UHPLC-Q/Orbitrap/MS/MS Fingerprinting, Free Radical Scavenging, and Antimicrobial Activity of Tessaria absinthiodes (Hook. & Arn.) DC. (Asteraceae) Lyophilized Decoction from Argentina and Chile.

The decoction of Tessaria absinthioides is used in traditional medicine of South America as hypocholesterolemic, balsamic, and expectorant; but it is also useful for the prevention of hepatitis, renal insufficiency, and diabetes, and is used as digestive. A lyophilized decoction from the aerial parts of this plant (TLD) collected in San Juan (TLDSJ) and Mendoza (TLDM) provinces (Argentina) and one collection from Antofagasta, Chile (TLDCH) were characterized regarding antioxidant and antibacterial activities, phenolics and flavonoids content, and ultrahigh resolution liquid chromatography Orbitrap MS analysis UHPLC-PDA-OT-MS/MS metabolite profiling. The antioxidant properties were carried out "in vitro" using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and trolox equivalent antioxidant activity (TEAC) methods, ferric-reducing antioxidant power (FRAP), and lipoperoxidation in erythrocytes (LP). The antibacterial activity was evaluated following the Clinical and Laboratory Standards Institute (CLSI) rules. TLDSJ, TLDM, and TLDCH displayed a strong DPPH scavenging activity (EC50 = 42, 41.6, and 43 µg/mL, respectively) and inhibition of lipoperoxidation in erythrocytes (86-88% at 250 µg TLD/mL), while a less effect in the FRAP and TEACantioxidant assays was found. Additionally, the decoctions showed a content of phenolics compounds of 94 mg gallic acid equivalents (GAE)/g, 185 GAE/g, and 64 GAE/g, for TLDSJ, TLDM, and TLDCH samples, respectively. Regarding the flavonoid content, the Chilean sample was highlighted with 19 mg quercetin equivalents (QE)/g. In this work, several phenolic compounds, including sesquiterpenes, flavonoids, and phenolic acids, were rapidly identified in TLDSJ, TLDM, and TLDCH extracts by means UHPLC-PDA-OT-MS/MS for the first time, which gave a first scientific support to consider this medicinal decoction from both countries as a valuable source of metabolites with antioxidant effects, some with outstanding potential to improve human health.

Antiproliferative effect and ultrastructural alterations induced by 5-O-methylembelin on Trypanosoma cruzi.

BACKGROUND: Embelin (EMB), obtained from Oxalis erythrorhiza Gillies ex Hooker et Arnott (Oxalidaceae), was reported against Trypanosoma cruzi and Leishmania spp. Additionally, antiprotozoan activity against Plasmodium falciparum was reported for its methylated derivative (ME). PURPOSE: To evaluate the potential anti-Trypanosoma cruzi activity of EMB, ME and 2,5-di-O-methylembelin (DME) and analyze the possible mechanism of action. STUDY DESIGN/METHODS: EMB was isolated by a chromatographic method from the air-dried ground whole plant. To evaluate the effects of methylation, ME and DME were synthesized and tested against T. cruzi epimastigotes and trypomastigotes. The most active compound ME was evaluated against amastigotes. Ultrastructural alterations, ROS generation and the effect on mitochondrial activity of ME were measured. RESULTS: Compounds inhibited the proliferation of epimastigotes. ME was also active against intracellular amastigotes. Mitochondrial alterations were observed by TEM. Additionally, ME modified the mitochondrial activity, and induced an increase in ROS levels. These evidences postulate the mitochondrion as a possible target of ME. CONCLUSION: ME inhibited amastigotes proliferation, thus being a potential lead compound for the treatment of Chagas' disease.

Green Synthesis of Potential Antifungal Agents: 2-Benzyl Substituted Thiobenzoazoles.

A series of benzyl-substituted thiobenzoazoles were synthesized by an environmentally friendly approach, to search for new antifungal agrochemicals. Compounds were prepared starting from 2-mercaptobenzoazoles, using KOH, benzyl halides, and water, resulting in a simple and ecological method. New antifungals were tested against a group of phytopathogenic fungi. Two compounds showed an interesting activity against Botrytis cinerea, Fusarium oxysporum, and Aspergillus spp.: 2-((4-(trifluoromethyl)benzyl)thio)benzo[d]thiazole, 3ac, and 2-((4-methylbenzyl)thio)benzo[d]thiazole, 3al. Thus, 3ac and 3al can be considered as broad spectrum antifungal agents. Furthermore, two new compounds, 2-((4-iodobenzyl)thio)benzo[d]thiazole, 3aj, and 2-(benzylthio)benzo[d]oxazole, 3ba, showed better inhibitory effect against Botrytis cinerea and Fusarium oxysporum when compared to the commercial fungicide Captan. Thus, 3aj and 3ba can be considered reduced-spectrum antifungals.

Synthesis, characterization and biological studies of a cobalt(III) complex of sulfathiazole.

The emergence of old and new antibiotic resistance created in the last decades revealed a substantial medical need for new classes of antimicrobial agents. The antimicrobial activity of sulfa drugs is often enhanced by complexation with metal ions, which is in concordance with the well-known importance of metal ions in biological systems. Besides, sulfonamides and its derivatives constitute an important class of drugs, with several types of pharmacological agents possessing antibacterial, anti-carbonic anhydrase, diuretic, hypoglycemic, antithyroid, antiviral and anticancer activities, among others. The purpose of this work has been the obtainment, characterization and determination of biological properties (antibacterial, antifungal, mutagenicity and phytotoxicity) of a new Co(III)-sulfathiazole complex: Costz, besides of its interaction with bovine serum albumin (BSA). The reaction between sodium sulfathiazole (Nastz) and cobalt(II) chloride in the presence of H2O2 leads to a brown solid, [CoIII(stz)2OH(H2O)3], (Costz). The structure of this compound has been examined by means of elemental analyses, FT-IR, 1H NMR, UV-Visible spectrometric methods and thermal studies. The Co(III) ion, which exhibits a distorted octahedral environment, could coordinate with the N thiazolic atom of sulfathiazolate. The complex quenched partially the native fluorescence of bovine serum albumin (BSA), suggesting a specific interaction with the protein. The Costz complex showed, in vitro, a moderate antifungal activity against Aspergillus fumigatus and A. flavus. As antibacterial, Costz displayed, in vitro, enhanced activity respective to the ligand against Pseudomonas aeruginosa. Costz did not show mutagenic properties with the Ames test. In the Allium cepa test the complex showed cytotoxic properties but not genotoxic ones. These results may be auspicious, however, further biological studies are needed to consider the complex Costz as a possible drug in the future.

Argentinian pistachio oil and flour: a potential novel approach of pistachio nut utilization.

In order to searching a potential novel approach to pistachio utilization, the chemical and nutritional quality of oil and flour from natural, roasted, and salted roasted pistachios from Argentinian cultivars were evaluated. The pistachio oil has high contents of oleic and linoleic acid (53.5 - 55.3, 29 - 31.4 relative abundance, respectively), tocopherols (896 - 916 µg/g oil), carotenoids (48 - 56 µg/g oil) and chlorophylls (41 - 70 µg/g oil), being a good source for commercial edible oil production. The processing conditions did not affect significantly the fatty acid and minor composition of pistachio oil samples. The content of total phenolic (TP) and flavonoids (FL) was not significantly modified by the roasting process, whereas free radical scavenging (DPPH radical) and antioxidant power decreased in a 20% approximately. Furthermore, salted roasted pistachio flour (SRPF) showed a significant decrease in TP and FL content in comparison to others samples. The phenolic profile of pistachio flours evaluated by LC-ESI-QTOF-MS. The major compounds identified were (+)-catechin (38 - 65.6 µg/g PF d.w.), gallic acid (23 - 36 µg/g PF d.w.) and cyanidin-3-O-galactoside (21 - 23 µg/g PF d.w.). The treatments effects on the phenolics constituents of pistachio flour. Roasting caused a significant reduction of some phenolics, gallic acid and (+)- catechin, and increased others, naringenin and luteolin. Salting and roasting of pistachio increased garlic acid and naringenin content.

In order to searching a potential novel approach to pistachio utilization, the chemical and nutritional quality of oil and flour from natural, roasted, and salted roasted pistachios from Argentinian cultivars were evaluated. The pistachio oil has high contents of oleic and linoleic acid (53.5­55.3, 29­31.4 relative abundance, respectively), tocopherols (896­916 µg/g oil), carotenoids (48­56 µg/g oil) and chlorophylls (41­70 µg/g oil), being a good source for commercial edible oil production. The processing conditions did not affect significantly the fatty acid distribution and minor components of pistachio oil samples. The roasting process not diminish total phenolic (TP) and flavonoids (FL) content significantly compared to natural pistachio flour (NPF), even so reduced the DPPH antioxidant capacity (approximately 20 %) in the roasted pistachio flour (RPF). Furthermore, salted roasted pistachio flour (SRPF) showed a slight and significant decrease on TP and FL content in relation to the others samples. The phenolic profile of pistachio flours were evaluated by LC-ESI-QTOF-MS. The major compounds were (+)-catechin (38­65.6 µg/g PF d.w.), gallic acid (23­36 µg/g PF d.w.) and cyanidin-3-O-galactoside (21­23 µg/g PF d.w.). The treatments have different effects on the phenolics constituents of pistachio flour. Roasting caused a significant reduction of some phenolics, gallic acid and (+)-catechin, and increased others, naringenin and luteolin. Otherwise, salting and roasting of pistachio increased levels of gallic acid and naringenin. These results suggest that Argentinian pistachio oil and flour could be considered as ingredients into applications that enhance human health.

Antibacterial Activity, Antioxidant Effect and Chemical Composition of Propolis from the Región del Maule, Central Chile.

Propolis is commercialized in Chile as an antimicrobial agent. It is obtained mainly from central and southern Chile, but is used for the same purposes regardless of its origin. To compare the antimicrobial effect, the total phenolic (TP), the total flavonoid (TF) content and the phenolic composition, 19 samples were collected in the main production centers in the Región del Maule, Chile. Samples were extracted with MeOH and assessed for antimicrobial activity against Gram (+) and Gram (-) bacteria. TP and TF content, antioxidant activity by the DPPH, FRAP and TEAC methods were also determined. Sample composition was assessed by HPLD-DAD-ESI-MS/MS. Differential compounds in the samples were isolated and characterized. The antimicrobial effect of the samples showed MICs ranging from 31.5 to > 1000 µg/mL. Propolis from the central valley was more effective as antibacterial than those from the coastal area or Andean slopes. The samples considered of interest (MIC ≤ 62.5 µg/mL) showed effect on Escherichia coli, Pseudomonas sp., Yersinia enterocolitica and Salmonella enteritidis. Two new diarylheptanoids, a diterpene, the flavonoids pinocembrin and chrysin were isolated and elucidated by spectroscopic and spectrometric means. Some 29 compounds were dereplicated by HPLC-MS and tentatively identified, including nine flavones/flavonol derivatives, one flavanone, eight dihydroflavonols and nine phenyl-propanoids. Propolis from the Región del Maule showed large variation in antimicrobial effect, antioxidant activity and composition. So far the presence of diarylheptanoids in samples from the coastal area of central Chile can be considered as a marker of a new type of propolis.

Essential oil of Azorella cryptantha collected in two different locations from San Juan Province, Argentina: chemical variability and anti-insect and antimicrobial activities.

The essential oils (EOs) of two populations of Azorella cryptantha (Clos) Reiche, a native species from San Juan Province, were obtained by hydrodistillation in a Clevenger-type apparatus and characterized by GC-FID and GC/MS analyses. The compounds identified amounted to 92.3 and 88.7% of the total oil composition for A. cryptantha from Bauchaceta (Ac-BAU) and Agua Negra (Ac-AN), respectively. The EO composition for the two populations was similar, although with differences in the identity and content of the main compounds and also in the identity of minor components. The main compounds of the Ac-BAU EO were α-pinene, α-thujene, sabinene, δ-cadinene, δ-cadinol, trans-ß-guaiene, and τ-muurolol, while α-pinene, α-thujene, ß-pinene, γ-cadinene, τ-cadinol, δ-cadinene, τ-muurolol, and a not identified compound were the main constituents of the Ac-AN EO, which also contained 3.0% of oxygenated monoterpenes. The repellent activity on Triatoma infestans nymphs was 100 and 92% for the Ac-AN and Ac-BAU EOs, respectively. Regarding the toxic effects on Ceratitis capitata, the EOs were very active with LD(50) values lower than 11 µg/fly. The dermatophytes Microsporum gypseum, Trichophyton rubrum, and T. mentagrophytes and the bacterial strains Escherichia coli LM(1), E. coli LM(2), and Yersinia enterocolitica PI were more sensitive toward the Ac-AN EO (MIC 125 µg/ml) than toward the Ac-BAU EO. This is the first report on the composition of A. cryptantha EO and its anti-insect and antimicrobial properties.

Argentinean Andean propolis associated with the medicinal plant Larrea nitida Cav. (Zygophyllaceae). HPLC-MS and GC-MS characterization and antifungal activity.

The chemical profile and botanical origin of Andean Argentinian propolis were studied by HPLC-ESI-MS/MS and GC-MS techniques as well as the antifungal activity according to CLSI protocols. Dermatophytes and yeasts tested were strongly inhibited by propolis extracts (MICs between 31.25 and 125 µg/mL). The main antifungal compounds were: 3'methyl-nordihydroguaiaretic acid (MNDGA) 1, nordihydroguaiaretic acid (NDGA) 2 and a NDGA derivative 3, showing strong activity against Trichophyton mentagrophytes, T. rubrum and Microsporum gypseum (MICs between 15.6 and 31.25 µg/mL). The lignans 1 and 2 showed activities against clinical isolates of Candidas spp., Cryptococcus spp., T. rubrum and T. mentagrophytes (MICs and MFCs between 31.25 and 62.5 µg/mL). The lignan and volatile organic compounds (VOCs) profiles from propolis matched with those of exudates of Larrea nitida providing strong evidences on its botanical origin. These results support that Argentinian Andean propolis are a valuable natural product with potential to improve human health. Six compounds (1-6) were isolated from propolis for the first time, while compounds 1 and 3-6 were reported for first time as constituents of L. nitida Cav.

Antifungal activity of extracts and prenylated coumarins isolated from Baccharis darwinii Hook & Arn. (Asteraceae).

The petroleum ether extract of Baccharis darwinii showed activity against Cryptococcus neoformans and dermatophytes. Bioactivity-guided fractionation of Baccharis darwinii has resulted in the isolation of three coumarins: 5'-hydroxy aurapten (anisocoumarin H, 1), aurapten (7-geranyloxycoumarin, 2) and 5'-oxoaurapten (diversinin, 3). The structures of these compounds were characterized by spectroscopic methods. These compounds were evaluated for their antimicrobialactivity against a panel of each, bacteria and fungi. Compound 3 showed the best activities against Microsporum gypseum, Trichophyton rubrum and Trichophyton mentagrophytes with MICs = 15.6 microg/mL, followed by compound 1 whose MICs against the same fungi were 62.5 microg/mL. In addition they showed fungicidal rather than fungistatic activity. Both compounds showed moderate activity (MICs = 125 microg/mL) against Cryptococcus neoformans. This is the first report of the presence of compound 1 in B. darwinii.

Argentinean propolis from Zuccagnia punctata Cav. (Caesalpinieae) exudates: phytochemical characterization and antifungal activity.

This paper reports the in vitro antifungal activity of propolis extracts from the province of Tucuman (Argentina) as well as the identification of their main antifungal compounds and botanical origin. The antifungal activity was determined by the microdilution technique, using reference microorganisms and clinical isolates. All dermatophytes and yeasts tested were strongly inhibited by different propolis extracts (MICs between 16 and 125 microg mL(-1)). The most susceptible species were Microsporum gypseum, Trichophyton mentagrophytes, and Trichophyton rubrum. The main bioactive compounds were 2',4'-dihydroxy-3'-methoxychalcone 2 and 2',4'-dihydroxychalcone 3. Both displayed strong activity against clinical isolates of T. rubrum and T. mentagrophytes (MICs and MFCs between 1.9 and 2.9 microg mL(-1)). Additionally, galangin 5, pinocembrin 6, and 7-hydroxy-8-methoxyflavanone 9 were isolated from propolis samples and Zuccagnia punctata exudates, showing moderate antifungal activity. This is the first study matching the chemical profile of Z. punctata Cav. exudates with their corresponding propolis, giving strong evidence on the botanical origin of the studied propolis.

A new antifungal and antiprotozoal depside from the Andean lichen Protousnea poeppigii.

Extracts from the Andean lichens Protousnea poeppigii and Usnea florida displayed antimicrobial activity against the pathogenic fungi Microsporum gypseum, Trichophyton mentagrophytes and T. rubrum with MIC values between 50 and 100 microg/mL. From the active extracts, four main metabolites were isolated and identified as the new depside, isodivaricatic acid, and the known metabolites 5-propylresorcinol, divaricatinic acid and usnic acid. Isodivaricatic acid and divaricatinic acid presented antifungal effect towards M. gypseum with a MIC of 50 microg/mL and against T. mentagrophytes and T. rubrum and with MIC values of 50 and 100 microg/mL, respectively. The new isodivaricatic acid was active towards Leishmania amazonensis, Leishmania brasiliensis and Leishmania infantum promastigotes with 100% lysis at 100 microg/mL.The activity of the new compound decreased on acetylation of the hydroxy groups as well as on methylation of the acid function. The structures were elucidated by spectroscopic means. The spectroscopic data of isodivaricatic acid are presented here for the first time.

Free radical scavengers and antioxidants from Baccharis grisebachii.

The exudate and seriated extracts from the aerial parts of Baccharis grisebachii (Asteraceae) which is recommended as a digestive and to relieve gastric ulcers in Argentina, showed activity as free radical scavengers and inhibited lipoperoxidation in erythrocytes. Assay-guided isolation led to seven p-coumaric acid derivatives and six flavonoids as the main active constituents of the crude drug. The activity towards the superoxide anion was mainly due to the flavonoid constituents. 5,7,4'-Trihydroxy-6-methoxyflavone and quercetin presented high activity (64 and 79%) even at 12.5 microg/ml. The xanthine oxidase inhibitory effect of the extracts can be related with the p-coumaric acid derivatives drupanin, 4-acetyl-3,5-diprenylcinnamic acid and trans-ferulic acid O-hexan-3-onyl-ether which showed IC(50) values in the range 28-40 microg/ml. Both p-coumaric acid derivatives and flavonoids inhibited lipoperoxidation in erythrocytes. The highest activity was found for the p-coumaric acid derivatives 4-acetyl-3-prenyl-ethoxycinnamate, 3-prenyl-4-(4'-hydroxydihydrocinnamoyloxy)-cinnamate and trans-ferulic acid O-hexan-3-onyl-ether (69-82%) and the flavonoids 5,7,4'-trihydroxy-6-methoxyflavone, quercetin, 5,7,4'-trihydroxy-6,3'-dimethoxyflavone and 5,7,4'-trihydroxy-6,8-dimethoxyflavone (64-84%) at 100 microg/ml. The most active free radical scavengers measured by the DPPH decoloration assay were the p-coumaric acid derivatives drupanin and trans-ferulic acid O-hexan-3-onyl-ether (27-35% at 10 microg/ml) and the flavonoid quercetin (97 and 23% at 10 and 1 microg/ml, respectively). The results support the use of Baccharis grisebachii in Argentinian traditional medicine.

Free radical scavengers and antioxidants from Tagetes mendocina.

Tagetes mendocina (Asteraceae) is a medicinal plant widely used in the Andean provinces of Argentina. Preliminary assays showed free radical scavenging activity in the methanol extract of the aerial parts, measured by the decoloration of a methanolic solution of the 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) and scavenging of the superoxide anion. Assay-guided isolation led to 4'-hydroxyacetophenone (1), protocatechuic acid (2), syringic acid (3), patuletin (4), quercetagetin 7-O-beta-D-glucoside (5), patuletin 7-O-beta-D-glucoside (6) and axillarin 7-O-beta-D-glucoside (7) as the free radical scavengers and antioxidant compounds from Tagetes mendocina. On the basis of dry starting material, the total phenolic content of the crude drug was 3.00% with 0.372% of flavonoids. The content of compounds 1-7 in the crude drug was 0.008, 0.015, 0.010, 0.029, 0.238, 0.058 and 0.017%, respectively. Quercetagetin 7-O-beta-D-glucoside proved to be the main free radical scavenger of the extracts measured by the DPPH decoloration test as well as for quenching the superoxide anion and inhibition of lipoperoxidation in erythrocytes. In the lipid peroxidation assay the percentual inhibition was related with the number of methoxy groups in the molecule, ranging from 86% for the quercetagetin glucoside to 67% for the monomethoxylated and 31% for the dimethoxylated derivative. The compounds showed low cytotoxicity towards human lung fibroblasts with IC50 > 1 mM for compounds 1-3 and 0.24 to 0.52 mM for the flavonoids 4-7.

Constituents of the Argentinian medicinal plant Baccharis grisebachii and their antimicrobial activity.

The resinous exudate of Baccharis grisebachii which is used to treat ulcers, burns, and skin sores in Argentina showed activity towards dermatophytes and bacteria. Two diterpenes, eight p-coumaric acid derivatives, and two flavones were isolated from the exudate and the structures elucidated by spectroscopic methods. 3-Prenyl-p-coumaric acid and 3,5-diprenyl-p-coumaric acid were active towards Epidermophyton floccosum and Trichophyton rubrum with MICs of 50 and 100-125 microg/ml, respectively. The diterpene labda-7,13E-dien-2beta,15-diol was active towards Epidermophyton floccosum and Trichophyton rubrum with MICs of 12.5 microg/ml while the MIC against Microsporum canis and Trichophyton mentagrophytes was 25 microg/ml. The diterpene was also active towards Microsporum gypseum with a MIC of 50 microg/ml, and showed inhibition in both Staphylococcus aureus (methicilline resistant and sensible strains) with MICs of 125 microg/ml. The results support the use of Baccharis grisebachii in Argentinian traditional medicine.

Bioactive alkyl phenols and embelin from Oxalis erythrorhiza.

The benzoquinone embelin and four alkyl phenols were isolated from an Argentinean collection of Oxalis erythrorhiza. 3-Heptadecyl-5-methoxy-phenol is reported for the first time. The structures were determined by spectroscopic methods. Embelin presented inhibitory effect on methicillin-resistant Staphylococcus aureus, Escherichia coli and the dermatophytic fungi Epidermophyton floccosum, Microsporum canis, Microsporum gypseum, Trichophyton mentagrophytes and Trichophyton rubrum with MICs ranging between 50 and 100 microg/ml. Furthermore, embelin was active against Trypanosoma cruzi trypomastigotes with 100% lysis at 100 microg/ml and cytotoxicity below the trypanocidal concentration. The new alkyl phenol 3-heptadecyl-5-methoxy-phenol was active towards Leishmania amazonensis and Leishmania donovani promastigotes with 100% lysis at 100 microg/ml. The cytotoxicity (IC50) of embelin and the new alkyl phenol on human lung fibroblasts were 739 and 366 microM, respectively. The plant is used to treat heart complains, a symptomatology related to Chagas' disease which is endemic in the San Juan Province, Argentine.

Free radical scavengers, anti-inflammatory and analgesic activity of Acaena magellanica.

Extracts of the whole plant Acaena magellanica (Rosaceae) were assessed for anti-inflammatory, antipyretic and analgesic activity in animal models. At 600 mg kg(-1), the global ethanolic extract (GEE), dichloromethane (DCM) and defatted methanol (MeOH) fractions showed a mild anti-inflammatory effect in the carrageenan-induced guinea-pig paw oedema. The GEE, DCM and defatted MeOH fractions significantly reduced inflammation by 43.2, 40.5 and 42.1%, respectively. The GEE did not showed any significant antipyretic activity in doses up to 600 mg kg(-1). A 20% w/v infusion administered orally at 16 mL kg(-1) presented analgesic effect in the acetic acid-induced abdominal constriction test in mice. The GEE and MeOH extract of A. magellanica showed free radical scavenging activity in the diphenylpicrylhydrazyl decolouration assay. Assay-guided isolation led to quercetin, Q-3-O-beta-glucoside, Q-3-O-beta-D-galactoside, ellagic acid and catechin as the free radical scavengers. The saponins tormentic acid 28-O-beta-D-galactopyranoside and 28-O-beta-D-glucopyranoside were isolated from the polar extract. The structures were determined by spectroscopic methods.