Content Of Verbascoside Research Articles

Wound healing and photoprotection properties of Acanthus ebracteatus Vahl. extracts standardized in verbascoside

Acanthus spp. have been documented in traditional Thai herbal medicine and are applicable for the treatment of inflamed skin with wound healing property. Nonetheless, the scientific evidence necessary to prove the herb’s doctrine has not yet been revealed. Verbascoside-rich extracts of the herbal medicine A. ebracteatus Vahl., were therefore prepared. The extracts and verbascoside were examined for their wound healing abilities using a scratch assay with fibroblasts. The anti-inflammatory effect suppressing MMP-9 was assessed in cocultures of keratinocyte (HaCaT cells) and fibroblasts. The extracts significantly improved wound healing compared with the control (p < 0.001). The wound healing effect of the extracts significantly (p < 0.01) increased with increasing verbascoside content. It should be noted that the extract was significantly (p < 0.05) better than verbascoside at the same test concentration. The extracts were capable of protecting cocultures of HaCaT cells and fibroblasts from photodamage. The extracts significantly (p < 0.001) suppressed cellular MMP-9 secretion following UV exposure, showing a better effect than that of verbascoside (p < 0.01). A. ebracteatus extract is promising for wound healing and photoprotection, and a prominent source of verbascoside. Verbascoside-rich A. ebracteatus could be utilized for the development of innovative skin-care products.

Highly Efficient Verbascoside Production from Olive (Olea europea L. var. Cellina di Nardò) In Vitro Cell Cultures.

Olive (Olea europea L.) is one of the oldest and most important fruit tree species cultivated in the Mediterranean region. Various plant tissues, drupes, and olive oil contain several phenolics (including verbascoside, although it is present in the plant at a low level) that are well-known for their highly beneficial effects on human health. An in vitro olive cell suspension culture (cultivar Cellina di Nardò, "CdN") was established, characterized for its growth and morphological features. Furthermore, a vital and relatively uniform population of protoplasts was generated from the olive suspension culture to investigate their cellular characteristics during growth. The polyphenolic extract of the in vitro "CdN" olive cells contained almost exclusively verbascoside, as revealed by the UPLC-ESI-MS analysis. The content of verbascoside reached up to 100 mg/g DW, with an average production rate of approximately 50 mg/g DW over one year of culture. This level of production has not been previously reported in a limited number of previous studies. This remarkable production of verbascoside was associated with an exceptionally high antioxidant capacity. The high level of verbascoside production and purity of the extract make this system a promising tool for secondary metabolite production.

Cytoprotective Effect of Acid-Hydrolyzed Sesame Leaves against Oxidative Stress in HepG2 Cells

In this study, we investigated the cytoprotective effects of acid-treated sesame leaf extracts against oxidative damage in HepG2 cells. Treatment with 0.1 M HCl of sesame leaves significantly increased their verbascoside content (4.398 g/100 g) compared to non-acid-treated leaves (3.950 g/100 g). Acid-treated sesame leaf extract (ASLE) showed no cytotoxicity in HepG2 cells. ASLE conferred a greater cytoprotective effect against oxidative insult than a methanol extract of sesame leaves (SLE), verbascoside, and a vehicle control group. ASLE treatment also significantly inhibited reactive oxygen species generation in response to oxidative stress. Treatment with tert-butyl hydroperoxide increased malondialdehyde (MDA) levels, and depleted reduced glutathione (GSH). However, ASLE treatment significantly ameliorates this MDA and GSH depletion. Moreover, ASLE increased the activities of antioxidant enzymes including catalase, glutathione reductase, and glutathione peroxidase. Phenolic compounds in ASLE and SLE were characterized using UPLC-Q-TOF/MS analysis. A total of 29 iridoid and phenol compounds were tentatively identified in ASLE, and 27 compounds were observed in SLE. These results suggest that acid treatment of sesame leaves enhances the protective effects of their extract against oxidative stress by modulating antioxidant enzymes in HepG2 cells.

A comparative study on chemical profiles and biological activities of different extracts of three Verbascum species from Turkey: In vitro, in silico and network pharmacological approaches

The present study aimed to evaluate the antioxidant and enzyme inhibition properties of aerial part of Verbascum cheiranthifolium Boiss. V. myriocarpum Boiss. & Heldr. and V. pyroliforme (Boiss. & Heldr.) Kuntze as well as their chemical profiles. Results showed that MeOH and hydromethanol recovered the highest total polyphenolic content with highest content obtained from V. cheiranthifolium (27.61and 27.69 mg GAE/g respectively) and V. myriocarpum (24.24 and 25.34 GAE/g respectively). The estimated content of verbascoside was quite higher in V. cheiranthifolium (987.05–6780.07 mg/100 g) and V. myriocarpum (1506.47–6433.73 mg/100 g) than V. pyroliforme (49.54–121.97 mg/100 g). All extracts of V. cheiranthifolium (274.17–713.79 mg/100 g) and polar ones of V. myriocarpum (185.67–304.51 mg/100 g) accumulated higher chlorogenic acid content than V. pyroliforme extracts (36.33–112.56 mg/100 g). The latter species was dominated by luteolin-7-O-glucoside and apigenin-7-O-glucoside with highest content recorded from the hydromethanolic extract (639.70 and 196.14 mg/100 g respectively). Extracts of V. cheiranthifolium (46.15–48.74 mg TE/g) and V. myriocarpum (46.05–48.50 mg TE/g) exerted significantly higher scavenging activity than those from V. pyroliforme (12.77–34.64 mg TE/g). Highest significant anti-acetylcholinesterase activity was obtained by MeOH extract of V. pyroliforme (2.65 mg GALAE/g) and V. myriocarpum (2.57 mg GALAE/g). In conclusion, extracts from the three investigated Verbascum spp. can be a potential source of bioactive metabolites with interesting antioxidant and enzyme inhibition properties.

Response of human blood platelets to preparations from leaves of Paulownia Clon in Vitro 112

Paulownia Clon in Vitro 112, also called the Oxytree, is a fast-growing hybrid of two trees belonging to the Paulowniaceae family – P. elongata and P. fortunei. It demonstrates a wide range of biological effects (including antioxidant, anti-inflammatory, antibacterial, and neuroprotective) due to the high concentration of secondary metabolites. Our previous results showed an in vitro antioxidant and antiplatelet activity of the extract and four fractions (A–D) from the leaves of Paulownia Clon in Vitro 112 in human plasma and washed blood platelets. Here, we used a microchip flow chamber-based thrombus formation analysis system (T-TAS) and flow cytometry to assess the anticoagulant and antiplatelet activity of the extract and four fractions with different chemical content (A-D) from Paulownia Clon in Vitro 112 leaves in human whole blood. Two tested fractions: fraction C and D (at the concentrations of 5 and 50 μg/mL) inhibited the exposition of the active form of GPIIb/IIIa (integrin αIIbβ3) on the surface of blood platelets stimulated by ADP and collagen. The antiplatelet activity of fraction C is likely due to its high verbascoside content and the presence of apigenin’s derivatives. Fraction D contains triterpenoids, including ursolic, pomoleic, and maslinic acid, which could be responsible for decreased activation of ADP- and collagen-stimulated blood platelets. These results suggest that fractions C and D might be promising sources of phytochemicals with antiplatelet activity, which are important for prophylaxis and treatment of cardiovascular diseases associated with hyperactivation of blood platelets. However, further research is needed to ascertain which exact compounds and mechanisms are responsible for this phenomenon.

Secondary Metabolite Production in Callus Culture of &lt;i&gt;Verbascum scamandri&lt;/i&gt; Murb.

&lt;i&gt;Verbascum scamandri&lt;/i&gt; Murb. known as "Kazdağı Mullein" is an endemic species in Türkiye and is classified as an endangered (EN) species. The aim of this study is to establish an efficient callus culture for &lt;i&gt;V. scamandri&lt;/i&gt; and analyze the amounts of verbascoside, luteolin, and aucubin metabolites of calli samples. Leaf explants were cultured on MS medium with cytokinin (BAP, Kin, 0, 0.5, 1, 2, 3 mg/L) and auxin (NAA, 2,4-D, 0, 0.1, 0.5, 1 mg/L), 1 g/L PVP, 3% sucrose, and 0.7% agar for callus induction. Callus tissue in MS with 2 mg/L Kin, 0.5 mg/L Kin+0.5 mg/L 2,4-D, 2 mg/L Kin+0.5 mg/L 2,4-D, and 3 mg/L Kin+0.5 mg/L 2,4-D was proliferated in MS basal medium containing PGR at the same concentrations and combinations as the callus induction media. Verbascoside, luteolin, and aucubin were quantified in leaf samples of the&lt;i&gt; in vivo&lt;/i&gt; collected plants, leaf samples of &lt;i&gt;in vitro&lt;/i&gt; growing plants, and calli using HPLC-DAD. According to the results, the verbascoside content in the leaf of collected plants was 7.03 mg/g, luteolin was 0.66 mg/g, and aucubin was 2.99 mg/g. The leaf of &lt;i&gt;in vitro&lt;/i&gt; plants had 1.62 mg/g verbascoside, 0.18 mg/g luteolin, and 1.32 mg/g aucubin. Whereas, the maximum content of secondary metabolites in the callus samples was observed 13.77 mg/g verbascoside in MS medium with 2 mg/L Kin, 0.51 mg/g luteolin in MS medium with 2 mg/L Kin+0.5 mg/L 2,4-D, and 9.32 mg/g aucubin in 0.5 mg/L Kin+0.5 mg/L 2,4-D.

Genetic diversity and verbascoside content in natural populations of Pyrostegia venusta (Ker Gawl.) Miers.

Pyrostegia venusta (Ker Gawl.) Miers occurs in threatened biodiversity hotspots of Cerrado and Atlantic forest biomes in Brazil and is used in traditional medicine to treat various respiratory and skin diseases. This study (i) examined the genetic diversity and structure of six natural populations of P. venusta from different Brazilian regions using sequence-related amplified polymorphism (SRAP) markers; and (ii) compared the intra- and inter-populational levels of the bioactive component verbascoside using high-performance liquid chromatography. The population from Nova Mutum, Mato Grosso, presented the highest genetic variability (Nei index H = 0.2759; Shannon index I = 0.4170; 85.14% polymorphic loci), whereas the population from Araxá, Minas Gerais, presented the lowest genetic variability (H = 0.1811; I = 0.2820; 70.27% polymorphic loci). The intra-populational variability (79%) was significantly higher (p = 0.001) than the inter-populational variability (21%). The populations were clustered into two groups but their genetic differentiation was not associated with geographical origin (Mantel test, r = 0.328; p > 0.05). The verbascoside content significantly differed (p > 0.05) among the six populations and between the individuals from each population. The highest verbascoside levels (> 200µg/mg extract) were detected in populations from Araxá and Serrana, while the lowest verbacoside levels were detected in populations from Paranaíta and Sinop. This is the first report on the use of SRAP markers to analyze genetic variability in the family Bignoniaceae. Our findings shall help to better understand the genetic and chemical diversity of P. venusta populations, as well as provide useful information to select the most appropriate individuals to prepare phytomedicines.

Evolution of Oleuropein and Other Bioactive Compounds in Arbequina Olive Leaves under Different Agronomic Conditions

Oleuropein and other phenolic compounds contained in olive leaves give it the potential to be transformed from residue to co-product in an oil mill. However, the moment of the agronomic cycle in which their potential transformation is higher is not known in detail. Therefore, for the first time, a monthly study of the evolution of such compounds throughout an agronomic cycle is made (November 2019 to October 2020). Arbequina olive leaves were collected from three plots and the interactive effects of agronomic conditions were investigated, such as crop management (conventional and ecological), plantation framework (intensive and super-intensive) and location under different climatic conditions. The results showed that the main compound throughout the cycle was oleuropein and the highest levels occurred around the pruning season (February/March). Crop management and location affected the content of verbascoside and hydroxytyrosol, while plantation framework only influenced the flavonoid content. All compounds were affected by relative humidity and differential temperature, although hydroxytyrosol showed the highest correlation with the maximum temperature. The absorbance measurements by ultraviolet-visible spectrophotometry showed trends parallel to the oleuropein concentration measured by high-performance liquid chromatography, which suggests that this method could be useful to easily study the evolution of oleuropein in the oil mill.

Determination of the Variability of Bioactive Compounds and Minerals in Olive Leaf along an Agronomic Cycle

Olive leaves are still considered waste in the oil industry; however, the leaves have a content of oleuropein and other bioactive compounds that gives them great potential to be transformed into by-products. The most appropriate moment along an agronomic cycle (November 2019 to October 2020) has been evaluated to take advantage of this potential of the olive leaves varieties of Picual, Cornicabra and Manzanilla. In addition, factors that affect the content of phenolic compounds such as absolute maximum and minimum temperatures, relative humidity, sunshine hours, rainfall, differential of temperatures and mineral nutrition have been studied. The results show that the pruning season was the best time to take advantage of the olive leaf due to its high oleuropein content, especially in Picual and Manzanilla. The variety was the factor that most affected all the phenolic compounds studied, while the absolute minimum temperature factor notably affected verbascoside. Particular mineral nutrients, such as Mg and Fe, turned out to be most useful for differentiating locations. The content of verbascoside and hydroxytyrosol was also affected by agronomic conditions (location/conventional or ecological).

Exploitation of virgin olive oil by-products (Olea europaea L.): phenolic and volatile compounds transformations phenomena in fresh two-phase olive pomace ('alperujo') under different storage conditions.

BACKGROUNDMuch effort has recently been spent for re‐using virgin olive oil by‐products as nutraceutical ingredients for human diet thanks to their richness in bioactive phenols, but their management is not easy for producers. We aimed to provide useful information for a better management of fresh olive pomace before drying, by studying the phenolic and volatile compounds transformations phenomena of fresh olive pomace stored under different conditions planned to simulate controlled and uncontrolled temperature conditions in olive oil mills.RESULTSThe evolution of the phenolic and volatile compounds was studied by high‐performance liquid chromatography‐diode array detector mass spectrometry (HPLC‐DAD‐MS) and headspace solid‐phase microextraction gas chromatography‐mass spectrometry (HS‐SPME‐GC‐MS). The phenolic profile varied rapidly during storage: the verbascoside content decreased about 70% after 17 days even at 4 °C, while the content of simple phenols such as hydroxytyrosol and caffeic acid increased over time. The low temperature was able to slow down these phenomena. A total of 94 volatile organic compounds (VOCs) were detected in the fresh olive pomace, with a prevalence of lipoxygenase (LOX) VOCs (78%), mainly aldehydes (19 490.9 μg kg−1) despite the higher number of alcohols. A decrease in LOX volatiles and a quick development of the ones linked to off‐flavors (carboxylic acids, alcohols, acetates) were observed, in particular after 4 days of storage at room temperature. Only storage at 4 °C allowed these phenomena to be slowed down.CONCLUSIONTo preserve the natural phenolic phytocomplex of fresh olive pomace before drying and to avoid off‐flavors development, storage in open containers must be avoided and a short storage in cold rooms (7–10 days) is to be preferred. © 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Rapid quality evaluation of Plantaginis Semen by near infrared spectroscopy combined with chemometrics

ObjectiveTo establish a rapid and comprehensive method for the quality evaluation of Plantaginis Semen (PS) by using near-infrared spectroscopy combined with chemometrics to determine the content of geniposidic acid (GEA), verbascoside (VE), isoverbascoside (IVE) and total flavonoids (TF) in PS, as well as the antioxidant activity (AOA). MethodsThe content of GEA, VE and IVE in PS were determined by HPLC, the content of TF in PS was determined by UV-Vis spectrophotometry, and the AOA of PS was characterized by the DPPH, ABTS and FRAP, respectively. Then, the measured values of each item were used as reference values and were correlated with the near infrared spectra of PS. Seven quantitative models were established by the partial least squares regression. A variety of spectral preprocessing, such as standard normal variation (SNV), multiplicative scatter correction (MSC), Savitzky-Golay smoothing (SG), derivative and their combination methods, were investigated. In addition, genetic algorithm (GA), particle swarm optimization (PSO), as well as competitive adaptive reweighted sampling (CARS) were also compared. All samples were divided into a calibration set and a prediction set at the ratio of 3:1 by the descending order of reference values. The coefficients of determination (R2), root mean square error (RMSE) and residual predictive deviation (RPD) were calculated to evaluate model performance. ResultsAfter optimization, the performance of each model was greatly improved, where the R2 for calibration and prediction were both greater than 0.8, the RPD were both greater than 2. Such satisfactory results indicated that the present models had good prediction accuracy. ConclusionQuantitative models based on near infrared spectroscopy were herein established, which proved to be able to quickly and accurately determine the content of GEA, VE and IVE and TF, as well as AOA in PS, and which might provide a new method for rapid and comprehensive quality evaluation of PS.

Antioxidant Activity, Polyphenolic Profiles and Antibacterial Properties of Leaf Extract of Various Paulownia spp. Clones

Paulownia spp. are widely distributed ornamental trees with leaves abundant in secondary metabolites of high medicinal potential. Eighteen breeding clones of Paulownia spp. were tested in terms of their antioxidant activity and total polyphenolic contents. The 50% ethanolic extracts (2 g/30 mL) of leaves and petioles were compared in the screening step. Eight paulownia clones were selected for detailed analyses including HPTLC polyphenolic profile, verbascoside content and antibacterial activity against five bacteria species (S. aureus, B. cereus, E. coli, Y. enterocolitica, S. enterica). The species-specific differences in terms of antioxidant activity correlated with phenolic compounds were found mainly in the case of leaf blade extracts, the highest for P. tomentosa × P. fortunei and the lowest for P. elongata × P. fortunei clones. The P. tomentosa clones varied greatly in this regard. In the HPTLC polyphenolic profile, the occurrence of some polyphenols was proved and the specific verbascoside content was quantified (70 to 225 mg/g DW). The P. tomentosa × P. fortunei hybrids had the highest inhibitory activity, mainly against Gram-positive bacteria, whereas only slight inhibition of S. aureus growth was observed for P. elongata × P. fortunei clones. The obtained results indicate diverse suitability of paulownia clones as a source of active ingredients.

Changes in the Harpagide, Harpagoside, and Verbascoside Content of Field Grown Scrophularia lanceolata and Scrophularia marilandica in Response to Season and Shade.

Scrophularia lanceolata Pursh and Scrophularia marilandica L. are two common species within the Scrophulariaceae family that are endemic to North America. Historically, these species were used by indigenous peoples and colonialists to treat sunburn, sunstroke, frostbite, edema, as well as for blood purification, and in women’s health. Several iridoid and phenylethanoid/phenylpropanoid glycosides detected in these species, such as harpagoside and verbascoside, possess anti-inflammatory and anti-nociceptive properties. Due to the presence of anti-inflammatory metabolites and the historical uses of these species, we performed a two-year field study to determine the optimal production of these important compounds. We subjected the plants to shade treatment and analyzed differences in the metabolite composition between the two species and each of their leaves, stems, and roots at various times throughout the growing seasons. We determined that S. lanceolata plants grown in full sun produced 0.63% harpagoside per dried weight in their leaves compared to shade-grown plants (0.43%). Furthermore, S. lanceolata accumulated more harpagoside than S. marilandica (0.24%). We also found that verbascoside accumulated in the leaves of S. lanceolata and S. marilandica as the growing season progressed, while the production of this metabolite remained mostly seasonally unchanged in the roots of both species.

Production of Verbascoside, Isoverbascoside and Phenolic Acids in Callus, Suspension, and Bioreactor Cultures of Verbena officinalis and Biological Properties of Biomass Extracts.

Callus, suspension and bioreactor cultures of Verbena officinalis were established, and optimized for biomass growth and production of phenylpropanoid glycosides, phenolic acids, flavonoids and iridoids. All types of cultures were maintained on/in the Murashige and Skoog (MS) media with 1 mg/L BAP and 1 mg/L NAA. The inoculum sizes were optimized in callus and suspension cultures. Moreover, the growth of the culture in two different types of bioreactors—a balloon bioreactor (BB) and a stirred-tank bioreactor (STB) was tested. In methanolic extracts from biomass of all types of in vitro cultures the presence of the same metabolites—verbascoside, isoverbascoside, and six phenolic acids: protocatechuic, chlorogenic, vanillic, caffeic, ferulic and rosmarinic acids was confirmed and quantified by the HPLC-DAD method. In the extracts from lyophilized culture media, no metabolites were found. The main metabolites in biomass extracts were verbascoside and isoverbascoside. Their maximum amounts in g/100 g DW (dry weight) in the tested types of cultures were as follow: 7.25 and 0.61 (callus), 7.06 and 0.48 (suspension), 7.69 and 0.31 (BB), 9.18 and 0.34 (STB). The amounts of phenolic acids were many times lower, max. total content reached of 26.90, 50.72, 19.88, and 36.78 mg/100 g DW, respectively. The highest content of verbascoside and also a high content of isoverbascoside obtained in STB (stirred-tank bioreactor) were 5.3 and 7.8 times higher than in extracts from overground parts of the parent plant. In the extracts from parent plant two iridoids—verbenalin and hastatoside, were also abundant. All investigated biomass extracts and the extracts from parent plant showed the antiproliferative, antioxidant and antibacterial activities. The strongest activities were documented for the cultures maintained in STB. We propose extracts from in vitro cultured biomass of vervain, especially from STB, as a rich source of bioactive metabolites with antiproliferative, antioxidant and antibacterial properties.

Genotype-Related Differences in the Phenolic Compound Profile and Antioxidant Activity of Extracts from Olive (Olea europaea L.) Leaves.

The phenolic compound contents and antioxidant activities of the leaf extracts of nine olive genotypes were determined, and the obtained data were analysed using chemometric techniques. In the crude extracts, 12 compounds belonging to the secoiridoids, phenylethanoids, and flavonoids were identified. Oleuropein was the primary component for all genotypes, exhibiting a content of 21.0 to 98.0 mg/g extract. Hydroxytyrosol, verbascoside, luteolin 7-O-glucoside, and luteolin 4′-O-glucoside were also present in noticeable quantities. Genotypes differed to the greatest extent in the content of verbascoside (0.45–21.07 mg/g extract). The content of hydroxytyrosol ranged from 1.33 to 4.03 mg/g extract, and the aforementioned luteolin glucosides were present at 1.58–8.67 mg/g extract. The total phenolic content (TPC), DPPH• and ABTS•+ scavenging activities, ferric reducing antioxidant power (FRAP), and ability to inhibit the oxidation of β-carotene-linoleic acid emulsion also varied significantly among genotypes. A hierarchical cluster analysis enabled the division of genotypes into three clusters with similarity above 60% in each group. GGE biplot analysis showed olive genotypes variability with respect to phenolic compound contents and antioxidant activities. Significant correlations among TPC, FRAP, the values of both radical scavenging assays, and the content of oleuropein were found. The contents of 7-O-glucoside and 4′-O-glucoside correlated with TPC, TEAC, FRAP, and the results of the emulsion oxidation assay.

Genetic diversity and quality analysis of Rehmannia glutinosa in different germplasm

The study aims at evaluating genetic diversity and medicinal quality of cultivated germplasm in Rehmannia glutinosa, and providing theoretical guidance for screening excellent germplasm. The genetic diversity of 21 species of R. glutinosa were analyzed by SRAP molecular markers, and the catalpol and verbascoside was determined by HPLC. The mass fraction of catalpol and verbascoside in R. glutinosa germplasm were respectively in the range of 2.393%-6.519% and 0.063%-0.478%, the germplasm 14, 16, 15 and 20 germplasm, witch catalpol and verbascoside content was higher. A total of 57 bands were produced by 10 primer, among which 40 polymorphic bands were polymorphic bands, and the percentage of polymorphic loci was 8.77%-54.39%, the Nei's genetic diversity index (H) was 0.374 1, Shannon's polymorphism information index (I) was 0.546 6. Gst and gene flow Nm were 0.608 8 and 0.321 3, respectively. Based on the genetic uniformity, 21 species of germplasm were grouped into 2 categories. The genetic diversity level of R. glutinosa was medium low. The comprehensive consideration of the genetic diversity and the content inculde catalpol and verbascoside, germplasm 7 and germplasm 18 could be used as the preferred materials for the cultivation of reticulum. Germplasm 15 and 16 can be used as the preservation and breeding object of rhubarb germplasm.

Callus production and analysis of some secondary metabolites in Globularia trichosantha subsp. trichosantha

This study describes the production of calli from Globularia trichosantha subsp. trichosantha and the quantitative determination of catalpol, aucubin, and verbascoside in the calli and the plant by HPLC. The seeds of the plant were sterilized and germinated in Murashige and Skoog (MS) medium without in vitro plant growth regulator. Hypocotyl, cotyledon, first leaf, epicotyl, apical meristem, and root explants were taken from the 30-day-old aseptic seedlings germinated in vitro. Explants were then transferred to MS media for callus production together with varying concentrations of plant growth regulators. The best callus production occurred in the media containing 6 mg L?1 IAA and 0.2 mg L?1 2,4-D + 0.1 mg L?1 BAP. The highest catalpol, aucubin, and verbascoside contents were determined in the roots (1.277 mg kg?1), stem (0.775 mg kg?1), and roots (0.290 mg kg?1), respectively. The study is important in the sense that, 1) it is the first tissue culture study on G. trichosantha subsp. trichosantha that provides basic information for callus production; 2) that catalpol, aucubin, and verbascoside were quantitatively determined in calli and plant for the first time; and 3) all the secondary metabolites studied here were produced in the callus culture in a quantity that is approximately 7 times higher than the amount that naturally occurs in the plant.

HPLC-DAD-ESI-Q-TOF-MS/MS profiling of Verbascum ovalifolium Donn ex Sims and evaluation of its antioxidant and cytogenotoxic activities.

Plant species of Verbascum genus have been intensively investigated in the last decades but most studies focused on evaluation of their biological activities; there are only few studies dealing with their chemical characterisation. Detailed investigation of the qualitative and quantitative chemical composition, antioxidant and cytogenotoxic activities of a previously non-studied Verbascum species (V. ovalifolium Donn ex Sims). Qualitative analysis of secondary metabolites was performed by HPLC-DAD-ESI-Q-TOF-MS/MS, whereas quantitative data were obtained through HPLC-DAD. Antioxidant activity was evaluated using in vitro assays; cytotoxic and genotoxic effects were assessed by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium (MTT) and comet assays, respectively. More than 50 secondary bioactive metabolites belonging to various classes (iridoids, phenylethanoids, flavonoids, phenolic acids) were detected in the methanolic extract of V. ovalifolium and its fractions. The fragmentation pathways of acylated catalpol-type iridoid diglycosides are thoughtfully described herein. The extracts showed good free radical scavenging and ferric ion reducing properties correlated with phenolic, flavonoid, chlorogenic acid and verbascoside contents. Moreover, 24h treatment of SK-MEL-2 cells with V. ovalifolium extracts produced significant changes in terms of tumour cell viability. The crude extract and the ethyl acetate fraction showed no important signs of cytogenotoxicity in non-tumour cells. The performed phytochemical and biological analyses contribute to the preclinical knowledge about V. ovalifolium and they could help exploiting it in novel herbal medicinal products.

Ionic liquid-based microwave-assisted extraction of verbascoside from Rehmannia root

Rehmannia root is widely used as drug and functional food in East Asia and verbascoside content is the main quality control index of Rehmannia root because of its pharmacological activities; thus a fast, and environmentally-benign extraction method of verbascoside from Rehmannia root is a much-needed project. In this work, an ionic liquid (IL)-based microwave-assisted method for the extraction of verbascoside from Rehmannia root was proposed and the driving forces underlying the extraction processes were also explored. Several ILs with different cations and anions and the parameters of microwave-assisted extraction (MAE), including extraction time, the IL concentration, liquid-solid ratio, and microwave irradiation power, were investigated. Experimental results indicated that hydrogen bonding and hydrophobicity of ILs, π–π stacking between ILs and verbascoside, and steric hindrance effect were the fundamental parameters affecting the extraction efficiency. The IL, 1-benzyl-3-methylimidazolium chloride ([Bemim]Cl) showed the highest extraction ability, thus it was selected as the optimal solvent. Meanwhile the optimized extraction conditions were 30 g g–1 of liquid-solid ratio, 0.3 mol L–1 of [Bemim]Cl, 210 W of microwave irradiation power, and 10 s of extraction time. Under the optimal extraction conditions, the extraction yields of verbascoside from fresh, dried, and processed Rehmannia roots were 0.92, 0.34, and 0.22 mg g–1, respectively. Compared with conventional heating reflux extraction and Soxhlet extraction, the IL-based MAE was an efficient and rapid method (extraction time was only 10 s) with simple sample preparation.

A Novel Integrative Processing Technology for the Preparation of Rehmanniae Radix Slices.

The traditional processing method for the slices preparation of Rehmanniae roots is time- and energy-consuming and is prone to result in loss of active components during twice water-treatment (once for wash and the other for softening) and drying steps. In this study, we firstly explored an integrative processing technique for Rehmanniae Radix by 2x3 factorial experiment based on the contents of catalpol and verbascoside as measured by HPLC. The potential differences between the traditional stepwise processing technique and the integrative processing technique for catalpol and verbascoside in the prepared slices were investigated. To further confirm the effectiveness of drugs using the integrative processing technique, some pharmacological variables, such as rectal temperature, hematologic parameters (RBC, HGB, HCT, and blood viscosity), and coagulation parameters (TT, APTT, PT and FIB), were detected in a blood-heat and hemorrhage syndrome rat model. Two-way ANOVA analysis showed that drying for 18 h at 50°C was considered as the best combination of process conditions. The mean catalpol and verbascoside contents in the integrative method-processed samples (4.30% and 0.33%, respectively) were higher than those in the traditional method-processed samples (2.61% and 0.21%, respectively). Significant increases in rectal temperature, and hematologic parameters, TT, APTT, and FIB, were observed in the model group rats, compared to the blank group animals (P<0.01). Both in the integrative groups and traditional groups, the extracts caused significant decreases in rectal temperature, RBC, HGB, and HCT with increased concentration compared to the model group animals. All coagulation parameters tested were shortened in model rats received two kind prepared slices. There were no significant therapeutic differences between the integrative and the traditional method-processed slices on the hemostasis and hemorheological parameters in this blood-heat and hemorrhage syndrome rat model, indicating that our integrative method may be a feasible technique for processing Rehmanniae Radix slices.