Rapid quantitative enrichment of carnosic acid from rosemary (Rosmarinus officinalis L.) by isoelectric focused adsorptive bubble chromatography

For the first time, the potent but unstable antioxidative diterpene carnosic acid could be enriched from an aqueous extract of rosemary (Rosmarinus officinalis L.) by isoelectric focused adsorptive bubble chromatography. Enrichment of carnosic acid in the foam was influenced by the pH value and the flow rate of the foam-forming gas. Efficiency was highest with diluted samples at pH 4. Under these conditions, the conversion of carnosic acid to carnosol was negligible. Transfer of carnosic acid to the foam from a standard solution in the presence of saponin as surfactive substance was similar to that from the aqueous rosemary extract.     

Antimicrobial and antioxidant properties of rosemary and sage (Rosmarinus officinalis L. and Salvia officinalis L., Lamiaceae) essential oils

The essential oils of rosemary ( Rosmarinus officinalis L.) and sage ( Salvia officinalis L.) were analyzed by means of gas chromatography-mass spectrometry and assayed for their antimicrobial and antioxidant activities. Antimicrobial activity was tested against 13 bacterial strains and 6 fungi, including Candida albicans and 5 dermatomycetes. The most important antibacterial activity of both essential oils was expressed on Escherichia coli, Salmonella typhi, S. enteritidis, and Shigella sonei. A significant rate of antifungal activity, especially of essential oil of rosemary, was also exhibited. Antioxidant activity was evaluated as a free radical scavenging capacity (RSC), together with the effect on lipid peroxidation (LP). RSC was assessed by measuring the scavenging activity of essential oils on 2,2-diphenyl-1-picrylhydrazil (DPPH) and hydroxyl radicals. Effects on LP were evaluated following the activities of essential oils in Fe(2+)/ascorbate and Fe(2+)/H2O2 systems of induction. Investigated essential oils reduced the DPPH radical formation (IC50 = 3.82 microg/mL for rosemary and 1.78 microg/mL for sage) in a dose-dependent manner. Strong inhibition of LP in both systems of induction was especially observed for the essential oil of rosemary.     

Recovery mechanism of the antioxidant activity from carnosic acid quinone,an oxidized sage and rosemary antioxidant

A solution of carnosic acid quinone, which is a radical chain-termination product having no antioxidant activity in the antioxidant reaction of carnosic acid, recovers potent antioxidant activity upon standing. The HPLC analysis of an aged solution of carnosic acid quinone revealed that several antioxidants are produced in the solution. From the time-course and quantitative analyses of the formation of the products and their structural analysis, an antioxidant mechanism from carnosic acid quinone is proposed that includes a redox reaction of carnosic acid quinone in addition to the isomerization to lactone derivatives. In the first stage of antioxidation, carnosic acid, the reduction product from carnosic acid quinone, contributes to the potent antioxidant activity of the solution. This proposed mechanism can explain one of the reasons for the strong antioxidant activity of the extract of the popular herbs sage and rosemary.     

Influence of organic mulching and nitrogen application on essential oil yield and nitrogen use efficiency of rosemary (Rosmarinus officinalis L.)

In a field study conducted at Bangalore, India for two years (2003–2004 and 2004–2005), eight treatment combinations consisting of two variables, organic mulch (lemongrass spent material as mulch at 7.5 t ha^?1 and no mulch) and four levels of nitrogen fertilizer (0, 100, 200 and 300 kg ha^?1) were examined to observe the effect of organic mulching on nitrogen use-efficiency, and herb and essential oil yield in a multiharvested rosemary crop. The results revealed that application of lemongrass spent material as mulch increased the herb and essential oil yields in rosemary by 16.2 and 24.2%, respectively, over the non-mulched control at first harvest. Corresponding values for the regenerated crop harvest were 18.8 and 16.8%. A significant response to N was observed with 300 kg N ha^?1 in non-mulched plots compared with 200 kg N ha^?1 in mulched plots. Using lemongrass spent material as mulch, nitrogen uptake by the rosemary crop increased by 15.1% over the non-mulched control. At 200 kg N ha^?1, apparent recoveries by the crop were estimated to be 33.64% for the non-mulched control, and 37.79% with mulch. The quality of the rosemary essential oil, in terms of the concentrations of its major constituents, α-pinene, 1 : 8 cineole, camphor and verbenone, was not affected by the use of organic mulching and nitrogen fertilization, and these constituents were found to be of astandard acceptable in international trade.     

Azo dye-mediated regulation of total phenolics and peroxidase activity in thyme (Thymus vulgaris L.) and rosemary (Rosmarinus officinalis L.) clonal lines

Thyme (Thymus vulgaris L.) and rosemary (Rosmarinus officinalis L.) clonal lines, which were previously isolated from a heterogeneous seed population by plant tissue culture techniques, have been targeted as potential plants for phytoremediation of organic pollutants such as azo dyes and related aromatic compounds. Three thyme clonal lines and three rosemary clonal lines were tested for the ability to grow on hormone-free medium containing 0.01% of azo dye Poly S-119. The results showed that dye tolerance was associated with reduced phenolics and enhanced peroxidase activity in these clonal lines. There was a clear inverse correlation between total phenolics and peroxidase activity in these plants in response to Poly S-119. The tolerance of these clonal lines showed variations at different growing stages. These observations suggested that the peroxidase activity was inducible. Because peroxidases are involved in lignification, wound healing, aromatic compound degradation, pathogen defense, and stiffening, the results suggest that azo dye stimulated the defense response of thyme and rosemary clonal plants by increasing the peroxidase activity. Stereomicroscopic observations revealed that the azo dye was sequestered within the growing axis of the plant roots, which may also enhance the polymerization of azo dye onto the cell wall with the help of enhanced peroxidase activity.     

Polyphenolic transmission to Segureno lamb meat from ewes' diet supplemented with the distillate from rosemary (Rosmarinus officinalis) leaves

The aim of the present work is to study whether the introduction of rosemary plant byproduct, from plant steam distillation, in daily Segurena sheep feeding allows the transfer of active antioxidant components to lamb meat, without detriment to the animal productivity. For this, 36 Segurena ewes were assigned randomly to three homogeneous groups. One group was fed a basal diet as a control and the diet of the other two groups was modified by substituting 10 or 20% of the control diet (respectively) with distilled rosemary leaves. Chromatographic analysis allowed the identification of 11 polyphenolic components previously identified in the rosemary and basal diet pellets, respectively. Among them, rosmarinic acid, carnosol, and carnosic acid were the phenolic components that had a significantly increased presence ( P < 0.05) in the lamb meat from sheep mothers fed this aromatic herb, when compared to the control group. The incorporation of this byproduct into the animal diet favored the antioxidant capacity of these lamb meat samples. Fresh meat produced on rosemary had higher total ferric reducing antioxidant power (FRAP) ( P < 0.05), greater ability to reduce ABTS*+, and lower IC50 (DPPH*) ( P < 0.05) values when compared to the control group. Because no statistically significant differences were detected among the results obtained from the lamb meat belonging to the different ewe groups fed rosemary leaf extract (10 or 20%), it can be concluded that the incorporation of distilled rosemary leaves at a rate of 10% of the ewes' diet should be enough to improve the lamb meat antioxidant status.     

Combined use of supercritical fluid extraction, micellar electrokinetic chromatography, and reverse phase high performance liquid chromatography for the analysis of antioxidants from rosemary (Rosmarinus officinalis L.)

Antioxidants from rosemary were determined by the combined use of supercritical fluid extraction (SFE) prior to reverse-phase high-performance liquid chromatography (RP-HPLC) or micellar electrokinetic chromatography (MEKC). The separation of antioxidants found in the SFE fractions was achieved by using a new MEKC method and a published HPLC procedure, both with diode array detection. The characterization of the different antioxidants was further done by HPLC-mass spectrometry. Advantages and drawbacks of HPLC and MEKC for analyzing the antioxidants found in the different extracts are discussed. From the results it is concluded that HPLC renders higher peak area and is better in its reproducibility than MEKC; both techniques provide similar analysis time reproducibility. The main advantage of MEKC is its much higher separation speed, which is demonstrated to be useful for the quick adjustment of SFE conditions, allowing rosemary fractions of higher antioxidative power to be obtained quickly. Moreover, the possibilities of this approach for following the degradation of antioxidants are discussed.     

Variation of the chemical profile and antioxidant behavior of Rosmarinus officinalis L. and Salvia fruticosa Miller grown in Greece

In this study, the essential oil and the phenolic composition along with the antioxidant activity of R. officinalis L. and S. fruticosa Miller, collected in Zakynthos island (Ionian Sea, Greece), were investigated. The essential oil composition of the plants was characterized by the presence of 1,8-cineole. Mean values of the antioxidant activities of rosemary and sage essential oils indicated slight differences. The antioxidant activity of sage oil was correlated with the oxygenated sesquiterpenes and diterpenes concentrations. Concerning the methanolic extracts, a close relationship between the phenolic content and the development stage during vegetative cycle of these plants was observed. The identified flavonoids, except rutin, seemed to increase with the advancement of developmental stages, while phenolic acids followed an opposite pattern. The antioxidant activity was correlated with the amount of total phenolic content.     

Combined effect of modified atmosphere packaging and addition of rosemary (Rosmarinus officinalis), ascorbic acid, red beet root (Beta vulgaris), and sodium lactate and their mixtures on the stability of fresh pork sausages

The effects of rosemary, in combination with ascorbic acid, red beet root, and sodium lactate, as well as their mixtures, on the inhibition of both lipid and pigment oxidation of fresh pork sausages packaged in a modified atmosphere were studied. Sausages (240) were packaged in a 80% O2 + 20% CO2 gas mixture and analyzed for CIE a, metmyoglobin, TBARS, psychrotrophic aerobes, and sensory discoloration and off-odor throughout 20 days of storage at 2 +/- 1 degrees C. The mixture of rosemary + ascorbic acid + sodium lactate + red beet root extract extended the shelf life of fresh pork sausages from 8 to 16 days. Results demonstrated that all of the components of the mixture contributed to obtaining the maximum delay in color and/or odor decay, due to a combined inhibitory action on both pigment and lipid oxidation, as well as on microbial growth.     

Degradation Study of Carnosic Acid, Carnosol, Rosmarinic Acid, and Rosemary Extract (Rosmarinus officinalis L.) Assessed Using HPLC

Rosemary, whose major caffeoyl-derived and diterpenoid ingredients are rosmarinic acid, carnosol, and carnosic acid, is an important source of natural antioxidants and is being recognized increasingly as a useful preservative, protectant, and even as a potential medicinal agent. Understanding the stability of these components and their mode of interaction in mixtures is important if they are to be utilized to greatest effect. A study of the degradation of rosmarinic acid, carnosol, carnosic acid, and a mixture of the three was conducted in ethanolic solutions at different temperatures and light exposure. As expected, degradation increased with temperature. Some unique degradation products were formed with exposure to light. Several degradation products were reported for the first time. The degradation products were identified by HPLC/MS/MS, UV, and NMR. The degradation of rosemary extract in fish oil also was investigated, and much slower rates of degradation were observed for carnosic acid. In the mixture of the three antioxidants, carnosic acid serves to maintain levels of carnosol, though it does so at least in part at the cost of its own degradation.     

chemical composition, plant genetic differences, antimicrobial and antifungal activity investigation of the essential oil of Rosmarinus officinalis L

The chemical composition of the essential oil of the Sardinian Rosmarinus officinalis L. obtained by hydro distillation and steam\hydro distillation was studied using GC-FID and MS. Samples were collected at different latitude and longitude of Sardinia (Italy). The yields ranged between 1.75 and 0.48% (v/w, volume/dry-weight). A total of 30 components were identified. The major compounds in the essential oil were alpha-pinene, borneol, (-) camphene, camphor, verbenone, and bornyl-acetate. Multivariate analysis carried out on chemical molecular markers, with the appraisal of chemical, pedological, and random amplified polymorphic DNA data, allows four different clusters to be distinguished. The antimicrobial and antifungal tests showed a weak activity of Sardinian rosemary. On the other hand, an inductive effect on fungal growth, especially toward Fusarium graminearum was observed.     

Patatin,the tuber storage protein of potato (Solanum tuberosum L.), exhibits antioxidant activity in vitro

The potato (Solanum tuberosum L.) tuber storage protein, patatin, was purified to homogeneity with a molecular mass of 45 kDa. The purified patatin showed antioxidant or antiradical activity by a series of in vitro tests, including 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical (half-inhibition concentration, IC(50), was 0.582 mg/mL) scavenging activity assays, anti-human low-density lipoprotein peroxidation tests, and protections against hydroxyl radical-mediated DNA damages and peroxynitrite-mediated dihydrorhodamine 123 oxidations. Using electron paramagnetic resonance (EPR) spectrometry for hydroxyl radical detections, it was found that the intensities of the EPR signal were decreased by the increased amounts of patatin added (IC(50) was 0.775 mg/mL). Through modifications of patatin by iodoacetamide or N-bromosuccinimide, it was found that the antiradical activities of modified patatin against DPPH or hydroxyl radicals were decreased. It was suggested that cysteine and tryptophan residues in patatin might contribute to its antioxidant activities against radicals.     

Phenolic diterpenes,flavones, and rosmarinic acid distribution during the development of leaves,flowers, stems,and roots of Rosmarinus officinalis. Antioxidant activity

The distribution of six compounds with three different polyphenol skeletons have been studied in Rosmarinus officinalis: phenolic diterpenes (carnosic acid, carnosol, and 12-O-methylcarnosic acid), caffeoyl derivatives (rosmarinic acid), and flavones (isoscutellarein 7-O-glucoside and genkwanin), each showing a characteristic behavior and distribution during the vegetative cycle. Only in leaves were all six compounds present, and the highest accumulation rate was related with the young stages of development. Rosmarinic acid showed the highest concentrations of all the polyphenols in all organs. The distribution of this acid in leaves, flowers, and stems suggests that in the first stages of flower growth, levels were due to in situ biosynthesis, and in the last stages, the contribution of transport phenomena was increased. The antioxidant activity of six extracts with different polyphenolic composition was evaluated in aqueous and lipid systems. The results clearly suggest that rosemary extracts are excellent antioxidants in both aqueous and lipid systems.     

New selection strategies for determining the traits contributing to increased grain yield in wheat (Triticum aestivum L.) under aluminum stress

Genetic Resources and Crop Evolution - Finding the suitable traits that mediate grain yield under stress condition represent an important challenge to plant breeders in order to improve production....     

Comparison of yield, composition, and antioxidant activity of turmeric (Curcuma longa L.) extracts obtained using various techniques

Turmeric extracts were obtained from two lots of raw material (M and S) using various techniques: hydrodistillation, low pressure solvent extraction, Soxhlet, and supercritical extraction using carbon dioxide and cosolvents. The solvents and cosolvents tested were ethanol, isopropyl alcohol, and their mixture in equal proportions. The composition of the extracts was determined by gas chromatography-flame ionization detection (GC-FID) and UV. The largest yield (27%, weight) was obtained in the Soxhlet extraction (turmeric (S), ethanol = 1:100); the lowest yield was detected in the hydrodistillation process (2.1%). For the supercritical extraction, the best cosolvent was a mixture of ethanol and isopropyl alcohol. Sixty percent of the light fraction of the extracts consisted of ar-turmerone, (Z)-gamma-atlantone, and (E)-gamma-atlantone, except for the Soxhlet extracts (1:100, ethanol), for which only ar-turmeronol and (Z)-alpha-atlantone were detected. The maximum amount of curcuminoids (8.43%) was obtained using Soxhlet extraction (ethanol/isopropyl alcohol). The Soxhlet and low pressure extract exhibited the strongest antioxidant activities.     

Physiological and biochemical responses of Melissa officinalis L. to nickel stress and the protective role of salicylic acid

The present study investigated the mediatory effects of salicylic acid (SA) in alleviating nickel (Ni) toxicity in Melissa officinalis L. One-month-old plants were exposed to different levels of Ni and SA concentrations in sand culture under greenhouse conditions. Excess Ni significantly inhibited the growth indices and dramatically increased accumulation of Ni in the leaves and roots. Exogenously SA applications (1.0 mM) led to a substantial improvement in the shoot and root fresh and dry weights. Foliar application of SA mitigated the deleterious effects of Ni and decreased its transport to the shoots. The results showed a significant loss in chlorophylls and carotenoids contents only at 500 µM of Ni. The impact of SA was not significant in terms of chlorophyll contents, while carotenoid contents of the Ni-stressed plants were significantly affected by SA. Exposure to Ni did not modify proline accumulation. Hydrogen peroxide accumulation was observed under Ni stress, while lipid peroxidation significantly decreased at the same conditions. Application of SA caused a significant decrease in electrolyte leakage of Ni-stressed plants. Due to the high potential for Ni accumulation in the roots and translocation factor values lower than 1, M. officinalis could be introduced as an excluder medicinal plant.     

Ferulic acid from aleurone determines the antioxidant potency of wheat grain (Triticum aestivum L.)

Grain is an important source of phytochemicals, which have potent antioxidant capacity. They have been implicated in the beneficial health effect of whole grains in reducing cardiovascular disease and type 2 diabetes. The aim of the present study was to identify the most important antioxidant fractions of wheat grain. It was found that the aleurone content of these fractions was highly correlated with the antioxidant capacity of the fractions (r = 0.96, p < 0.0001). Ferulic acid appeared to be the major contributor to the antioxidant capacity in fractions with higher antioxidant capacity. The contribution of protein was rather limited. It was concluded that the antioxidant potency of wheat grain fractions is predominantly determined by aleurone content, which can be attributed to the presence of relatively large amounts of phenolic compounds, primarily ferulic acid.     

Effects of nitrogen,boron and potassium on boron deficiency,leaf mineral concentrations,and yield of table beets (Beta vulgaris L.)

Abstract

Application of 11.2 kg B/ha and/or of 224 kg N/ha reduced the number of table beet (Beta vulgarisL. cv Detroit Dark Red) roots with B deficiency compared to low rates or no N or B application in three field experiments. In two of the three experiments, the 224 kg/ha N rate decreased B concentration in leaves when no B was added yet B deficiency of roots was reduced compared to the 56 kg/ha N rate. B application increased B concentration in leaves. Calcium concentration in leaves was higher at 224 kg N/ha than for 56 kg N/ha. Nitrogen concentration in leaves was increased by increasing the rate of N while K concentration was decreased at the higher N rate. Average number of roots with B deficiency was significantly higher for the Morse strain than for the Harris strain of ‘Detroit Dark Red’ in one experiment where the two strains were compared. There were no differences between the two strains in leaf B concentration but Ca concentration was higher in the Morse strain than for the Harris strain. Yields were higher at the 224 kg/ha rate of N than at 56 kg N/ha but were not consistently affected by B and K applications. There was no clear relationship of leaf N, Ca and K concentrations to B deficiency of roots     

Growth and antioxidant responses in mustard (Brassica juncea L.) plants subjected to combined effect of gibberellic acid and salinity

The effects of salt stress on plant growth parameters, lipid peroxidation and some antioxidant enzyme activities [superoxide dismutase (SOD; EC 1.15.1.1), catalase (CAT, EC 1.11.1.6), peroxidase (POD, EC 1.11.1.7), glutathione reductase (GR; EC EC 1.6.4.2) and ascorbate peroxidase (APX, EC 1.11.1.11) activity] were studied in the leaves of mustard. Plants were exposed to two different concentrations of NaCl stress (100 and 150 mM) for 45 days and were sprayed with GA₃ (75 ml pot^?1, conc. 75 mg l^?1) once a week. Salt stress resulted in decrease in the growth and biomass yield of mustard but the exogenous application of GA₃ enhanced these parameters significantly. Application of GA₃ counteracted the adverse effects of NaCl salinity on relative water content, electrolyte leakage and chlorophyll (Chl) content. GA₃ was sufficient to attenuate partially the stimulatory effect of NaCl supply on proline and glycinebetaine biosynthesis. GA₃ reduced lipid peroxidation in the leaves, which was increased during salt stress. The activity of all the antioxidant enzymes was increased significantly during salt stress in mustard. The exogenous application of GA₃ decreased the enzyme activity. The results of the present study indicate that usage of GA₃ reduces the harmful effects of salinity and increases resistance to salinity in mustard plant.     

Changes in phenolic concentrations during recurrent selection for resistance to the Mediterranean corn borer (Sesamia nonagrioides Lef.)

Recurrent selection has been reported as successful for improving maize resistance against corn borers. This study was conducted to determine if phenolics concentration in maize changes during recurrent selection to improve stalk resistance to the Mediterranean corn borer. Three cycles of selection [EPS12(S)C0, ESP12(S)C2, and EPS12(S)C3] from the maize synthetic population EPS12 and test crosses to inbred lines A639, B93, and EP42 were field grown and artificially infested with Mediterranean corn borer larvae, and the pith tissues were sampled for biochemical analyses. Two major simple phenolic acids [p-coumaric (p-CA) and trans-ferulic (E-FA) acids] were identified in free and cell-wall fractions, whereas four isomers of diferulic acid (DFA) (8-5'l, 5-5', 8-o-4', and 8-5' benzofuran form) were present in the cell-wall bound fraction. The selection cycles EPS12(S)C0 and EPS12(S)C3 showed less damage and higher cell wall phenolics concentrations than the cycle EPS12(S)C2. In addition, higher concentrations of total DFAs were associated with shorter tunnel length and lower numbers of larvae per stem. The current study shows new and concrete evidence that the cell-wall bound phenolics could have a determinative role in the resistance to the Mediterranean corn borer, although future development of recurrent and divergent selection cycles will clarify this point.