Fatty Acid Analysis, Chemical Constituents, Biological Activity and Pesticide Residues Screening in Jordanian Propolis.

Propolis is a resinous natural product collected by honeybees (Apis mellifera and others) from tree exudates that has been widely used in folk medicine. The present study was carried out to investigate the fatty acid composition, chemical constituents, antioxidant, and xanthine oxidase (XO) inhibitory activity of Jordanian propolis, collected from Al-Ghour, Jordan. The hexane extract of Jordanian propolis contained different fatty acids, which are reported for the first time by using GC-FID. The HPLC was carried out to identify important chemical constituents such as fatty acids, polyphenols and α-tocopherol. The antioxidant and xanthine oxidase inhibitory activities were also monitored. The major fatty acid identified were palmitic acid (44.6%), oleic acid (18:1∆9cis, 24.6%), arachidic acid (7.4%), stearic acid (5.4%), linoleic acid (18:2∆9-12cis, 3.1%), caprylic acid (2.9%), lignoceric acid (2.6%), cis-11,14-eicosaldienoic acid (20:2∆11-14cis, 2.4%), palmitoleic acid (1.5%), cis-11-eicosenoic acid (1.2%), α-linolenic acid (18:3∆9-12-15cis, 1.1%), cis-13,16-docosadienoic acid (22:2∆13-16cis, 1.0%), along with other fatty acids. The major chemical constituents identified using gradient HPLC-PDA analysis were pinocembrin (2.82%), chrysin (1.83%), luteolin-7-O-glucoside (1.23%), caffeic acid (1.12%), caffeic acid phenethyl ester (CAPE, 0.79%), apigenin (0.54%), galangin (0.46%), and luteolin (0.30%); while the minor constituents were hesperidin, quercetin, rutin, and vanillic acid. The percentage of α-tocopherol was 2.01 µg/g of the lipid fraction of propolis. Antioxidant properties of the extracts were determined via DPPH radical scavenging. The DPPH radical scavenging activities (IC50) of different extracts ranged from 6.13 to 60.5 µg/mL compared to ascorbic acid (1.21 µg/mL). The xanthine oxidase inhibition (IC50) ranged from 75.11 to 250.74 µg/mL compared to allopurinol (0.38 µg/mL). The results indicate that the various flavonoids, phenolic compounds, α-tocopherol, and other constituents which are present in propolis are responsible for the antioxidant and xanthine oxidation inhibition activity. To evaluate the safety studies of propolis, the pesticide residues were also monitored by LC-MS-MS 4500 Q-Trap. Trace amounts of pesticide residue (ng/mL) were detected in the samples, which are far below the permissible limit as per international guidelines.

Salvianolic acid A attenuates steroid resistant nephrotic syndrome through suPAR/uPAR-αvß3 signaling Inhibition.

ETHNOPHARMACOLOGICAL RELEVANCE: Salvianolic acid A (SAA) is extracted from traditional Chinese medicine Salvia miltiorrhiza and is the main water-soluble and the biologically active ingredient. SAA possesses a variety of pharmacological activities and has an excellent protective effect on kidney disease, especially steroid resistant nephrotic syndrome (SRNS), and has advantages in improving the efficacy of glucocorticoids, but its mechanism needs to be further explored. PURPOSE: The study was designed to explore the effect of suPAR and uPAR in SRNS patients and evaluate the potential effect of SAA in improving podocyte steroid resistance and explore its mechanism. METHODS AND MATERIALS: The ELISA kits were used to detect the levels of suPAR in the blood and urine of subjects. The levels of uPAR, GRα, and GRß expression in renal tissues of SRNS patients was detected by immunohistochemistry and analyzed using the Pearson method. In vitro studies, steroid resistance model was induced by the TNF-α and IFN-γ. The protein and mRNA expression of Nephrin, GR, GRα and GRß were analyzed using western blot and qRT-PCR. The activity of GR-DNA binding was detected by using TransAM™ GR kits. Adriamycin further induced steroid resistance podocyte. Flow cytometry was used to detect the effect of SAA on podocyte apoptosis. ELISA assay was used to detect the suPAR expression in the podocyte supernatant. Western blot and qRT-PCR were used to detect the protein and mRNA expression of uPAR and Nephrin in podocytes. RESULTS: The serum and urine levels of suPAR were conspicuously higher in SRNS patients than healthy volunteers and SSNS patients, and the expression of uPAR in renal tissue of SRNS patients is negatively correlated with GRα, but positively correlated with GRß. The combination of TNF-α and IFN-γ could conspicuously increase the GRß expression and reduce GRα/GRß, and induce steroid resistance in podocytes. Moreover, we found that SAA could reduce the apoptosis of podocytes and suppress the expression of suPAR/uPAR, and increase the expression of Nephrin. CONCLUSION: The level of suPAR and uPAR expression may have important value in predicting glucocorticoids resistance in patients with idiopathic nephrotic syndrome (INS). The combination of TNF-α and IFN-γ induce podocytes can establish steroid resistance model in vitro. SAA could improve glucocorticoids resistance of podocyte which can be attributed in part to regulate the suPAR/uPAR-αvß3 signaling pathway.

Bioactivity-Guided Isolation of Phytochemicals from Vaccinium dunalianum Wight and Their Antioxidant and Enzyme Inhibitory Activities.

Vaccinium dunalianum Wight, usually processed as a traditional folk tea beverage, is widely distributed in the southwest of China. The present study aimed to investigate the antioxidant, α-glucosidase and pancreatic lipase inhibitory activities of V.dunalianum extract and isolate the bioactive components. In this study, the crude extract (CE) from the buds of V. dunalianum was prepared by the ultrasound-assisted extraction method in 70% methanol and then purified with macroporous resin D101 to obtain the purified extract (PM). Five fractions (Fr. A-E) were further obtained by MPLC column (RP-C18). Bioactivity assays revealed that Fr. B with 40% methanol and Fr. D with 80% methanol had better antioxidant with 0.48 ± 0.03 and 0.62 ± 0.01 nM Trolox equivalent (TE)/mg extract for DPPH, 0.87 ± 0.02 and 1.58 ± 0.02 nM TE/mg extract for FRAP, 14.42 ± 0.41 and 19.25 ± 0.23 nM TE/mg extract for ABTS, and enzyme inhibitory effects with IC50 values of 95.21 ± 2.21 and 74.55 ± 3.85 for α-glucosidase, and 142.53 ± 11.45 and 128.76 ± 13.85 µg/mL for pancreatic lipase. Multivariate analysis indicated that the TPC and TFC were positively related to the antioxidant activities. Further phytochemical purification led to the isolation of ten compounds (1-10). 6-O-Caffeoylarbutin (7) showed significant inhibitory effects on α-glucosidase and pancreatic lipase enzymes with values of 38.38 ± 1.84 and 97.56 ± 7.53 µg/mL, and had the highest antioxidant capacity compared to the other compounds.

Methyl Caffeate Isolated from the Flowers of Prunus persica (L.) Batsch Enhances Glucose-Stimulated Insulin Secretion.

Phenolic compounds from natural products are considered effective enhancers of insulin secretion to prevent and treat type 2 diabetes (T2DM). The flowers of Prunus persica (L.) Batsch also contain many phenolic compounds. In this study, the extract of flowers of P. persica (PRPE) exhibited an insulin secretion effect in a glucose-stimulated insulin secretion (GSIS) assay, which led us to isolate and identify the bioactive compound(s) responsible for these effects. Compounds isolated from PRPE were screened for their efficacy in INS-1 rat pancreatic ß-cells. Among them, caffeic acid (5), methyl caffeate (6), ferulic acid (7), chlorogenic acid (8), naringenin (11), nicotiflorin (12), and astragalin (13) isolated from PRPE increased GSIS without inducing cytotoxicity. Interestingly, the GSIS effect of methyl caffeate (6) as a phenolic compound was similar to gliclazide, an antidiabetic sulfonylurea drug. Western blot assay showed that methyl caffeate (6) enhanced the related signaling proteins of the activated pancreatic and duodenal homeobox-1 (PDX-1) and peroxisome proliferator-activated receptor-γ (PPAR-γ), but also the phosphorylation of the total insulin receptor substrate-2 (IRS-2), phosphatidylinositol 3-kinase (PI3K), and Akt, which influence ß-cell function and insulin secretion. This study provides evidence that methyl caffeate (6) isolated from PRPE may aid in the management of T2DM.

Biotechnologically-Produced Myconoside and Calceolarioside E Induce Nrf2 Expression in Neutrophils.

The pathological manifestation of various diseases can be suppressed by the activation of nuclear factor erythroid 2 p45-related factor 2 (Nrf2), a transcriptional regulator of the cellular redox balance. Haberlea rhodopensis Friv. is a resurrection plant species endemic for Bulgaria, containing biologically active phenylethanoid glycosides that might possess antioxidant or redox activity. This study aimed to analyze the metabolic profile of in vitro cultured H. rhodopensis and to identify molecules that increase Nrf2 expression in bone marrow neutrophils. Fractions B, D, and E containing myconoside, or myconoside and calceolarioside E in ratios 1:0.6 and 0.25:1 were found to be the most active ones. Fraction B (200 µg/mL) improved neutrophil survival and strongly increased the Nrf2 intracellular level, while D and E, as well as, myconoside and calceolarioside E at the same ratios had a superior effect. Calceolarioside E (32 µg/mL) had stronger activity than myconoside, the effect of which was very similar to that of 2-cyano-3,12-dioxo-oleana-1,9(11)-dien-28-oic acid methyl ester (CDDO-Me), used as a positive control. These data indicate that both molecules, used alone or in combination have stimulatory activity on the endogenous Nrf2 level, indicating their therapeutic potential to regulate the cellular redox homeostasis oxidative stress-associated pathologies.

Caffeate and piperidine-3-ol derivatives from the stem bark of Cassia sieberiana.

A new caffeate derivative from the ethanol extract of the stem bark of Cassia sieberiana DC. is described herein along with the known secondary metabolites spectaline (2), iso-6-cassine (3), 3-O-methyl-chiro-inositol (4), monobehenin (5), octyl nonadecyloate (6), ß-sitosterol (7), stigmasterol (8) and sitosterol 3-O-ß-D-glucopyranoside (9). The chemical structures were elucidated by means of various spectroscopic and spectrometric techniques. Extract and isolated compounds were devoid of inhibitory action against the herein selected bacterial strains (MICs > 256 µg/mL) but showed capacities to reduce 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical (EC50 < 3 µg/mL) considerably better than the "gold standard" trolox (EC50 6.47 ± 0.48 µg/mL).

Use of asparaginase for acrylamide mitigation in coffee and its influence on the content of caffeine, chlorogenic acid, and caffeic acid.

A factorial design with a duplicate in the central point was used to investigate the effect of treating arabica coffee beans with asparaginase. The investigated factors were enzymatic load (1000 and 5000 ASNU/Kg), water percentage (30 and 90%), and hydrolysis time (1 and 3 h). The acrylamide content was determined by UPLC-MS/MS, and the caffeic acid, chlorogenic acid and caffeine concentrations were determined by HPLC-DAD. The statistical analysis was carried out in the R platform using RStudio graphical interface. The results indicated the importance of coffee bean pretreatment with steam, and that the enzyme load reduced the acrylamide content to 65 mg/kg in coffee beans. The predicted reduction was obtained with hydrolysis time of 2 h, water content of 90%, and asparaginase load of 5000 ASNU/kg. The asparaginase treatment did not influence the major bioactive compounds in coffee.

Symphytum officinale L.: Liquid-liquid chromatography isolation of caffeic acid oligomers and evaluation of their influence on pro-inflammatory cytokine release in LPS-stimulated neutrophils.

ETHNOPHARMACOLOGICAL RELEVANCE: Symphytum officinale L. (comfrey, Boraginaceae) has been traditionally used for millennia in joint distortions, myalgia, bone fractures and hematomas. However, key activity-determining constituents and molecular mechanisms underlying its use have not been completely elucidated. AIM OF THE STUDY: The objective of this study was to isolate and identify the major compounds from a hydroethanolic root extract of S. officinale and evaluate their antioxidant potential, alongside their effect on the cytokine production of ex vivo stimulated neutrophils, thus providing scientific support for the traditional use of comfrey root. MATERIAL AND METHODS: Four caffeic acid oligomers were isolated from comfrey roots by liquid-liquid chromatography, their structures being established by MS and NMR analyses. In vitro antioxidant evaluation was performed by DPPH and ABTS assays. The cytotoxicity of isolated compounds was established by flow cytometry. The effect on cytokine release, such as interleukin (IL)-1ß, IL-8 and tumor necrosis factor alpha (TNF-α), in lipopolysaccharide (LPS)-stimulated neutrophils was determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: The main constituents found in comfrey root were represented by four caffeic acid oligomers, namely globoidnan B (1), rabdosiin (2), rosmarinic acid (3) and globoidnan A (4). Rabdosiin, globoidnans A and B were isolated for the first time from S. officinale. In the in vitro antioxidant tests, compound 2 was the most active, with EC50 values in DPPH and ABTS assays of 29.14 ± 0.43 and 11.13 ± 0.39, respectively. Neutrophils' viability over the tested concentration domain of 12.5-50 µM was not altered. At 50 µM, all compounds significantly inhibited IL-1ß release, with compound 3 (45.60% release vs. LPS stimulated neutrophils) being the most active, followed by compounds 1 (53.85%), 2 (69.89%) and 4 (60.68%). CONCLUSIONS: The four caffeic acid oligomers reported in S. officinale root may contribute to the overall anti-inflammatory activity for which comfrey preparations are used in traditional medicine.

Caffeic acid oligomers from Mesona chinensis and their In Vitro antiviral activities.

The phytochemical study of the aerial part of Mesona chinensis led to the isolation of five new caffeic acid oligomers (1-5), as well as four known analogues (6-9). The structures of the new compounds including their absolute configurations were elucidated by comprehensive spectroscopic analysis, chemical method, and quantum-chemical electronic circular dichroism (ECD) calculation. Among the isolates, compound 7 showed significant in vitro antiviral activity on respiratory syncytial virus (RSV).

Iridoid, phenylethanoid and flavonoid glycosides from Forsythia suspensa.

As part of our continuing efforts to explore bioactive compounds from natural resources, a new iridoid glycoside, adoxosidic acid-6'-oleuroperic ester (1), together with one known phenylethanoid glycoside (2) and two known flavonoid glycosides (3-4) were isolated from the fruit of Forsythia suspensa. The structure of the new compound (1) was elucidated through 1D and 2D NMR spectroscopic data and HR-ESIMS. Interestingly, compound 1 was a monoterpene ester of one iridoid glycoside. Compounds 2-4 were identified as calceolarioside A (2), kaempferol-3-O-rutinoside (3), kampferol-3-O-robinobioside (4) on the basis of NMR spectroscopic data analyses and comparison with the data reported in the literature. The antiviral activity aganisist influenza A (H5N1) virus of compound 1 was studied as well.

Chemosensitizing Effect of Cernumidine Extracted from Solanum cernuum on Bladder Cancer Cells in Vitro.

Cernumidine (CER) is a guanidinic alkaloid isolated from Solanum cernuum leaves. In this work, we investigated the cytotoxicity, chemosensitizing effect of cernumidine to cisplatin (cDDP) and the possible mechanism of action of the combination on bladder cancer cells. Cernumidine showed cytotoxicity and could sensitize bladder cancer cells to cisplatin. The combination of CER+cDDP inhibited cell migration on T24 cells. CER+cDDP down-regulated MMP-2/9 and p-ERK1/2, while it increased EGFR activity corroborating the observed cell migration inhibition. Down-regulation of Bcl-2 and up-regulation pro-apoptotic Bax and further depletion of the mitochondrial membrane potential (ΔΨm) indicates that mitochondria play a central role in the combination treatment inducing the mitochondrial signaling pathway of apoptosis in T24 cells. Our data showed that the alkaloid cernumidine is worthy of further studies as a chemosensitizing agent to be used in complementary chemotherapy.

Liquid chromatography fingerprint analysis and antioxidant activity of selected lavender species with chemometric calculations.

The extracts of seven Lavandulae species (Lavandula stoechas, Lavandula lanata, Lavandula viridis, Lavandula angustifolia "Rosea", Lavandula angustifolia "Afropurpurea", Lavandula angustifolia and one unknown) were analyzed using the reversed-phase-high performance liquid chromatography-diode array detection (RP-HPLC-DAD) with gradient elution technique to obtain the chromatographic fingerprint profiles. The HPLC analysis was performed using the Kinetex RP18 chromatographic column and eluent consisting of methanol-water-0.1% formic acid (5-100% (v/v)) at 30 °C with the run time of 60 min. and the detection wavelength 280 nm. The chromatograms were preliminary processed with the smoothing, noise reduction, background subtraction and alignment using the SpecAlign program (version 2.4.1). The presence of selected standards (apigenin, myricetin, luteolin, luteolin 7-glucoside, chlorogenic acid, caffeic acid, ferulic acid) in the extracts was confirmed. The chemical similarity between studied plants was evaluated using the Cluster Analysis (Pearson correlation coefficient, r, and Euclidean) and PCA. The preliminary antioxidant activity of studied extracts was evaluated based on the total phenolic content (Folin-Ciocalteu method), ferric ion reducing antioxidant parameter (FRAP) and α,α-diphenyl-ß-picrylhydrazyl (DPPH) free radical scavenging method using the spectrophotometric technique.

Optimization of ultrasound assisted extraction method for phytochemical compounds and in-vitro antioxidant activity of New Zealand and China Asparagus cultivars (officinalis L.) roots extracts.

A two-level Plackett-Burman design with 8 variables was used to evaluate ultrasonic treatment variables influencing the total phenolic content (TPC) extracted from asparagus roots. Steepest ascent method was conducted to identify the significance of parameters such as extraction temperature, stirring speed, intermission time, extraction time, ultrasonic frequency, and ultrasonic power. Ethanol and methanol aqueous solutions were used as extraction solvents and solvent's concentration, extraction time, ultrasonic power and solid: liquid ratio were optimized in this study. A predicted value of TPC (71.1 mg/g) was obtained under the optimum conditions of extraction time 120 min, ultrasonic power 550 W, ethanol concentration of 20% and a solid: liquid ratio of 1:100. Central composite design was employed to further analyse the common interactions between the extraction variables and to further determine the optimal values that would generate the maximum TPC, total flavonoids content, total saponins content, caffeic acid and in vitro antioxidant activities. The optimal variables for ethanol extraction (80 min, 50% of ethanol, 360 W and 1:40) generated higher than methanol (410 W for 114.9 min using 73.7% methanol at 1:24).

Biosynthesis of gold nanoparticles using Caffeoylxanthiazonoside, chemical isolated from Xanthium strumarium L. fruit and their Anti-allergic rhinitis effect- a traditional Chinese medicine.

This study significantly aims to analyze the effect of CFX coated gold nanoparticles on the allergic rhinitis in mouse model. Female BALB/C mice were intraperitoneally injected with ovalbumin and aluminum hydroxide in order to sensitize on 0th, 7th, 14th, and 21st day. Nano­gold was also nasally inoculated earlier to the intranasal administration of OVA. The acuteness of allergic rhinitis was evaluated based on the nasal indications, interleukin IL-4, and interferon (INF)-γ levels present in the NLF of mice. Also, Hematoxylin-eosin as well as Schiff-periodic acid stain assays were carried out to briefly investigate the histological modifications in the sensitized mice. Nano­gold reduced the incidence of nasal symptoms as well as minimized the IL-4 production, no impact was showed over the levels of INF-γ. Further, the extent of inflammatory cell intrusion was also influenced by the CFX/AuNPs. These experimental findings manifested that AuNPs may considerably inhibit the allergic inflammation in mice models and may be expected to serve as anti-allergic rhinitis agents.

Identification of Six Phytochemical Compounds from Asparagus officinalis L. Root Cultivars from New Zealand and China Using UAE-SPE-UPLC-MS/MS: Effects of Extracts on H2O2-Induced Oxidative Stress.

A simple, rapid, specific, and sensitive method was developed for the simultaneous identification and quantification of six major bioactive compounds, namely, caffeic acid, quercetin, apigenin, ferulic acid, baicalein, and kaempferol, from Asparagus officinalis roots (ARs) native to New Zealand (green and purple cultivars) and China (yellow, green, purple, and white cultivars) using ultrasound-assisted, solid-phase extraction (UASE-SPE) coupled with ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The method was validated in terms of linearity, limit of detection (LOD), limit of quantification (LOQ), accuracy (expressed as recovery %), and precision (expressed as relative standard deviation (%RSD)). The retention times, ultraviolet visible (UV-vis) data, and mass spectral patterns of the detected peaks matched those of commercial standards, allowing characterization of the target compounds. The LODs and LOQs were 23 ng/mL and 70 ng/mL, 50 ng/mL and 150 ng/mL, 10 ng/mL and 30 ng/mL, 18 ng/mL and 54 ng/mL, 14.4 ng/mL and 43.6 ng/mL, and 7.5 ng/mL and 22.5 ng/mL for caffeic acid, quercetin, apigenin, ferulic acid, baicalein, and kaempferol, respectively, and the mean recovery rates were 85.8%, 73.0%, 90.2%, 80.6%, 76.7%, and 74.5% for the six compounds, respectively. The levels of the target compounds were significantly different (p < 0.05) among the six cultivars. The Chinese yellow AR had the highest levels of bioactive compounds: 6.0, 3.9, 0.4, 1.0, 0.86, and 0.8 mg/g for caffeic acid, quercetin, apigenin, ferulic acid, baicalein, and kaempferol, respectively. The AR extracts showed protective effects against oxidative stress in the HepG2 and L929 cell lines. The results indicate that AR extracts contain high flavonoid levels that provide protective functions against oxidative stress and support the potential commercial application of AR extracts.

Isolation, identification of secondary metabolites from Salvia absconditiflora and evaluation of their antioxidative properties.

Salvia absconditiflora Greuter & Burdet (Synonym Salvia cryptantha Montbret & Aucher ex Benth) has been used extensively for traditional medicine. The aerial of plant material was boiled in water then filtrated. The filtrate was partitioned with ethyl acetate and n-butanol sequentially to yield the ethyl acetate and n-butanol extract. A sample of water was lyophilized to yield the water extract. Ethyl acetate extract revealed the highest antioxidant activity and included the most phenolic compounds among the extracts. Hence, ethyl acetate extract was subjected to chromatographic techniques. Ursolic acid (1), crismaritin (2), luteolin (3), rosmarinic acid methyl ester (4), 3,4-dihydroxyl benzaldehyde (protocatechuic aldehyde) (5), caffeic acid (6), apigenin-7-O-ß-glucoside (7), rosmarinic acid (8) and luteolin-7-O-ß-glucoside (9) were isolated and the structures were elucidated by spectroscopic methods including 1D, 2D NMR, and LC-TOF/MS. Cirsimaritin (2), luteolin (3), rosmarinic acid methyl ester (4), rosmarinic acid (8), luteolin-7-O-ß-glucoside (9) displayed the considerable antioxidant activity.

Clinopodium vulgare L. (wild basil) extract and its active constituents modulate cyclooxygenase-2 expression in neutrophils.

Clinopodium vulgare L. (wild basil) has a wide range of ethnopharmacological applications and accumulates a broad spectrum of phenolic compounds, recognized for their anti-inflammatory and anticancer properties. The triggered cyclooxygenase-2 (COX-2) expression is creating an immunosuppressive microenvironment in the inflamed tissue and considered to be the main cause of failure of even new anticancer-/immune-therapies. Nowadays, selective and novel plant-derived COX-2 inhibitors with safe profile are subject of profound research interest. This study aimed to analyze the metabolic profile of C. vulgare and search for phenolic molecules with potential biological properties. By application of 1H and 2D-NMR (Nuclear Magnetic Resonance) profiling, caffeic, chlorogenic acids and catechin were identified along with a bunch of primary and secondary metabolites. Further, the biological effect of C. vulgare extract (CVE) and its constituents on zymosan-induced COX-2 expression and apoptosis of murine neutrophils have been studied. The CVE, caffeic and chlorogenic acids inhibited zymosan-induced COX-2 expression in bone marrow neutrophils, in vitro and in vivo activated. The obtained data indicate that CVE may have a good potential to manipulate neutrophil functions, however, its action may depend on the cellular state, the inflammatory milieu and the relative content of caffeic and chlorogenic acid in the extract.

Highly selective separation and purification of chicoric acid from Echinacea purpurea by quality control methods in macroporous adsorption resin column chromatography.

Chicoric acid is the main phenolic active ingredient in Echinacea purpurea (Asteraceae), best known for its immune-enhancing ability, as well as used as a herbal medicine. To achieve further utilization of medicinal ingredients from E. purpurea, an efficient preparative separation of chicoric acid was developed based on macroporous adsorption resin chromatography. The separation characteristics of several different typical macroporous adsorption resins were evaluated by adsorption/desorption column experiments, and HPD100 was revealed as the optimal one, which exhibited that the adsorbents fitted well to the pseudo-second-order kinetics model and Langmuir isotherm model, and the optimal process parameters were obtained. The breakthrough curves could be predicted and end-point could be determined early. Besides, the optimal elution conditions of chicoric acid can be achieved using the quality control methods. As a result, the purity of chicoric acid was increased 15.8-fold (from 4 to 63%) after the treatment with HPD100. The process of the enrichment and separation of chicoric acid is considerate, because of its high efficiency and simple operation. The established separation and purification method of chicoric acid is expected to be valuable for further utilization of E. purpurea according to product application in pharmaceutical fields in the future.

Harvest Strategies for Optimization of the Content of Bioactive Alkamides and Caffeic Acid Derivatives in Aerial Parts and in Roots of Echinacea purpurea.

Aerial parts and roots of Echinacea purpurea were harvested consecutively in order to find the best strategy for harvest of both types of plant material for an optimal content of bioactive alkamides and caffeic acid derivatives. Four caffeic acid derivatives and 15 alkamides were identified and quantified. The aerial parts were harvested in bud, bloom, and wilting stage and the roots were harvested 1 week, 1 month, and 3 months after each harvest of aerial parts. The highest yield per area of both alkamides and caffeic acid derivatives is achieved when the aerial parts are harvested late (wilting stage). To obtain an optimal content of alkamides and caffeic acid derivatives it is not recommendable to harvest the aerial parts and the roots in the same year. If the aerial parts must be harvested, the roots should be harvested 1 week after because this will result in the most optimal concentration of bioactive compounds in both products.

Metabolites and Biological Activities of Thymus zygis, Thymus pulegioides, and Thymus fragrantissimus Grown under Organic Cultivation.

Thymus plants are marketed for diverse usages because of their pleasant odor, as well as high nutritional value and wealth of health-promoting phytochemicals. In this study, Thymuszygis, Thymuspulegioides, and Thymusfragrantissimus grown under organic cultivation regime were characterized regarding nutrients and phenolic compounds. In addition, the antioxidant and antibacterial properties of these species were screened. The plants were particularly notable for their high K/Na ratio, polyunsaturated fatty acids content and low omega-6/omega-3 fatty acids ratios, which are valuable features of a healthy diet. Caffeic acid and/or its derivatives, mainly rosmarinic acid and caffeoyl rosmarinic acid, represented the majority of the phenolic constituents of these plants, although they were less representative in T. pulegioides, which in turn was the richest in flavones. The latter species also exhibited the highest antioxidant capacity (DPPH● EC50 of 9.50 ± 1.98 µg/mL and reducing power EC50 of 30.73 ± 1.48 µg/mL), while T. zygis was the most active towards Gram-positive and Gram-negative bacteria. Overall, the results suggest that the three thyme plants grown in organic farming are endowed with valuable metabolites that give them high commercial value for applications in different industries.