Abstract: The variation in the chemical composition of the essential oil of Salvia officinalis , growing in different habitats, was studied. GC-MS analysis revealed 57 compounds representing 94.68-96.80% of total oils. The major components were alpha-thujone (11.55-19.23%), viridiflorol (9.94-19.46%), 1,8-cineole (8.85-15.60%), camphor (5.08-15.06%), manool (5.52-13.06%), beta-caryophyllene (2.63-9.24%), alpha-humulene (1.93-8.94%), and beta-thujone (5.45-6.17%), showing significant differences between different collection sites. Analysis of some representative polyphenolic compounds and antioxidant activity was performed using postdistilled dry samples. Rosmarinic acid, carnosol, and carnosic acid were the prevalent compounds of S. officinalis methanolic extracts. The results revealed differences in the polyphenolic composition and also exhibited antioxidant and radical-scavenging activities at different magnitudes of potency. However, within the used methods, only the DPPH(*) assay showed significant differences (p < 0.05) in free radical scavenging activity among samples collected in different regions. Plants collected in the coastal regions Soliman and Kelibia accumulate more polyphenolic compounds, known to be responsible for the main antioxidant activity of sage (rosmarinic acid, carnosol, and carnosic acid), than those growing inland at Bou Arada and Sers. Moreover, the former presented a higher radical-scavenging activity. The methanolic extracts of postdistilled S. officinalis might be valuable antioxidant natural sources and seemed to be applicable in both the health medicine and food industries.

Abstract: Reduction or elimination of chemically synthesized additives from foods is a current demand in food industry. A new approach to prevent the proliferation of microorganisms or protect food from oxidation is the use of essential oils or plant extracts as natural additives in foods. We have studied antimicrobial activity of rosemary extracts (Rosmarinus officinalis L.) against different species of Listeria and against different strains of L. monocytogenes. We used two extracts of rosemary, VivOX 20 and VivOX 40 (Vitiva d.d., Slovenia) containing different levels of carnosic acid. We wanted to proof an antimicrobial activity of selected rosemary extracts with two most commonly used methods: disc diffusion method and broth dilution method. With the disc diffusion method we have obtained the inhibition zone and at the lowest concentrations, where no visible bacterial growth was recorded, were assumed as minimal inhibitory concentration values (MIC). We determined MIC values in the ranges from 625 μg extract/ml EtOH to 5000 μg extract/ml EtOH for VivOX 20 and from 312.5 μg extract/ml EtOH do 2500 μg extract/ml EtOH for VivOX 40 in the medium. We have established that the resistance of Listeria species against rosemary extracts depends on: selected extract, selected concentration, various species and strain of Listeria. With broth dilution method we have determined minimal bactericidal concentration (MBC), as the concentration giving 0.1 % bacterial survival. With this method we have tested two strains of L. monocytogenes and in determinate MBC values in the range from 15.63 µg/ml TSB to 98.5 µg/ml TSB for both tested extracts. Results have confirmed our assumption that resistance of Listeria against rosemary extracts depended on the selected strain.

Abstract: Amyotrophic lateral sclerosis (ALS) is a late-onset progressive neurodegenerative disease affecting motor neurons. About 2% of patients with the disease are associated with mutations in the gene encoding Cu/Zn superoxide dismutase (SOD1). The purpose of this study is to assess the effect of rosemary extract and its major constituents, rosmarinic acid (RA) and carnosic acid (CA), in human SOD1 G93A transgenic mice, which are well-established mouse models for ALS. The present study demonstrates that intraperitoneal administration of rosemary extract or RA from the presymptomatic stage significantly delayed motor dysfunction in paw grip endurance tests, attenuated the degeneration of motor neurons, and extended the life span of ALS model mice. In addition, RA administration significantly improved the clinical score and suppressed body weight loss compared with a vehicle-treated group. In conclusion, this study provides the first report that rosemary extract and, especially, RA have preventive effects in the mouse model of ALS.

Abstract: A sensitive and reproducible high-performance liquid chromatography method with ultraviolet detection (UV) was developed for the determination of carnosic acid (CA) in rat plasma. After simple acidification and liquid-liquid extraction of plasma samples using gemfibrozil as an internal standard, the supernatant was evaporated to dryness under a gentle stream of nitrogen. The residue was reconstituted in 200 microL before being injected into the chromatographic system. The analysis was performed on a C(18) column protected by an ODS guard column using acetonitrile-0.1% phosphoric acid (55:45, v/v) as mobile phase, and the wavelength of the UV detector was set at 210 nm. The calibration curve was linear over the range of 0.265-265.0 microg/mL with a correlation coefficient of 0.9997. The recovery for plasma samples of 0.530, 13.25, 132.5 and 265.0 microg/mL was 72.2, 87.9, 90.4 and 94.7%, respectively. The intra-day and inter-day relative standard deviations for the measurements of quality control samples were less than 3.1%. The stability of the plasma samples was also validated. This method was successfully used to study the pharmacokinetics and bioavailability of CA in rats.

Abstract: Rosmarinus officinalis L. is receiving increasing attention due to its anti-inflammatory and antioxidative constituents. Our recent studies showed that R. officinalis extract, containing 31.7% of carnosic acid, was able to counteract the deleterious effects of UV-R, by protecting plasmid DNA from hydroxyl radicals generated by UV-A. In this work, we evaluated the effects of this extract on pBR322 DNA cleavage induced by nitric oxide, and the growth inhibitory activity against two human melanoma cell lines, M14 and A375. The extract showed a protective effect on plasmid DNA damage, and at concentrations of 10-80 microg/mL was able to reduce significantly (p<0.001) the growth (MTT assay) of both melanoma cell lines. In addition, our results indicate that apoptotic cell demise is induced in M14 and A375 cells. No statistically significant increase in LDH release was observed in melanoma cells, correlated to a fragmentation of genomic DNA, determined by COMET assay.

Abstract: Reduction or elimination of chemically synthesized additives from foods is a current demand in food industry. A new approach to prevent the proliferation of microorganisms or protect food from oxidation is the use of essential oils or plant extracts as natural additives in foods. We have studied antimicrobial activity of rosemary extracts (Rosmarinus officinalis L.) against different species of Listeria and against different strains of L. monocytogenes. We used two extracts of rosemary, VivOX 20 and VivOX 40 (Vitiva d.d., Slovenia) containing different levels of carnosic acid. We wanted to proof an antimicrobial activity of selected rosemary extracts with two most commonly used methods: disc diffusion method and broth dilution method. With the disc diffusion method we have obtained the inhibition zone and at the lowest concentrations, where no visible bacterial growth was recorded, were assumed as minimal inhibitory concentration values (MIC). We determined MIC values in the ranges from 625 g extract/ml EtOH to 5000 g extract/ml EtOH for VivOX 20 and from 312.5 g extract/ml EtOH do 2500 g extract/ml EtOH for VivOX 40 in the medium. We have established that the resistance of Listeria species against rosemary extracts depends on: selected extract, selected concentration, various species and strain of Listeria. With broth dilution method we have determined minimal bactericidal concentration (MBC), as the concentration giving 0.1 % bacterial survival. With this method we have tested two strains of L. monocytogenes and in determinate MBC values in the range from 15.63 ?g/ml TSB to 98.5 ?g/ml TSB for both tested extracts. Results have confirmed our assumption that resistance of Listeria against rosemary extracts depended on the selected strain.

Abstract: Disclosed are methods of alleviating oxidative stress, regulating blood glucose levels, and controlling pancreatic lipase activity by administering a Rosemary extract. Such methods may be useful to help body weight and body fat, prevent obesity, and treat hyperglycemia, hyperinsulinemia, cardiovascular disease and Type-2 diabetes. Preferred extracts contain 20% or 50% carnosic acid.

Abstract: OBJECTIVE To establish a HPLC method to determine the carnosic acid in the stomach and intestine of rats and study its tissue distribution characteristics. METHOD After intragastric administration of carnosic acid (90 mg x kg(-1)), rats for each time-point were sacrificed by decapitation. After removal of the blood, various tissues were rapidly removed and weighted, all tissues were treated with a series of pretreatment before HPLC. Chromatographic separation was achieved on a Kromasil C18 column (4.6 mm x 150 mm, 5 microm) protected by an ODS guard column at 25 degrees C, using acetonitrile-0.1% phosphoric acid solution (55:45) as mobile phase, at a flow rate of 1 mL x min(-1). The wavelength of the UV detector was set at 210 nm for carnosic acid and internal standard. RESULT Good linearities were obtained in every tissue over a range of 0.3212-160.6 mg x L(-1). The recovery, intra-day and inter-day precision and accuracy of three concentrations of carnosic acid in tissues met the requirements of methodology. And the stability of the tissue samples were also validated. The results of distribution in stomach and intestine showed that the highest concentration was (307.1 +/- 119.2) microg x g(-1) in stomach and (33.32 +/- 17.70) microg x g(-1) in intestine after intragastric administration of carnosic acid. CONCLUSION The HPLC method was established to determine the concentration of carnosic acid in tissues. This method is quick, precise, and reproducible. It is the first time to study the tissue distribution of carnosic acid in rats after intragastric administration.

Abstract: The present study is aimed to the effect of a Tunisian aromatic plant Rosmarinus officinalis extract on - a neuronal cell model - PC12 cells and on HSP47. The extract of R. officinalis and two pure compounds rosmarinic acid and carnosic acid promote markedly the neurite outgrowth of PC12 cells and enhanced the acetylcholinesterase activity same as NGF (neurite outgrowth factor). The investigation of phosphorylation marker illustrates that Rosemary extract treatment showed ERK1/2 phosphorylation same as NGF. Moreover, Rosmarinus extracts and its pure compounds have shown a stress recovery activity on HSP47.

Abstract: Both supercritical CO2 and methanolic extracts from the leaves of rosemary (Rosmarinus officinalis) harvested from three different locations of Turkey at four different times of the year were added at a concentration of 100 mg/kg to wheat germ oil. Wheat germ oil samples were stored in an incubator for 10 days at 50C in order to promote oxidation and for the efficacy of the extracts for stabilization purposes to be examined. Degree of oxidation was determined by peroxide and p-anisidine values, which were performed every 2 days. Extracts from Mersin and Canakkale regions performed better results. Additionally, June and September harvests had lower peroxide values. According to the descriptive sensory analysis, both locations and extraction methods were found to effect flavor. Some flavor attributes, such as wheatlike/starchy, fishy and rubbery/metallic changed during storage regardless of locations and extraction methods. Supercritical CO2 extracts performed better results in terms of both oxidation and sensory properties. PRACTICAL APPLICATIONS From the perspective of the food industry, wheat germ oil stabilized with a natural antioxidant such as rosemary can be marketed as a functional product that can create a niche. Rosemary extracts containing higher amounts of rosmarinic acid and carnosic acid should be preferred in order to provide a better shelf life of an edible oil such as wheat germ oil.

Abstract: Microspheres loaded with carnosic acid (CA) as an active ingredient which was extracted from various species of Salvia L., were prepared by a spray drying technology. The matrix of the microspheres consisted of 85% deacetylated chitosan (average MW 55000 kDa), poloxamer P407 and carnosic acid. Briefly, chitosan and poloxamer P407 were dissolved in 1% v/v acetic acid followed by dispersion of carnosic acid under vigorous stirring. The dispersion was then fed into a spray dryer (190 Mini, Buchi, Flawil, Switzerland) and processed under the following operative conditions: T inlet 180°C, T outlet 120°C, air aspiration 70% and peristaltic pump 15% (feeding rate 4 mL/min). This process yielded microspheres with 5-8 μm in diameter containing 17% active ingredients corresponding to an encapsulation efficiency of 45% and a practical yield of 34%.

Abstract: PROBLEM TO BE SOLVED: To provide arachidonic acid oil and fat excellent in oxidation stability. SOLUTION: A composition containing an arachidonic acid is used for adding arachidonic acid or enhancing its content. The composition contains tocopherol and a rosemary extract together with arachidonic acid. The arachidonic acid may be an alcohol ester of arachidonic acid or triglyceride, phospholipid, or glycolipid arachidonic acid whose part or whole of the constituent fatty acid is arachidonic acid. It is desirable that the composition contain 1 wt.% or above of arachidonic acid. The rosemary extract is preferably an oil soluble extract, more preferably carnosic acid. The composition is added to food and drink-favorite food, bait and feed, health care-medicine, or cosmetics so as to add arachidonic acid or enhance its content. COPYRIGHT: (C)2010,JPO&INPIT

Abstract: PURPOSE: A food composition is provided to activate and proliferate osteoblast cell and to prevent the bone metabolism disorder such as osteoporosis by using a rosemary extract. CONSTITUTION: A food composition comprises a rosemary extract as an active ingredient. The rosemary extract comprises any one selected from carnosic acid and rosmarinic acid, or their mixture. The rosemary extract contains 102 mg/g or more of a phenolic compound. The rosemary extract contains 9.8 mg/g or more of flavonoids. The rosemary extract is obtained by extracting rosemary with an organic solvent or distilled water. The organic solvent is at least one selected from methanol, ethanol and acetone.

Abstract: This study aimed at demonstrating the stabilizing effect of up to 0.4% (wt/wt flour) of rosemary spice (Rosemarinum Officinalis L.) in a flour-based protein-rich product intended for young children. The flour was made of full-fat soya flour, meat (beef) powder, and carrot flour as β-carotene source. Analysis was done for proximate composition, β-carotene content, microbial load and sensory evaluation for rancidity over time. Different levels of rosemary spice salvaged a net of 3.42 - 3.83 mg/100 g of β-carotene within a storage period of 7 weeks at 35°C accounting for up to 18% of β-carotene sparing as compared to the non-spicy sample. There was, however, no evidence of increased protection of β-carotene with increase in rosemary spice concentration. Rancid odors and flavour were detected in samples with spice, latter than in samples with no spice. Rosemary spice exhibited up to a net of 38% reduction in microbial load in spicy samples as compared to the non-spicy sample. In a protein, fat and β-carotene rich flour-based product, rosemary spice exhibits triple stabilizing action. The phenolic compounds (rosemarinic, carnosol and carnosic acid) in rosemary spice limits β-carotene degradation and decelerates the production of secondary products of lipid oxidation while the terpene fractions are implicated for halting the proliferation of micro-organisms. Key words: Rosemary spice, high protein β-carotene-rich flour, antioxidant effect, antimicrobial effect.

Abstract: Disclosed is a process for producing carnosic acid (the major antioxidant active substance contained in a rosemary plant) with high efficiency by carrying out the ortho-position oxidation of a phenol with an oxidizing agent (e.g., 2-iodoxybenzoic acid) and the subsequent ester removal reaction by using, as a starting material, pisiferic acid (the main component of Chamaecyparis pisifera which is a plant which has been planted in great numbers as wood resources). Specifically disclosed is a process for producing an antioxidant active substance contained in a rosemary plant and represented by general formula (5). The process is characterized by comprising: a first step of oxidizing a pisiferic acid derivative represented by general formula (1) with an oxidizing agent represented by general formula (2) or (4); and a second step of reducing or hydrolyzing a pisiferic acid derivative intermediate produced in the first step.

Abstract: The shoots of Salvia officinalis growing in MS liquid medium supplemented with IAA 0.1 mg l-1) and BAP (0.45 mg l-1) were treated with methyl jasmonate (MeJA) to increase production of compounds with antioxidant activity (carnosic acid, carnosol and rosmarinic acid). The increase in metabolite production depended on MeJA concentration, the period of exposure to elicitor and type of compound. The MeJA action was observed 24 h after elicitation. It was found that the maximum level of diterpenoids, calculated as the sum of CA and Car (about 8 mg g-1 dry wt) was achieved at 3 days after elicitation with 20 μM methyl jasmonate. The highest amount of rosmarinic acid (about 41 mg g-1 dry wt) was achieved with 50 or 100 μM methyl jasmonate on the 5th day after elicitation. It was almost 2-fold higher compared to the control (cultures treated with only ethanol).

Abstract: Expression of cell adhesion molecules on the endothelium and the attachment of monocytes to endothelium may play a major role in the early atherogenic process We investigated the effects of carnosic acid on the adhesion of U937 cells to IL-1β-treated human umbilical vein endothelial cells (HUVECs), as well as on the expression of adhesion molecules Our data showed that pretreatment with 10 and 20 µmol/l carnosic acid significantly reduced the number of U937 cells adhering to IL-1β-treated HUVECs In addition, we found that 20 µmol/l carnosic was more effective than 10 µmol/l carnosic acid at inhibiting expression of cell adhesion molecules (ICAM-1, VCAM-1, and E-selectin), the nuclear translocation of NF-κB subunits p65 and p50, and the production of ROS in IL-1β-stimulated HUVECs We conclude that carnosic acid inhibits IL-1β-induced ICAM-1, VCAM-1 and E-selectin expression in HUVECs through a mechanism that involves NFκB We propose that the reduction in binding of human monocytic cell line U937 to IL-1β-treated HUVECs is due to the anti-inflammatory properties of carnosic acid

Abstract: BACKGROUND: Garden sage (Salvia officinalis L., Lamiaceae) has gained importance during the last decade as a natural antioxidant, mainly owing to the antioxidant secondary plant metabolites rosmarinic acid (RA) and diterpenes such as carnosic acid. The aim of this work was to study the biomass production, the concentrations of total phenolics and RA and the essential oil of garden sage in response to phosphorus (P) supply. The treatments included P fertilisation and inoculation with arbuscular mycorrhizal fungi (AMF), since AMF are the most efficient biotic factor promoting the P uptake of plants. RESULTS: The P concentration in the plant was highest in P-fertilised plants, intermediate in AMF-inoculated plants and lowest in non-inoculated/non-P-supplied plants. The leaf biomass increased only in P-fertilised plants. AMF-inoculated plants, in contrast, did not show an increased leaf biomass, but their root biomass was enhanced. The total phenolic and RA concentrations were affected differently in leaves and roots, with the highest concentration and yield being found in leaves of P-fertilised plants. In none of the treatments were the essential oil concentration and composition affected. CONCLUSION: Optimal P fertilisation improves the antioxidant potential of garden sage and increases its biomass yield, thus optimising agronomic production of this plant for antioxidant use. c � 2009 Society of Chemical Industry