Abstract: Rosemary (Rosmarinus officinalis L.) extracts (RE) are natural antioxidants that are used in food, food supplements and cosmetic applications; exert anti-inflammatory and anti-hyperglycaemic effects; and promote weight loss, which can be exploited to develop new preventive strategies against metabolic disorders. Therefore, the aim of the present study was to evaluate the preventive effects of rosemary leaf extract that was standardised to 20 % carnosic acid (RE) on weight gain, glucose levels and lipid homeostasis in mice that had begun a high-fat diet (HFD) as juveniles. The animals were given a low-fat diet, a HFD or a HFD that was supplemented with 500 mg RE/kg body weight per d (mpk). Physiological and biochemical parameters were monitored for 16 weeks. Body and epididymal fat weight in animals on the HFD that was supplemented with RE increased 69 and 79 % less than those in the HFD group. Treatment with RE was associated with increased faecal fat excretion but not with decreased food intake. The extract also reduced fasting glycaemia and plasma cholesterol levels. In addition, we evaluated the inhibitory effects of RE in vitro on pancreatic lipase and PPAR-γ agonist activity; the in vitro findings correlated with our observations in the animal experiments. Thus, the present results suggest that RE that is rich in carnosic acid can be used as a preventive treatment against metabolic disorders, which merits further examination at physiological doses in randomised controlled trials.

Abstract: The absorption, distribution and elimination of carnosic acid, the main antioxidant found in rosemary was studied, in vivo, in rats. Therefore, carnosic acid was administrated in a single dose, intravenously (20.5 ± 4.2 mg/kg) and orally (64.3 ± 5.8 mg/kg), to four and nine rats, respectively. Blood samples were collected at different time points, and plasma concentrations of carnosic acid were determined using LC-MS. Furthermore, total collection of urine and feces was done during 4 h and 24 h for the intravenous and oral administrations, respectively. After euthanizing the rats, intestinal content, liver and muscle tissue were sampled to determine carnosic acid concentrations. The bioavailability of carnosic acid, after 360 min, was 40.1%. Traces of carnosic acid were found in the rats intestinal content, liver and muscle tissue of abdomen and legs. The recovery of carnosic acid in the feces, 24 h after oral administration, was 15.6 ± 8.2%. Carnosic acid is absorbed into the bloodstream after oral administration in rats and is therefore bioavailable. It was found that carnosic acid in vivo is present in its free form and that its main elimination route is the fecal route.

Abstract: 1α,25-dihydroxyvitamin D3 (1,25D) is a powerful differentiation agent, which has potential for treatment of acute myeloid leukemia (AML), but induces severe hypercalcemia at pharmacologically active doses. We have previously shown that carnosic acid (CA), the polyphenolic antioxidant from rosemary plant, markedly potentiates differentiation induced by low concentrations of 1,25D in human AML cell lines. Here, we demonstrated similar enhanced differentiation responses to the 1,25D/CA combination in primary leukemic cells derived from patients with AML, and determined the role of the Nrf2/antioxidant response element (Nrf2/ARE) pathway in these effects using U937 human monoblastic leukemia cells as the model. CA strongly transactivated the ARE-luciferase reporter gene, induced the ARE-responsive genes, NADP(H)-quinone oxidoreductase and the γ-glutamylcysteine synthetase heavy subunit, and elevated cellular glutathione levels. Interestingly, 1,25D potentiated the effects of CA on these activities. Stable tra...

Abstract: Carnosic acid (CA), a rosemary phenolic compound, has been shown to display anti-cancer activity. We examined the apoptotic effect of CA in human neuroblastoma IMR-32 cells and elucidated the role of the reactive oxygen species (ROS) and mitogen-activated protein kinase (MAPK) associated with carcinogenesis. The result indicated that CA decreased the cell viability in a dose-dependent manner. Further investigation in IMR-32 cells revealed that cell apoptosis following CA treatment is the mechanism as confirmed by flow cytometry, hoechst 33258, and caspase-3/-9 and poly(ADP-ribose) polymerase (PARP) activation. Immunoblotting suggested a down-regulation of anti-apoptotic Bcl-2 protein in the CA-treated cells. In flow cytometric analysis, CA caused the generation of reactive oxygen species (ROS); however, pretreatment with the antioxidant N-acetylcysteine (NAC) attenuated the CA-induced generation of ROS and apoptosis. This effect was accompanied by increased activation of p38 and by decreased activation of extracellular signal-regulated kinase (ERK) as well as activation of c-Jun NH2-terminal kinase (JNK). Moreover, NAC attenuated the CA-induced phosphorylation of p38. Silencing of p38 by siRNA gene knockdown reduced the CA-induced activation of caspase-3. In conclusion, ROS-mediated p38 MAPK activation plays a critical role in CA-induced apoptosis in IMR-32 cells.

Abstract: Supercritical fluid antisolvent fractionation was used to obtain antioxidant compounds, mainly carnosic acid (CA), from high viscous oleoresins derived from dried rosemary leaves ( Rosmarinus officinalis ) extracted with ethanol. Due to the high viscosity of the starting material, which may hinder the mass transfer between the phases, a special nozzle was designed to blend the SCCO 2 stream with the high viscous oleoresin. Experiments were conducted at 50 °C and six different pressures in the first separation vessel, ranging from 150 to 400 bar; the best separation was achieved at 300 bar. As a result of the oleoresin two-stage fractionation, the starting material was separated in two fractions. The first one was an insoluble dark green powder, with low concentration of CA ( 2 , with a high concentration of CA (33 g/100 g extract at 300 bar). The antioxidant effect of this extract was higher to that of BHT when added to soybean oil. Industrial relevance The present study adds a possibility for the purification of carnosic acid from rosemary by using SCCO 2 antisolvent fractionation. Since the starting material employed (oleoresin) is a fluid phase, the process might be carried out in a continuous or semi-continuous way instead of discontinuous as it should be done if the starting material were a solid (leaves). This feature makes the procedure very promising toward its application at the industrial scale.

Abstract: The concentrations of carnosic acid, carnosol and rosmarinic acid in different materials from differentiated (multiple shoot cultures and regenerated plants) and undifferentiated (callus and cell suspension) in vitro cultures of Salvia officinalis were determined by HPLC. The results suggested that diterpenoid (carnosic acid and carnosol) production is closely related to shoot differentiation. The highest diterpenoid yield (11.4 mg g -1 for carnosic acid and 1.1 mg g -1 for carnosol) was achieved in shoots of 10-week-old micropropagated plants. The levels were comparable to those found in shoots of naturally growing plants. Undifferentiated callus and cell suspension cultures produced only very low amounts of carnosol (ca. 0.05 mg g -1 of dry weight). In contrast, content of rosmarinic acid in callus and suspension cultures as well as shoots growing in vitro and in vivo was similar and ranged between 11.2 and 18.6 mg g -1 of dry weight.

Abstract: The effects of carnosic acid (CA) of different concentrations (0.05, 0.1, and 0.2 mg/g) and two common antioxidants (butylated hydroxytoluene and α-tocopherol) on oxidative stability in pine nut oil at different accelerated conditions (heating and ultraviolet radiation) were compared. The investigation focused on the increase in peroxide and conjugated diene values, as well as free fatty acid and thiobarbituric acid-reactive substances. The changes in trans fatty acid and aldehyde compound contents were investigated by Fourier transform infrared spectroscopy, while the changes in pinolenic acid content were monitored by gas chromatography–mass spectrometry. The results show that CA was more effective in restraining pine nut oil oxidation under heating, UV-A and UV-B radiation, in which a dose–response relationship was observed. The antioxidant activity of CA was stronger than that of α-tocopherol and butylated hydroxytoluene. Pine nut oil supplemented with 0.2 mg/g CA exhibited favorable antioxidant effec...

Abstract: This report describes the effect of triacontanol on shoot multiplication and production of antioxidant compounds (carnosic acid, carnosol and rosmarinic acid) in S. officinalis cultures grown on MS basal medium (agar solidified medium supplemented with 0.1 mg l -1 IAA, 0.45 mg l -1 BAP). It was found that shoot proliferation significantly increased when triacontanol at concentrations of 5, 10 or 20 µg l -1 was added to the medium. HPLC analysis of acetone and methanolic extracts of sage shoots showed that the production of diterpenoids, carnosic acid/carnosol ratio, as well as, contents of rosmarinic acid were also affected by the treatment with triacontanol. The highest stimulation effect of triacontanol was observed on the production of carnosol, where the treatment with 20 µg l l -1 increased the content of this diterpenoid 4.5-fold compared to that in the control (sage shoots growing on MS basal medium, only).

Abstract: Natural polyphenols found in the leaves of rosemary (Rosmarinus officinalis L.) have potential therapeutic benefits, because of their potent antioxidant activity and their anticancerogenic and antiviral properties, observed in vitro and in human liver (Aruoma et al., 1996; Offord et al., 1997). Main active components in rosemary extract are carnosol, carnosic acid, rosmarinic acid and rosmanol (Pearson et al., 1997)...

Abstract: The yield of phenolic acids; rosmarinic acid (RA), caffeic acid (CAF) and phenolic diterpenes; carnosic acid (CA), carnosol (CAR), rosmanol (ROL) from rosemary (Rosmarinus officinalis L.) was investigated. Aqueous extraction was conducted for leaf and callus tissues, the chemical analysis for extracts was carried out using high performance liquid chromatography (HPLC). Data showed that RA and CAF production was 4.5, 2.7 µg/ml respectively. CA and CAR production reached 3.3, 2.8 µg/ml when the fungal elicitor; Fusarium oxysporum was added at the concentration 2.0 mg/l (2×10 4 CFU/ml), except ROL which was found at high levels (4.3, 4.2 and 4.6 µg/ml) only from leaf extracts, untreated callus and when callus treated with 0.4 mg/l of CaCl2 respectively.

Abstract: The invention discloses a production technology employing a ultrasonic countercurrent extraction process and a membrane separation (molecular sieve) process to acquire two antioxidants from Rosmarinus officinalis L. The production technology comprises adding 30 to 85% of edible alcohol into fresh rosemary branches and leaves, carrying out a ultrasonic counter current extraction process for a mixture of the edible alcohol and the fresh rosemary branches and leaves to obtain an extracted liquid with a solid-to-liquid ratio of from 1:10 to 1:30, and carrying out a membrane separation (molecular sieve) process to separate two antioxidant agents consisting of a rosemary fat-soluble antioxidant agent (comprising carnosic acid as a representative) and a rosemary water-soluble antioxidant agent (comprising rosmarinic acid as a representative) from the extracted liquid, wherein a heating time of the extracted liquid can be reduced through the membrane separation (molecular sieve) process thus a conversion ratio of rosmarinic acid to carnosol is reduced. A content of carnosic acid in the rosemary fat-soluble antioxidant agent is increased through a CO2 supercritical extraction process. The rosemary water-soluble antioxidant agent (comprising rosmarinic acid as a representative) is separated from a liquid obtained through a concentration process under reduced pressure and a solid-liquid separation process, and a content of rosmarinic acid in the rosemary water-soluble antioxidant agent is increased through recrystallization from ethanol. The production technology has the advantages of simple processes, low energy consumption, high production efficiency, production safety, and no harmful residual solvent due to single edible alcohol usage during all production processes.

Abstract: Liver: Carnosic acid could be a new treatment option for patients with NAFLD or the metabolic syndrome

Abstract: Aim of the present study was to focus the protective effect of carnosic acid on 7,12-dimethylbenz(a)anthracene (DMBA) induced cell surface glycoconjugates (Protein bound hexose, Hexosamine, Lipid bound sialic acid, Total sialic acid and Fucose) abnormalities in the plasma and buccal mucosa of golden Syrian hamsters. Oral squamous cell carcinoma was developed in the buccal pouch of hamsters by painting with 0.5% DMBA three times a week for 14 weeks. Glycoconjugates status was assessed both histologically and biochemically in the buccal mucosa of DMBA treated hamsters. The levels of glycoconjugates were increased in both plasma and buccal mucosa of DMBA treated hamsters. Oral administration of carnosic acid at a dose of 10mg/kg bw brought back the status of glycoconjugates to near normal concentrations in DMBA treated hamsters. Present results thus suggest that carnosic acid protected DMBA induced cell surface abnormalities during DMBA induced hamster buccal pouch carcinogenesis.

Abstract: Carnosic acid C20H28O4, the major compound in Rosmarinus officinalis L. (Lamiaceae) leaves was isolated and purified through silica gel column chromatography and detected on TLC plates in comparison with standard carnosic acid that served as positive control. Moreover, FTIR spectrometry and HPLC analysis were used to confirm the purity and identity of carnosic acid. The separated acid was investigated for its antimicrobial activity against six selected pathogenic microorganisms: Staphylococcus aureus, Bacillus cereus, Escherichia coli, Salmonella typhimurium, Pseudomonas aeruginosa and the yeast Candida albicans. Carnosic acid at different concentrations (5, 2.5, 1.25 and 0.62 mg/ml) was active against all tested microorganisms. High inhibitory effects were observed against Bacillus cereus and Staphylococcus aureus (zone of inhibition: 21.7 and 20.3 mm respectively) using the disc diffusion method. The minimal inhibitory concentration (MIC) of carnosic acid was determined using a broth microdilution method in 96-well microtiter plates. MIC values ranged from 15.6 to 125.0 μg/ml, and the highest value was recorded against Bacillus cereus and Staphylococcus aureus (MIC 15.6 μg/ml), followed by Escherichia coli, Salmonella typhimurium and Pseudomonas aeruginosa respectively. Moreover, carnosic acid observed good antifungal activity against the yeast Candida albicans (zone of inhibition range: 11.2-17.4mm; MIC: 125.0 μg/ml).

Abstract: The invention relates to application of carnosic acid in preparing a medicament for suppressing angiogenesis. The invention has the advantages that: the novel application of the carnosic acid discovers the effect of the carnosic acid in resisting angiogenesis for the first time, and the carnosic acid can effectively suppress proliferation, migration and canaliculization of vascular endothelial cells for the first time, which proves that the carnosic acid has the activity of resisting the angiogenesis, can be used as an angiogenesis suppressant, and can be applied to preparation of angiogenesis medicaments as the angiogenesis suppressant to treat neovascularization dependent and neovascularization related diseases, such as tumors, arthritis, psoriasis, ocular diseases, atherosclerosis, and the like.

Abstract: PURPOSE: An antibacterial composition containing carnosic acid is provided to ensure excellent brain protection and antioxidation CONSTITUTION: An antibacterial composition contains 05-1 wt% of carnosic acid as an active ingredient The carnosic acid is isolated from the extract The antibacterial composition is manufactured in the form of antibacaterial agent, feed additive, cosmetic additive, or antiseptic The carnosic acid is prepared from the rosemary extract with water or organic solvent

Abstract: The invention discloses a method for preparing a carnosic acid-rich liquid extract, wherein the extract can be extracted from an acidic ethanol solution; and an active substance is transferred into a vegetable oil phase, then subjected to a series of high-speed refrigerated centrifuge and then dried, thus preparing the extract. The extract prepared by the method is rich in the main anti-oxidationactive substance of rosemary, and can be added directly to various oil and fat type foods when being utilized as a food antioxidant. The extract extracted by the method provided by the invention has the characteristics of good oil solubility, light color, good storage stability and the like, and can be applied to various oil and fat products without additionally adding an emulsion stabilizer.

Abstract: Objective: Alzheimer’s disease is the most common type of neurodegenerative disorder. It has been suggested that oxidative stress can be one of the pathological mechanisms of this disease. Carnosic acid (CA) is an effective antioxidant substance and recent studies have shown that its electrophilic compounds play a role in reversing oxidative stress. Thus we tried to find out whether CA administration protects hippocampal neurons, preventing neurodegeneration in rats. Materials and Methods: Animals were divided into four groups: Sham-operated (sham), CA-pretreated sham-operated (sham+CA), untreated lesion (lesion) and CA-pretreated lesion (lesion+CA). Animals in all groups received vehicle or vehicle plus CA (CA: 10mg/ kg) intra-peritoneally one hour before surgery, again the same solution injected 3-4 hours after surgery (CA: 3 mg/kg) and repeated each afternoon for 12 days. A lesion was made by bilateral intra-hippocampal injection of 4 μl of beta amyloid protein (1.5 nmol/μl) or vehicle in each side. 14 days after surgery, the brains were extracted for histochemical studies. Data was expressed as mean ± SEM and analyzed using SPSS statistical software. Results: Results showed that pretreatment with carnosic acid can reduce cellular death in the cornu ammonis 1 (CA1) region of the hippocampus in the lesion+CA group, as compared with the lesion group. Conclusion: Carnosic acid may be useful in protecting against beta amyloid-induced neurodegeneration in the hippocampus.

Abstract: Tubular cellulose-based food casing with a fungicidal coating or impregnation comprising carnosol, carnosic acid, rosemary, rosemary extract, sage oil, or sage extract as fungicidal active ingredient, is claimed. Independent claims are also included for: (1) a sausage comprising the food casing and a sausage filling enclosed by the food casing; (2) protecting the tubular cellulose-based food casing from mildew comprising providing fungicidal coating or impregnation comprising the fungicidal active ingredient; and (3) use of carnosol, carnosic acid, rosemary, rosemary extract, sage oil, or sage extract for fungicidal treatment of a food casing or sausage.

Abstract: A study to track and detect the variation of bioactive components in rosemary extraction solution and extracts, by adding organic acid synergists, such as citric acid, oxalic acid and tartaric acid, was performed using high performance liquid chromatography (HPLC). Ultrasound was also used to increase the extraction efficiency, improve the yields of rosemary extracts and shorten the extraction time. The experimental results showed that the content of the key bioactive component carnosic acid was easier to reduce during the storage of extraction solution and extracts, meanwhile, adding citric acid, oxalic acid and tartaric acid during the extraction could actually slow down the trend of the reduction of the bioactive components, especially for carnosic acid.

Abstract: The variation regulation of five bioactive components including carnosic acid (CA), carnosol (CO), rosmarinic acid (RA), oleanolic acid (OA) and ursolic acid (UA) in rosemary leaves and extraction procedure was studied. The effects of rosemary sources, steam distillation time, activated carbon dosage and storage time of extraction solution on variation regulation of bioactive components were discussed in detail.