Carnosic Acid Content Research Articles

Relative molar sensitivities of carnosol and carnosic acid with respect to diphenylamine allow accurate quantification of antioxidants in rosemary extract

ABSTRACTWe have been developing a high-performance liquid chromatography/photodiode array (HPLC/PDA) employing relative molar sensitivities (RMSs) and adopted it to the accurate quantification of carnosol (CL) and carnosic acid (CA) which are the antioxidants in rosemary extract. The method requires no references of CL or CA and instead uses RMSs with respect to diphenylamine (DPA) whose certified reference material is available from a reagent manufacturer. The molar and response ratios of the analytes to the reference in an artificial mixture of them were determined using 1H-quantitative nuclear magnetic resonance spectroscopy (1H-qNMR) and HPLC/PDA at a wavelength of 284 nm under isocratic condition, respectively, and then RMSs were calculated to be 0.111 for CL/DPA and 0.0809 for CA/DPA as averaged values in three HPLC–PDA instruments. The RMS values varied by up to 1.1% as relative standard deviation. To evaluate the performance of HPLC/PDA with the RMSs, the CL and CA contents in rosemary extracts were determined using DPA as a reference. The CL and CA contents were compared with those determined using calibration curves of CL and CA obtained by HPLC measurement of standard solutions prepared from their reagents whose absolute purities were determined using 1H-qNMR. The differences between the two methods for CL and CA were ≤3% as relative error. This chromatographic method with RMSs allows a simple and reliable quantification when reference of the analyte is unavailable.

Extraction of Carnosic Acid and Carnosol from Sage (Salvia officinalis L.) Leaves by Supercritical Fluid Extraction and Their Antioxidant and Antibacterial Activity.

Sage (Salvia officinalis L.) is a good source of antioxidant compounds, carnosic acid and carnosol being the prominent ones. Both are soluble in CO2, and our goal was to investigate the application of supercritical CO2 extraction to obtain sage extracts rich in these compounds. The effect of pressure, temperature, and CO2 flow rate on the carnosic acid and carnosol yield was studied. These variables were optimized by response surface methodology (RSM). The pressure significantly affected carnosol extraction, while the extraction of carnosic acid was affected by the pressure, temperature, and CO2 flow rate. Carnosic acid content varied from 0.29–120.0 µg mg−1, and carnosol content from 0.46–65.5 µg mg−1. The optimal conditions according to RSM were a pressure of 29.5 MPa, a temperature of 49.1 °C, and a CO2 flow rate of 3 kg h−1, and the sage extract yield was calculated to be 6.54%, carnosic acid content 105 µg mg−1, and carnosol content 56.3 µg mg−1. The antioxidant activities of the sage extracts were evaluated by the scavenging activities of 2,2-diphenyl-1-picrylhydrazyl (DPPH). Sage extract obtained at 30 MPa and 40 °C with 2 kg h−1 CO2 flow rate with a carnosic acid content of 72 µg mg−1 and carnosol content of 55 µg mg−1 exhibited the highest antioxidant activity (80.0 ± 0.68%) amongst the investigated supercritical fluid extracts at 25 µg mL−1 concentration. The antimicrobial properties of extracts were tested on four bacterial strains: Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, and Staphylococcus aureus. The extract with a carnosic acid content of 116 µg mg−1 and a carnosol content of 60.6 µg mg−1 was found to be the most potent agent against B. subtilis.

Bioaccessibility and inhibitory effects on digestive enzymes of carnosic acid in sage and rosemary

In this study, the aim was to determine the bioaccessibilities of carnosic acid in sage and rosemary and in vitro inhibitory effects of these samples on lipid and starch digestive enzymes by evaluating the lipase, α-amylase and α-glucosidase enzyme inhibition activities. The content of carnosic acid in rosemary (18.72 ± 0.33 mg/g) was found to be higher than that content of that in sage (3.76 ± 0.13 mg/g) (p < 0.05). The carnosic acid bioaccessibilities were found as 45.10 ± 1.88% and 38.32 ± 0.21% in sage and rosemary, respectively. The tested sage and rosemary showed inhibitory activity against α-glucosidase (Concentration of inhibitor required to produce a 50% inhibition of the initial rate of reaction - IC50 88.49 ± 2.35, 76.80 ± 1.68 μg/mL, respectively), α-amylase (IC50 107.65 ± 12.64, 95.65 ± 2.73 μg/mL, respectively) and lipase (IC50 6.20 ± 0.63, 4.31 ± 0.62 μg/mL, respectively). Furthermore, to the best of our knowledge, this is the first work that carnosic acid standard equivalent inhibition capacities (CAEIC50) for these food samples were determined and these values were in agreement with the IC50 values. These results show that sage and rosemary are potent inhibitors of lipase, α-amylase and α-glucosidase digestive enzymes.

Characterization of rosemary and thyme extracts for incorporation into a whey protein based film

Active packaging is designed to interact with foods by releasing active components with biological properties, allowing the extension of foods shelf-life, being aromatic plants a source of active compounds. The main goal of this research was to develop a biodegradable whey protein based film, incorporated with an aromatic plant extract. Ethanolic extracts from rosemary and thyme were obtained from three commercial brands. The DPPH• scavenging activity assay showed that the rosemary extracts presented the highest antioxidant capacity. Thyme acetone extract had the highest antioxidant activity in the β-carotene bleaching assay. Three different mixtures of rosemary and thyme extracts with the highest antioxidant capacity showed a lower antioxidant capacity than the single extracts. The content of carnosic acid, carnosol and rosmarinic acid was determined in the three rosemary extracts, having the extract from the brand C shown the highest phenolics content. The extract showed antimicrobial activity against Staphylococcus aureus, Listeria monocytogenes and Clostridium perfringens. For the active film, 1% of the rosemary extract was incorporated into the whey protein film. The film presented antimicrobial activity against L. monocytogenes and S. aureus.

Optimization of antioxidant efficacy of a deflavored and decolorized rosemary extract: effect of carnosol content on the oxidative stability of paprika colored beef patties.

Considering the significance of natural antioxidants to preserve meat, the present study was undertaken to evaluate the efficacy of a deflavored and decolorised extract of rosemary (StabilRose™) for the production and preservation of naturally colored fresh meat. Oxidative rancidity of meat and color degradation of paprika oleoresin were exploited as model systems and compared with butylated hydroxyanisole (BHA). The results showed similar efficacy for 3% carnosic acid extract and BHA, with further enhancement in efficacy with respect to the carnosic acid content. A synergetic antioxidant effect of carnosol on carnosic acid content was also noticed to an extent of 1:1 (w/w) ratio, and further increase in carnosol content showed no improvement in the antioxidant efficacy. Finally, stabilized paprika and optimized rosemary extract containing carnosic acid and carnosol in 1:1 (w/w) ratio was successfully applied to produce naturally colored meat suitable for storage at 4±1°C.

Carnosic acid as a component of rosemary extract stimulates skeletal muscle cell glucose uptake via AMPK activation.

Compounds that increase the activity of the energy sensor AMP-activated kinase (AMPK) have the potential to regulate blood glucose levels. Although rosemary extract (RE) has been reported to activate AMPK and reduce blood glucose levels in vivo, the chemical components responsible for these effects are not known. In the present study, we measured the levels of the polyphenol carnosic acid (CA) in RE and examined the effects and the mechanism of action of CA on glucose transport system in muscle cells. High performance liquid chromatography (HPLC) was used to measure the levels of CA in RE. Parental and GLUT4myc or GLUT1myc overexpressing L6 rat myotubes were used. Glucose uptake was assessed using [3 H]-2-deoxy-d-glucose. Total and phosphorylated levels of Akt and AMPK were measured by immunoblotting. Plasma membrane GLUT4myc and GLUT1myc levels were examined using a GLUT translocation assay. Statistics included analysis of variance (ANOVA) followed by Tukey's post-hoc test. At concentrations found in rosemary extract, CA stimulated glucose uptake in L6 myotubes. At 2.0μmol/L CA a response (226±9.62% of control, P=.001), similar to maximum insulin (201±7.86% of control, P=.001) and metformin (213±10.74% of control, P=.001) was seen. Akt phosphorylation was not affected by CA while AMPK and ACC phosphorylation was increased and the CA-stimulated glucose uptake was significantly reduced by the AMPK inhibitor compound C. Plasma membrane GLUT4 or GLUT1 glucose transporter levels were not affected by CA. Our study shows increased muscle cell glucose uptake and AMPK activation by low CA concentrations, found in rosemary extract, indicating that CA may be responsible for the antihyperglycemic properties of rosemary extract seen in vivo.

Carnosic acid acts synergistically with gentamicin in killing methicillin-resistant Staphylococcus aureus clinical isolates

BackgroundMethicillin-resistant Staphylococcus aureus (MRSA) is resistant to different commonly used antibiotics, stressing the need for further strategies to treat this human pathogen with worldwide prevalence. The use of phytochemicals within the current pharmacology is a promising approach to enhance the antimicrobial activity of common antibiotics in the battle against these bacteria. PurposeThe purpose of this study was to determine the antimicrobial effectiveness of carnosic acid, the major constituent of Rosmarinus officinalis L. leaves, in combination with gentamicin against multi-drug resistant MRSA clinical isolates obtained from pediatric patients with bacteremia. Materials and methodsAnti-MRSA activity was studied using the broth microdilution assay and time–kill method. Combinations of subinhibitory concentrations of carnosic acid and gentamicin were examined using the checkerboard method. ResultsCarnosic acid exhibited a good antibacterial activity against all multidrug-resistant MRSA clinical isolates tested, which are resistant to four up to nine antibiotics. In addition, the combination of carnosic acid with gentamicin not only decreased the minimal inhibitory concentration (MIC) of both by 4- to 5-fold, but also improved the bactericidal potency of the common antibiotic by 32- to 40-fold against both gentamicin-susceptible and gentamicin-resistant MRSA clinical isolates. A clear bactericidal synergistic activity between carnosic acid and gentamicin in killing multidrug-resistant MRSA clinical isolates with a fractional bactericidal concentration index (FBCI) of 0.28–0.35 was demonstrated. ConclusionsOur findings show the potential use of carnosic acid in combination with gentamicin as a promising alternative for the control of healthcare-associated infections caused by multidrug-resistant MRSA.

Selective recovery of rosmarinic and carnosic acids from rosemary leaves under ultrasound- and microwave-assisted extraction procedures

Ultrasound-assisted extraction (UAE) and microwave-assisted extraction (MAE) are currently amongst the foremost green techniques for accelerating extraction processes. Several methods for the efficient recovery of the phenolic compounds from rosemary leaves have so far been proposed, but little data are available on the use of UAE and MAE. The aim of this work is to investigate the efficiency and selectivity of these techniques in recovering fractions of specific phenolic compounds from dried rosemary leaves using solvents that are suitable for food applications. UAE has been carried out by means of a probe system equipped with a titanium horn working at 19.5 kHz (140 W). MAE has been performed in a closed multimode reactor under N 2 (20 bar) at 100 °C. All obtained extracts were dissolved in a defined solvent volume and the solutions were directly analyzed using a combination of the HPLC-DAD-MS and TOF techniques. MAE and UAE in ethanol and acetone dramatically increased phenol yield (more than three times) as compared to more traditional solid–liquid extraction processes. In terms of selectivity, remarkably high rosmarinic acid content (6.8% of the dried extract) was obtained in ethanol under ultrasound (US). Even more impressive is the selectivity of UAE in n -hexane which gave the highest carnosic acid content, up to 13% of the dried extract. In conclusion, non-conventional energy sources and, in particular, high-intensity US have proven themselves to be rapid, efficient, and selective techniques for rosemary leaf extraction and provide fractions with high rosmarinic and carnosic acid contents.

Antioxidant activities from different rosemary clonal lines

Rosemary extract is widely used in food industry and carnosic acid is reported to be the major component that is responsible for its antioxidant activities. However, it is unclear how the numerous plant metabolites interact and contribute to the overall antioxidant activity. In this study, with poultry fat as the model food system, rosemary extract from six clonal lines were evaluated that each represented a different genetic variant. As expected, rosemary extract with higher carnosic acid content had higher antioxidant activity. However, rosemary extract which had carnosic acid removed retained a significant amount of activity. Furthermore, when the individual contributions of carnosic acid and the portion without carnosic acid were evaluated separately, neither was shown to be responsible for the overall level of its stabilization effect from rosemary extract as a whole entity. The interactions among different plant metabolites have a major impact on the overall antioxidant capabilities of rosemary extract.

Callicarpa longissima extract, carnosol-rich, potently inhibits melanogenesis in B16F10 melanoma cells.

Cosmetic industries focus on developing materials and resources that regulate skin pigmentation. Melanin, the major pigment in human skin, protects the skin against damage from ultraviolet light. An ethanolic extract of the leaves of Callicarpa longissima inhibits melanin production in B16F10 mouse melanoma cells by suppressing microphthalmia-associated transcription factor (MITF) gene expression. Following purification and analysis using liquid chromatography-mass spectrometry (LC-MS), NMR, and biochemical assays, carnosol was determined to be responsible for the major inhibitory effect of the C. longissima extract on melanin production. Carnosol is an oxidative product of carnosic acid, whose presence in the extract was also confirmed by an authentic reference. The carnosol and carnosic acid content in the extract was approximately 16% (w/w). These results suggest that C. longissima is a novel, useful, and attractive source of skin-whitening agents.

Improved Bioavailability of Supercritical Rosemary Extract Through Encapsulation in Different Delivery Systems After In Vitro Digestion

Rosmarinus officinalis L. is an aromatic plant with components known to confer health-promoting properties. The present study aimed to enhance the bioavailability of the main compounds, carnosic acid and carnosol, by encapsulation. The rosemary extracts were obtained by supercritical fluid extraction, and then different delivery systems were tested, namely oil emulsion droplets and casein micelles, native protein particles present in milk. Soybean oil in water emulsions and casein dispersions were digested in vitro with simulated gastric and intestinal fluids, and the bioavailability of the extract was tested on Caco-2 cell monolayers. The apical and basolateral compartments and the cell lysates were further analyzed for carnosic acid and carnosol content by means of ultra-performance liquid chromatography-tandem mass spectrometry (UPLC/MS/MS). The encapsulation in both casein micelles and emulsion droplets significantly protected the rosemary extract from degradation during digestion, resulting in 81 and 97 % recovery of carnosic acid, respectively. This high protection rate prompted an increased bioavailability of carnosic acid and carnosol in Caco-2 monolayers, with up to 10 % carnosic acid recovered in the bioavailable fraction. Encapsulation in delivery systems like casein micelles or emulsion droplets opens new possibilities for successful incorporation of rosemary extracts in healthy food formulations with increased functionality.

Carnosic acid as a major bioactive component in rosemary extract ameliorates high-fat-diet-induced obesity and metabolic syndrome in mice.

In this study, we investigated the preventive effects of carnosic acid (CA) as a major bioactive component in rosemary extract (RE) on high-fat-diet-induced obesity and metabolic syndrome in mice. The mice were given a low-fat diet, a high-fat diet or a high-fat diet supplemented with either 0.14% or 0.28% (w/w) CA-enriched RE (containing 80% CA, RE#1L and RE#1H), or 0.5% (w/w) RE (containing 45% CA, RE#2), for a period of 16 weeks. There was the same CA content in the RE#1H and RE#2 diets and half of this amount in the RE#1L diet. The dietary RE supplementation significantly reduced body weight gain, percent of fat, plasma ALT, AST, glucose, insulin levels, liver weight, liver triglyceride, and free fatty acid levels in comparison with the mice fed with a HF diet without RE treatment. RE administration also decreased the levels of plasma and liver malondialdehyde, advanced glycation end products (AGEs), and the liver expression of receptor for AGE (RAGE) in comparison with those for mice of the HF group. Histological analyses of liver samples showed decreased lipid accumulation in hepatocytes in mice administrated with RE in comparison with that of HF-diet-fed mice. Meanwhile, RE administration enhanced fecal lipid excretion to inhibit lipid absorption and increased the liver GSH/GSSG ratio to perform antioxidant activity compared with HF group. Our results demonstrate that rosemary is a promising dietary agent to reduce the risk of obesity and metabolic syndrome.

Two-step sequential supercritical fluid extracts from rosemary with enhanced anti-proliferative activity

Previously, carnosic acid and carnosol have demonstrated anti-proliferative activity against different types of cancer. To obtain extracts enriched in these two key phenolic compounds, two different processes have been developed in the present work based on the use of two-step sequential supercritical fluid extraction (SFE). By removing the interfering, less active fractions in a first step (150 or 300 bar, 40°C, neat CO2, 60 min), suitable enrichment is achieved in the second step (150 bar, 40°C, CO2 + 7% ethanol, 120 min), and this leads to carnosic acid concentrations in the extract as high as 40% of total dry weight, which are among the highest concentrations that have been described with this type of process. The enriched extracts were tested against the HT-29 human adenocarcinoma cell line, showing enhancement of their antiproliferative activity by approximately 3-fold compared to previously reported SFE rosemary extracts and higher inhibitory effects at lower concentrations (30 µg mL−1 of extract). Thus, the proposed two-step SFE process effectively improves the carnosic acid and carnosol recovery in shorter processing times (180 min vs. 300 min). Moreover, the obtained extracts possess higher anti-proliferative activity and consume less solvent.

Variations of Carnosic acid and Carnosol Concentrations in Ethanol Extracts of Wild Lepechinia salviae in Spring (2008–2011)

Ethanol extracts from dried leaves of wild Lepechinia salvia (Lindl) Epling, collected during the flowering period (September-November), contained 15% to 25% carnosic acid and 2 to 8% carnosol, depending on the month of collection. The highest concentration of carnosic acid in extracts was in October, while carnosol concentration had a peak in September, which suggests that it is not a product of carnosic acid oxidation. A comparison of extracts obtained in September 2008 to 2011 shows that the production of both abietanes increased in years with less winter rainfall and higher temperatures, which induced an early blooming. EC50 values in DPPH radical scavenging and antiproliferative (CCRF-CEM tumor cells) bioassays confirm that the high bioactivity of the extracts of rosemary, sage and L. salviae does not arise only from carnosol and carnosic acid. The cytotoxic activity was significantly higher in extracts of L. salviae, probably due to water stress differences between the cultivars and the wild species. These results correlate well with the close phylogenetic relationship between the three species, and their similar medicinal uses.

The use of rosemary extracts in vegetable oil-based lubricants

The use of vegetable oils as basestocks represents an important alternative to mineral lubricants. However, vegetable oil oxidation is a serious disadvantage for its use as a lubricant. In this sense, the use of natural antioxidant additives, which were obtained by different extraction processes from the rosemary plant, has been investigated to prevent the oxidation process in two selected potential vegetable oil-based lubricants (high oleic sunflower (HOSO) and castor (CO) oils). A variety of rosemary extracts with a different distribution of phenolic diterpene compounds have been obtained depending on the solvent used (n-hexane, methanol or 50% (v/v) methanol–water). The antioxidant resistance of these oils containing rosemary extract has been analyzed under accelerated conditions using both the ASTM D942-02 and ASTM E2009-08 standards. The results showed that the rosemary extracts are better solubilized in CO than in HOSO, obtaining the highest oxidation onset temperatures in CO. In addition to this, the rosemary extracts obtained with polar solvents had the highest concentration of rosmarinic acid, and showed a better antioxidant activity in CO; whereas the extract obtained with a non polar solvent had the highest concentration of carnosic acid, showing a better antioxidant activity in HOSO than the rest of rosemary extracts studied.

Anti‐inflammatory activity of rosemary extracts obtained by supercritical carbon dioxide enriched in carnosic acid and carnosol

SummaryThe in vitro anti‐inflammatory activity of supercritical rosemary (Rosmarinus officinalis L.) extracts (rosemary A and B) is been reported in this study. To achieve that, THP‐1 macrophages were activated using lipopolysaccharide or human ox‐LDL and secretion and gene expression of TNF‐α, IL‐1β, IL‐6 and IL‐10 were evaluated, as well as COX‐2 gene expression. Results indicated that both rosemary extracts (A &amp; B) exhibit high anti‐inflammatory activity although at a higher extent in case of rosemary B extract (5 μg mL−1), representing a higher quantity of carnosic acid and carnosol than rosemary A. When comparing the activity of the extract to the standard itself, the anti‐inflammatory activity of standards of carnosic acid and carnosol was not as intense as that obtained with rosemary B. These data indicated that although carnosic acid content in the extracts is considered as the main anti‐inflammatory compound, a synergistic interaction with other compounds may play a significant role in enhancing its activity. Results provided the grounds for possible increase in the application of supercritical rosemary extracts in food formulations for mitigation or prevention of inflammatory diseases.

Effect of different format-solvent rosemary extracts (Rosmarinus officinalis) on frozen chicken nuggets quality

Three kinds of Rosmarinus officinalis extract (powder-acetone, liquid-methanol, liquid-acetone) were used to examine the effects of format-solvent on the active compounds extracted (total phenolic, carnosol and carnosic acid content) and antioxidant activity (FRAP, ABTS). The results showed that both, as the format but also the solvent used, had significant effect on the parameters analyzed (p<0.05). The highest antioxidant activity was found for the powder-acetone extract followed by the liquid methanol and liquid acetone extracts (p<0.05). The effect of the three different extracts on the physical–chemical and sensory quality of frozen chicken nuggets was evaluated. At the dose proposed by the European Union Directive 2010/69/EU for the carnosic and carnosol compounds [150ppm (mg/kg fat basic)], the format-solvent combination of the rosemary extracts used did not modify the chicken nuggets quality characteristics (pH, colour, sensory quality) and still underlines the effectiveness of these extracts.

NMR-based quantification of rosmarinic and carnosic acids, GC–MS profile and bioactivity relevant to neurodegenerative disorders of Rosmarinus officinalis L. extracts

A comparative study of phytochemicals content and biological properties of eight Rosmarinus officinalis (rosemary) populations (RO1–RO8) collected in different areas of Tunisia was carried out. Two of the main rosemary constituents, rosmarinic and carnosic acids, were quantified by an NMR technique. Carnosic acid content was higher than that of rosmarinic acid. The non-polar constituents were examined by GC and GC–MS. Total phenols and flavonoids content were also determined in order to discuss the possible correlation between these phytochemicals and bioactivity. Antioxidant activity was investigated through different in vitro assays. Sample RO3 from a sub-humid area showed the highest potency in 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay (IC50 of 6.74μg/mL) while RO5 from arid area exhibited the highest radical 2,2′-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) scavenging activity (IC50 of 5.92μg/mL). The most active sample in Ferric Reducing Antioxidant Power (FRAP) assay was RO4 (62.21μM Fe(II)/g), followed by RO1–RO3 and RO8 harvested in sub-humid areas with values in the range 49.61–58.72μM Fe(II)/g. Extracts were able to inhibit acethylcholinesterase (AChE) and butyrylcholinesterase (BChE). Sample RO5 exhibited a promising AChE inhibitory activity (IC50 of 4.47μg/mL). Thus, we suggest rosemary extracts as a functional food ingredient for preventing AD.

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 (50wt.% 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 30MPa). 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.

Isolation of carsonic acid from rosemary extracts using semi-preparative supercritical fluid chromatography

Supercritical fluid chromatography (SFC) in analytical and semi-preparative scale was studied to separate rosemary (Rosmarinus officinalis) supercritical extracts to produce fractions with high antioxidant activity. Carnosic acid, the major antioxidant present in rosemary supercritical extracts, was the target compound to be isolated in the fractions produced. First, rosemary extracts were produced in a supercritical CO2 pilot-plant employing different extraction conditions, in order to obtain samples with diverse composition. The content of carnosic acid was determined and the DPPH test was accomplished in order to assess the antioxidant activity of the different samples produced. Then, using some of these samples, experiments were carried out in a semi-preparative SFC system testing the performance of four different chromatographic columns to isolate and recover a fraction with high concentration of carnosic acid. The four chromatographic columns employed were silica and 2-ethylpyridine (especially packed for SFC) and silica and diol columns (packed for HPLC). The SFC fractionation conditions explored were in the ranges 10–20MPa, 313–333K and 5–20% of cosolvent (ethanol). Using the first three columns, it was possible to isolate fractions with concentrations of carnosic acid greater than 80% mass.