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      Comparative Study of Green Sub- and Supercritical Processes to Obtain Carnosic Acid and Carnosol-Enriched Rosemary Extracts with in Vitro Anti-Proliferative Activity on Colon Cancer Cells

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          Abstract

          In the present work, four green processes have been compared to evaluate their potential to obtain rosemary extracts with in vitro anti-proliferative activity against two colon cancer cell lines (HT-29 and HCT116). The processes, carried out under optimal conditions, were: (1) pressurized liquid extraction (PLE, using an hydroalcoholic mixture as solvent) at lab-scale; (2) Single-step supercritical fluid extraction (SFE) at pilot scale; (3) Intensified two-step sequential SFE at pilot scale; (4) Integrated PLE plus supercritical antisolvent fractionation (SAF) at pilot scale. Although higher extraction yields were achieved by using PLE (38.46% dry weight), this extract provided the lowest anti-proliferative activity with no observed cytotoxic effects at the assayed concentrations. On the other hand, extracts obtained using the PLE + SAF process provided the most active rosemary extracts against both colon cancer cell lines, with LC 50 ranging from 11.2 to 12.4 µg/mL and from 21.8 to 31.9 µg/mL for HCT116 and HT-29, respectively. In general, active rosemary extracts were characterized by containing carnosic acid (CA) and carnosol (CS) at concentrations above 263.7 and 33.9 mg/g extract, respectively. Some distinct compounds have been identified in the SAF extracts (rosmaridiphenol and safficinolide), suggesting their possible role as additional contributors to the observed strong anti-proliferative activity of CA and CS in SAF extracts.

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          Most cited references57

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          Feasibility of a high-flux anticancer drug screen using a diverse panel of cultured human tumor cell lines.

          We describe here the development and implementation of a pilot-scale, in vitro, anticancer drug screen utilizing a panel of 60 human tumor cell lines organized into subpanels representing leukemia, melanoma, and cancers of the lung, colon, kidney, ovary, and central nervous system. The ultimate goal of this disease-oriented screen is to facilitate the discovery of new compounds with potential cell line-specific and/or subpanel-specific antitumor activity. In the current screening protocol, each cell line is inoculated onto microtiter plates, then preincubated for 24-28 hours. Subsequently, test agents are added in five 10-fold dilutions and the culture is incubated for an additional 48 hours. For each test agent, a dose-response profile is generated. End-point determinations of the cell viability or cell growth are performed by in situ fixation of cells, followed by staining with a protein-binding dye, sulforhodamine B (SRB). The SRB binds to the basic amino acids of cellular macromolecules; the solubilized stain is measured spectrophotometrically to determine relative cell growth or viability in treated and untreated cells. Following the pilot screening studies, a screening rate of 400 compounds per week has been consistently achieved.
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            Antioxidant activity of essential oils.

            Essential oils (EOs) are liquid mixtures of volatile compounds obtained from aromatic plants. Many EOs have antioxidant properties, and the use of EOs as natural antioxidants is a field of growing interest because some synthetic antioxidants such as BHA and BHT are now suspected to be potentially harmful to human health. Addition of EOs to edible products, either by direct mixing or in active packaging and edible coatings, may therefore represent a valid alternative to prevent autoxidation and prolong shelf life. The evaluation of the antioxidant performance of EOs is, however, a crucial issue, because many commonly used "tests" are inappropriate and give contradictory results that may mislead future research. The chemistry explaining EO antioxidant activity is discussed along with an analysis of the potential in food protection. Literature methods to assess EOs' antioxidant performance are critically reviewed.
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              Oleanolic acid and ursolic acid: research perspectives.

              Jie Liu (2005)
              Oleanolic acid and ursolic acid are ubiquitous triterpenoids in plant kingdom, medicinal herbs, and are integral part of the human diet. During the last decade over 700 research articles have been published on their research, reflecting tremendous interest and progress in our understanding of these triterpenoids. This included the isolation and purification of these tritepernoids from various plants and herbs, the chemical modifications to make more effective and water soluble derivatives, the pharmacological research on their beneficial effects, the toxicity studies, and the clinical use of these triterpenoids in various diseases including anticancer chemotherapies. A briefly commentary is attempted here for their research perspectives.
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                Author and article information

                Contributors
                Role: Academic Editor
                Journal
                Int J Mol Sci
                Int J Mol Sci
                ijms
                International Journal of Molecular Sciences
                MDPI
                1422-0067
                07 December 2016
                December 2016
                : 17
                : 12
                : 2046
                Affiliations
                Laboratory of Foodomics, Institute of Food Science Research, Instituto de Investigación en Ciencias de la Alimentación, Consejo Superior de Investigaciones Científicas, Nicolas Cabrera 9, 28049 Madrid, Spain; andreap.sanchez@ 123456csic.es (A.d.P.S.-C.); virginia.garcia@ 123456csic.es (V.G.-C.); m.herrero@ 123456csic.es (M.H.); a.cifuentes@ 123456csic.es (A.C.)
                Author notes
                [* ]Correspondence: elena.ibanez@ 123456csic.es ; Tel.: +34-910-017596; Fax: +34-910-017905
                Article
                ijms-17-02046
                10.3390/ijms17122046
                5187846
                27941607
                c6a4b380-027c-448e-81b5-6d2f4ddf3345
                © 2016 by the authors; licensee MDPI, Basel, Switzerland.

                This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 13 October 2016
                : 29 November 2016
                Categories
                Article

                Molecular biology
                rosemary,supercritical fluid extraction (sfe),process intensification,subcritical fluids,supercritical fluids,anti-proliferative,colon cancer cell,ht-29,hct116

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