Search

Background and aim: Echinodorus macrophyllus (Kunth.) Micheli is popularly used for acute and chronic inflammatory conditions. The anti-inflammatory activity was previously demonstrated for its flavonoid-enriched fractions. The aim of this work assessed the antinociceptive properties of both aqueous extract and its fractions. Experimental procedure: The antinociceptive activity was determined by acetic acid-induced writhing, formalin test, tail immersion test, hot-plate test, xylene-induced ear edema methods, and the evaluation of its mechanism was performed in the writhing model. The aqueous extract of Echinodorus macrophyllus (AEEm) was fractionated, yielding Fr20, and Fr40. Fr40 composition was determined by HPLC-DAD-ESI-MS. Results and conclusion: Fr20 (all doses) and Fr40 (100 mg/kg) reduced the nociception in the tail-flick model. Both fractions increased the percentage of maximum possible effect with 25 mg/kg, in the hot-plate assay, at 60 min, while AEEm reduced pain only with 50 and 100 mg/kg. There was a reduction in xylene-edema index, with Fr40 (25 mg/kg), AEEm (50 mg/kg) and Fr20 (50 mg/kg). All doses of AEEm, Fr20, and Fr40 reduced both phases of the formalin model. In the abdominal contortion model, Fr40 presented the highest activity, reducing 96% of contortions and its antinociceptive mechanism was evaluated. The results indicated the involvement of NO and adrenergic activation pathways. The main components of Fr40 are swertisin, swertiajaponin, isoorientin 7,3′-dimethyl ether, swertisin-O-rhamnoside, isoorientin, isovitexin, isovitexin-Orhamnoside, and isovitexin-7-O-glucoside. The aqueous extract of E. macrophyllus leaves and its fractions exhibited significant analgesic effect, mediated through both peripheral and central mechanisms being considered a potentially antinociceptive drug.

Cissus rotundifolia has been reported to possess various biological activities such as anti-diabetic, anti-fertility, anti-hyperlipidemic, anti-malarial, anti-osteoporotic, and anti-parasitic activities. Therefore in the present study, eleven selected constituents of Cissus rotundifolia which includes aconitic acid, astragalin, acteoside, aliospiroside A, beta amyrin, bergenin, formononetin, gallic acid, isovitexin, isoorientin, and isoquercitrin were studied on the docking behavior of human neutrophil elastase (HNE), matrix metalloproteinases (MMP 2 and MMP 9), and tyrosinase by using PatchDock method. Furthermore, molecular physicochemical, bioactivity score/drug-likeness, ADME (absorption, distribution, metabolism, and excretion), and toxicity analyses were also carried out using Molinspiration, Swiss ADME, and ProTox-II methods, respectively. The molecular physicochemical investigation showed that three ligands such as acteoside, aliospiroside A, and isoorientin have three violations for Lipinski’s rule of five. Similarly, ADME analysis one ligand (formononetin) predicated to have high blood-brain barrier (BBB) permeability effect. The docking studies showed that isovitexin exhibited the highest atomic contact energy (−341.61 kcal/mol) for human neutrophil elastase (HNE), more over alliospiroside A has shown maximum atomic contact energy for both matrix metalloproteinases (MMP 2 [−618.00 kcal/mol] and MMP 9 [−634.73 kcal/mol]). Furthermore, isoquercitrin has exhibited the highest atomic contact energy (−145.70 kcal/mol) for tyrosinase. Thus, the present investigation outcome provides new knowledge in understanding eleven Cissus rotundifolia constituents as possible novel inhibitors against HNE, MMP 2, MMP 9, and tyrosinase.

There is increasing interest in evaluating anti-inflammatory activities of plant substances such as extracts and flavonoid rich fractions. A promising source of new medicinal drugs may be species from the Passifloraceae family. The most interesting group of principal chemical substances in Passiflora species are polyphenolic compounds, including flavonoids (quercetin, rutin, apigenin, luteolin, chrysin, and C-glycosylflavones i.e., vitexin, isovitexin, orientin, isoorientin), due to their antioxidant activity demonstrated in various studies. However, each extract from Passiflora spp. as multi-component mixtures should be estimated for chemical composition (in the standardization process) and its activity using in vitro and in vivo tests. The current standard for drug discovery and development from plants indicates that only collective assessment allows estimating plant substances by definition of the origin of raw materials and their quality, methods of extractions, and metabolite profiles. Increasingly, due to complex phytochemical procedures to obtain extracts, individual flavonoid compounds are also tested for anti-inflammatory action. However, it should be emphasized that various sources of potential new drugs from plant origin are not mutually exclusive, but are complementary. A review of bibliographic data includes the following information about Passiflora species, such as distribution, classification, phytochemical compounds, the anti-inflammatory activity of the extracts, the anti-inflammatory activity of flavonoids, and antioxidant potential. The review concluded that extracts and flavonoids (mainly quercetin, apigenin, and vitexin) from Passiflora spp. can be a valuable source of anti-inflammatory and anti-oxidative medications for the prevention and treatment of many diseases, which occur with complex inflammatory processes.

An analytical method for the simultaneous determination of ten major functional flavonoids (isoorientin, orientin, vitexin, isovitexin, apigenin, luteolin, tricin, quercetin, rutin, and kaempferol) in different bamboo species was developed by liquid chromatography-tandem mass spectrometry. Chromatographic separation was carried out on a reversed-phase C-18 column with acetonitrile and water as the mobile phases. Detection was performed in negative ion electrospray ionization mode using multiple reaction monitoring mode. The correlation coefficients for the calibration curves ranged from 0.9955 to 0.9997. The limit of detection ranged from 1 to 45 ng/ml. The applicability of this analytical approach was confirmed by the successful analysis of real leaf samples of four bamboo species, family Poaceae: Pleioblastus amarus (Keng) Keng f., Phyllostachys glauca McClure, Phyllostachys edullis (Carrière) J.Houz, and Indocalamus latifolius (Keng) McClure. The total flavonoid contents were 3321.09, 3095.96, 4037.33, and 2808.42 mg/kg for P. amarus, P. glauca, P. edullis, and I. latifolius, respectively. Graphical abstract

The present work realized the chemical study of Silene sibirica (L.) Pers. (Caryophyllaceae) high-performance liquid chromatography with diode array and mass spectrometric detection (electrospray ionization) (HPLC-DAD-ESI-MS). As a result, 25 compounds were found, including nine ecdysteroids and sixteen flavonoids. Ecdysteroids components were podecdysone C, integristerone A, turkesterone, polypodine B, 20-hydroxyecdysone and its 2-O-cinnamate, ecdysone, 2-deoxy-20-hydroxyecdysone, and 2-deoxyecdysone. Flavonoids were the glycosylflavones and derivatives of luteolin and apigenin. Luteolin glycosides included luteolin-7-O-rutinoside, lucenin-2, carlinoside, isoorientin and its 2″-O-arabinoside, as well as isoscoparin. The largest group of apigenin glycosides included O-glycosides as 7-O-glucoside and 7-O-rutinoside, C-glycosides as isovitexin, schaftoside, vicenin-2, and mixed C,O-glycosides as isovitexin-2′′-O-arabinoside and 2′′-O-rhamnoside. Three unidentified derivatives of apigenin were pre studied and their structural features discussed. Quantitative data about the content of selected compounds indicated that ecdysteroids accumulated in S. sibirica flowers (7.14–14.92 mg/g) and glycosylflavones were found predominantly in leaves (7.88–18.55 mg/g). The major ecdysteroid compound was 20-hydroxyecdysone, while flavonoids predominants were shaftoside and isovitexin-2″-O-ramnoside. A comparative analysis of the chemical composition of wild-growing and cultivated S. sibirica samples showed the stability of the metabolic profile of the plants during the introduction. The biological studies revealed the antiradical and antiglucosidase activity of the extracts. Thus, it was shown that the studied plant species (S. sibirica) is a source of ecdysteroids and glycosylflavones, and S. sibirica extracts have biological potency.

Objective: phytochemical investigation of the ethyl acetate fraction (EAF) of 80% aqueous methanol extract (AME) of Dypsis leptocheilos leaves, in addition to evaluation of the antioxidant, cytotoxic and antimicrobial activities of the AME and EAF. Docking was used to predict and understand cytotoxicity of the isolated compounds. Methods: The ethyl acetate fraction (EAF) of Dypsis leptocheilos leaves was subjected to different chromatographic separation techniques. Structures of the isolated compounds were established by different spectroscopic techniques (1H/13C NMR). Antioxidant activity was evaluated by DPPH assay, while cytotoxicity was evaluated by MTT cell viability assay. Antimicrobial activity was evaluated by agar diffusion method. The docking study was conducted using Auto Dock Vina; the estrogen receptor (PDB 5t92) was used as a receptor for the docking. Results: Chromatographic separation techniques were led to the isolation of five phenolic compounds; these compounds were identified to be apigenin 8-C-β-D-glucopyranoside (Vitexin) (1), apigenin 6-C-β-D-glucopyranoside (Isovitexin) (2), luteolin 7-O-β-D-glucopyranoside (3), luteolin 8-C-β-D-glucopyranoside (Orientin) (4), luteolin 6-C-β-D-glucopyranoside (Isoorientin) (5). They were isolated and identified for the first time from this plant species. The AME and EAF showed moderate activity against Gram positive and Gram negatvie bacteria, while both of them showed similar and powerful antioxidant activity with SC50 = 12.8±0.56 µg/ml and SC50 = 17±0.77 µg/ml respectively, compared to ascorbic (reference drug) SC50 = 14.2±0.35 µg/ml. The EAF showed higher cytotoxic activity on the MCF-7 cells (human breast cancer cell line), with IC50 = 12.3 ± 1.82 µg/ml, compared to Vinblastine Sulfate (reference drug). All isolated compounds showed good binding affinity to the estrogen receptors existed in the MCF-7 cell. Conclusion: Five phenolic compounds were isolated for the first time from the EAF of Dypsis leptocheilos leaves. The AME and EAF extracts showed variable antioxidant, antimicrobial and cytotoxic activities.

V magistrskem delu smo ovrednotili spremembe v spektru fenolnih spojin med procesom priprave ajdovega slada. V poskusu smo uporabili semena dveh sort (pyra in darja) navadne ajde (Fagopyrum esculentum Moench), ki smo jih namakali, kalili, posušili in v različnih fazah procesa odvzemali vzorce za analize vsebnosti vode in fenolnih spojin (nekaljena ajda, 8 ur namočena ajda, kaljena ajda po 16-ih, 40-ih, 64-ih in 88-ih urah, ajdov slad sušen 22 ur pri 60 °C in 18 ur pri 80 °C) Vsebnost vode smo določali s sušenjem, sestavo in vsebnost fenolnih spojin po ekstrakciji z metanolom pa smo določili s tekočinsko kromatografijo z detekcijo na masnem spektrometru (LC-MS). V vzorcih ajde smo določili 12 fenolnih spojin (rutin, orientin, viteksin, izoviteksin, kvercetin, izoorientin, kavno kislino, luteolin, kamferol, naringenin, apigenin in galno kislino). Med kaljenjem se je vsebnost vseh fenolnih spojin v obeh sortah ajde značilno povečala. Najpomembnejša fenolna spojina v nekaljeni ajdi je rutin, ki smo ga tudi v kaljeni ajdi določili največ (pri nekaljeni ajdi v sorti pyra 91,56 μg/g in v sorti darja 103,07 μg/g pri kaljeni ajdi 88 ur v sorti pyra 355,60 μg/g in v sorti darja 299,95 μg/g). Največje povečanje vsebnosti med kaljenjem smo določili pri orientinu (za faktor 29,2 pri pyri in 21,3 pri darji), viteksinu (za faktor 15,2 pri pyri in 14,0 pri darji) in izoviteksinu (za faktor 17,7 pri pyri in 15,7 pri darji). Pri obeh režimih sušenja se je vsebnost vseh fenolnih spojin v obeh sortah ajde značilno zmanjšala, vendar je bila skupna vsebnost fenolnih spojin v ajdovem sladu še vedno za faktor 2,2 do 3,4 (odvisno od sorte in načina sušenja) večja od skupne vsebnosti fenolnih spojin v nekaljeni ajdi. V ajdi sorte pyra smo po zaključenem kaljenju in sušenju določili nekoliko večjo vsebnost fenolnih spojin kot v ajdi sorte darja. Kaljena ajda in ajdov slad sta dober vir prehransko pomembnih fenolnih spojin. In the master thesis we evaluated the changes in the spectrum of phenolic compounds during the process of buckwheat malting. In the experiment we used the seeds of two varieties (Pyra and Darja) of common buckwheat (Fagopyrum esculentum Moench), which were soaked, germinated and dried. At different stages of the process, samples were taken for the analysis of water content and determination of various phenolic compounds (unmalted buckwheat, buckwheat soaked for 8 hours, germinated buckwheat after 16, 40, 64 and 88 hours, dried buckwheat for 22 hours at 60 °C and 18 hours at 80 °C). Water content was determined by drying and the content of phenolic compounds after extraction with methanol was determined by liquid chromatography with mass spectrometer detection (LC-MS). Twelve phenolic compounds (rutin, orientin, vitexin, isovitexin, quercetin, isoorientin, caffeic acid, luteolin, kaempferol, naringenin, apigenin and gallic acid) were determined in buckwheat samples. During germination, the content of all phenolic compounds increased significantly in both buckwheat cultivars. The most important phenolic compound in buckwheat is rutin, which was also the most abundand in germinated buckwheat (91.56 μg/g in unmalted buckwheat in Pyra cultivar and 103.07 μg/g in Darja cultivar in 88 hours germinated buckwheat in Pyra cultivar 355.60 μg/g and 299.95 μg/g in Darja cultivar). The greatest increase in content during germination was found for orientin (factor 29.2 in Pyra and 21.3 in Darja), vitexin (factor 15.2 in Pyra and 14.0 in Darja) and isovitexin (factor 17.7 in Pyra and 15.7 in Darja). Under both drying regimes, the content of all phenolic compounds in both buckwheat cultivars decreased significantly, but the sum of phenolic compound content in the buckwheat malt was still a factor 2.2 to 3.4 (depending on cultivar and drying regime) higher than the sum of phenolic compound content in the unmalted buckwheat. After germination and drying, the buckwheat cultivar Pyra was found to have a slightly higher content of phenolic compounds than the buckwheat cultivar Darja. Germinated buckwheat and buckwheat malt are good sources of nutritionally important phenolic compounds.

Several medical plants belonging to the genera Passiflora, Viola, and Crataegus accumulate flavonoid C-glycosides, which likely contribute to their efficacy. Information regarding their phase I and II metabolism in the liver are lacking. Thus, in vitro liver metabolism of orientin, isoorientin, schaftoside, isoschaftoside, vitexin, and isovitexin, all of which accumulated in Passiflora incarnata L., was investigated by incubation in subcellular systems with human liver microsomes and human liver S9 fraction. All metabolite profiles were comprehensively characterized using HPLC-DAD and UHPLC–MS/MS analysis. Mono-glycosylic flavones of the luteolin-type orientin and isoorientin showed a broad range of mono-glucuronidated and mono-sulfated metabolites, whereas for mono-glycosylic flavones of the apigenin-type vitexin and isovitexin, only mono-glucuronidates could be detected. For di-glycosylic flavones of the apigenin-type schaftosid and isoschaftosid, no phase I or II metabolites were identified. The main metabolite of isoorientin was isolated using solid-phase extraction and prep. HPLC-DAD and identified as isoorientin-3′-O-α-glucuronide by NMR analysis. A second isolated glucuronide was assigned as isoorientin 4′-O-α-glucuronide. These findings indicate that vitexin and isovitexin are metabolized preferentially by uridine 5′-diphospho glucuronosyltransferases (UGTs) in the liver. As only orientin and isoorientin showed mono-sulfated and mono-glucuronidated metabolites, the dihydroxy group in 3′,4′-position may be essential for additional sulfation by sulfotransferases (SULTs) in the liver. The diglycosylic flavones schaftoside and isoschaftoside are likely not accepted as substrates of the used liver enzymes under the chosen conditions.

In this study, a valid and comprehensive evaluation method for assessing the quality of Desmodium styracifolium (Osb.) Merr has been established, based on analysis of high-performance liquid chromatography fingerprint combined with the similarity analysis (SA), hierarchical cluster analysis (HCA), principal component analysis (PCA), discriminant analysis (DA) and the quantitative analysis multi-components by single marker (QAMS) method. Eleven peaks of the common model were obtained and analyzed using SA, HCA, PCA and DA analysis. These methods indicated a similar conclusion that 31 batches of D. styracifolium samples were categorized into two clusters basically coincident with their geographical regions of origin. Four peaks were identified as schaftoside, isoorientin, isoschaftoside and isovitexin. Schaftoside was selected as the internal standard, and the relative correction factors between schaftoside and the other three flavonoids were calculated using the QAMS method. The accuracy of the QAMS method was verified by comparing with the results calculated by the external standard method. No significant difference between the two methods was found. In conclusion, the established methods were scientifically applied in the quality evaluation of D. styracifolium.

Isoorientin and isovitexin, kinds of flavone C-glycosides, exhibit a number of biological properties. In this work, The C-glucosyltransferase (Gt6CGT) gene from Gentiana triflora was cloned and expressed in Escherichia coli BL21(DE3). The optimal activity of Gt6CGT was at pH 7.5 and 50° C. The enzyme was stable over pH range of 6.5–9.0, and had a 1-h half-life at 50° C. The Vmax for luteolin and apigenin was 21.1 nmol/min/mg and 31.7 nmol/min/mg, while the Km was 0.21 mM and 0.22 mM, respectively. Then, we developed an environmentally safe and efficient method for isoorientin and isovitexin production using the coupled catalysis of Gt6CGT and Glycine max sucrose synthase (GmSUS). By optimizing coupled reaction conditions, the titer of isoorientin and isovitexin reached 3820 mg/L with a corresponding molar conversion of 94.7% and 3772 mg/L with a corresponding molar conversion of 97.1%, respectively. The maximum number of UDP-glucose regeneration cycles (RCmax) reached 28.4 for isoorientin and 29.1 for isovitexin. The coupled catalysis reported herein represents a promising method to meet industrial requirements for large-scale isoorientin and isovitexin production in the future.

Otholobium pubescens (Poir.) J.W. Grimes (O. pubescens) is a commercial and multiple therapeutic medicinal plant in South America, yet its phytochemicals are rarely studied. In the present study, a combination of affinity-based ultrafiltration, HSCCC, and preparative HPLC was applied for the efficient screening and isolation of ten tyrosinase inhibitors from the O. pubescens methanol extract and eight of them were successfully identified as daidzin, isoorientin, vitexin, isovitexin, isoorientin 3′-methyl ether, daidzein, genistein, and a novel compound 3-(5-hydroxybenzofuran-6-yl) propanoic acid by EI-MS and NMR; all of these compounds are first reported in O. pubescens and possessed tyrosinase inhibitory activities by in vitro tyrosinase inhibition assay. Among the inhibitors, vitexin, isovitexin, 3-(5-hydroxybenzofuran-6-yl) propanoic acid, and daidzein revealed higher tyrosinase inhibitory activities than the positive control β-arbutin, with IC50 values of 0.35 mM, 1.73 mM, 1.33 mM, 1.57 mM, and 1.83 mM, respectively. Moreover, a long time tyrosinase inhibition duration, up to 150 min, was observed for compounds vitexin, daidzein, and particularly 3-(5-hydroxybenzofuran-6-yl) propanoic acid, which was assessed as a special mixed-type tyrosinase inhibitor because of its capability of inhibiting tyrosinase almost in an irreversible inhibition manner, through tyrosinase inhibition duration, inhibition kinetics, and dialysis studies. In addition, the major nonpolar compound from O. pubescens methanol extract was also purified by silica gel column chromatography and identified as bakuchiol. Results suggested that vitexin, isovitexin, daidzein, and particularly the novel compound 3-(5-hydroxybenzofuran-6-yl) propanoic acid are promising tyrosinase inhibitors with the potential for pharmaceutical, cosmetic, and food industrial applications.

Purpose: Discovering new antimyocardial ischemia drug candidates that are highly efficient, have low toxicity, and originate from natural products is a popular trend for new cardiovascular drug development at present. The ethanol extract of Livistona chinensis leaves showed a favorable antioxidant activity in our preliminary screening test. This study aims to screen out antioxidants from the herb leaves further and evaluate their efficacy in acute myocardial ischemia treatment at the cellular level. Materials and methods: Guided with online 1, 1-diphenyl-2-picrylhydrazyl (DPPH)-high-performance liquid chromatography (HPLC) screening, antioxidants were first separated and isolated from the ethanol extract of L. chinensis leaves by preparative-HPLC. Subsequently, offline DPPH approach was used to validate the free radical scavenging activity of the components. Ultimately, the resulting antioxidants were evaluated against the hypoxia/reoxygenation (H/R)-, H2O2-, or adriamycin (ADM)-induced injury in H9c2 cells to verify their cardioprotective effects in vitro. Results: Five antioxidant ingredients, namely, orientin, isoorientin, vitexin, isovitexin, and tricin, were quickly distinguished and isolated from L. chinensis leaves. The IC50 values of these ingredients were further examined by offline DPPH assay, as follows: 15.51±0.22, 6.64±0.38, 11.86±0.24, 8.89±0.66, and 31.86±0.24 μg/mL, respectively. Out of these ingredients, isoorientin showed the strongest antioxidation, which was equivalent to that of the positive control drug (vitamin C, IC50: 6.99±0.62 μg/mL). Using H/R-, H2O2-, and ADM-induced H9c2 cell injury models, the five ingredients had different extents of cardioprotective effects in vitro. In particular, isoorientin showed the strongest protection. All the five ingredients also showed insignificant cytotoxic effect to normal H9c2 cells. Conclusion: The ethanol extract of L. chinensis leaves contained five antioxidants with low cardiac cytotoxicity. Isoorientin possessed the strongest antioxidation, which can predominantly account for the myocardial protection effects within the extract.

Introduction Plant medicine/herbal extracts are typically complex, encompassing a wide range of flavonoid diversity and biological benefits. Combined with a lack of standards; species authentication profiling is a challenge. A non-targeted screening strategy using two-dimensional (2D) separation and specificity of ultra-high-performance liquid chromatography ion mobility collision-induced dissociation mass spectrometry (UHPLC-IM-CID-MS) has been investigated, to identify the 6-C and 8-C-glycosylflavone isomer orientin/isoorientin and vitexin/isovitexin pairs in Passiflora species. Utilising available standards and "known-unknowns" a reference CCS (collision cross-section) speciation finger print for Passiflora extracts could be generated to illustrate species profiling. Material and methods SPE was performed to extract flavonoids of interest from powdered and ground Passiflora leaf. Chromatographic separation was achieved via UHPLC and analysis performed using positive/negative ion electrospray coupled with linear T-wave IM-MS (calibrated to perform accurate mass and CCS measurements). Results Comparative phytochemical screening of Passiflora alata, P. edulis, P. incarnata and P. caerulea leaf extracts has generated CCS, CID IM product ion spectra, 2D separation with UHPLC-IM-MS, enabling the unequivocal identification of flavone C-glycosides in complex extracts. A phytochemical reference CCS library was generated comprised of "knowns" and "known-unknowns". Isomers have been differentiated using a CCS metric enabling novel CCS specific isomeric quantitation of co-eluting isomers. Conclusions The screening approach illustrated has the potential to play an important role in the profiling of medicinal plants to determine phytochemical make-up and improve consumer safety through generation of highly specific speciation profiles.

Background Himalayan Columbine (Aquilegia pubiflora Wall. Ex Royle) is a medicinal plant and have been used as traditional treatments for various human diseases including skin burns, jaundice, hepatitis, wound healing, cardiovascular and circulatory diseases. Till now there is no report available on phytochemical investigation of Himalayan Columbine and to the best of our knowledge, through present study we have reported for the first time, the phytochemical analysis and pharmacological potentials of different leaf extracts of Aquilegia pubiflora. Methods Four types of extracts were prepared using solvent of different polarities (Distilled water APDW, Methanol APM, Ethanol APE and Ethyl acetate APEA), and were evaluated to determine the best candidate for potent bioactivity. Phytochemical constituents in prepared extracts were quantified through HPLC analysis. Subsequently, all four types of leaf extracts were then evaluated for their potential bioactivities including antimicrobial, protein kinase inhibition, anti-inflammatory, anti-diabetic, antioxidant, anti-Alzheimer, anti-aging and cytotoxic effect. Results HPLC analysis demonstrated the presence of dvitexin, isovitexin, orientin, isoorientin, ferulic acid, sinapic acid and chlorogenic acid in varied proportions in all plant extracts. Antimicrobial studies showed that, K. pneumonia was found to be most susceptible to inhibition zones of 11.2 ± 0.47, 13.9 ± 0.33, 12.7 ± 0.41, and 13.5 ± 0.62 measured at 5 mg/mL for APDW, APM, APE and APEA respectively. A. niger was the most susceptible strain in case of APDW with the highest zone of inhibition 14.3 ± 0.32, 13.2 ± 0.41 in case of APM, 13.7 ± 0.39 for APE while 15.4 ± 0.43 zone of inhibition was recorded in case of APEA at 5 mg/mL. The highest antioxidant activity of 92.6 ± 1.8 μgAAE/mg, 89.2 ± 2.4 μgAAE/mg, 277.5 ± 2.9 μM, 289.9 ± 1.74 μM for TAC, TRP, ABTS and FRAP, respectively, was shown by APE. APM, APE and APEA extracts showed a significant % cell inhibition (above 40%) against HepG2 cells. The highest anti-inflammatory of the samples was shown by APE (52.5 ± 1.1) against sPLA2, (41.2 ± 0.8) against 15-LOX, followed by (38.5 ± 1.5) and (32.4 ± 0.8) against COX-1 and COX-2, respectively. Conclusions Strong antimicrobial, Protein Kinase potency and considerable α-glucosidase, α-amylase, and cytotoxic potential were exhibited by plant samples. Significant anti-Alzheimer, anti-inflammatory, anti-aging, and kinase inhibitory potential of each plant sample thus aware us for further detailed research to determine novel drugs.

Natural populations of Gentiana asclepiadea L., located at two mountainous sites, were HPLC-analyzed regarding the contents of six representative secondary metabolites. The contents of swertiamarin (SWM), gentiopicrin (GP), sweroside (SWZ), mangiferin (MGF), isoorientin (ISOOR), and isovitexin (ISOV) were determined in six populations (three per study site), and separately for aboveground and belowground plant parts. PCA showed a clear separation of four groups according to the contents of the analyzed secondary metabolites. Out of six analyzed compounds, five were present in all samples and only one (SWZ) was found in Golija populations (belowground parts) but not in Vlasina populations, and its presence can be indicative of the geolocation of populations. Clear separation of groups was mostly affected by the different contents of chemical compounds in plant parts (aboveground versus belowground) and by the differences related to population origin (higher content of SWM and GP in belowground parts of individuals from Vlasina populations and higher content of MGF and ISOOR of individuals from Golija populations). The results of this study contribute to the spatiochemical profiling of G. asclepiadea populations and a better understanding of inter- and intrapopulation variability of pharmacologically important compounds.

Passiflora caerulea L. (Passifloraceae) (“mburucuya”, “maracuja-laranja”), a widely distributed species in South America, has been traditionally considered as a medicinal, ornamental, and edible plant. Its fruits, besides being well appreciated for their flavor and sweetness, have valuable nutritional properties. The ethnopharmacological use as anxiolytic, antistress and sedative of the non-reproductive aerial parts (leaves and tendrils) from P. caerulea is the main reason for its commercialization as an infusion herb. In general, its preparations are rich in flavonoids (chrysin and C-glycosides such as vitexin, isovitexin, orientin, isoorientin, and vicenin-2, among others), which were confirmed as responsible for the anxiolytic effect. The Argentine Pharmacopeia includes a monograph on P. caerulea, while other Pharmacopeias (Brazilian, European, and British) consider it as a potential adulterant of P. incarnata and P. edulis. Harmala alkaloids from P. caerulea might also contribute to the anxiolytic effect, but no preclinical assays have been conducted to confirm it. Pharmacological research confirmed other bioactivities associated with the aerial parts of P. caerulea, such as antimicrobial, gastroprotective, analgesic, and anticonvulsant. Even though, no systematic toxicological assessments have been conducted in order to evaluate the safety in the consumption, especially regarding the presence of cyanogenic glycosides both in the vegetal drug and the unripe fruits.

Luteolin and apigenin derivatives present in oil palm (Elaeis guineensis) leaves (OPL) are reported to possess excellent antioxidant properties relating to numerous health benefits. To meet the global demand for flavonoids, OPL, which is plentifully generated as an agricultural by-product from oil palm plantations, can be further exploited as a new source of natural antioxidant compounds. However, to produce a standardized herbal preparation, validation of the quantification method for these compounds is required. Therefore, in this investigation, we developed and validated an improved and rapid analytical method, ultra-high-performance liquid chromatography equipped with ultraviolet/photodiode array (UHPLC-UV/PDA) for the quantification of 12 luteolin and apigenin derivatives, particularly focusing on flavonoid isomeric pairs: orientin/isoorientin and vitexin/isovitexin, present in various OPL extracts. Several validation parameters were assessed, resulting in the UHPLC-UV/PDA technique offering good specificity, linearity, accuracy, precision, and robustness, where the values were within acceptable limits. Subsequently, the validated method was employed to quantify luteolin and apigenin derivatives from OPL subjected to different drying treatments and extraction with various solvent systems, giving total luteolin (TLC) and apigenin content (TAC) in the range of 2.04–56.30 and 1.84–160.38 µg/mg extract, respectively. Additionally, partial least square (PLS) analysis disclosed the combination of freeze dry-aqueous methanol yielded OPL extracts with high TLC and TAC, which are strongly correlated with antioxidant activity. Therefore, we provide the first validation report of the UHPLC-UV/PDA method for quantification of luteolin and apigenin derivatives present in various OPL extracts, suggesting that this approach could be employed in standardized herbal preparations by adopting orientin, isoorientin, vitexin, and isovitexin as chemical markers.

Objectives This study was carried out to evaluate the effects of flavonoids present in leaves of Passiflora edulis fruit on complications induced by diabetes in rats. Methods The extract of P. edulis leaf was obtained by 70% ethanol maceration. From the dry extract, the fractions were obtained by consecutive liquid–liquid partition with hexane, ethyl acetate and n-butanol. The content of isoorientin of ethyl acetate and n-butanol fractions was determined by ultra-performance liquid chromatography coupled with electrospray and triple quadrupole ionization (TQD) analysis in tandem mass spectrometry (UPLC–ESI-Tq-MS). Only Fr-BuOH was used to treat diabetic or not Wistar rats. Biochemical parameters, platelet aggregation and production of reactive species were evaluated. Key findings The UPLC–ESI-Tq-MS analysis revealed the presence of several flavonoids, among which we identified five possible flavonoids c-heterosides (luteolin-7-O-pyranosyl-3-O-glucoside, apigenin-6-8-di-C-glycoside, apigenin-6-C-arabinoside-8-C-glycoside, isoorientin, isovitexin). The diabetic rats (treated intraperitoneally with alloxan, 150 mg/kg) treated with Fr-BuOH (20 mg/kg/day for 90 days) presented improvement in blood glucose, serum levels of fructosamine, lipid profile and urea. Furthermore, the Fr-BuOH reduced both platelet aggregation and the production of oxidant species in diabetic animals. Conclusions These results suggested that flavonoid C-glycosides present in the Fr-BuOH may be beneficial for the diabetic state, preventing complications induced by diabetes.

Brazilian biodiversity and favourable environmental conditions open up possibilities not yet explored, showing potential to shift the country’s monochromatic economy into an emancipated, diversified and sustainable economic environment. This can be made possible through the integral use of its resources, exploring every functional fraction to create novel solutions to modern problems. Biorefineries present an interesting strategy to fully use the potential of agricultural feedstocks and together with green separation methods can contribute to the generation of sustainable processes and products. Passion Fruit (Passiflora edulis Sims f. flavicarpa Deg species) is produced on a large scale in Brazil and in other tropical countries, and its processing plants generate tons of residues that basically consist of peel, seeds and bagasse, which account for around 75% of its mass. These fractions of P. edulis can contain significant amounts of flavonoids, secondary metabolites that are the main compounds responsible for the fruit’s bioactivity (antioxidant, anti-inflammatory, pesticide and biocide, in general). Therefore, this work aims to develop, apply and compare the best conditions for the extraction of isoorientin, orientin and isovitexin from passion fruit applying solid–liquid methodologies, followed by analyte quantification using UHPLC-PDA. Homogenizer-assisted (HAE), ultrasound-assisted (UAE) and microwave-assisted (MAE) extraction techniques were used, as well as a full factorial design to reach optimal parameters concerning the extraction yield and energy and solvent efficiencies. According to the results, the procedure based on HAE presented the best conditions for the extraction of selected flavonoids (1.07, 0.90 and 0.33 mg g−1 of isoorientin, orientin and isovitexin, respectively) and was considered the best method according to the green and sustainable described factors.

Cissus erosa (Vitaceae), popularly known in Brazil as Cipó-fogo, is a medicinal plant used in the treatment of warts and external ulcers. The present study aimed to evaluate the activity of stems and leaves ethanol extracts of C. erosa against the Dengue and Zika virus by the MTT colorimetric method and to carry on the phytochemical characterization of active extracts by high performance liquid chromatography coupled to mass spectrometry (UPLC-MS). Only the leaves ethanol extract showed anti-Dengue virus activity EC50 18.2 µg/ml (SI > 27.5) and low cytotoxicity for LLCMK2 cells (CC50 > 500 mg/ml). Both extracts (stems and leaves) showed anti-Zika virus activity with EC50 of 45.8 mg/ml and 82.8 mg/ml, respectively. These extracts presented CC50 of 309.2 µg/ml (leaves) and 387.6 µg/ml (stems) to Vero cells. Analysis by CCD and HPLC-DAD detected the presence of triterpenes, steroids, flavonoids and tannins. UPLC-MS analyses of these extracts, allowed the identification of the majority of flavonoids present known as vitexin, isovitexin, orientin, isoorientin and two flavones derivatives, methoxyluteolin-6(8)-C-hexosyl and luteolin-7,4’-di-O-glycosylflavone. The results of the phytochemical studies here described suggest that flavonoids and terpenoids are the substances that contribute to the antiviral activity of the ethanol extracts within this species. Resumo A espécie Cissus erosa (Vitaceae) conhecida popularmente, no Brasil, como Cipó fogo é uma planta medicinal empregada no tratamento de verrugas e úlceras externas. O presente estudo objetivou a avaliação da atividade antiviral frente a Dengue e Zika virus dos extratos etanólicos de caules e de folhas desta espécie pelo método colorimétrico do MTT e determinar a caracterização fitoquímica por cromatografia líquida de ultra eficiência acoplada a espectrometria de massa destes extratos (CLUE-EM). Os extratos apresentaram atividade antiviral e baixa citotoxicidade. O extrato de folhas apresentou CC50 > 500 µg/ml para células LLCMK2 e CE50 = 18,2 µg/mL, IS > 27,5 contra o Dengue virus 2 (DENV 2). Ambos os extratos (caules e folhas) apresentaram atividade anti-Zika virus com CE50 de 45,8 µg/ml e 82,8 µg/ml, respectivamente. Os extratos apresentaram CC50 de 309,2 µg/ml (folhas) e 387,6 µg/ml (caules) para células Vero. Nas análises por CCD e CLAE-DAD detectou-se a presença de triterpenos, esteroides, flavonoides e taninos. Análises por CLUE-EM dos extratos permitiram a identificação parcial dos flavonoides majoritárias destes extratos como sendo, vitexina, isovitexina, orientina, isoorientina, luteolina-7,4’-di-O-glicosilflavona, metoxiluteolia-6(8)-C-hexosil. Os resultados dos estudos fitoquímicos até o momento sugerem que flavonoides e terpenoides são os constituintes majoritários dos extratos etanólicos destas espécies e devem contribuírem para a atividade antiviral observada in vitro.

Angiotensin-converting enzyme (ACE) is an important therapeutic target in the regulation of high blood pressure. This study was conducted to investigate the alterations in blood pressure associated with ACE inhibition activity of the polyphenols (1–10), including 3-O-feruloylquinic acid (1), lutonarin (2), saponarin (3), isoorientin (4), orientin (5), isovitexin (6), isoorientin-7-O-[6-sinapoyl]-glucoside (7), isoorientin-7-O-[6-feruloyl]-glucoside (8), isovitexin-7-O-[6-sinapoyl]-glucoside (9), and isovitexin-7-O-[6-feruloyl]-glucoside (10), isolated from barley seedlings (BS). All the isolated polyphenols exhibited comparable IC50 values of ACE inhibition activity (7.3–43.8 µM) with quercetin (25.2 ± 0.2 µM) as a positive control, and their inhibition kinetic models were identified as noncompetitive inhibition. Especially, compound 4 was revealed to be an outstanding ACE inhibitor (IC50 = 7.3 ± 0.1 µM, Ki = 6.6 ± 0.1 µM). Based on the compound structure–activity relationships, the free hydroxyl groups of flavone-moieties and glucose connections at the A ring of the flavone moieties were important factors for inhibition of ACE. The alcohol extract of BS also demonstrated potent ACE inhibition activity (66.5% ± 2.2% at 5000 µg mL−1). The polyphenols from BS had strong inhibitory activity on ACE and this study results suggest that BS can be used as an effective blood pressure regulator through ACE inhibition.

Oil palm (Elaeis guineensis Jacq.) leaves (OPL) are widely available at zero cost in Southeast Asia countries, especially in Malaysia and Indonesia due to large scale oil palm plantations. OPLs contain a large amount of flavonoids in particular flavonoid C-glycosides, which are known to possess useful biological properties including antioxidant and wound healing properties. The present study aimed at evaluating the wound healing efficacy of OPL in various solvent extracts and flavonoid enriched fractions and to determine the contribution of flavonoid C-glycosides (orientin, isoorientin, vitexin and isovitexin) using in-vitro scratch assay on 3T3 fibroblast cells. Solvent crude extracts with different polarity were screened and the most active extract was subjected to acid hydrolysis. The crude and acid hydrolysed extracts were further enriched using macroporous resins, XAD7HP. UHPLC-UV/PDA and LC-MS/MS analysis were applied for identification and confirmation of flavonoid C-glycosides. The wound healing properties comprised of cell viability, cell proliferation and cell migration were studied. Allantoin was used as a positive control to compare the efficacy among the tested samples. The results revealed all OPL crude extracts, flavonoid enriched fractions and flavonoid C-glycosides were non-toxic at concentrations below 25 &micro, g/mL and showed better cell proliferation and migration activities at low concentrations than higher concentrations.. This study also demonstrated orientin, isoorientin, vitexin and isovitexin presented in OPL extracts and flavonoid enriched fractions stimulated proliferation and migration of 3T3 fibroblast cells. Hence, these findings may pose potential therapeutic bioactive agents for wound healing by enhancing fibroblast proliferation and migration.

Trigonella foenum-graecum (fenugreek), a legume has been used as spice throughout the world to enhance the sensory quality of foods, is considered to have a high nutraceutical value. The antidiabetic actions of Trigonella seeds have been speculated due to the presence of steroid saponins, alkaloids, and fiber content. Five bioactive compounds, namely, trigonelline, isoorientin, orientin, vitexin, and isovitexin, were detected and quantified by HPLC system. Further rapid detection of bioactive compounds was achieved using high-performance liquid chromatography hybrid electrospray quadrupole time-of-flight mass spectrometric (HPLC-ESI-QTOF-MS/MS) method. Twenty-five compounds were tentatively identified based on their retention time and measured accurate mass. The fragmentation patterns of known compounds were utilized to elucidate their structures. Six of these identified compounds, that is, 4-hydroxyisoleucine, trigonelline, isoorientin, isovitexin, pinitol, and sarsasapogenin, were simultaneously quantified by UHPLC-ESI-MS/MS method under the multiple reactions monitoring (MRM) mode using ultrahigh-performance liquid chromatography hybrid electrospray triple quadrupole linear ion trap mass spectrometer. The analytical method was validated and successfully applied for simultaneous determination of compounds in Fenugreek seeds, which indicated the suitability of the validated method for its quality control.

A variety of neuroactive flavonoids can be found in the species of the Passiflora genus, however, their difficulty in crossing the blood&ndash, brain barrier limits their in vivo neuropharmacological activity. In this study, cationic nanoparticles were developed as a novel nanocarrier for improving the antidepressant activity of Passiflora edulis f. flavicarpa leaf extract. Formulations obtained using Eudragit E PO polymethylmethacrylate copolymer, as polymeric matrix had their physicochemical properties investigated. The analytical content of the flavonoids vicenin-2, orientin, isoorientin, vitexin, and isovitexin was determined in the plant extract. Small-sized and spherical nanoparticles loaded with Passiflora edulis f. flavicarpa were obtained with positive zeta potential and high encapsulation efficiency. In addition, the nanosystems were shown to be stable for at least 6 months. The antidepressant activity of P. edulis extract (50 and 100 mg/kg) as well as the extract-loaded nanoparticles (5 mg/kg) were investigated in mice using the forced swimming test, where the latter increased the potency of the former by 10-fold. In addition, histopathological and biochemical analysis confirmed the biocompatibility of the extract-loaded nanoparticles. This study demonstrated that the Eudragit cationic nanoparticles were able to improve the antidepressant activity of P. edulis in the central nervous system of mice.

Willow gentian (Gentiana asclepiadea L., Gentianaceae) is a medicinal plant that is primarily used in the treatment of gastrointestinal and liver diseases in folk medicine. As a bitter raw material, it is also used in the food industry for liquors flavoring. In order to optimize the ultrasound-assisted extraction (UAE) of gentiopicroside, isovitexin, isoorientin, and isogentisin, as well as total polyphenols from the underground parts of willow gentian, the influence of process parameters was investigated. Extraction time (from 5 to 65 min), ethanol concentration (from 10 to 90% v/v), solid to liquid ratio (from 1:10 to 1:50 w/v), and extraction temperature (from 20 to 80 °C) were estimated at five levels as independent variables. The response surface methodology (RSM) and the artificial neural network (ANN) approach were employed to develop a predictive extraction model and to establish optimal extraction conditions to simultaneously maximize the extraction yield of all target compounds. Established optimal values of process parameters were: extraction time 50 min, ethanol concentration 53% (v/v), solid to liquid ratio 1:40 (w/v), and extraction temperature 65 °C. The obtained extraction yields under predicted optimal conditions for gentiopicroside (49.23 mg/g DW), isoorientin (1.00 mg/g DW), isovitexin (4.04 mg/g DW), isogentisin (1.14 mg/g DW), and TPC (10.03 mg of GAE/g DW) confirmed the adequacy of both RSM and ANN models. Compared with the RSM model, the ANN model fitted better with the experimental values of each compound (higher coefficient of determination and lower mean absolute deviation) indicating its higher predictive capacity. The developed high-accuracy extraction model provides the basis for further separation and purification, as well as for studying the bioactivity of selected compounds from underground parts of G. asclepiadea.

A two-year field experiment was conducted to investigate the effects of different intercropping ratios and fertilizer types on antioxidant activity, total phenolic content (TPC), total flavonoids content (TFC) and content of main flavonoids of buckwheat seeds (Fagopyrum esculentum Moench). The treatments consisted of sole cropping of buckwheat (B), fenugreek (F), and three intercropping ratios (F:B = 1:2, 1:1 and 2:1) under three fertilizer types: chemical fertilizer (CF), integrated fertilizer (IF) and broiler litter (BL). The buckwheat seeds grown in intercropping had higher antioxidant activity measured by DPPH (20.2% in 2014 and 29.6 in 2015) and FRAP (16.9% in 2014 and 29.9% in 2015) assay, TPC (13.6% in both years), TFC (22.9% in 2014 and 11.3% in 2015), flavonoids such as rutin (12.4% in 2014 and 10.8% in 2015), vitexin (10.4% in 2014 and 14.2% in 2015), isovitexin (18.1% in 2014 and 7.8% in 2015), orientin (7.3% in 2014 and 7.07% in 2015), isoorientin (6.6% in 2014 and 9.6% in 2015) and hyperoside (25.1% in 2014 and 27.4% in 2015) than with sole cropping. The intercropping ratio of F:B (2:1) was the most suitable for promoting the tested antioxidant activity and bioactive compounds. Overall, the IF and BL showed significant benefits compared to CF for all detected compounds in both sole and intercrops.

Background: Clinacanthus nutans, also known as ‘Sabah snake grass’ or ‘Belalai gajah’, is a herb well known locally for its medicinal values. The primary chemical constituents of the leaves are schaftoside, vitexin, isovitexin, orientin and isoorientin, and antiviral activity is shown by two glycoglycerolipids. Despite the importance of C. nutans, complete information with respect to commercial production and postharvest handling of the herb in the local herbal industry is still lacking. Thus, the objective of this study was to determine the optimum postharvest handling processes that could retain the phytochemicals quality of C. nutans. Materials and Methods: The flavonoid compounds of C. nutans were analysed by using ultra fast liquid chromatography (UFLC). Total phenolic content and antioxidant activity were determined using a spectrophotometer. Results: The total phenolic compounds and antioxidant activity in C. nutans were found to be higher in the young vegetative stage than in the mature vegetative stage. Flavonoid compounds (schaftoside, isovitexin, vitexin and orientin) were also found to be highest in the young vegetative plant compared to the mature vegetative plant. All of the assayed phytochemicals and flavonoid compounds levels were found to be highest in oven dried samples compared to the sun, air and solar dried samples. Conslusion: This study suggests that oven-drying young vegetative C. nutans plant material is the optimum method to retain postharvest quality.

The secondary metabolites of the aerial parts of Zornia brasiliensis Vogel, Fabaceae, and the biological activity of one of these secondary metabolites were characterized in this study. A phytochemical investigation was performed using chromatographic techniques including analytical and preparative reverse-phase HPLC column sequences, which resulted in the isolation of fourteen compounds: one previously undescribed C-glycosylated dihydrochalcone (zornioside), one cyclitol (D-pinitol), one glycosylated megastigmane (roseoside) and eleven phenolic compounds: 7-methoxyflavanone, 7,4′-dimethoxyisoflavone, medicarpin, 2′-4′-dihydroxychalcone, onionin, isoorientin-3′-O-methyl ether, isovitexin, glycosylated (Z)-O-coumaric acid, glycosylated (E)-O-coumaric acid, dihydromelilotoside, and isoorientin. The structures of the isolated compounds were determined based on 1D and 2D-NMR, HRESIMS, IR and CD spectroscopic analyses. The cytotoxic activity of zornoside was assessed against tumor cell lines (MCF-7, HCC1954, T-47D, 4T1, HL60), and a non-tumor cell line (RAW264.7) using MTT assay. The compound zornioside was selectively cytotoxic for HL60 leukemia cells (IC50: 37.26 μM). Keywords: Phenolic compounds, Dihydrochalcone, Pterocarpan, Isoflavonoid, Glycosylated megastigmane, Cytotoxic activity

A liquid chromatography-tandem mass spectrometry method was developed and validated for simultaneous determination of six bioactive constituents including vitexin, orientin, isoorientin, 2″-O-β-d-xylopyranosyl isoorientin, 2″-O-β-d-xylopyranosyl isovitexin, and 6-C-l-α-arabipyranosyl vitexin in rats’ various tissues using isoquercitrin as the internal standard. Biological samples were pretreated by protein precipitation with acetonitrile. Chromatographic separation was carried out on a C18 column with a gradient mobile phase consisting of acetonitrile and 0.1% aqueous formic acid. All analytes and internal standard were quantitated through electrospray ionization in negative ion selected reaction monitoring mode. The mass transitions were as follows: m/z 431 → 311 for vitexin, m/z 447 → 327 for orientin or isoorientin, m/z 579 → 459 for 2″-O-β-d-xylopyranosyl isoorientin, m/z 563 → 293 for 2″-O-β-d-xylopyranosyl isovitexin, m/z 563 → 353 for 6-C-l-α-arabipyranosyl vitexin, and m/z 463 → 300 for the internal standard, respectively. The lower limits of quantification for the six analytes in different tissue homogenates were 0.8–2.2 ng/ml. The validated assay was successfully applied to a tissue distribution study of the six components in rats after intravenous administration of total flavonoids of Scorzonera austriaca Willd; Asteraceae. The results of the tissue distribution study showed that the high concentrations of six components were mainly in the kidney. Keywords: Scorzonera austriaca, LC-MS/MS, Rat plasma, Tissue distribution

Flavonoids are naturally occurring phenolic compounds with potential health-promoting activities. Although anthocyanins and phenolic acids in coloured rice have been investigated, few studies have focused on flavonoids. Herein, we analysed flavonoids in a yellow grain rice mutant using UHPLC-DAD-ESI-Q-TOF-MS, and identified 19 flavonoids by comparing retention times and accurate mass measurements. Among them, six flavonoids, isoorientin, isoorientin 2″- O -glucoside, vitexin 2″- O -glucoside, isovitexin, isoscoparin 2″- O -glucoside and isoscoparin, were isolated and fully identified from the yellow grain rice mutant, and the levels were significantly higher than wild-type, with isoorientin particularly abundant in mutant embryo. Significant differences in total phenolic compounds and antioxidant activity were observed in mutant rice by DPPH, FRAP and TEAC assays. The results suggest that the representative six flavonoids may play an important role in colouration and antioxidant activity of embryo and endosperm tissue. The findings provide insight into flavonoid biosynthesis and the possibility of improving functionality in rice.

Background: Dissotis rotundifolia, commonly referred to as pink lady, has several medicinal uses including peptic ulcer. This study investigated the inhibitory effects of D. rotundifolia extract on H+/K+-ATPase and also assessed its antiradical activity. In silico study of some isolated compounds of this plant was also carried out to affirm the suspected binding properties of extract to H+/K+-ATPase enzyme. Materials and Methods: D. rotundifolia whole plant extract was obtained after extraction process and then assessed for its ability to scavenge free radicals in four in vitro test models. Its ability to inhibit the activity of H+/K+-ATPase enzyme was also evaluated. Molecular docking was carried out on phytoconstituents, namely, vitexin, isovitexin, orientin, and isoorientin reported to be present in the whole plant extract. Results: Data obtained indicated that D. rotundifolia extract (DRE) exhibits strong antioxidant activity. DRE also showed inhibitory effects on H+/K+-ATPase enzyme activity. Docking studies affirmed the in vitro binding effect of the extract to H+/K+-ATPase. Conclusion: These findings suggest that the plant extract possess antioxidant and antipeptic ulcer activity.

Ethnopharmacological relevance Numerous Alphonsea species including Alphonsea elliptica (mempisang) leaves and fruits are indigenously used in inflammatory conditions such as postpartum swelling and rheumatism in southeast Asian countries. In our previous in-vitro findings, A. elliptica methanol extract exhibited platelet-activating factor inhibition, suggesting the presence of phyto-constituents with anti-inflammatory potential. Aim of the study However, so far there is no literature available on the anti-inflammatory activity of this species. Henceforth, based on the above background and our previous laboratory findings, we hypothesize that phytoconstituents of A. elliptica could possess anti-inflammatory potential against inflammatory mediators including prostaglandin-E2 (PGE2), cyclooxegenase-2 (COX-2) and cytokines (IL-1β and IL-6). Materials and methods Vacuum and column chromatography techniques were employed for the isolation of phytoconstituents. The structure elucidation was carried out using HRESI-MS, 1H and 13C-NMR analysis and compared with the published literature. For cytotoxicity analysis, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed on peripheral blood mononuclear cells. In-vitro anti-inflammatory activities were evaluated against the levels of PGE2, COX-2, IL-1β and IL-6 in lipopolysaccharide (LPS)-induced human plasma using enzyme-linked immunosorbent assay and radioimmunoassay. Results Unprecedentedly, chromatographic purification of methanolic leaves extract afforded five flavones namely vitexin, isovitexin, orientin, isoorientin, schaftoside with three flavanols; kaempferol, myricetin and rutin from A elliptica. In cell viability analysis, isolates did not present cytotoxicity up to 50 μM. In anti-inflammatory evaluation, orientin and isoorientin exhibited strong (≥70%), while isovitexin and vitexin produced strong to moderate (50–69%) PGE2, COX-2, IL-1β and IL-6 inhibition at 25 and 50 μM. Isoorientin, orientin, isovitexin, and vitexin showed significant (p Conclusion Conclusively, isolated phytoconstituents are reported for the first time from the A. elliptica. Moreover, isovitexin, vitexin orientin and isoorientin abrogated LPS-induced inflammatory responses in human plasma at non-cytotoxic concentrations.

Echinodorus scaber Rataj and Echinodorus grandiflorus (Cham. & Schltdl.) Micheli, Alismataceae, are popularly used to relieve inflammatory complaints and as diuretic. A study on the antinociceptive effect and selected marker compounds in eleven extracts from different locations was undertaken and their antinociceptive effect was assessed. The fingerprints were compared by HPLC-DAD and the content of vitexin, isovitexin, isoorientin and vitexin-2-O-rhamnoside were determined. All samples presented antinociceptive activity reducing the writhes by 36.4–62.5% and 47.4–79.8% at 10 and 50 mg/kg, respectively; indomethacin (5 mg/kg) reduced writhes by 82.6–90.1%. The content of the flavonoids C-glycosides, however, presented a strong variation. Isovitexin and isoorientin were found in all the samples, with content ranging from traces to 14.70 μg/mg and 2.12–84.27 μg/mg extract, respectively, while vitexin-2-O-rhamnoside occurred in quantifiable amounts only in 3 out of 11 samples ranging from 5.43 to 33.13 μg/mg extract; vitexin was not detected at all or detected in trace amounts. According to the fingerprints, the samples could be arranged in four main groups. All eleven extracts showed antinociceptive activity. Isovitexin was the only flavonoid present in all samples and can be regarded, acting in synergy with the other compounds or not, as the responsible for the antinociceptive activity. Therefore, isovitexin is a good choice as chemical marker when the antinociceptive activity of E. scaber and E. grandiflorus is investigated. Keywords: Chapéu-de-couro, Flavonoids, Isovitexin marker, Fingerprint, Antinociceptive activity

Passiflora plants are strategic in the context of biodiversity for food and nutrition. We applied the procedures of a systematic review protocol to study the state of the art on identification of phenolic compounds from Passiflora plants. An automated literature search was conducted using six databases and a combination of seven keywords. All the analytical, chromatographic, and spectroscopic methods were included. The studies were classified according to their method of identification, phenolic classes, and method of extraction. In total, 8,592 abstracts were found, from which 122 studies were selected for complete reading and 82 were selected for further analysis. Techniques of extraction, evaluated parts of the plant and methods of identification were systematized. Studies with leaves were most conspicuous (54.4%), 34 species of Passiflora were evaluated and orientin, isoorientin, vitexin, isovitexin were commonly found structures. A High Performance Liquid Chromatography-diode array detector was the technique most applied, with which the same structures were identified all through the studies, although other unknown structures were detected, but not elucidated. The use of Nuclear Magnetic Resonance and Mass Spectrometry, which are more sensitive techniques, needs to be intensified, to identify other unconventional compounds detected in Passiflora, to enhance the comprehension of the bioactive compounds in these plants.

A novel method of simultaneous extraction and determination of vitexin, isovitexin, orientin and isoorientin in Crotalaria sessiliflora L. was developed by microwave-assisted aqueous two-phase extraction (MAATPE) coupled with high performance liquid chromatography (HPLC). With an aqueous two-phase system (ATPS) of ethanol/(NH 4 ) 2 SO 4 as extracting agent, the effects of the composition of the ATPS, extraction temperature, extraction time, particle size and solvent-to-material ratio on extraction yields of the flavonoids were investigated by single-factor experiment and response surface methodology (RSM). Optimum conditions were concluded as follows: ethanol concentration 32% and (NH 4 ) 2 SO 4 concentration 22% for the ATPS, extraction temperature 80 °C, extraction time 8 min, particle size 80 mesh and solvent-to-material ratio 50:1. With the help of MAATPE, HPLC analysis of four flavonoids above has been successfully conducted with UV detection at 340 nm with linear ranges (0.20–20.00 mg/L), correlation coefficients ( R 2 > 0.9996) and limits of detection (0.0309–0.0492 mg/L) using acetonitrile-water containing 0.5% phosphoric acid solution (14:86) as the mobile phase. Under the optimum conditions, MAATPE coupled with HPLC was applied to simultaneous determination of vitexin, isovitexin, orientin and isoorientin in Crotalaria sessiliflora L. Their extraction yields and recoveries were in range of 162.7–240.0 μg/g and 94.14%–105.5%, respectively. The extraction process was explored by comparative study. The results demonstrated that addition of (NH 4 ) 2 SO 4 in the water-ethanol mixture improved not only demixing effect but also microwave action, and extraction yields of the ATPS were superior to that of mono-phase solvent such as water, ethanol and the combination of both. Thus, MAATPE was a rapid and efficient alternative to the conventional extraction approaches for herb analysis and quality control.

Popularly known as passion fruit, some species of the genus Passiflora are widely used in folk medicines, such as sedatives and tranquilizers in many countries. Although these plants are employed for the same purposes, research with different species of Passiflora has indicated their heterogeneous chemical compositions. Since different chemical compositions can result in varying degrees of therapeutic efficiency, quality control based on the chemical constituents of each species is essential. To that end, the aim of this study was to compare pharmacognostically species of Passiflora in order to establish a chromatographic profile for the quality control of drugs in herbal medicines containing passion fruit. The study was conducted by collecting samples of leaves from twelve Passiflora taxa (i.e., ten species and two forms of P. edulis) – P. actinia, P. alata, P. amethystina, P. capsularis, P. cincinnata, P. edulis f. flavicarpa, P. edulis f. edulis, P. incarnata, P. morifolia, P. urnifolia, P. coccinea, and P. setacea – from different locations and obtaining their chromatographic profiles via thin-layer chromatography and high-performance liquid chromatography. Both methods used the flavonoid C-glycosides isoorientin, orientin, vitexin, and isovitexin as reference compounds and could ultimately establish specific profiles for each species. The chromatographic analyses discussed here can be used to assist in determining the quality and authenticity of herbal drugs derived from Passiflora species. Keywords: Passiflora, Passion fruit, Quality control, Thin-layer chromatography, High-performance liquid chromatography, Flavonoid C-glycosides

Buckwheat sprouts that are synthesized during the germination process are rich in flavonoids, including orientin, vitexin, rutin, and their isomers (isoorientin, isovitexin, and quercetin-3-O-robinobioside, respectively). The purpose of this study was to optimize and validate an analytical method for separating flavonoid isomers in common buckwheat sprout extract (CSE). Factors, such as range, linearity, precision, accuracy, limit of detection, and limit of quantification, were evaluated for each standard using high-performance liquid chromatography (HPLC). On the basis of resolution and symmetry, a column temperature of 40 &deg, C with 0.1% (v/v) acidic water and acetonitrile as mobile phases, at a flow rate of 1 mL min&minus, 1 were determined to be the optimal analytical conditions. Calibration curves for orientin, isoorientin, vitexin, isovitexin, and rutin exhibited good linearity with correlation coefficients of 0.9999 over the 6.25&ndash, 100.00 &mu, g mL&minus, 1 range. Recovery values of 96.67&ndash, 103.60% confirmed that the method was accurate for all flavonoids. The relative standard deviations of intra-day repeatability and inter-day reproducibility confirmed method preciseness, with values of less than 5.21% and 5.40%, respectively. The developed method was used to analyze flavonoids in CSE, with isomers satisfactorily separated and simultaneously quantified. We demonstrated that the developed HPLC method can be used to monitor flavonoids in buckwheat sprouts.

Apigenin (4′,5,7-trihydroxyflavone) and luteolin (3′,4′,5,7-tetrahydroxyflavone) are among the most widely distributed flavone aglycones in flowering plants. These metabolites often occur in the form of O- and C-glycosides. In this group, four C-glucosides pay special attention: vitexin (apigenin 8-C-glucoside) and isovitexin (apigenin 6-C-glucoside) as well as orientin (luteolin 8-C-glucoside) and isoorientin (luteolin 6-C-glucoside). The above-mentioned compounds show various biological activities, including antioxidant, anti-inflammatory, immunomodulatory, anticancer as well as neuro-, cardio-, and hepatoprotective effects. The aim of the present work was to determine the antibacterial activity of these flavones. In the in vitro tests, there were investigated clinical strains of two Gram-positive (Staphylococcus aureus, Enterococcus faecalis) and Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa). The antimicrobial activity of chosen substances was determined by the micro-dilution method according to recommendations of the Clinical and Laboratory Standards Institute (CLSI). Curcumin was used as a positive control. Our results exhibited a relatively low sensitivity of the tested strains to the plant metabolites. In the case of curcumin – natural compound with known strong antibacterial effect, the minimal inhibitory concentration (MIC) for all species of bacteria was 500 μg/mL. The same level of biological activity was observed for apigenin, luteolin, and their glucosides against E. coli and P. aeruginosa. Among Gram-positive bacteria, the obtained results showed significant variability. Strains of S. aureus demonstrated a weak sensitivity to apigenin (MIC = 500-1000 μg/mL), and were resistant to its derivatives: vitexin and isovitexin (>1000 μg/mL). Additionally, these compounds poorly inhibited the growth of E. faecalis (1000 μg/mL). In turn, luteolin and its C-glucosides (orientin, isoorientin) reached the same values of the MICs: moderate against S. aureus (500 μg/mL) and weak for E. faecalis (1000 μg/mL). Our research points to the problem of varied sensitivity and even resistance of some clinical strains of common pathogens to the widespread natural plant compounds, such as flavonoids and other phenolics (e.g., curcumin). It is interesting that apigenin, luteolin, and their C-glucosides were generally more potent against Gram-negative bacteria than Gram-positive ones. A pair of analysed flavone aglycones has a very similar chemical structure, and they did not differ significantly in the antibacterial activity. Similarly, the presence and location of the sugar group in the flavone glucosides usually did not affect the values of the MICs.

Fluorescence sensors prepared from natural polymers have received increasing attention based on their luminescence characteristics for bioimaging, cell imaging, and intracellular detection of inorganic metabolites. In this work, flavonoids isolated from bamboo residues (BRF) were applied as fluorescence sensors for different metal cations&rsquo, detection in vitro. Results showed the optimal flavonoids extraction condition of solid to liquid ratio, ethanol concentration, extraction time and temperature were determined at 1:25, 50%, 240 min and 90 &deg, C, respectively, resulting in an extraction yield with 104.7 mg/100 g bamboo residues. The BRF is mainly composed of isoorientin, isovitexin, pinosylvin, tricin and isorhamnetin by liquid chromatography&ndash, mass spectrometry (LC-MS) analysis. It is found that the BRF displayed strong blue-green emission as well as notable excitation, which can selectively and sensitively detect Fe3+ with the limit of detection (LOD) as low as 38.0 nM. In the Fe3+ detection was no obvious interference by other cations except for Al3+. In addition, the BRF displayed excellent biocompatibility that can be applied to bioimages of the intracellular detection of Fe3+ in L02 cells. Finally, it is found that the BRF possessed significant antioxidant properties in scavenging H2O2-induced endogenous reactive oxygen species (ROS) in a zebrafish module (in vivo) and L02 cells (in vitro). These results showed that the flavonoid products sustainably isolated from an abundant lignocellulosic waste appear to be effective fluorescent sensors for Fe3+ detection in biological systems with excellent biocompatibility and antioxidant activity.

Antioxidants and flavonoids are beneficial compounds used in the pharmaceutical and food industries that are derived from natural sources. A two-year field experiment was undertaken to investigate the effect of agricultural management practices on the trigonelline content, antioxidant activity measured with DPPH (2,2-Diphenyl-1-Picrylhydrazyl) and FRAP (Fe+++-Reduction, Ferric reducing antioxidant power), total phenolic content, total flavonoids content, and specific flavonoid contents of fenugreek seeds. The treatments examined were sole fenugreek (Sole F) and three intercropping ratios with buckwheat (B) (F:B = 2:1, 1:1, and 1:2), each with three types of fertilizer (chemical fertilizer, integrated fertilizer, and broiler litter). The highest trigonelline content of fenugreek seeds was obtained in F:B = 2:1 fertilized with broiler litter. Compared to Sole F, the fenugreek seeds harvested in the intercropped treatments had higher antioxidant activity, total phenolic content, vitexin, isovitexin, orientin, and isoorientin. The treatment F:B = 2:1 enhanced the antioxidant activity and the content of bioactive compounds. Overall, the addition of organic manure to fenugreek seeds was highly effective at promoting all compounds analyzed in both the sole and the intercrop systems. Our study found that harvested fenugreek seeds from plants that were intercropped with buckwheat and with the application of organic fertilizer enhanced the seed content of antioxidants and flavonoids.

Bryonia alba L. is the only Bryonia species found in Romanian flora, being known as a remedy for inflammatory pathologies or for its hepatoprotective and adaptogen activities. The present investigation studied the flavonoid composition and antioxidant activities of the aerial parts of this species. Flavonoid profile was evaluated by HPLC coupled with Diode Array Detection (DAD), while antioxidant capacity was assessed by various methods, testing different antioxidant mechanisms: DPPH (2,2-diphenyl-1-picrylhydrazyl), CUPRAC (cupric reducing antioxidant capacity), FRAP (ferric reducing ability of plasma), TEAC (Trolox equivalent antioxidant capacity), EPR (electron paramagnetic resonance method) and SNPAC (silver nanoparticles antioxidant capacity). Cytotoxicity was tested on human cancerous and healthy cell lines. Anti-plasmodial tests were performed on two strains of Plasmodium falciparum. Whole organism toxicity was assessed on zebrafish larvae. The HPLC-DAD analysis proved the presence of lutonarin, saponarin, isoorientin, and isovitexin as the major flavonoids in the composition of tested samples. Significant results were obtained for all antioxidant capacity assays. The cytotoxicity tests proved the absence of cellular and parasitic toxicity and these results were confirmed by the lack of toxicity on the zebrafish larvae model. This study proves a promising potential of the aerial parts of Bryonia alba L. as antioxidant agents.