1-1-diphenyl-2-picrylhydrazyl and salvin

Topics: Abietanes; Biphenyl Compounds; Cell Line, Tumor; Cell Proliferation; Chromatography, Liquid; Colonic Neoplasms; Gas Chromatography-Mass Spectrometry; HCT116 Cells; HT29 Cells; Humans; Mass Spectrometry; Picrates; Plant Extracts; Rosmarinus

Ethanol extracts from dried leaves of wild Lepechinia salvia (Lindl) Epling, collected during the flowering period (September-November), contained 15% to 25% carnosic acid and 2 to 8% carnosol, depending on the month of collection. The highest concentration of carnosic acid in extracts was in October, while carnosol concentration had a peak in September, which suggests that it is not a product of carnosic acid oxidation. A comparison of extracts obtained in September 2008 to 2011 shows that the production of both abietanes increased in years with less winter rainfall and higher temperatures, which induced an early blooming. EC50 values in DPPH radical scavenging and antiproliferative (CCRF-CEM tumor cells) bioassays confirm that the high bioactivity of the extracts of rosemary, sage and L. salviae does not arise only from carnosol and carnosic acid. The cytotoxic activity was significantly higher in extracts of L. salviae, probably due to water stress differences between the cultivars and the wild species. These results correlate well with the close phylogenetic relationship between the three species, and their similar medicinal uses.

Topics: Abietanes; Antineoplastic Agents; Antioxidants; Biphenyl Compounds; Cell Line, Tumor; Cell Proliferation; Ethanol; Humans; Lamiaceae; Picrates; Plant Extracts; Plant Leaves

Supercritical fluid chromatography (SFC) in analytical and semi-preparative scale was studied to separate rosemary (Rosmarinus officinalis) supercritical extracts to produce fractions with high antioxidant activity. Carnosic acid, the major antioxidant present in rosemary supercritical extracts, was the target compound to be isolated in the fractions produced. First, rosemary extracts were produced in a supercritical CO2 pilot-plant employing different extraction conditions, in order to obtain samples with diverse composition. The content of carnosic acid was determined and the DPPH test was accomplished in order to assess the antioxidant activity of the different samples produced. Then, using some of these samples, experiments were carried out in a semi-preparative SFC system testing the performance of four different chromatographic columns to isolate and recover a fraction with high concentration of carnosic acid. The four chromatographic columns employed were silica and 2-ethylpyridine (especially packed for SFC) and silica and diol columns (packed for HPLC). The SFC fractionation conditions explored were in the ranges 10-20 MPa, 313-333 K and 5-20% of cosolvent (ethanol). Using the first three columns, it was possible to isolate fractions with concentrations of carnosic acid greater than 80% mass.

Topics: Abietanes; Biphenyl Compounds; Carbon Dioxide; Chromatography, High Pressure Liquid; Chromatography, Supercritical Fluid; Linear Models; Picrates; Plant Extracts; Plant Leaves; Rosmarinus

A solvent system consisting of ethyl acetate, ethyl alcohol and water, in the volume ratio of 4.5:4.5:1, was developed and used to extract, at room temperature, betulin from white birch bark and antioxidants from spices (rosemary, thyme, sage, and oregano) and white oak chips. In addition, under reflux conditions, trimyristin was extracted from nutmeg using the same solvent system, and eugenol from olives was extracted using a mixture of salt water and ethyl acetate. The protocol demonstrates the use of water in organic solvents to extract natural products from plants. Measurement of the free-radical scavenging activity using by 2,2-diphenyl-1-picrylhydrazyl (DPPH) indicated that the extraction of plant material using ethyl acetate, ethyl alcohol and water (4.5:4.5:1, v/v/v) was exhaustive when carried out at room temperature for 96 h.

Topics: Abietanes; Acetates; Betula; Biphenyl Compounds; Ethanol; Eugenol; Free Radical Scavengers; Free Radicals; Liquid-Liquid Extraction; Myristica; Origanum; Picrates; Plant Bark; Plant Extracts; Quercus; Rosmarinus; Salvia officinalis; Solvents; Thymus Plant; Triglycerides; Triterpenes; Water

The aim of this study was to examine the effect of ultrasound on the solvent extraction of anti-oxidants from the rosemary herb and to scale up the ultrasonic extraction process. The anti-oxidants of interest were identified using HPLC. Results indicated that, compared with conventional solvent extraction, the use of ultrasound gives a more effective extraction at lower temperatures with less dependence on the extraction solvent employed and that scale up of the process is possible.

Topics: Abietanes; Antioxidants; Biphenyl Compounds; Cinnamates; Depsides; Ethanol; Methanol; Picrates; Plant Extracts; Plant Leaves; Rosmarinic Acid; Rosmarinus; Solvents; Spectrophotometry, Ultraviolet; Ultrasonics

Rosmarinus officinalis extracts were investigated by a combination of bioassays and biochemical analysis to identify bioactive compounds. The 2,2-diphenyl-2-picrylhydracyl hydrate (DPPH) radical scavenging method, Folin-Ciocaulteau method and HPLC chromatography were used to study the distribution and levels of antioxidants (AOXs). Antimicrobial activity analysis was carried out using the disk diffusion and broth dilution techniques. A good correlation between the AOX activities and total phenol content in the extracts was found. Although all rosemary extracts showed a high radical scavenging activity, a different efficacy as antimicrobial agent was observed. Methanol extract containing 30% of carnosic acid, 16% of carnosol and 5% of rosmarinic acid was the most effective antimicrobial against Gram positive bacteria (minimal inhibition concentration, MIC, between 2 and 15 mug/ml), Gram negative bacteria (MIC between 2 and 60 mug/ml) and yeast (MIC of 4 mug/ml). By contrast, water extract containing only 15% of rosmarinic acid showed a narrow activity. MIC value of the methanol and water extracts is in a good correlation with the values obtained with pure carnosic acid and rosmarinic acid, respectively. Therefore, our results suggested that the antimicrobial rosemary extracts efficacy was associated with their specific phenolic composition. Carnosic acid and rosmarinic acid may be the main bioactive antimicrobial compounds present in rosemary extracts. From a practical point of view, rosemary extract may be a good candidate for functional foods as well as for pharmaceutical plant-based products.

Topics: Abietanes; Anti-Infective Agents; Antioxidants; Bacteria; Biphenyl Compounds; Chromatography, High Pressure Liquid; Cinnamates; Depsides; Flavonoids; Free Radicals; Indicators and Reagents; Phenanthrenes; Phenols; Picrates; Plant Extracts; Polyphenols; Rosmarinic Acid; Rosmarinus; Saccharomyces cerevisiae

A supercritical fluid extract of rosemary has been fractionated under supercritical conditions by using a preparative-SFC system. In this work, the optimum conditions have been evaluated to achieve a selective isolation of the compounds responsible for both, antioxidant and antimicrobial activities. A 25 cm x 10 mm i.d. LC-Diol packed column (dp=5 microm) has been used and the separation took place at 80 degrees C of column temperature, 130 bar of pressure, and 10% of ethanol as modifier of the mobile phase (CO(2)). Two cyclones were employed to collect the fractions which were subsequently characterized by HPLC-DAD, GC, and in vitro antioxidant and antimicrobial assays. By a careful selection of the separation conditions it is possible to obtain two different fractions, one enriched with antioxidant and antimicrobial compounds (with an improvement of about 20% and 40% of antioxidant and antimicrobial activity, respectively, compared to the original extract) collected in cyclone 2 and with no residual rosemary aroma and another one containing the essential oil.

Topics: Abietanes; Anti-Infective Agents; Antioxidants; Biphenyl Compounds; Chromatography, Gas; Chromatography, High Pressure Liquid; Dietary Supplements; Food Analysis; Free Radical Scavengers; Ledum; Mass Spectrometry; Microbial Sensitivity Tests; Odorants; Picrates; Plant Extracts; Plant Leaves; Staphylococcus aureus

Carnosol 1, rosmanol 2, carnosic acid 3 and 20-deoxocarnosol 4, the main phenolic abietanes present in aerial parts of the Chilean medicinal plant Sphacele salviae were tested for antioxidant activity of measuring the decay of the radical cation diphenyl-picrylhydrazyl (DPPH). All compounds displayed higher antioxidant levels than BHT under the same conditions and carnosic acid was more efficient than vitamin E.

Topics: Abietanes; Antioxidants; Biphenyl Compounds; Butylated Hydroxytoluene; Chile; Diterpenes; Dose-Response Relationship, Drug; Lamiaceae; Medicine, Traditional; Phenanthrenes; Picrates; Plant Extracts; Plant Leaves; Plants, Medicinal; Structure-Activity Relationship; Time Factors; Vitamin E

Preparative-scale incubations with Nocardia sp. NRRL 5646 were conducted to produce new derivatives of the abietane diterpene chemoprotectant and antioxidant carnosic acid (1). Reductive biotransformation of the C-20 carboxylic acid functional group followed by biological methylation at the C-11 phenol afforded 4. Oxidative cyclization of 1 to carnosol 5 followed by dihydroxylation at the isopropyl moiety afforded 6. Metabolites 4 and 6 are new carnosic acid derivatives whose structures were confirmed by mass spectrometry and NMR spectroscopic analysis. The radical quenching properties of 4-6 using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free-radical-scavenging assay showed activities similar to that of mixed tocopherols and carnosic acid.

Topics: Abietanes; Antioxidants; Biphenyl Compounds; Cyclization; Diterpenes; Free Radical Scavengers; Mass Spectrometry; Methylation; Molecular Structure; Nocardia; Nuclear Magnetic Resonance, Biomolecular; Oxidation-Reduction; Picrates; Plant Extracts; Rosmarinus; Structure-Activity Relationship