Brazil). Fatty acid and triacilglycerides compositions were analyzed by gas chromatography with flame ionization detector (GC-FID). Secondary metabolite profiles were analyzed by ultra-performance liquid chromatography coupled with electrospray ionization and quadrupole time-of-flight mass spectrometer (UPLC-ESI-QToF-MS), in both negative and positive ionization modes. Results: Results revealed thar capric and lauric acids contents of RESEX Medio Jurua region butter were different from Para’s Northeast and Marajo island regions. LaMM and MMM triacilglycerols were the majoritarian triacilglycerides and the distribution of RESEX Medio Jurua region was also distinct. Different metabolites profiles were observed for the samples. Primary and secondary metabolites differences between V. surinamensis seed butter from three different Amazonian regions suggests that the environment influences the metabolome. Conclusion: Advanced analytical techniques and statistical analysis have proved to be essential for the metabolomic studies and for better understanding of the global chemical composition of different vegetable samples.

Untargeted and targeted chemical profiling for efficacy-directed discrimination of Hedera helix L. subspecies using HPTLC- image analysis and HPTLC/MS

A strategy combining untargeted and targeted chemical profiling for discrimination of the different subspecies of Hedera helix L (Ivy) based on HPTLC fingerprints and antioxidant and antimicrobial activity testing followed by chemometric analysis has been implemented for the first time for the holistic profiling of bioactive secondary metabolites in the different subspecies. Principal component analysis (PCA) based on HPTLC-image analysis fingerprints of secondary metabolites loading plots were utilized for determination of chemical markers responsible for the classification which were then identified using HPTLC/MS technique as the six phenolics; caffeic acid, kaempferol-3-O-glucoside, quercetin-3-O-glucoside, kaempferol-3-O-rutinoside, rutin and chlorogenic acid and the three saponins; alpha-hederin, hederasaponin B and hederacoside C. The identified markers were then quantified using a HPTLC-high resolution plate imaging technique for targeted profiling. Biomarkers were determined from the coefficients plot of orthogonal projection to latent structures (OPLS) model based on the secondary metabolites quantitative data together with the biological activity data which revealed that the phenolic metabolites were strongly correlated to the antioxidant and antimicrobial activity data points compared to saponins. According to targeted hierarchical clustering analysis (HCA)-heat-map, samples could be divided into three main clusters; H. helix subsp. helix cluster, H. helix subsp. rhizomatifera and H. helix subsp. hiberinica sub-clusters and H. helix subsp. poetrum and H. helix subsp. canariensis sub-clusters, and such clustering pattern was directly related to the variation in the content of bioactive phenolics among the different subspecies rather than their saponins content, providing an insight into the unsettled taxonomy of the ivy subspecies and successful precise efficacy-directed discrimination of the different samples.