Antioxidant activity of lipid-soluble phenolic diterpenes from rosemary

A high-performance liquid chromatography method for analyzing the phenolic diterpenes present in rosemary (Rosmarinus officinalis L.) and commercial rosemary extracts is reported. Carnosic acid was the major phenolic diterpene present in rosemary leaves, with lesser amounts of 12-methoxycarnosic acid and carnosol. Several commercial rosemary extracts also were analyzed by this method, and in addition to these three compounds other phenolic diterpenes, such as 7-methoxyrosmanol, 7-methoxy-epirosmanol, and rosmanol, were found in some samples. These latter three compounds seem to be artifacts, produced from carnosic acid by oxidation and cyclization. The major phenolic diterpenes were isolated, and their relative antioxidatn activities in soybean oil were measured by the Rancimat. The potency of carnosic acid was more than twice that of any other compound. The antioxidant activity of pure carnosic acid was compared to butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), and tertiary butylhydroquinone (TBHQ) and was several times greater than BHT and BHA but less than TBHQ. Nuclear magnetic resonance data for several of the compounds that were incompletely characterized in previous literature are reported.     

Antioxidative activity and phenolic composition of pilot-plant and commercial extracts of sage and rosemary

Eight sage (Salvia officinalis) and twenty-four rosemary (Rosmarinus officinalis) extracts, originating from pilot-plant or commercial sources, had different antioxidative activities as measured by accelerated autoxidation of methyl linoleate. Twenty-seven compounds were characterized in the Labiatae family extracts by high-performance liquid chromatography (HPLC) coupled with mass spectrometry, equipped with an atmospheric pressure chemical ionization interface, and by HPLC coupled with a photodiode array spectrophotometer. Twenty-two compounds were identified, including phenolic acids, carnosol derivatives, and flavonoids. The extracts showed great variation in their HPLC profiles, and no correlation was apparent between their antioxidative efficiency and their composition, in twenty specific phenols. Data indicated that the most effective compounds were carnosol, rosmarinic acid, and carnosic acid, followed by caffeic acid, rosmanol, rosmadial, genkwanin, and cirsimaritin.     

Use optimization of natural antioxidants in refined, bleached, and deodorized palm olein during repeated deep-fat frying using response surface methodology

An optimization study on the use of oleoresin rosemary extract, sage extract, and citric acid added into refined, bleached, and deodorized (RBD) palm olein in deep-fat frying of potato chips was carried out using response surface methodology (RSM). Results showed that oleoresin rosemary extract was the most important factor affecting the sensory acceptability of potato chips. For taste and odor, its effects were highly significant (P<0.01), while for crispiness and overall acceptability, the effects were significant (P<0.05). As for sage extract, the level of this antioxidant had a highly significant (P<0.01) effect on appearance and taste and a significant effect (P<0.05) on odor and overall acceptability, but had no effect on crispiness. Although there was no significant synergistic correlation between citric acid and oleoresin rosemary extract or sage extract at the first order, its second order was significantly (P<0.05) related to taste, crispiness, and overall acceptability. An interaction between oleoresin rosemary and sage extracts also significantly (P<0.05) improved the score of overall acceptability of the potato chips. Contour maps of the sensory scores of potato chips indicated that the optimal points for appearance were achieved using 0.062% oleoresin rosemary extract, 0.066% sage extract, and 0.023% citric acid, while optimal task was achieved with 0.063% oleoresin rosemary extract, 0.075% sage extract, and 0.025% citric acid. With the same sequence of ingredients added into oil, the combinations required to achieve the optimal odor, crispiness, and overall acceptability scores were 0.058-0.046-0.026, 0.060-0.071-0.022, and 0.060-0.064-0.026%, respectively.     

Supercritical rosemary extracts,their antioxidant activity and effect on hepatic tumor progression

The use of supercritical fluid technology as an innovative technology to extract bioactive compounds has grown considerably in recent decades. Particularly, the recovery of antioxidants from different herbs is a matter of continuous research and development. Antioxidants can protect cells against the effects of free radicals and thus, play an important role in heart illness, cancer and other diseases.Rosemary (Rosmarinus officinalis L.) has been recognized as one of the Lamiaceae plant with many important biological activities. Particularly, large antioxidant power has been recognized in rosemary and main substances related with this activity were the phenolic diterpenes such as carnosol, rosmanol, carnosic acid, methyl carnosate, and phenolic acids such as the rosmarinic and caffeic acids. Moreover, carnosic acid and carnosol are recognized as the most abundant antioxidants present in rosemary.In this work, supercritical fluid technology was applied to produce rosemary extracts with different compositions and thus, with different bioactivity properties. Selected extracts, from the variety of samples obtained, were used to study the capability of rosemary supercritical extracts to inhibit the proliferation of human liver carcinoma cells. These extracts showed a dose-dependent effect on inhibiting the proliferation of human hepatoma cells. Moreover, observed citostaticity appeared to be significantly influenced by their different composition, suggesting a relevant role of the technology to produce the extracts and the consequently obtained compositions on the potential antitumoral activity of rosemary.     

Rosmarinus Officinalis Leaves as a Natural Source of Bioactive Compounds

In an extensive search for bioactive compounds from plant sources, the composition of different extracts of rosemary leaves collected from different geographical zones of Serbia was studied. The qualitative and quantitative characterization of 20 rosemary (Rosmarinus officinalis) samples, obtained by microwave-assisted extraction (MAE), was determined by high performance liquid chromatography coupled to electrospray quadrupole-time of flight mass spectrometry (HPLC–ESI-QTOF-MS). The high mass accuracy and true isotopic pattern in both MS and MS/MS spectra provided by the QTOF-MS analyzer enabled the characterization of a wide range of phenolic compounds in the extracts, including flavonoids, phenolic diterpenes and abietan-type triterpenoids, among others. According to the data compiled, rosemary samples from Sokobanja presented the highest levels in flavonoids and other compounds such as carnosol, rosmaridiphenol, rosmadial, rosmarinic acid, and carnosic acid. On the other hand, higher contents in triterpenes were found in the extracts of rosemary from Gložan (Vojvodina).     

Optimization of physicochemical changes of palm olein with phytochemical antioxidants during deep-fat frying

Response surface methodology (RSM) was used to optimize the amounts of rosemary and sage extracts together with citric acid as synergist antioxidants in stabilizing refined, bleached, and deodorized palm olein during repeated deep-fat frying of potato chips. For all physicochemical properties studied, these phytochemical antioxidant treatments significantly (P<0.05) reduced the oxidation rate of the oil. During 5 d of frying, anisidine value, peroxide value, free fatty acid, polymer content, color units, viscosity, and absorbances at 232 and 268 nm gradually increased, whereas iodine value and ratio of 18∶2/16∶0 decreased. Further statistical analyses, including coefficient of determination (R ²) and probability of F values, indicated that mathematical models for each physicochemical parameter could be developed confidently in this study, with R ² for all parameters greater than 0.90. These results suggested that an optimal mixture of phytochemical antioxidants derived from rosemary and sage together with citric acid could be produced using RSM for stabilizing thermally processed oil. For many physicochemical parameters examined, the use of moderate levels of antioxidants could result in optimal responses.     

Lipid oxidation inhibiting capacities of blackseed essential oil and rosemary extract

Lipid oxidation is the main deterioration process that occurs in vegetable oils containing lipid molecules with polyunsaturation. The aim of the present study was to investigate the possible effects of blackseed (Nigella sativa L.) essential oil (BEO) and rosemary (Rosmarinus officinalis L.) extract (RE) on stabilization of sunflower oil under accelerated storage conditions. RE was obtained by soxhlet extraction using methanol, whereas BEO was hydrodistilled from the blackseed extract obtained by extraction using petroleum ether (b.p. 40–60°C). The results indicate that both extracts stabilize sunflower oil to a certain extent, the extent being greater with RE than with BEO based on measurements of peroxide value, p‐anisidine value, amount of nonoxidized linoleic acid in saponified oil samples by HPLC‐DAD, concentration of conjugated diene hydroperoxide, and UV light absorption. The oil stabilizing effect of BEO against lipid oxidation, especially at concentrations of 0.06 and 0.1 g/100 g oil was found to increase as the temperature increased, according to principal component analysis results. Based on its strong lipid oxidation inhibiting capacity, RE can be used as a potential natural extract for stabilizing sunflower oil against oxidation. BEO, at carefully selected concentrations, can be suggested as an alternative supplement of plant origin for improving oil stabilization. Practical applications: Sunflower oil is a widely used vegetable oil for cooking and frying, and has a high linoleic acid content of about 40–70%. Here we show that sunflower oil can be stabilized by adding rosemary and blackseed extracts. The treatment can be useful especially in applications which require heating the oil to high temperatures.     

Synergies between plant antioxidant blends in preventing peroxidation reactions in model and food oil systems

A study was conducted to investigate the oxidative behavior of various mixtures of rosemary, sage, and citric acid in a linoleic acid model system by oxygen consumption measurement and in a palm olein system by differential scanning calorimetry (DSC) analysis. Response surface methodology was used to optimize the use of the mixtures. Results showed that rosemary and sage were two important factors for the protective index (PI). The two antioxidants were highly significantly (P<0.001) in influencing PI values. There was a significant (P<0.01) synergistic effect between rosemary and sage on PI values. Citric acid was also found to be significant (P<0.05) for PI. With respect to onset time (T _o ), all three antioxidants were significant (P<0.05). However, no significant interaction among antioxidants was found for T _o . Mathematical models for both PI and T _o could be developed with confidence. The R ² values for PI and T _o were 0.992 and 0.926, respectively. A combination of 0.078% rosemary, 0.067% sage and 0.037% citric acid was the optimal combination for PI, whereas a combination of 0.068% rosemary, 0.075% sage, and 0.039% citric acid was required to reach the optimal T _o value.     

The effect of primary antioxidants and synergists on the activity of plant extracts in lard

The antioxidant activities of combinations of plant extracts and primary antioxidants or citric acid were studied in lard stored at 75°C. Methanol extracts of oregano, thyme, marjoram, dittany, rosemary and sage were used. When combined with butylated hydroxyanisole, butylated hydroxytoluene or ascorbyl palmitate the extracts showed an additive antioxidant effect, while combinations with propyl gallate did not increase the induction period obtained by the phenolic antioxidant. Negative synergism was observed when the plant extracts were mixed with DL-α-tocopherol. Citric acid showed a high synergistic efficiency with marjoram extract and a minor one with thyme extract, but it had a negative effect with all the others. Ternary mixtures of the plant extracts with citric acid and ascorbyl palmitate had lower activity than the corresponding dual combinations with ascorbyl palmitate, except for the mixture of marjoram extract.     

Suitability of TBA method for the evaluation of the oxidative effect of non-water-soluble and water-soluble rosemary extracts

The antioxidative effects of rosemary and grape-seed extracts spiked in human plasma were examined using the thiobarbituric acid (TBA) method. The TBA values of plasma spiked with reagents to generate reactive oxygen species, such as singlet oxygen ((1)O(2)), hydroxyl radicals ((·)OH), peroxynitrite (ONOO(-)), and superoxide anions (O(2)(·-)), were measured by a flow injection analysis method with fluorescence (FL) detection. TBA values obtained by the addition of 50 mg/mL of rosemary extracts for (1)O(2), (·)OH, ONOO(-), and O(2)(·-) increased to 964 ± 65%, 1063 ± 61%, 758 ± 78%, and 698 ± 41%, respectively (n = 3, P < 0.01), whereas the values with 1 mg/mL of grape-seed extracts or tocopherol decreased (40.2 - 66.3%). Furthermore, the antioxidative effects of rosemary extract in rat plasma, spiked with reagents to generate (·)OH, were examined by high-performance liquid chromatography with FL detection. No peak, other than TBA-malondialdehyde, could be detected using wavelengths of 532 (λ(ex)) and 553 nm (λ(em)).     

Phenolic Metabolites in Leaves of the Invasive Shrub, Lonicera maackii, and Their Potential Phytotoxic and Anti-Herbivore Effects

Lonicera maackii is an invasive shrub in North America for which allelopathic effects toward other plants or herbivores have been suspected. We characterized the major phenolic metabolites present in methanol extracts of L. maackii leaves. In addition, we examined the effects of methanol–water extracts of L. maackii leaves on seed germination of a target plant species and on feeding preference and growth rate of a generalist insect herbivore. A total of 13 individual major and minor compounds were detected in crude leaf extracts by high-performance liquid chromatography coupled to electronspray ionization-tandem mass spectrometry (ESI-MS/MS). Extracts were dominated by two major flavones, apigenin and luteolin, and their glucoside derivatives, apigenin-7-glucoside and luteolin-7-glucoside. Quantities of these compounds, along with chlorogenic acid, varied between two sampling points. Leaf extracts that contained these compounds were inhibitory to seed germination of Arabidopsis thaliana. In addition, treatment of artificial diet with leaf extracts deterred feeding of the generalist herbivore, Spodoptera exigua, in choice experiments but had no effect on growth rate in short-term no-choice bioassays. Purified apigenin tended to deter feeding by S. exigua and inhibited seed germination of A. thaliana. We conclude that leaves of L. maackii contain phenolic compounds, including apigenin and chlorogenic acid, capable of having biological effects on other plants and insects.     

Stabilization of Camelina Oil with Synthetic and Natural Antioxidants

Camelina oil was found to have a much lower Oil Stability Index and higher p-anisidine rates in the oven storage test than either rapeseed or sunflower oils. Stabilization of camelina oil was evaluated with 21 food grade synthetic and natural antioxidants and antioxidant formulations, using both the Oil Stability Index (OSI) and the oven storage test. The Oil Stability Index of camelina oil was able to be increased above that of rapeseed oil with TBHQ and its formulation with citric acid, and above that of sunflower oil with EGC, EGCG, carnosic acid, propyl gallate, rosemary extract with ascorbyl palmitate or with gallic acid. para-Hydroxyphenols were found to be more effective than ortho-hydroxyphenols and monohydroxyphenols had no significant effect on the OSI. Good correlation (R ² = 0.96) was found between the stabilizing effect of ortho-hydroxyphenols and the molarity of the phenyl hydroxyl groups per weight of antioxidant. The oven storage test carried out with six of the evaluated antioxidants indicated that p-anisidine rates of camelina oil stabilized with commercial formulations of TBHQ with citric acid or rosemary extract with ascorbyl palmitate were about the same as that of sunflower oil, an almost 90% rate reduction when compared to camelina oil. Accordingly, camelina oils stabilized with TBHQ/citric acid and rosemary extract/ascorbyl palmitate formulations were more stable than rapeseed and sunflower oils, respectively in terms of OSI induction times and p-anisidine rates.     

Simultaneous extraction of rosemary and spinach leaves and its effect on the antioxidant activity of products

Mixed vegetal extracts are interesting target of new products as nutraceuticals, superior ingredients for the design of functional food, singular ingredients for cosmetics, etc. In this work the extraction of a mixture of spinach and rosemary leaves (50 wt.% of each plant) was investigated in terms of its antioxidant activity, and compared with the extraction of the separate species. Phenolic diterpenes of rosemary and carotenoids of spinach were target compounds due their recognized biological activities. Two different extraction techniques were applied, namely pressurized liquid extraction using hexane at two different temperatures (100 and 150 °C) and supercritical fluid extraction with pure carbon dioxide at 40 °C and two different pressures (20 and 30 MPa). For each extraction technique and conditions three different raw materials were employed: spinach leaves, rosemary leaves and the mixture 50:50 of spinach and rosemary leaves.The antioxidant activity of the samples produced was evaluated with the ABTS assay and showed to be enhanced when the species are simultaneously extracted, with antioxidant values around 20% higher than the values corresponding to mixing the extracts obtained by separate. A possible synergic effect between carotenoids and phenolic diterpenes was studied, although no specific synergic activity could be observed. However, the enhanced antioxidant activity could be attributed to a definite increase of the concentration of carnosic acid, which was observed in the samples produced by the simultaneous extraction.     

Repellency of Rosemary Oil Against Myzus persicae in a Laboratory and in a Screenhouse

The repellencies of 13 labiate essential oils against Myzus persicae were investigated with a linear track olfactometer. Rosemary, thyme, peppermint, lavender, and spearmint oils repelled aphids at a dose of 10 l. Rosemary and thyme oils repelled at a dose of 1 l. The repellent actions of 13 components of rosemary oil were also evaluated. Among these components linalool, d,l-camphor, and -terpineol had repellent action. The repellency of rosemary oil in a screenhouse was investigated. Aphids were released in a screenhouse and allowed to choose between tobacco plants in an area permeated with rosemary oil odor and plants in a control area. The number of aphids in the treatment area was about 70% of that in the control area. These results indicated that the landing of M. persicae on host plants was influenced by odors and that it may be possible to control aphids with repellents.     

Natural antioxidants from herbs and spices

There is a growing interest in natural antioxidants found in plants because of the world‐wide trend toward the use of natural additives in food and cosmetics. Herbs and spices are one of the most important targets to search for natural antioxidants from the point of view of safety. This review presents the results on stabilization of lipids and lipid‐containing foods with different herbs and spices (ground materials or extracts) and reports the structure of the main antioxidatively acting compounds isolated from them. The review presents information about the antioxidative effects of rosemary, sage, oregano, thyme, ginger, summer savory, black pepper, red pepper, clove, marjoram, basil, peppermint, spearmint, common balm, fennel, parsley, cinnamon, cumin, nutmeg, garlic, coriander, etc. Among the herbs of the Labiatae family, rosemary has been more extensively studied and its extracts are the first marketed natural antioxidants. Sage and oregano, which belong to the same family, have gained the interest of many research groups as potential antioxidants.     

Evaluation of antioxidants for cod liver oil by chemiluminescence and the rancimat method

Commercial blends of natural antioxidants,viz., tocopherol concentrates, rosemary extracts, sage extracts, and lecithins, were tested for their ability to stabilize cod liver oil. The antioxidants were tested by using the Rancimat apparatus at 80°C and by a method based on hypochlorite-activated chemiluminescence analysis of samples stored at 35°C for 24 h in light. In addition, a stability study at 5°C in the dark for 8 wk, under conditions realistic for normal consumption of cod liver oil was carried out. A low correlation (r=0.339) was found between Rancimat induction times and chemiluminescence data for the sixteen antioxidant systems tested, probably due to temperature differences, and different ways of detecting oxidation products. Based on Rancimat induction times, δ-tocopherol-rich antioxidants and lecithin had the best stabilizing effect. However, based on the chemiluminescence method, the tocopherols acted as prooxidants, while tocopherols with lecithin increased the stability. Both Racimat and chemiluminescence data showed stabilizing effects with rosemary and sage extracts, but no synergistic effect between the herbal extracts and lecithin or tocopherol was observed. Analyses of oil aged at 5°C for 8 wk showed the highest stability for cod liver oil containing rose-mary extracts. The tocopherol mixtures showed only a minor effect on the stability. Ranking of antioxidants varied considerably depending on the method used, and increasing the temperature seemed to decrease the usefulness of the method. Antioxidant evaluation has to be done by using as many evaluation methods as possible under conditions relevant for normal storage and use.     

Antioxidant activities of hot water extracts from various spices

Recently, the natural spices and herbs such as rosemary, oregano, and caraway have been used for the processing of meat products. This study investigates the antioxidant activity of 13 spices commonly used in meat processing plants. The hot water extracts were then used for evaluation of total phenolic content, total flavonoids content and antioxidant activities. Our results show that the hot water extract of oregano gave the highest extraction yield (41.33%) whereas mace (7.64%) gave the lowest. The DPPH radical scavenging ability of the spice extracts can be ranked against ascorbic acid in the order ascorbic acid > clove > thyme > rosemary > savory > oregano. The values for superoxide anion radical scavenging activities were in the order of marjoram > rosemary > oregano > cumin > savory > basil > thyme > fennel > coriander > ascorbic acid. When compared to ascorbic acid (48.72%), the hydroxyl radical scavenging activities of turmeric and mace were found to be higher (p < 0.001). Clove had the highest total phenolic content (108.28 μg catechin equivalent (CE)/g). The total flavonoid content of the spices varied from 324.08 μg quercetin equivalent (QE)/g for thyme to 3.38 μg QE/g for coriander. Our results indicate that hot water extract of several spices had a high antioxidant activity which is partly due to the phenolic and flavonoid compounds. This provides basic data, having implications for further development of processed food products.     

Application of organic solvent nanofiltration for concentration of antioxidant extracts of rosemary (Rosmarinus officiallis L.)

The present investigation revealed the potential benefits of nanofiltration application in herbal extracts processing. Nanofiltrations of modelling solution of caffeic and rosmarinic acid in ethanol and ethanolic rosemary extract were carried out in a laboratory cross-flow system. Duramem™ 200 nanofiltration membrane was selected based on screening experiments. The experimental data and model predictions demonstrated the efficacy of a semi-batch cross-flow diafiltration process for concentration of fresh rosemary extracts. The observed absence of significant loss of antioxidant capacity in the retentate during the process and the degree of extract concentration achieved may allow retentate direct application as preservative and functional ingredient in the foods, cosmetics, neutraceuticals and medicines. The capability of the selected membrane to separate monophenolic acids from higher molecular weight antioxidant compounds in the extracts was also discussed. The dry solids content in the permeate was found sufficiently low as to permit its direct re-use in the extraction process thus bringing additional economical benefits.     

Stabilisation of edible oils with natural antioxidants

Natural antioxidants have recently gained increased interest because of the belief that natural food ingredients are better and safer than synthetic ones. The review presents the results on stabilisation of the main edible oils with different types of natural antioxidants. Sources of natural antioxidants are spices, herbs, teas, oils, seeds, cereals, cocoa shell, grains, fruits, vegetables, enzymes, proteins. Researchers concentrate on ascorbic acid, tocopherols and carotenoids as well as on plant extracts containing various individual antioxidants such as flavonoids (quercetin, kaemferol, myricetin), catechins or phenols (carnosol, rosmanol, rosamaridiphenol) and phenolic acids (carnosic acid, rosmarinic acid). Ascorbyl palmitate is regarded as a 'natural' antioxidant because it is hydrolysed in the body to ascorbic and palmitic acids. Among the herbs of the Lamiaceae family, rosemary has been more extensively studied and its extracts are the first marketed natural antioxidants. Oregano, which belongs to the same family, has gained the interest of many research groups as a potent antioxidant in lipid systems. The review concerns the following main topics: stabilisation of oil with individual natural antioxidants, interaction of antioxidants with synergists, stabilisation of oil with extracts or dry materials from different plant sources (e.g. herbs and spices), stabilisation at frying temperatures and in emulsions.     

Changes in Quality Indices,Phenolic Content and Antioxidant Activity of Flavored Olive Oils during Storage

The chemical characteristics, phenolic content and antioxidant activity of olive oils flavored with garlic, lemon, oregano, hot pepper, and rosemary were evaluated during 9 months of storage. At the end of the storage period, the unflavored and the garlic-flavored oils maintained their chemical parameters within the limits fixed for extra-virgin olive oils. After 9 months of storage, a noticeable decrease in phenolic content was observed in all the oils. The highest (35.0 ± 3.9 mg/kg oil) and the lowest (6.3 ± 0.4 mg/kg) phenolic contents were detected in the unflavored and garlic-flavored oils, respectively. Compounds such as 3,4-DHPEA-EDA (3,4-dihydroxyphenylethyl 4-formyl-3-formylmethyl-4-hexenoate, the dialdehydic form of decarboxymethyl elenolic acid linked to hydroxytyrosol) and p-HPEA-EDA (dialdehydic form of the decarboxymethyl elenolic acid linked to tyrosol) were the most abundant in both unflavored and lemon-flavored oils up till 6 months of storage. At the end of the storage period, increases in 3,4-DHPEA (hydroxytyrosol) and p-HPEA (tyrosol) were measured in almost all the oils. During storage, the antioxidant activity coefficients of the phenolic extracts, calculated according to the β-carotene bleaching assay, significantly decreased and, after 9 months, were in a decreasing order: rosemary (51.3 ± 4.2), hot pepper, lemon, oregano, unflavored, and garlic (8.5 ± 0.7).