Database-aided ultrahigh-performance liquid chromatography Q-Exactive-Orbitrap tandem mass spectrometry putatively identifies 16 unexpected compounds and three anticounterfeiting pharmacopoeia quality markers for Perillae Fructus.

RATIONALE: Perillae Fructus (PF) is a common traditional Chinese medicine (TCM) for the treatment of asthma. It has not been effectively characterized by rosmarinic acid (RosA), which is currently designed as the sole quality indicator in the Chinese Pharmacopoeia. METHODS: This study introduced a database-aided ultrahigh-performance liquid chromatography equipped with quadrupole-Exactive-Orbitrap mass spectrometry (UHPLC/Q-Exactive-Orbitrap MS/MS) technology to putatively identify the compounds in PF, followed by literature research, quantum chemical calculation, and molecular docking to screen potential quality markers (Q-markers) of PF. RESULTS: A total of 27 compounds were putatively identified, 16 of which had not been previously found from PF. In particular, matrine, scopolamine, and RosA showed relatively high levels of content, stability, and drug-likeness. They exhibited interactions with the asthma-related target and demonstrated the TCM properties of PF. CONCLUSIONS: The database-aided UHPLC/Q-Exactive-Orbitrap MS/MS can identify at least 27 compounds in PF. Of these, 16 compounds are unexpected, and three compounds (matrine, scopolamine, and RosA) should be considered anticounterfeiting pharmacopoeia Q-markers of PF.

Quantitative analysis of three ingredients in Salvia miltiorrhiza by near infrared spectroscopy combined with hybrid variable selection strategy.

Rosmarinic acid (RA), Tanshinone IIA (Tan IIA), and Salvianolic acid B (Sal B) are crucial compounds found in Salvia miltiorrhiza. Quickly predicting these components can aid in ensuring the quality of S. miltiorrhiza. Spectral preprocessing and variable selection are essential processes in quantitative analysis using near infrared spectroscopy (NIR). A novel hybrid variable selection approach utilizing iVISSA was employed in this study to enhance the quantitative measurement of RA, Tan IIA, and Sal B contents in S. miltiorrhiza. The spectra underwent 108 preprocessing approaches, with the optimal method being determined as orthogonal signal correction (OSC). iVISSA was utilized to identify the intervals (feature bands) that were most pertinent to the target chemical. Various methods such as bootstrapping soft shrinkage (BOSS), competitive adaptive reweighted sampling (CARS), genetic algorithm (GA), variable combination population analysis (VCPA), successive projections algorithm (SPA), iteratively variable subset optimization (IVSO), and iteratively retained informative variables (IRIV) were used to identify significant feature variables. PLSR models were created for comparison using the given variables. The results fully demonstrated that iVISSA-SPA calibration model had the best comprehensive performance for Tan IIA, and iVISSA-BOSS had the best comprehensive performance for RA and Sal B, and correlation coefficients of cross-validation (R2cv), root mean square errors of cross-validation (RMSECV), correlation coefficients of prediction (R2p), and root mean square errors of prediction (RMSEP) were 0.9970, 0.0054, 0.9990 and 0.0033, 0.9992, 0.0016, 0.9961 and 0.0034, 0.9998, 0.0138, 0.9875 and 0.1090, respectively. The results suggest that NIR spectroscopy, along with PLSR and a hybrid variable selection method using iVISSA, can be a valuable tool for quickly quantifying RA, Sal B, and Tan IIA in S. miltiorrhiza.

Light spectra manipulation stimulates growth, specialized metabolites and nutritional quality in Anethum graveolens.

Light-Emitting Diodes (LED) play a major role in manipulating light spectra that helps in regulating the growth and specialized metabolite synthesis relevant to the plant defence system. In this study, we assessed photosynthetic performance, phytonutrients, and anatomical variations of an aromatic herb Anethum graveolens (also known as dill), grown under various combinations of LED lights viz. red (100R:0B), red:blue (50R:50B); blue (0R:100B) and warm white (WW, served as control). Exposure to 0R:100B LED lights led to the tallest stem height, whereas, the number of leaves were highest under 50R:50B LED lights. The photosynthetic performance was observed to be highest under 50R:50B LED lights. HPLC analysis revealed chlorogenic acid and rosmarinic acid as the major phenolic compounds accumulated under different spectral irradiations. The highest chlorogenic acid content was observed in 50R:50B LED treated dill plants, while 100R:0B light showed the highest accumulation of rosmarinic acid. Dill plants grown under 50R:50B light displayed a relatively higher content of volatile compounds including, myristicin (phenylpropene), psi-limonene, and α-phellandrene (monoterpenoids). Expression analyses of candidate genes of phenylpropanoid and monoterpenoid biosynthetic pathways showed good correlations with the enhanced phenolic compounds and monoterpenes detected under appropriate light treatments. Further, the stem anatomy revealed higher vascularization under the influence of 0R:100B LED lights, whereas, intense histochemical localization of specialized metabolites could be correlated with enhanced accumulation of phenolic compounds and terpenoids observed in this study. Taken together, these studies suggest that proper combinations of blue and red spectra of light could play important role to augment the growth and phytochemical characteristics of dill, thus improving its value addition in the food industry.

Comprehensive Lichenometabolomic Exploration of Ramalina conduplicans Vain Using UPLC-Q-ToF-MS/MS: An Identification of Free Radical Scavenging and Anti-Hyperglycemic Constituents.

In this study, we propose ultra-performance liquid chromatography coupled with quadrupole/time-of-flight mass spectrometry (UPLC-QToF-MS/MS)-guided metabolite isolation as a choice analytical approach to the ongoing structure−activity investigations of chemical isolates from the edible lichen, Ramalina conduplicans Vain. This strategy led to the isolation and identification of a new depside (5) along with 13 known compounds (1−4, 6−14), most of which being newly described in this lichen species. The structures of the isolates were established by detailed analysis of their spectral data (IR, NMR, and Mass). The acetone extract was further analyzed by UPLC-Q-ToF-MS/MS in a negative ionization mode, which facilitated the identification and confirmation of 18 compounds based on their fragmentation patterns. The antioxidant capacities of the lichen acetone extract (AE) and isolates were measured by tracking DPPH and ABTS free radical scavenging activities. Most isolates displayed marked radical scavenging activities against ABTS while moderate activities were observed against DPPH radical scavenging. Except for atranol (14), oxidative DNA damage was limited by all the tested compounds, with a marked protection for the novel isolated compound (5), as previously noted for the acetone extract (p < 0.001). Furthermore, compound (4) and acetone extract (AE) have inhibited intestinal α-glucosidase enzyme significantly (p < 0.01). Although some phytochemical studies were already performed on this lichen, this study provided new insights into the isolation and identification of bioactive compounds, illustrating interest in future novel analytical techniques.

Regioisomers Salviprolin A and B, Unprecedented Rosmarinic Acid Conjugated Dinorditerpenoids from Salvia przewalskii Maxim.

Salvia przewalskii Maxim is a perennial plant from the genus Salvia (family Lamiaceae). The roots of S. przewalskii were long used as a traditional herb to treat blood circulation related illnesses in China. As part of our continuing interest in polycyclic natural products from medicinal plants, two unprecedented adducts comprised of a dinor-diterpenoid and a 9'-nor-rosmarinic acid derivative, linked by a 1,4-benzodioxane motif (1 and 2), were isolated from the roots of S. przewalskii. Their structures were established by extensive spectroscopic approaches including 1D, 2D NMR, and HRFABMS. Their cytotoxic activities against five human tumor cell lines were evaluated.

The new rapid and accurate analytical HPLC-ECD method for the determination of rosmarinic acid in meat products.

A new rapid method has been developed for the determination of low levels of rosmarinic acid extracted from rosemary (Rosmarinus officinalis L.) and has been used as an antioxidant in meat and meat products after cold storage at 4°C. The method is a high performance liquid chromatography using a coulometric electrochemical detector. It provides a significant improvement on the limit of detection, which was 0.33 ppb, while the limit of quantification was 1 ppb of rosmarinic acid. The advantage of the method also lies in the simpler and faster sample preparation, which can quantify a very low concentration of rosmarinic acid (60 ppb), and is more than 40 ppb below the limits of previously existing methods. A coulometric method is well suited for determining low analyte concentrations and is one of the most sensitive analytical approaches available today, in addition to being time efficient and cost effective. PRACTICAL APPLICATION: A new method for determining low concentrations (60 ppb) of rosmarinic acid in meat and meat products is presented. The method is user-friendly, as it does not require complex sample preparation. It is a selective, precise, and accurate method that makes it useful for routine applications in the meat and other food industries.

Effect of Elicitation with (+)-Usnic Acid on Accumulation of Phenolic Acids and Flavonoids in Agitated Microshoots of Eryngium alpinum L.

The present work was aimed at studying the potential of elicitation on the accumulation of phenolic compounds in in vitro shoot cultures of Eryngium alpinum L., a protected plant from the Apiaceae family. The study examined the influence of (+)-usnic acid on the biomass growth as well as on the biosynthesis of the desired flavonoids and phenolic acids in the cultured microshoots. The phenolic compound content was determined by HPLC-DAD. The flavonoid of the highest concentration was isoquercetin, and the phenolic acids of the highest amount were rosmarinic acid, caffeic acid and 3,4-dihydroxyphenylacetic acid, both in the non-elicited and elicited biomass. Isoquercetin accumulation was efficiently increased by a longer elicitation with a lower concentration of lichenic compound (107.17 ± 4.67 mg/100 g DW) or a shorter elicitation with a higher concentration of acid (127.54 ± 11.34 and 108.37 ± 12.1 mg/100 g DW). Rosmarinic acid production generally remained high in all elicited and non-elicited microshoots. The highest content of this acid was recorded at 24 h of elicitation with 3.125 µM usnic acid (512.69 ± 4.89 mg/100 g DW). The process of elicitation with (+)-usnic acid, a well-known lichenic compound with allelopathic nature, may therefore be an effective technique of enhancing phenolic compound accumulation in alpine eryngo microshoot biomass.

The Contents of Polyphenols in Perilla frutescens (L.) Britton var. frutescens (Egoma) Leaves are Determined by Vegetative Stage, Spatial Leaf Position, and Timing of Harvesting during the Day.

The leaf of Perilla frutescens (L.) Britton var. frutescens (egoma) is a rich source of polyphenolic compounds, including rosmarinic acid. However, there is still a lack of detailed information concerning the content of phenolic compounds in these leaves. Since some flavonoids were found as a conjugated form, leaves were used untreated or hydrolyzed using ß-glucuronidase for analysis. Enzymatic hydrolysis method successfully identified some polyphenols, which have not been reported before. Scutellarin, a flavone glucuronide with a molecular mass similar to that of luteolin 7-O-glucuronide, was present in egoma leaves. Scutellarin was the second most abundant polyphenolic compound, after rosmarinic acid. Egoma leaves at the top of the plant contained a higher amount of rosmarinic acid and scutellarin compared to that in the leaves below. The difference in plant growth stage also influenced the rosmarinic acid and scutellarin contents, while the time of harvesting during the day did rosmarinic acid contents only. This is the first time that scutellarin, a traditional Chinese medicine, widely used for the treatment of cerebrovascular disease, was quantitatively determined in egoma leaves. The present study may help adding value to egoma leaves, developing dietary supplements, functional foods, and cosmetics.

Protective effect of Zataria multiflora Boiss. and its main compound, rosmarinic acid, against malathion induced oxidative stress and apoptosis in HepG2 cells.

Malathion (MT) is one of the most widely used organophosphorus insecticides which induces toxicity through oxidative stress induction, free radical production and acetylcholinesterase inhibition. In this work, HepG2 cells were used to determine the effect of Zataria multiflora methanolic extract (MEZM) and rosmarinic acid (RA) on MT-induced cytotoxicity, oxidative stress, and apoptosis. Total phenolic content (TPC) and total flavonoid content (TFC) were determined and plant was further standardized based on RA content using HPLC method. The cultured HepG2 cells were pretreated with MEZM (1 µg/ml) and RA (0.1 µg/ml) for 4 h and exposed to MT (100 µM). Cell viability, oxidative stress biomarkers, ROS production, and cell death were examined after 24 h. The amount of RA was determined 73.48 mg/g dried extract. IC50 values of MEZM and MT were 368.56 µg/ml and 99.43 µM, respectively. Pretreatment with MEZM and RA decreased the cytotoxicity, oxidative stress, and cell percentage in the late apoptosis and necrosis stages induced by MT. There was no significant difference between MEZM and RA effects. The present study showed the significant protective effects of MEZM against toxicity induced by MT in hepatocytes which can be attributed to the plant antioxidant constituents including RA.

Application of smartphone in detection of thin-layer chromatography: Case of salvia miltiorrhiza.

In this work, a smartphone-based device was constructed for thin-layer chromatography (TLC) detection and semi-quantitative analysis of the components of Salvia miltiorrhiza. The key construction and shooting parameters were investigated by the relative peak area and signal-to-noise ratio. The best conditions were as follows: shooting height, 17 cm; angle between the UV lamp and TLC plate, 58°; exposure compensation, 0~0.2 EV; and shutter speed under daylight and UV 365 nm, 1/50 s and 1/5 s, respectively. These ideal conditions could be replicated by smartphones from different brands with different versions of software. With good precision, repeatability and stability, the developed device was used for the semi-quantitative analysis of salvianolic acid B, rosmarinic acid, cryptotanshinone, tanshinone I, tanshinone IIA, and miltirone in the TLC analysis of 10 batches of S. miltiorrhiza. The results were compared with those obtained by a TLC densitometric scanner and two common types of image processing software, i.e., Gelanalyzer and ImageJ. Except for salvianolic acid B in the TLC densitometric scanner, all results were not significantly different among these methods, which suggested that smartphones might be a useful tool for the quality control of traditional Chinese medicines.

Chitosan stimulates secondary metabolite production and nutrient uptake in medicinal plant Dracocephalum kotschyi.

BACKGROUND: A wide variety of secondary metabolites are synthesized from primary metabolites by plants which have a vast range in pharmaceutical, food additive and industrial applications. In recent years, the use of elicitors has opened a novel approach for the production of secondary metabolite compounds. Dracocephalum kotschyi is a valuable herb due to pharmaceutical compounds like rosmarinic acid, quercetin and apigenin. In the current study, foliar application of chitosan (0, 100, 400 mg L-1 ) as an elicitor was used. RESULTS: After chitosan treatment, the amounts of hydrogen peroxide (H2 O2 ) increased and the plant was able to increase the activities of enzymatic (guaiacol peroxidase, catalase and phenylalanine ammonium lyase) and non-enzymatic (total phenols and flavonoids) defensive metabolites. Also, foliar spray of chitosan promoted nutrient absorption which led to the accumulation of macroelements in the plant. CONCLUSIONS: Chitosan was found to be a very effective elicitor for improving rosmarinic acid and quercetin content (up to 13-fold). Also, the content of apigenin (anticancer flavonoid) showed 16-fold enhancement compared to the control. Therefore, the treatment of D. kotschyi leaves with chitosan caused a very large increase in the induction and production of important pharmaceutical compounds such as rosmarinic acid and quercetin. © 2020 Society of Chemical Industry.

Protective effect of rosmarinic acid-rich trichodesma khasianum clarke leaves against ethanol-induced gastric mucosal injury in vitro and in vivo.

BACKGROUND: Although gastroprotective drugs have been used for peptic ulcer disease prevention and treatment, side effects have been observed. Finding a safe and effective treatment strategy is important. PURPOSE: Edible Trichodesma khasianum (T. khasianum) Clarke leaves are considered to protect against peptic ulcers. However, scientific evidence of this effect of T. khasianum Clarke leaves remains limited. STUDY DESIGN/METHODS: In this study, we aimed to evaluate the effect of T. khasianum Clarke leaves on ethanol-induced gastric injury and gut microbiota using RAW 264.7 cells, RGM-1 cells, and BALB/c mice, respectively. RESULT: The rosmarinic acid was identified as the major component of T. khasianum Clarke leaves extracted by 80% ethanol (80EETC). The results showed that 80EETC suppressed inflammatory mediator protein levels in LPS-induced RAW 264.7 cells. Additionally, heat shock protein expression, antiapoptotic ability, and wound healing migration capability were increased by 80EETC pretreatment in RGM-1 cells with the ethanol-induced injury. Remarkably, pretreatment with 80EETC (150 mg/kg b.w.) promoted gastric mucosal healing by decreasing oxidative stress, inflammatory response, proapoptotic protein expression, and gastric mucosa damage in ethanol-induced gastric injury in mice. Crucially, no liver or kidney toxicities were observed by 80EETC oral gavage. Moreover, 80EETC increased gut microbiota diversity and short-chain fatty acid production. CONCLUSION: Our results illustrated the remarkable gastroprotective effect by 80EETC treatment in vitro and in vivo. These findings are the first to demonstrate the powerful protective effect of T. khasianum Clarke leaves against gastric mucosal injury development.

Supercritical Fluid Chromatography development of a predictive analytical tool to selectively extract bioactive compounds by supercritical fluid extraction and pressurised liquid extraction.

Selective extraction is a great concern in the field of natural products. The interest is to apply specific conditions favouring the solubility of targeted secondary metabolites and avoiding the simultaneous extraction of unwanted ones. Different ways exist to reach selective extractions with suited conditions. These conditions can be determined from experimental studies through experimental design, but a full experimental design takes time, energy, and uses plant samples. Prediction from varied solubility models can also be applied allowing a better understanding of the final selected conditions and eventually less experiments. The aim of this work was to develop and use a chromatographic model to determine optimal extraction conditions without the need for numerous extraction experiments. This model would be applied on the selective extraction of the desired antioxidant compounds in rosemary leaves (rosmarinic and carnosic acids) vs chlorophyll pigments to limit the green colour in extracts. This model was achieved with Supercritical Fluid Chromatography (SFC) and then applied to Supercritical Fluid Extraction (SFE) and Pressurised Liquid Extraction (PLE) assays. SFC models predicted low solubility of chlorophylls for low (5%) and high (100%) percentage of solvent in carbon dioxide. Also, low solubility was predicted with acetonitrile solvent compared to methanol or ethanol. This was confirmed with different extractions performed using SFE with different percentages of solvent (5, 30, and 70%) and with the three solvents used in the SFC models (acetonitrile, methanol and ethanol). Also extractions using PLE were carried out using the same neat solvents in order to confirm the SFC models obtained for 100% of solvent. Globally, extractions validated the SFC models. Only some differences were observed between ethanol and methanol showing the complexity of plant extraction due to matrix effect. For all these extracts, the content of carnosic acid and rosmarinic acid was also monitored and selective extraction conditions of bioactive compounds could be determined.

Rindera graeca (Boraginaceae) Phytochemical Profile and Biological Activities.

Rindera graeca is a Greek endemic plant of the Boraginaceae family which has never been studied before. Consequently, this study attempted to phytochemically examine the aerial parts of this species. Nine phenolic secondary metabolites were identified, consisting of seven caffeic acid derivatives and two flavonol glucosides, namely rutin and quercetin-3-rutinoside-7-rhamnoside. These flavonoids, together with rosmarinic acid, were isolated via column chromatography and structurally determined through spectral analysis. Quercetin-3-rutinoside-7-rhamnoside is an unusual triglycoside, which is identified for the first time in Rindera genus and among Boraginaceae plants. This metabolite was further examined with thermal analysis and its 3D structure was simulated, revealing some intriguing information on its interaction with biological membrane models, which might have potential applications in microcirculation-related conditions. R. graeca was also analyzed for its pyrrolizidine alkaloids content, and it was found to contain echinatine together with echinatine N-oxide and rinderine N-oxide. Additionally, the total phenolic and flavonoid contents of R. graeca methanol extract were determined, along with free radical inhibition assays. High total phenolic content and almost complete inhibition at experimental doses at the free radical assays indicate a potent antioxidant profile for this plant. Overall, through phytochemical analysis and biological activity assays, insight was gained on an endemic Greek species of the little-studied Rindera genus, while its potential for further applications has been assessed.

LC-MS/MS profiles and interrelationships between the enzyme inhibition activity, total phenolic content and antioxidant potential of Micromeria nervosa extracts.

The objective of this study was to quantify the phenolic compounds and to evaluate and compare the biological activities of the ethyl acetate (EtOAc), methanolic (MeOH) and aqueous extracts from the Micromeria nervosa aerial parts, based on their antioxidant activity and enzymatic inhibition. Total phenolic and flavonoid contents were calculated and individual compo3unds were detected using LC-ESI-MS/MS. The antioxidant activity was determined using six different assays while enzymatic activity was determined by α-amylase and tyrosinase enzyme inhibition. The main phenolic constituents detected in the extracts were rosmarinic acid. In the antioxidant assays the aqueous extract was shown to be more efficient than the others. The EtOAc and MeOH extracts presented higher inhibitory activity with respect to α-amylase and tyrosinase. Regardless of the solvent, the results suggest M. nervosa aerial extracts present a biological potential due to their antioxidant activity and enzymatic inhibition.

Polyphenol composition and biological activity of Thymus citriodorus and Thymus vulgaris: Comparison with endemic Iberian Thymus species.

The polyphenol compositions of Thymus × citriodorus and Thymus vulgaris extracts as obtained by exhaustive hydroethanolic (HE) extraction and aqueous decoction (AD) were compared. In addition, their compositions and bioactivities were compared to those of Thymus pulegioides and Thymus mastichina, grown under the same edaphoclimatic conditions, and Thymus carnosus. Rosmarinic acid was the most abundant polyphenol followed by luteolin-hexuronide, salvianolic acids I and K. Cluster analysis suggests a similarity of the polyphenol composition of T. citriodorus and T. vulgaris. A significant antioxidant activity was observed and correlated with their polyphenol levels. The same being observed for the higher anti-proliferative activity/cytotoxicity of HE extracts on Caco-2 and HepG2 cells as compared to AD extracts. Significant association between the total phenolic compounds with the anti-proliferative activity, for both cell lines, was observed. These results support the importance of salvianolic acids levels in Thymus extracts and their in vitro anti-proliferative/cytotoxic activities.

Phytochemical, antioxidant, enzyme activity and antifungal properties of Satureja khuzistanica in vitro and in vivo explants stimulated by some chemical elicitors.

Context: Satureja khuzistanica Jamzad. (Lamiaceae), is known for its antifungal and antioxidant compounds, especially rosmarinic acid (RA).Objective: The study examines the effect of elicitors on RA production and phytochemical properties of S. khuzistanica.Materials and methods: In vitro plants were treated with methyl jasmonate (MeJA) and multi-walled carbon nanotubes (MWCNTs). In vivo plants were treated with MWCNTs and salicylic acid (SA). RA was measured by HPLC. Catalase (CAT), guaiacol peroxidase (POD) and ascorbate peroxidase (APX) were quantified. DPPH and ß-carotene were assayed in in vivo extracts. The antifungal effects of extracts were evaluated against Fusarium solani K (FsK).Results: The highest RA contents of in vitro plants were 50 mg/L MeJA (140.99 mg/g DW) and 250 mg/L MWCNTs (140.49 mg/g DW). The highest in vivo were 24 h MWCNTs (7.13 mg/g DW) and 72 h SA (9.12 mg/g DW). The maximum POD and APX activities were at 100 mg/L MeJA (5 and 4 mg protein, respectively). CAT had the highest activities at 50 mg/L MeJA (2 mg protein). DPPH and ß-carotene showed 50% and 80% inhibition, respectively. The FsK aggregation was the lowest for in vitro extract in number of conidia [1.82 × 1010], fresh weight (6.51 g) and dry weight (0.21 g) that proved RA inhibitory effects. The callus reduces FsK growth diameter to 2.75 on the 5th day.Discussion and conclusions: Application of MeJA, SA, and MWCNTSs could increase RA in S. khuzistanica and highlighted potential characteristics in pharmaceutical and antifungal effects.

Biocompatible chitosan-zinc oxide nanocomposite based dispersive micro-solid phase extraction coupled with HPLC-UV for the determination of rosmarinic acid in the extracts of medical plants and water sample.

In the present research, a procedure was described for the recovery of rosmarinic acid (RA) from medical extract samples using chitosan­zinc oxide nanoparticles as a biocompatible nanocomposite (CS-ZnO-NC). The dispersive micro-solid phase extraction (D-µ-SPE) of RA from the medical extract samples was investigated by using the prepared biocompatible composite as a solid phase. The HPLC-UV method was used for measuring the extracted RA. The important variables (pH, biocompatible composite mass, contact time, and volume of eluent) associated with the extraction process were analyzed by the application of central composite design (CCD). The achieved optimum values for the mentioned variables were 7.0, 10 mg, 4 min, and 180 µL, respectively. The extraction recovery (99.68%) obtained from the predicted model was in agreement with the experimental data (98.22 ± 1.33%). In addition, under the obtained optimum conditions and over the concentration in the range of 2-3500 ng mL-1, a linear calibration curve was obtained with R2 > 0.993. The limit of detection (LOD) and quantification (LOQ) values were computed, and the obtained ranges were respectively from 0.060 to 0.089 ng mL-1 and 0.201 to 0.297 ng mL-1. In addition, the enrichment factors were obtained in the range of 93.7-110.5 with preconcentration factor of 83.3. Therefore, the D-µ-SPE-HPLC-UV method could be used for analyzing RA in the samples of the extracts obtained from the medical plants and water with the recovery values of the analyte in the range of 96.6%-105.4% and the precision with relative standard deviation <5.7%.

In vitro bioaccessibility and activity of Greek oregano (Origanum vulgare L. ssp. hirtum (link) Ietswaart) compounds as affected by nitrogen fertilization.

BACKGROUND: Greek oregano is a culinary and medicinal herb native to the Mediterranean region; however nowadays it is cultivated in many regions all over the world. It is commonly used as a spice for flavoring food products and in various traditional medicine applications. This study investigated the effect of nitrogen fertilization at 0-150 kg N ha-1 on the potential bioaccessibility and activity of Greek oregano compounds. For this purpose the total phenolic content, rosmarinic acid content, and antioxidant activities of the raw material, as well as digested fractions, were determined. RESULTS: Nitrogen fertilization had a negative influence on the phenolic content and antioxidant activity of raw material; however, its effect on the potential bioaccessibility varied depending on the dose. The highest potential bioaccessibility and activity was determined for plants fertilized with 30 kg N ha-1 . For the gastric and intestinal phases of digestion, the potential bioaccessibility percentages were 39.5% and 29.6% for total phenolics, 53.1% and 11.2% for rosmarinic acid content, 45.2% and 44.4% for antiradical activity against 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 39.2% and 27.2% for antiradical activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 50.2% and 23.4% for reducing power, respectively. CONCLUSION: The results showed that nitrogen fertilization is an important factor determining the in vitro bioaccessibility of Greek oregano compounds. Furthermore, the nutraceutical potential of herbs, in term of the bioaccessibility of bioactive compounds, may be optimized during plant cultivation by applying an accurate nitrogen level. © 2020 Society of Chemical Industry.

Polyphenolic contents and antioxidant activities of aerial parts of Salvia multicaulis from the Iran flora.

Folk medicine sometimes involves the use of salvia species for therapeutic purposes. Polyphenols with the highest amounts included rosmarinic acid (7.358mg/g), catechin (1.5 mg/g), vanillin (1.00 mg/g), chlorogenic acid (0.53 mg/g), quercetin (0.16 mg/g), and p-coumaric acid (0.015). Furthermore, the results showed that S. multicaulis has a high content of total phenol (4.39 mg/g) and DPPH activity (8.44 mg/g). Salvia multicaulis could be potentially used as a medicinal plant because of its antioxidant activity and polyphenol content.