Abstract: Nuclear factor-erythroid 2-related factor 2 (Nrf2) is a redox-sensitive transcription factor, which plays an important role in the cellular defense against oxidative stress by induction of anti-oxidant and cytoprotective enzymes. In the current study, we sought to investigate the osteoprotective effect of carnosic acid (CA), a phenolic (catecholic) diterpene. It is widely identified for its electrophilic nature under oxidative stress conditions and thus anticipated to counter osteoporosis by facilitation of Nrf2 signalling. Osteoclast differentiation was induced by incubation of RAW 264.7 (mouse macrophage) cells and mouse bone marrow macrophages (BMMs) in the presence of receptor activator of NF-κB ligand (RANKL) (100 ng/ml). After treatment, osteoclastogenesis was assessed using tartrate-resistant acid phosphatase (TRAP) assay. We observed that 6 h pretreatment with CA (1.25, 2.5, 5 μM) decreased RANKL-induced osteoclast formation and abolished RANKL-induced ROS generation by activating Nrf2 and its transcriptional targets. Further, CA also inhibited RANKL-induced activation of NF-κB and MAPK signalling. RANKL-induced mRNA expression of osteoclast related genes and transcription factors was also diminished by CA. In vivo osteolysis was developed in C57BL/6 male mice using lipopolysaccharide (LPS). Consistent with in vitro results, in vivo μ-CT analysis of femurs showed that bone mineral density (BMD), bone mineral content (BMC), and bone architecture parameters such as trabecular thickness (Tb.Th) and trabecular space (Tb.Sp) were positively modulated by CA in LPS-injected mice. The results obtained in this study support that CA inhibits RANKL-induced osteoclastogenesis by maintaining redox homeostasis through modulation of Nrf2 and NF-κB pathways.

Abstract: Background and Purpose The diterpenoids carnosol (CS) and carnosic acid (CA) from Salvia spp. exert prominent anti-inflammatory activities but their molecular mechanisms remained unclear. Here we investigated the effectiveness of CS and CA in inflammatory pain and the cellular interference with their putative molecular targets. Experimental Approach The effects of CS and CA in different models of inflammatory pain were investigated. The inhibition of key enzymes in eicosanoid biosynthesis, namely microsomal prostaglandin E2 synthase-1 (mPGES-1) and 5-lipoxygenase (5-LO) was confirmed by CS and CA, and we determined the consequence on the eicosanoid network in activated human primary monocytes and neutrophils. Molecular interactions and binding modes of CS and CA to target enzymes were analyzed by docking studies. Key Results CS and CA displayed significant and dose-dependent anti-inflammatory and anti-nociceptive effects in carrageenan-induced mouse hyperalgesia 4 h post injection of the stimuli, and also inhibited the analgesic response in the late phase of the formalin test. Moreover, both compounds potently inhibited cell-free mPGES-1 and 5-LO activity and preferentially suppressed the formation of mPGES-1 and 5-LO-derived products in cellular studies. Our in silico analysis for mPGES-1 and 5-LO supports that CS and CA are dual 5-LO/mPGES-1 inhibitors. Conclusion and Implications In summary, we propose that the combined inhibition of mPGES-1 and 5-LO by CS and CA essentially contributes to the bioactivity of these diterpenoids. Our findings pave the way for a rational use of Salvia spp., traditionally used as anti-inflammatory remedy, in the continuous expanding context of nutraceuticals. Linked Articles This article is part of a themed section on Principles of Pharmacological Research of Nutraceuticals. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v174.11/issuetoc

Abstract: Enhanced removal of abnormal protein aggregates or injured organelles through autophagy is related to neuroprotection in Parkinson's disease. In this study, we explored whether the induction of autophagy is associated with the neuroprotection of rosemary carnosic acid (CA) against 6-hydroxydopamine (6-OHDA)-induced neurotoxicity in SH-SY5Y cells. The results indicated that cells treated with CA had increased protein levels of parkin and autophagy-related markers, including phosphatidylinositol 3-kinase p100, Beclin1, autophagy-related gene 7, and microtubule-associated protein 1 light chain 3-II, as well as enhanced formation of autophagic vacuoles. Treatment of cells with 6-OHDA decreased the levels of parkin and the autophagy markers, but CA pretreatment reversed these effects. However, wortmannin (an autophagosome formation blocker) pretreatment attenuated the effect of CA. After CA pretreatment, the induction of cleaved caspase 3, cleaved poly-ADP ribose polymerase, and nuclear condensation by 6-OHDA were alleviated. Both wortmannin and bafilomycin A1 (an autophagosome-lysosome fusion blocker) inhibited the anti-apoptosis effects of CA. Additionally, we performed immunoprecipitation with anti-parkin antibody and found that the interaction of parkin and Beclin1 protein was reduced by 6-OHDA but that this effect was reversed in cells pretreated with CA. Moreover, transfection of parkin siRNA in cells inhibited the ability of CA to alleviate 6-OHDA-decreased autophagy-related markers and nuclear condensation. In conclusion, CA protects against 6-OHDA-induced apoptosis by inducing autophagy through the interaction of parkin and Beclin1. These results provide a future strategy for use of CA in the prevention of Parkinson's disease.

Abstract: Carnosic acid (CA; C20H28O4), which is also called salvin, is a major phenolic diterpene found in Rosmarinus officinalis L. and exhibits antioxidant, anti-inflammatory, and antiproliferative properties. CA activates the nuclear factor erythroid 2-related factor 2 (Nrf2) transcription factor, leading to the upregulation of antioxidant and phase II detoxification enzymes, such as heme oxygenase-1 (HO-1), glutathione reductase (GR), γ-glutamate-cysteine ligase (γ-GCL), and glutathione S-transferase (GST), among others. We have previously demonstrated that CA upregulates the total and mitochondrial synthesis of glutathione (GSH), causing mitochondrial protection against paraquat (PQ) and methylglyoxal (MG). Nonetheless, the complete mechanism by which CA prevented mitochondrial dysfunction was not clear yet. Here, we examine whether HO-1 would be involved in the CA-induced mechanism of mitochondrial protection in SH-SY5Y-treated cells. SH-SY5Y cells were pretreated with CA (1 μM) for 12 h prior to a challenge with PQ at 100 μM for additional 24 h. Zinc protoporphyrin IX (ZnPP IX; a specific inhibitor of HO-1; 10 μM) was utilized prior to exposure to CA in order to investigate whether HO-1 was involved in the cytoprotective effects elicited by CA. We found that the CA-induced Nrf2-dependent HO-1 upregulation ameliorated, at least in part, the mitochondrial function in PQ-treated cells. Therefore, CA protected mitochondria of SH-SY5Y cells and exerted anti-apoptotic effects by activating the Nrf2/HO-1 axis.

Abstract: Rosmarinus officinalis L., a medicinal herb from the labiates family, has been reported to have potential benefit in the treatment and prevention of several diseases. In particular its phenolics have demonstrated protective effects on various types of cancer through several mechanisms. The present study aimed to determine the effects of rosemary phenolic extracts on human cell functions, with particular regard to their anti-proliferative properties in three cell types U937, CaCo-2 and the peripheral blood mononuclear cells (PBMCs). The radical scavenging and Ferric reducing abilities of the extracts have been assessed as well as their cyto-toxicity and effects on cell cycle distribution and apoptosis. About 13 compounds were identified with dominance of rosmarinic acid in the methanolic extract and phenolic diterpens in the ethyl acetate fraction (Carnosol, Carnosic acid and methyl Carnosate). The total polyphenolic content was important in the first extract with 2.589 ± 0.005 g/100 g in gallic acid equivalent compared to 0.763 ± 0.005 g/100 g. The methanolic fraction displayed higher antioxidant activity (DPPHIC50: 0.510 mg/mL and FRAP: 1.714 ± 0.068 mmol Fe2+/g) while ethyl acetate showed pronounced antiproliferative effects (IC50: 14.85 ± 0.20µg/mL and 14.95 ± 2.32 µg/mL respectively for U937 and CaCo-2 cells). The anti-proliferative effect was associated with a cell cycle arrest in S phase for U937 (62% of the population at 5 µg/mL) with a concomitant decrease in G1 and G2/M phases. Tested extracts displayed in addition early apoptotic effects in U937 and late apoptosis in CaCo-2 cells. The obtained data indicate that the identified phenolics are at least partially responsible for the observed cytotoxicity.

Abstract: 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 [3H]-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 2.0 μM CA a response (226±9.62 % of control, p=0.001), similar to maximum insulin (201±7.86 % of control, p=0.001) and metformin (213±10.74 % of control, p=0.001) 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. This article is protected by copyright. All rights reserved.

Abstract: Whole blood consists of plasma (~54%) and various types of circulating cells (~46%) with a very low percentage of cells other than erythrocytes (Jones and Allison, 2007). The main ester-lipid components of blood are triacylglycerols, cholesterol esters and glycerophospholipids. The concentrations of essential elements (like Se, Zn, Cu, Co, Fe or Mg), fatty acids (FAs), vitamins or cholesterol in plasma and circulating blood cells as well as in other mammal tissues vary due to different physiological factors (i.e. dietary intake, intestinal absorption, metabolism and exchange among compartments) (Kincaid, 1999; Risé et al., 2007; Juniper et al., 2008; Niedźwiedzka et al., 2008; Czauderna et al., 2010). ABSTRACT. The concentration of macro and trace elements, fatty acids (FAs), vitamins, total cholesterol (TCh) in blood as well as in other tissues can be modulated by diet composition. Thus, the purpose of the present study was to evaluate the effects of fish oil (FO), carnosic acid (CA) and selenized-yeast (SeY) or selenate (SeVI) on concentration of FAs, TCh, α-tocopherol (αT) and selected elements in whole blood of lambs. Thirty male lambs were allocated into 5 groups of 6 animals each and fed for 35 days the following diets: control – basal diet (BD) with 3% rapeseed oil (RO), ROFO – BD with 2% RO and 1% FO, CA – BD with 2% RO, 1% FO and 0.1% CA, CASeY – BD with 2% RO, 1% FO, 0.1% CA and 0.35 mg Se as selenized-yeast (SeY) per kg of BD and CASeVI – BD with 2% RO, 1% FO, 0.1% CA and 0.35 mg Se as sodium selenate (SeVI) per kg of BD. In animals fed CASeVI diet the levels of saturated (SFAs), monoand polyunsaturated FAs, thrombogenic-SFAs and atherogenicSFAs decreased in comparison to the control group. On the other hand, in lambs fed CASeY diet the concentration of TCh in blood increased in comparison to lambs fed CA and CASeVI diets. Moreover, feeding CASeY diet also enhanced the concentration of αT in blood as compared to the animals fed ROFO and CASeVI diets. The lowest αT concentration in blood was noted in blood of lambs fed CASeVI diet. Feeding diets supplemented with SeY or SeVI increased the concentrations of Se and malondialdehyde in blood in comparison to other diets. So, the whole blood can be treated as the valuable non-invasive marker for evaluation of ruminant health status and nutritional quality of ruminant feeds. Received: 2 March 2017 Revised: 5 June 2017 Accepted: 8 September 2017

Abstract: Myocardial ischemia and reperfusion occurs in myocardial infarction. Timely reperfusion will exacerbate the injury through the mitochondria-mediated apoptosis in cardiomyocytes due to the accumulation of excessive reactive oxygen species (ROS). In order to identify novel therapeutic approaches, the cardioprotective effects of carnosic acid and the underlying mechanisms were investigated in the present study in H9c2 cardiomyocytes injured by hypoxia/reoxygenation in vitro. The viability of H9c2 cardiomyocytes was detected by MTT assay and further confirmed by the detection of intracellular lactate dehydrogenase (LDH) release. The mitochondrial function in H9c2 cardiomyocytes was evaluated using fluorescence methods. The proteins related to apoptosis, including caspase-3, B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (Bax) were analyzed by western blot analysis, and the activity of caspase-3 was determined using a colorimetric method. As a result, carnosic acid was demonstrated to improve the viability of H9c2 cardiomyocytes and suppress the leakage of cytosolic lactate dehydrogenase under hypoxia/reoxygenation. In addition, the overproduction of intracellular ROS and intracellular calcium overload were ameliorated in the presence of carnosic acid. The dysfunction of mitochondria in H9c2 cardiomyocytes was also attenuated by carnosic acid through blocking the collapse of mitochondrial membrane potential (MMP) and mitochondrial permeability transition pore (mPTP) opening. Furthermore, the apoptosis of H9c2 cardiomyocytes resulted from hypoxia/reoxygenation was inhibited by carnosic acid through the upregulation of Bcl-2 and the downregulation of Bax and caspase-3 levels. These results provided evidence for further investigation that would assist in the development of novel therapeutic approaches for myocardial infarction.

Abstract: Plants synthesize numerous specialized metabolites (also termed natural products) to mediate dynamic interactions with their surroundings. The complexity of plant specialized metabolism is the result of an inherent biosynthetic plasticity rooted in the substrate and product promiscuity of the enzymes involved. The pathway of carnosic acid-related diterpenes in rosemary and sage involves promiscuous cytochrome P450s whose combined activity results in a multitude of structurally related compounds. Some of these minor products, such as pisiferic acid and salviol, have established bioactivity, but their limited availability prevents further evaluation. Reconstructing carnosic acid biosynthesis in yeast achieved significant titers of the main compound but could not specifically yield the minor products. Specific production of pisiferic acid and salviol was achieved by restricting the promiscuity of a key enzyme, CYP76AH24, through a single-residue substitution (F112L). Coupled with additional metabolic engineering interventions, overall improvements of 24 and 14-fold for pisiferic acid and salviol, respectively, were obtained. These results provide an example of how synthetic biology can help navigating the complex landscape of plant natural product biosynthesis to achieve heterologous production of useful minor metabolites. In the context of plant adaptation, these findings also suggest a molecular basis for the rapid evolution of terpene biosynthetic pathways.

Abstract: Purpose Light-induced photoreceptor cell degeneration and disease progression in age-related macular degeneration (AMD) involve oxidative stress and visual cell loss, which can be prevented, or slowed, by antioxidants. Our goal was to test the protective efficacy of a traditional Age-related Eye Disease Study antioxidant formulation (AREDS) and AREDS combined with non-traditional antioxidants in a preclinical animal model of photooxidative retinal damage. Methods Male Sprague-Dawley rats were reared in a low-intensity (20 lux) or high-intensity (200 lux) cyclic light environment for 6 weeks. Some animals received a daily dietary supplement consisting of a small cracker infused with an AREDS antioxidant mineral mixture, AREDS antioxidants minus zinc, or zinc oxide alone. Other rats received AREDS combined with a detergent extract of the common herb rosemary, AREDS plus carnosic acid, zinc oxide plus rosemary, or rosemary alone. Antioxidant efficacy was determined by measuring retinal DNA levels 2 weeks after 6 h of intense exposure to white light (9,000 lux). Western blotting was used to determine visual cell opsin and arrestin levels following intense light treatment. Rhodopsin regeneration was determined after 1 h of exposure to light. Gene array analysis was used to determine changes in the expression of retinal genes resulting from light rearing environment or from antioxidant supplementation. Results Chronic high-intensity cyclic light rearing resulted in lower levels of rod and cone opsins, retinal S-antigen (S-ag), and medium wavelength cone arrestin (mCAR) than found for rats maintained in low cyclic light. However, as determined by retinal DNA, and by residual opsin and arrestin levels, 2 weeks after acute photooxidative damage, visual cell loss was greater in rats reared in low cyclic light. Retinal damage decreased with AREDS plus rosemary, or with zinc oxide plus rosemary whereas AREDS alone and zinc oxide alone (at their daily recommended levels) were both ineffective. One week of supplemental AREDS plus carnosic acid resulted in higher levels of rod and cone cell proteins, and higher levels of retinal DNA than for AREDS alone. Rhodopsin regeneration was unaffected by the rosemary treatment. Retinal gene array analysis showed reduced expression of medium- wavelength opsin 1 and arrestin C in the high-light reared rats versus the low-light rats. The transition of rats from low cyclic light to a high cyclic light environment resulted in the differential expression of 280 gene markers, enriched for genes related to inflammation, apoptosis, cytokine, innate immune response, and receptors. Rosemary, zinc oxide plus rosemary, and AREDS plus rosemary suppressed 131, 241, and 266 of these genes (respectively) in high-light versus low-light animals and induced a small subset of changes in gene expression that were independent of light rearing conditions. Conclusions Long-term environmental light intensity is a major determinant of retinal gene and protein expression, and of visual cell survival following acute photooxidative insult. Rats preconditioned by high-light rearing exhibit lower levels of cone opsin mRNA and protein, and lower mCAR protein, than low-light reared animals, but greater retention of retinal DNA and proteins following photooxidative damage. Rosemary enhanced the protective efficacy of AREDS and led to the greatest effect on the retinal genome in animals reared in high environmental light. Chronic administration of rosemary antioxidants may be a useful adjunct to the therapeutic benefit of AREDS in slowing disease progression in AMD.

Abstract: Carnosic acid and carnosol are the main bioactive components responsible for the significant antioxidant activity of Rosmarinus officinalis. Nevertheless, they are known for their instability in solutions. Separation of both compounds from crude rosemary extract was successfully achieved by one-step centrifugal partition chromatography without any degradation. A two-phase solvent system, hexane/ethyl acetate/methanol/water (3:2:3:2 v/v) was run on a preparative scale applying the elution-extrusion technique in descending mode. A 900 mg quantity of the crude extract containing 39.7% carnosic acid and 12.3% carnosol was loaded onto a 500 mL column, rotating at 1800 rpm. Carnosic acid and carnosol were obtained at purities of 96.1 ± 1% and 94.4 ± 0.9%, with recoveries of 94.3 ± 4.4% and 94.8 ± 2.3%, respectively. The compounds were identified by mass spectrometry, tandem mass spectrometry, and comparison with authentic standards.

Abstract: 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.

Abstract: Purpose Oral vitamin and mineral supplements reduce the risk of visual loss in age-related macular degeneration (AMD). However, the pathways that mediate this beneficial effect are poorly understood. Macrophages may exert oxidative, inflammatory, and angiogenic effects in the context of AMD. We aim to assess if oral supplements can modulate the macrophage phenotype in this disease. Methods Monocytes were isolated from patients with neovascular AMD (nvAMD), cultured, matured to macrophages, and polarized to classical [M1 (stimulated by IFNγ and lipopolysaccharide (LPS))] and alternative [M2 (stimulated with IL-4 and IL-13)] phenotypes. Combinations of antioxidants including lutein+zeaxanthin (1 μM; 0.2 μM), zinc (10 µM), carnosic acid (2 µM), beta-carotene (2 µM), and standardized tomato extract containing lycopene and other tomato phytonutrients were added to the culture media. Levels of anti-inflammatory, antioxidant, and pro-angiogenic gene and protein expression were then evaluated. Results Combinations of lutein and carnosic acid with zinc and standardized tomato extract or with beta-carotene yielded an antioxidative, anti-inflammatory, and antiangiogenic effect in M1 and M2 macrophages. These effects manifested in the upregulation of antioxidative genes (HMOX1, SOD1) and the downregulation of pro-angiogenic genes and pro-inflammatory genes (SDF-1, TNF-alpha, IL-6, MCP-1). Lutein monotherapy or a combination of lutein and zinc had less effect on the expression of these genes. Conclusions Combinations of supplements can modify the expression of genes and proteins that may be relevant for the involvement of macrophages in the pathogenesis of AMD. Further studies are required to evaluate if the modulation of the macrophage phenotype partially accounts for the beneficial effect of oral supplements in AMD and if modification of the AREDS formula can improve its effect on macrophages.

Abstract: The equine arteritis virus (EAV) is responsible by an important respiratory and reproductive disease in equine populations and there is no specific antiviral treatment available. The objective of this study was to investigate the activity of an ethanolic crude extract of Origanum vulgare (EEO) and of isolated compound caffeic acid, p-coumaric acid, rosmarinic acid, quercetin, luteolin, carnosol, carnosic acid, kaempferol and apigenin against EAV. The assays were performed using non-cytotoxic concentrations. The antiviral activity was monitored initially by cytopathic effect inhibition (CPE) assay in RK13 cells in the presence or absence of EEO. Pre-incubated cells with EEO were also examined to show prophylactic effect. Direct viral inactivation by EEO and isolated compounds was evaluated by incubation at 37°C or 20°C. After the incubation period, the infectivity was immediately determined by virus titrations on cell cultures and expressed as 50% tissue culture infective dose (TCID50)/100 µL. There was significant virucidal activity of EEO and of the compounds caffeic acid, p-coumaric acid, quercetin, carnosic acid and kaempferol. When EEO was added after infection, EEO inhibited the virus growth in infected cells, as evidenced by significant reduction of the viral titre. The results provide evidence that the EEO exhibit an inhibitory effect anti-EAV. Among the main compounds evaluated, caffeic acid, p-coumaric acid, carnosic acid, kaempferol and mainly quercetin, contributed to the activity of EEO. EEO may represent a good prototype for the development of a new antiviral agent, presenting promising for combating arteriviruses infections.

Abstract: In this study, the sage (Salvia officinalis) plant collected from the Mediterranean region was extracted with different solvents and methods. The extract yields were compared. The quantities of rosmarinic acid, carnosic acid and carnosol, which are responsible for the antioxidant capacity of the sage plant, were analyzed qualitatively and quantitatively by an HPLC. Ground sage plant were sieved with 70 mesh sieves. Maceration were done for powder sage plant at 45°C with 70% ethyl alcohol in “1:6” (w/v) ratio at different time (3; 6; 8; 10 hours); 100% methanol and 100% ethanol extractions; Soxhlet extraction efficiencies were compared. The proportions of rosmarinic acid and the amounts of carnosic acid and carnosol were analyzed in the extracted at UV detector and 280 nm wavelength by a Thermo Scientific Ulimate HPLC instrument. As a result of the experiments, a maceration method with 70% ethanol with 25% extraction yield was chosen. As a result of calculations on the standard graphic of rosmarinic acid in analytical standards. It was determined that 7.45 mg "rosmarinic acid"; 3.42 mg “carnosol+carnosic acid” in gram 70% ethanol extract of sage.

Abstract: The invention relates to a compound antioxidant, which is prepared from vitamin E, tea polyphenol-palmitate, ascorbyl palmitate and a rosemary extract. According to the compound antioxidant, the content of the vitamin E is smaller than or equal to 3,000ppm and greater than or equal to 5ppm, the content of the tea polyphenol-palmitate is smaller than or equal to 600ppm and greater than or equal to 5ppm, the content of the ascorbyl palmitate is smaller than or equal to 200ppm and greater than or equal to 5ppm and the content of the rosemary extract is smaller than or equal to 700ppm and greater than or equal to 5ppm; the content of the vitamin E is calculated according to the total content of mixed tocopherol; and the content of the rosemary extract is calculated according to the content of carnosic acid in the rosemary extract. The compound antioxidant is high in stability and high in antioxidant activity, and can be applied to vegetable fat, animal fat, margarine, oleomargarine or shortening.

Abstract: Taking into account the bioactive and healthy properties of alkylglycerols (AKG) and the antioxidant rosemary extract (RE), the production of a cooked meat product with partial replacement of animal fat by AKG-rich oil and inclusion of RE was performed. Six batches were manufactured: control, control+RE (0.045%RE); low-AKG (0.75%AKG-oil), low-AKG+RE (0.75%AKG-oil+0.045%RE); high-AKG (4.3%AKG-oil) and high-AKG+RE (4.3%AKG-oil+0.045%RE). Fatty acid and lipid profile, changes on lipid oxidation (TBARS), antioxidant ability (TEAC) and carnosic acid were determined during refrigerated storage (0, 30, 60 days). A relevant replacement of animal fat by AKG-rich oil was obtained (around 20% for the high-AKG samples), together with healthier indexes of the fatty acid profile. A slight trend to worse lipid oxidation due to addition of AKG-rich oil was measured, but being only significantly higher for the high-AKG treatment in absence of RE. A significant better stability was measured due to inclusion of RE for all the treatments. Therefore, the oxidation of both low-AKG+RE and high-AKG+RE treatments were significantly comparable to the control batch. After storage, TBARS values increased for all the batches. Samples containing RE showed better TEAC values after the storage. In agreement, the amount of carnosic acid kept constant during the self-life of the products. Practical application: Cooked meat products combining the bioactive ingredients AKG-rich oil and RE might be produced, without compromising the oxidative stability and with potential healthier and functional interest due to AKG enrichment, healthier fatty acid indexes, and antioxidant activity. Furthermore, the amount of one of the main bioactive compounds of RE, carnosic acid, would be preserved during the storage of the meat products, regardless of the addition of labile oils such as AKG-rich oil.

Abstract: Carnosol and carnosic acid in Rosmarinus officinalis L. were separated by high-performance capillary electrophoresis using a 50 cm capillary, a 12 mmol/L 80:20 pH 9.9 borax:methanol buffer using 25 kV of applied voltage at 30°C with detection at 280 nm. Rosmarinus officinalis L. was extracted with methanol with ultrasound at 170 W and 50 kHz for 20 min. The precision, repeatability, and recovery of the method were evaluated. The calibration relationships for carnosol and carnosic acid were y = 2.331x + 5.92 from 0.01 to 0.160 mg/mL and y = 7.980x + 4.32 from 0.005 to 0.080 mg/mL, respectively. The established method allows the separation and quantification of carnosic acid and carnosol in R. officinalis from various locations in China.

Abstract: A procedure is developed for the quantitative determination of diterpene acids in garden sage leaves by UV spectrophotometry at the wavelength 285 nm. The target group of compounds was selectively extracted by petroleum ether 40/70. It was shown that the completeness of extraction is determined mainly by the number of portions of the pure solvent: at the optimum ratio of the mass of the weighed portion to the volume of solvent 1 g/200 mL, double extraction is sufficient. The duration of each extraction is 20 min. The procedure was used in the analysis of samples of garden sage leaves from various producers. It was found that the concentration of diterpene acids in samples varied from 2.1 to 3.6 wt % (in terms of carnosic acid). The error of a single determination of the sum of diterpene acids in garden sage leaves is ±2.38% (P = 0.95).

Abstract: The invention discloses a preparation method of a rosemary extract. The method comprises the steps of S1, taking rosemary branches and leaves as raw materials and extracting carnosol, carnosic acid and ursolic acid at the same time to obtain an extracting solution I; S2, concentrating the extracting solution I to obtain a concentrated solution; S3, adding water into the concentrated solution for dilution to obtain turbid liquid; S4, treating the turbid liquid by an alkali dissolution process to obtain an extracting solution II and an extract I containing the ursolic acid; S5, treating the extracting solution II by adopting an acid sediment process to obtain an extract II containing the carnosol and the carnosic acid, thus realizing the synchronous extraction and separation of the extract containing the carnosol and the carnosic acid and the extract containing the ursolic acid.

Abstract: The invention provides a sodium alginate antibacterial spray and a preparation method thereof. the sodium alginate antibacterial spray is composed of, by weight, 4-6 parts of sodium alginate, 3-5 parts of water-soluble chitosan, 1-3 parts of carnosic acid, 2-4 parts of antibacterial agent, 0.5-5 parts of medical glycerin, 0.05-0.2 part of borneol, 0.5-2 parts of vitamin A, 0.5-2 parts of vitamin B, 0.5-2 parts of emulsifier, 1-3 parts of solubilizer and 95-100 parts of purified water. By adding the water-soluble chitosan and the carnosic acid, synergism of the sodium alginate and the water-soluble chitosan is utilized, and under the assistant action of the carnosic acid, so that the sodium alginate antibacterial spray is has good sterilizing effect, has high film forming durability and can meet long-acting bactericidal needs.