In vitro antioxidant and antiproliferative activity of three rosemary (Rosmarinus officinalis L.) extract formulations

Antioxidant and antiproliferative activities of three rosemary extract formulations (VivOX 20, VivOX 40 and Inolens 50) with different contents of carnosic acid, carnosol and methylcarnosol were tested in vitro. Electron spin resonance measurements revealed that Inolens 50 extract that contained highest amount of carnosic acid was the most potent scavenger of hydroxyl (concentration of extract where 50% of its maximal scavenging activity is observed, that is, EC₅₀, 109.54 μg mL^?1), superoxide anion (EC₅₀ = 7.94 μg mL^?1) and 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) (EC₅₀ = 27.4 μg mL^?1)‐free radicals. Comparison of the radar charts of standard antioxidants and rosemary extracts showed similarity between antioxidant characteristics of Inolens 50 and chlorogenic and caffeic acids. Tested rosemary extracts exhibited significant (P ≤ 0.01) antiproliferative effect in cervix epitheloid carcinoma (HeLa), breast adenocarcinoma (MCF7) and colon adenocarcinoma (HT‐29) cell lines. In both MCF7 and HeLa cell lines, the extracts yielded very low IC₅₀ values (concentration of extract needed to inhibit cell growth by 50%), the most pronounced being for Inolens 50 in MCF7 (IC₅₀ = 9.95 μg mL^?1) and VivOX 20 in HeLa cell line (IC₅₀ = 10.02 μg mL^?1). The obtained results may provide support for the use of tested rosemary extracts as nutraceuticals and phytopharmaceuticals.     

Phenolic diterpene and α‐tocopherol contents in leaf extracts of 60 Salvia species

BACKGROUND: Carnosic acid and carnosol, the two major phenolic diterpenes present in rosemary and sage extracts, have received attention in food science and biomedicine because of their potent antioxidant properties. In plants, these compounds have been identified as being present in some species of the family Lamiaceae, but there is still little information about their distribution within the plant kingdom. The aim of the present study was to gain a better understanding of the occurrence of these compounds within leaf extracts of the genus Salvia and also to examine to what extent leaf senescence may influence their accumulation in relation to that of α‐tocopherol (vitamin E). RESULTS: Methanolic leaf extracts of 60 species of the genus Salvia were tested for the presence of carnosic acid and carnosol. These two diterpenes were detected in 48 and 27 species respectively. In contrast, α‐tocopherol was present in all species examined. Leaf senescence in Salvia officinalis resulted in α‐tocopherol increases by up to 5.5‐fold, while carnosic acid and carnosol increased by up to 18 and 290% respectively. Isorosmanol, an oxidation product of carnosic acid, increased by up to 2.1‐fold in senescing leaves. CONCLUSION: It was found that, while α‐tocopherol is ubiquitous in the genus Salvia, phenolic diterpenes are present in several but not all species. It was also shown that leaf senescence may increase the yield of both phenolic diterpenes and α‐tocopherol in sage extracts. Copyright © 2008 Society of Chemical Industry     

Inhibition of Heterocyclic Amine Formation in Beef Patties by Ethanolic Extracts of Rosemary

ABSTRACT: Heterocyclic amines (HCAs) are mutagenic compounds formed during cooking muscle foods at high temperature. Inhibition of HCAs by rosemary extracts were evaluated with beef patties cooked at 191 °C (375 °F) for 6 min each side and 204 °C (400 °F) for 5 min each side. Five rosemary extracts extracted with different solvents were used in this study: extract 100W (100% water), 10E (10% ethanol), 20E (20% ethanol), 30E (30% ethanol), and 40E (40% ethanol). The 5 extracts were directly added to beef patties at 3 levels (0.05%, 0.2%, and 0.5%) before cooking and HCA contents were extracted and quantified. All of the patties contained 2-amino-3,8-dimethylimidazo [4,5-f]quinoxaline (MeIQx), and 2-amino-1-methyl-6-phenylimidazo [4,5-b]pyridine (PhIP). There was no statistical difference in the inhibition of HCAs in the 0.05%, 0.2%, and 0.5% rosemary extracts. All rosemary extracts significantly decreased the levels of MeIQx and PhIP at both cooking conditions. When cooking at 204 °C (400 °F) for 5 min each side, rosemary extracts 10E and 20E were superior to rosemary extracts 100W, 30E, and 40E in inhibiting HCA formation. Rosemary extract 20E showed the greatest inhibition of MeIQx (up to 91.7%) and PhIP (up to 85.3%). The inhibiting effect of rosemary extracts on HCA formation corresponded to their antioxidant activity based on a DPPH scavenging assay. Rosemary extract 10E and 20E contain a mixture of rosmarinic acid, carnosol, and carnosic acid. It is possible that these compounds might act synergistically in inhibiting the formation of HCAs.     

In vitro antioxidant activities of Rosmarinus officinalis extracts treated with supercritical carbon dioxide

The leaves of Rosmarinus officinalis (rosemary) were subjected to supercritical CO₂ extraction (SFE). Different sources of variability, including location (Izmir, Canakkale and Mersin) and harvesting time (December, March, June and September), were considered. Among active constituents of rosemary, carnosic acid, carnosol and rosmarinic acid were analyzed by HPLC. Variability of the amounts of active constituents appears to be due to different geographical locations of growth and seasonal variations. The levels of the constituents were higher in the months of December 2003 and September 2004. In addition to this, 12 SFE extracts were screened for their radical-scavenging capacities and antioxidant activities by various in vitro assays, namely total phenol assay, DPPH radical-scavenging activity and trolox equivalent antioxidant capacity (TEAC).     

Extraction of lipid-soluble antioxidants from rosemary leaves using vegetable oils

The objective of this research was to extract carnosic acid and carnosol, natural lipid-soluble antioxidants, from rosemary leaves without the use of chemical aid. Three different vegetable oils [high oleic soybean oil (HOSO), peanut oil (PO) and cottonseed oil (CO)] were used as the medium for extracting rosemary's lipid-soluble antioxidants. Dried rosemary leaves were ground into fine particles, added two of the selected vegetable oils, mixed and heated. The vegetable oil containing the extracted antioxidants was filtered from the rosemary particles and analysed using high-performance liquid chromatography (HPLC). As a control, a common extraction method using ethanol was performed to compare the oil extraction efficiency for carnosic acid and carnosol. HPLC analysis confirmed the presence of carnosic acid and carnosol in all three vegetable oil solutions. Compared to the ethanol extraction, a higher level of the carnosic acid was found in the vegetable oil solutions. Among the vegetable oils, high oleic soybean oil contained the most carnosic acid; however, there were similar amounts of carnosol in all three vegetable oil solutions. The extractions' efficacy was tested on soybean oil's (SO) oxidative stability by adding 2% (w/w) of the antioxidant–vegetable oil solution. The results obtained from peroxide value (PV) and p-anisidine value (AnV) analysis documented a significantly lower degree of oxidation in the soybean oil containing the extracted antioxidant solutions, compared to the pure soybean oil.     

UV-B radiation effects on foliar concentrations of rosmarinic and carnosic acids in rosemary plants

The influence of two different dosages of UV-B radiation on foliar concentrations of rosmarinic and carnosic acids was studied. The results showed that UV-B radiation significantly increased the concentrations of both rosmarinic and carnosic acids, as well as other rosemary compounds, such as naringin and carnosol. However, the increase of rosmarinic and carnosic acid levels were not concomitant with an increase in the DPPH radical-scavenging activity of rosemary extracts from UV-B-treated plants when compared with control plants. The significance of these results is considered.     

Antioxidant effect rosemary (Rosmarinus officinalis L.) and oregano (Origanum vulgare L.) extracts on TBARS and colour of model raw pork batters

Herbs and spices are traditional used as food ingredients as well as for their antioxidant properties. The objective of this work was to study the concentration of carnosol, rosmarinic and carnosic acids in rosemary (Rosmarinus officinalis L.) and oregano leaves (Origanum vulgare L.), and their effect on the oxidation and colour of model pork batters. Extracts were obtained by maceration with ethanol and reflux with chloroform. Rosemary extracts showed higher antioxidant activity, even more than the phenol compounds separately. These extracts also showed the highest antioxidant capacity, possibly due to the presence of high concentrations of carnosic acid and carnosol and unidentified active compounds. However, ethanol oregano extracts containing high concentrations of phenols, mainly rosmarinic acid, efficiently prevented colour deterioration. The antioxidant effect of the studied extracts depends, not only on the concentration of phenol compounds (rosmarinic acid, carnosol and carnosic acid), but also on the extraction method and solvent.     

Cross‐linking of bovine and caprine caseins by microbial transglutaminase and their use as microencapsulating agents for n‐3 fatty acids

Bovine and caprine caseins were cross‐linked with microbial transglutaminase (mTG). The mTG‐cross‐linked bovine or caprine casein dispersion, mixed with 14.5% maltodextrin (DE = 40), was used to prepare emulsions with 10.5% algae oil. Oxidative stability of emulsions was evaluated by peroxide values (PVs) and anisidine values. Adding liposoluble rosemary extract rich in carnosic acid and δ‐tocopherol lowered the formation of hydroperoxides and their subsequent decomposition products in emulsions. Emulsions stabilised with liposoluble rosemary extract rich in carnosic acid and δ‐tocopherol were spray‐dried at 180/95 °C. Algae oil microencapsulated with mTG‐cross‐linked bovine casein reduced PV by ≈ 34%, while the algae oil microencapsulated with mTG‐cross‐linked caprine casein with low levels of α_s1‐casein reduced PV by ≈ 42% at 4 weeks of storage at 30 °C. The investigation suggests that liposoluble rosemary extract rich in carnosic acid and δ‐tocopherol effectively protected algae oil during the coating process with mTG‐cross‐linked bovine and caprine caseins. The above results clearly indicated that the choice of milk caseins (bovine vs. caprine) cross‐linked with mTG impacts the oxidative stability of spray‐dried algae oil emulsions (microcapsules) enriched with n‐3 fatty acids.     

超临界二氧化碳萃取天然迷迭香抗氧化剂有效成分工艺的研究

利用水蒸气蒸馏法提取迷迭香精油后的迷迭香残渣,通过超临界二氧化碳工艺提取残渣中抗氧化剂的有效成分,以鼠尾草酸、鼠尾草酚含量作为产物的主要质量指标,以产品的物性和得率衡量工艺参数的优劣,通过L9(33)正交实验发现,在压力40MPa、温度80℃、萃取时间2.5h状况下可以达到较好的得率和有效成分含量,产品总得率7.2%,鼠尾草酸含量32%,鼠尾草酚含量8.1%,产品呈粉末状、气味淡,有利于工业化生产和应用。     

In vitro antioxidant analysis of supercritical fluid extracts from rosemary (Rosmarinus officinalis L.)

Different supercritical fluid extraction conditions were tested on rosemary leaves using a pilot-plant-scale extractor. Each of them provided two separated fractions which were characterized chemically by liquid chromatography–diode-array detection–mass spectrometry using electrospray in the positive mode. Twelve compounds were identified, including phenolic diterpenes and flavonoids. Owing to the lack of available standards, only carnosic acid could be quantified. The antioxidant activity of the extracts was determined by the 2,2-diphenyl-1-picrylhydrazyl hydrate (DPPH) test and the -carotene bleaching test was used to study the differences in behavior of antioxidants in an emulsified medium for some of the extracts obtained. The results of the correlation studies to examine the relationship between the antioxidant activity (EC₅₀), obtained by using the DPPH test, and the concentration of all the compounds detected in the extracts showed that carnosic acid was the more correlated compound, with a coefficient of correlation of –0.87. Using forward stepwise multiple linear regression, carnosic acid, methyl carnosate and carnosol were the compounds selected to predict the mentioned activity, with a value of 0.95 for the coefficient of determination.Área de Tecnología de Alimentos, Facultad de Ciencias, Universidad Autónoma de Madrid is an associated unit of IFI-CSIC.     

Oxidation of cholesterol in butter during storage – effects of light and temperature

 Rosemary is one of the herb spices and it exhibits antioxidative activity. The phenolic diterpene carnosic acid is mainly responsible for these antioxidant effects in rosemary and rosemary extracts. A modified high-performance liquid chromatography method for the determination of carnosic acid in rosemary extract was established. The analysis can be accomplished under isocratic conditions using electrochemical detection. The effect of dietary rosemary extract on the bioavailability of the antioxidative compound carnosic acid in eggs was evaluated. Using this method carnosic acid could be detected in 20 ng/g of egg yolk. Results showed that carnosic acid is bioavailable in egg yolk but not in albumen. Received: 22 January 1999     

Determination of the Most Bioactive Phenolic Compounds from Rosemary Against Listeria Monocytogenes: Influence of Concentration, pH, and NaCl

ABSTRACT: The anti-Listeria monocytogenes effects of 8 phenolic compounds, carnosol, carnosic acid, 12-methoxy carnosic, ferulic and caffeic acid, rosmarinic acid, luteolin and luteolin-7-glucoside were evaluated with a Plackett and Burman design in mixtures mimicking the phenolic composition of rosemary extract without essential oils. At 30°C carnosic acid was the most efficient compound during 24 h, whereas luteolin became more active after 72 h. The antibacterial effect of pure carnosic acid was modeled under a range of different pH and NaCl concentration, using a Doehlert design. Under moderately acidified conditions, carnosic acid displayed a bactericidal effect at low concentration (5.5 μg/ml). Its activity was not greatly influenced by NaCl.     

富含鼠尾草酸液态提取物的制备工艺研究

通过响应面分析优化了富含鼠尾草酸液态提取物制备工艺条件.结果表明,其最优工艺条件为:乙醇浓度为60%,油水比例为7.6,旋转浓缩温度为65℃,上述工艺条件下提取鼠尾草酸含量为14.91%.抗氧化实验结果表明该提取物具有良好的油脂抗氧化活性.     

Oxidative stability of butter with added phenolics from Lamiaceae herbs and in vitro evaluation of potential cytotoxicity of rosemary (Rosmarinus officinalis L.) extract

This study evaluated the oxidative stability of butter with added phenolics from Lamiaceae herbs. Initially antioxidant activity of crude rosemary, oregano, sage, thyme and marjoram extracts were screened through methods of inhibition of the 1,1‐Diphenyl‐2‐picrylhydrazyl (DPPH) radical, malondialdehyde (MDA) quantification and ferric reducing antioxidant power test. The alcoholic extract of rosemary showed the highest antioxidant activity in both assays for DPPH radical inhibition and MDA quantification. The alcoholic rosemary extract showed no cytotoxicity when assayed using MTT [3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide] reduction method and had dose‐dependent cytoprotective activity at 50 and 250 μg mL^?1. These results suggest that the use of rosemary as natural antioxidant is safe in the concentrations tested. The highest oxidative stability of butter with added alcoholic rosemary extract at temperatures of 60 and 110 °C was obtained with the concentration of 400 mg of phenolic compounds per kg of butter and was reflected at the lowest formation of degraded peroxides from lipids.     

Anti‐inflammatory properties of high pressure‐assisted extracts of Grifola frondosa in lipopolysaccharide‐activated RAW 264.7 macrophages

The aim of this study was to determine the in vitro anti‐inflammatory properties of the shake extract (SE) and the high pressure‐assisted extract (PE) of the mycelia of Grifola frondosa in a lipopolysaccharide‐stimulated RAW 264.7 macrophage model. The content of total polysaccharides and β‐glucans of PE at 600 MPa (PE‐600) was 41.2 and 6.2 mg g^?1 dry weight, respectively, which were significantly higher than SE extracts. The results showed that treatment with 500 μg mL^?1 of PE by 600 MPa (PE‐600) did not reduce RAW 264.7 cell viability but did significantly inhibit the production of LPS‐induced NO, PGE₂ and intracellular ROS. The PE‐600 inhibited the activation of NF‐kB and then reduced the production of LPS‐induced TNF‐α, IL‐6 and IL‐1β in a dose‐dependent manner. Thus, the PE could be used as an alternative extraction method for improving the extraction efficacy of G. frondosa and serve as an alternative source of anti‐inflammatory agents.     

Screening of free radical scavenging capacity and antioxidant activities of Rosmarinus officinalis extracts with focus on location and harvesting times

Methanolic extracts from the leaves of Rosmarinus officinalis (rosemary) harvested from different locations of Turkey at four different times of the year were analyzed by HPLC, and their radical scavenging capacities and antioxidant activities were studied by various assays. The amounts of carnosol, carnosic acid and rosmarinic acid, active constituents of rosemary, varied in different geographical regions of growth, and also showed a seasonal variation. The levels of the constituents were higher in the warm months of June 2004 and September 2004. The antioxidant activities of 12 extracts were determined by in vitro DPPH radical scavenging activity, by Trolox equivalent antioxidant capacity (TEAC), and by reversing H₂O₂-induced erythrocyte membrane lipid peroxidation (EMLP). The two antioxidant enzyme activities of human erythrocyte, namely superoxide dismutase (SOD) and catalase (CAT), after in vitro incubation with the extracts, were also examined in order to see whether the observed effects are related to altered enzymatic efficiency. The resulting values were correlated with active metabolite and total phenol contents of the extracts. The results indicated that the plants harvested in September possessing higher levels of active constituent had superior antioxidant capacities compared to the ones collected at other times. With respect to the location, plants harvested from the Izmir region had lower total phenol and active constituent levels resulting in poorer antioxidant activity.     

Improving the Sensory and Oxidative Stability of Cooked and Chill‐Stored Lamb Using Dietary Rosemary Diterpenes

Two dietary rosemary extracts (DREs) containing diterpenes (carnosic acid and carnosol at 1:1 and 2:1 w:w) were tested in fattening lambs to stabilize the sensory quality of cooked and chill‐stored patties. A total of 63 lambs were fed freely for 80 ± 5 d with a basal diet supplemented or not with DRE. Minced leg meat from each lamb was used to make patty batches. The patties were cooked at 72 ºC for 2 min, aerobically packed, kept at 2 ºC for up to 4 d and then reheated. Sensory traits (color, odor, flavor, and texture), CIELab color, and lipid oxidation (assessed as TBARS) were determined. In a first experiment, the lamb diet was supplemented with 600 mg of 1:1‐DRE or 2:1‐DRE kg^?1 feed. The 1:1‐DRE diet delayed discoloration, flavor deterioration, and rancidity, while the 2:1‐DRE diet was ineffective in this respect. In a second experiment, 4 supplementation levels of 1:1‐DRE (0, 200, 400, and 600 mg kg^?1 feed) were compared. Flavor deterioration was delayed when the lamb diet was supplemented with at least 400 mg 1:1‐DRE kg^?1 feed. The effects of the diet on the odor, flavor, and color were corroborated by differences in TBARS and CIELab. The results obtained suggest that rosemary diterpenes and/or their active secondary compounds deposited in muscle can act as endogenous antioxidants in cooked lamb. The carnosol intake seems crucial in the antioxidant actions achieved through DRE. The use of rosemary antioxidants in animal feeding would allow meat‐based dishes to be preserved longer without adding preservatives.     

Synergistic antimicrobial activity of galangal (Alpinia galanga), rosemary (Rosmarinus officinalis) and lemon iron bark (Eucalyptus staigerana) extracts

BACKGROUND: In this study the synergistic antimicrobial activities of combinations of extracts from galangal (Alpinia galanga), rosemary (Rosmarinus officinalis) and lemon iron bark (Eucalyptus staigerana) were evaluated against Staphylococcus aureus, Listeria monocytogenes, Escherichia coli, Salmonella typhimurium and Clostridium perfringens. Chemical compositions of these extracts were also determined to provide further insight into antimicrobial constituents and their potential mechanisms of action. RESULTS: Combinations of galangal with either rosemary or lemon iron bark showed synergistic antimicrobial activity. Specifically, galangal and rosemary showed synergistic activity against S. aureus and L. monocytogenes only, while galangal and lemon iron bark showed synergistic activity against E. coli and S. typhimurium. Chemical compositions of the extracts were determined by gas chromatographic–mass spectrometric analysis. The major chemical components of the galangal and lemon iron bark extracts were 1′‐acetoxy‐chavicol acetate (1′ACA) (63.4%) and neral (15.6%), respectively, while 1,8‐cineole (26.3%) and camphor (20.3%) were identified as major chemical components of the rosemary extract. CONCLUSION: The results of this study show that galangal, rosemary and lemon iron bark extracts contain components that may have different modes of antimicrobial action and combinations of these extracts may have potential as natural antimicrobials to preserve foods. Copyright © 2010 Society of Chemical Industry