Llana Ruíz-Cabello, María

Profesor/a Contratado Doctor
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First Name
Last Name
Llana Ruíz-Cabello
Universidad Pablo de Olavide
Biología Molecular e Ingeniería Bioquímica
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Research Group
Ciencias Forenses y Toxicología
Ciencia y Técnicas de la Salud
PhD programs
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Now showing 1 - 10 of 11
  • Publication
    A subchronic 90-day oral toxicity study of Origanum vulgare essential oil in rats
    (Elsevier, 2017-01-03) Llana Ruíz-Cabello, María; Maisanaba, Sara; Puerto, Maria; Pichardo, Silvia; Jos, Angeles; Moyano, Rosario; Cameán, Ana María
    Oregano essential oil (Origanum vulgare L. virens) (OEO) is being used in the food industry due to its useful properties to develop new active packaging systems. In this concern, the safety assessment of this natural extract is of great interest before being commercialized. The European Food Safety Authority requests different in vivo assays to ensure the safety of food contact materials. One of these studies is a 90 days repeated-dose oral assay in rodents. In the present work, 40 male and 40 female Wistar rats were orally exposed to 50, 100 and 200 mg/kg body weight (b.w.) OEO during 90 days following the OECD guideline 408. Data revealed no mortality and no treatment-related adverse effects of the OEO in food/ water consumption, body weight, haematology, biochemistry, necropsy, organ weight and histopathology. These findings suggest that the oral no-observed-adverse-effect level (NOAEL) of this OEO is 200 mg/kg b.w. in Wistar rats, the highest dose tested. In conclusion, the use of this OEO in food packaging appears to be safe based on the lack of toxicity during the subchronic study at doses 330-fold higher than those expected to be in contact consumers in the worst scenario of exposure.
  • Publication
    Cytotoxic and mutagenic in vitro assessment of two organosulfur compounds derived from onion to be used in the food industry
    (Elsevier, 2014-06-16) Llana Ruíz-Cabello, María; Maisanaba, Sara; Gutiérrez-Praena, Daniel; Prieto, Ana Isabel; Pichardo, Silvia; Jos, Angeles; Moreno, Francisco Javier; Cameán, Ana María
    Edible members of the Allium family are widely used since they exhibit antioxidant and antibacterial related to the organosulphur compounds. One the most promising use of Allium species, hence, onion essential oil, could be in the packaging food industry. The present work aims to assess the safety of two organosulphur compounds present in onion essential oil; dipropyl disulphide, dipropyl sulphide and their mixture. For this purpose, cytotoxicity, reactive oxygen species and glutathione contents, and ultrastructural cellular damages were studied in the human intestinal cells, Caco-2, exposed to these organosulphur compounds. Moreover, their potential mutagenicity was also assessed. The results revealed no significant adverse effects. Additionally, reactive oxygen species scavenger activity was observed for both compounds. Therefore, they could be a good natural alternative to other synthetic antioxidant and antibacterial substances used in the food industry.
  • Publication
    New Method for Simultaneous Determination of Microcystins and Cylindrospermopsin in Vegetable Matrices by SPE-UPLC-MS/MS
    (MDPI, 2018-10-08) Diez-Quijada, Leticia; Guzmán-Guillén, Remedios; Prieto, Ana Isabel; Llana Ruíz-Cabello, María; Campos, Alexandre; Vasconcelos, Vitor; Jos, Angeles; Cameán, Ana María
    Cyanotoxins are a large group of noxious metabolites with different chemical structure and mechanisms of action, with a worldwide distribution, producing effects in animals, humans, and crop plants. When cyanotoxin-contaminated waters are used for the irrigation of edible vegetables, humans can be in contact with these toxins through the food chain. In this work, a method for the simultaneous detection of Microcystin-LR (MC-LR), Microcystin-RR (MC-RR), Microcystin-YR (MC-YR), and Cylindrospermopsin (CYN) in lettuce has been optimized and validated, using a dual solid phase extraction (SPE) system for toxin extraction and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for analysis. Results showed linear ranges (5–50 ng g􀀀1 f.w.), low values for limit of detection (LOD) (0.06–0.42 ng g􀀀1 f.w.), and limit of quantification (LOQ) (0.16–0.91 ng g􀀀1 f.w.), acceptable recoveries (41–93%), and %RSDIP values for the four toxins. The method proved to be robust for the three variables tested. Finally, it was successfully applied to detect these cyanotoxins in edible vegetables exposed to cyanobacterial extracts under laboratory conditions, and it could be useful for monitoring these toxins in edible vegetables for better exposure estimation in terms of risk assessment.
  • Publication
    Gold(I) metallocyclophosphazenes with antibacterial potency and antitumor efficacy. Synergistic antibacterial action of a heterometallic gold and silver-cyclophosphazene
    (Royal Society of Chemistry, 2022-08-22) Gascón, Elena; Otal, Isabel; Maisanaba, Sara; Llana Ruíz-Cabello, María; Valero Blanco, Eva María; Repetto, Guillermo; Jones, Peter G; Oriol, Luis; Jiménez, Josefina
    One of the most important uses of phosphazenes today involves its biomedical applications. They can also be employed as scaffolds for the design and construction of a variety of ligands in order to coordinate them to metallic drugs. The coordination chemistry of the (amino)cyclotriphosphazene ligand, [N3P3(NHCy)6], towards gold(I) complexes has been studied. Neutral complexes, [N3P3(NHCy)6{AuX}n] (X = Cl or C6F5; n = 1 or 2) (1–4), cationic complexes, [N3P3(NHCy)6{Au(PR3)}n](NO3)n (PR3 = PPh3, PPh2Me, TPA; n = 1, 2 or 3) (6–12) [TPA = 1,3,5-triaza-7-phosphaadamantane] and a heterometallic compound [N3P3(NHCy)6{Au(PPh3)}2{Ag(PPh3)}] (NO3)3 (13) have been obtained and characterized by various methods including single-crystal X-ray diffraction for 7, which confirms the coordination of gold atoms to the nitrogens of the phosphazene ring. Compounds 1, 4, 6–13 were screened for in vitro cytotoxic activity against two tumor human cell lines, MCF7 (breast adenocarcinoma) and HepG2 (hepatocellular carcinoma), and for antimicrobial activity against five bacterial species including Gram-positive, Gram-negative, and Mycobacteria. Both the median inhibitory concentration (IC50) and minimum inhibitory concentration (MIC) values are among the lowest found for any gold or silver derivatives against the cell lines and particularly against the Gram-positive (S. aureus) strain and the mycobacteria used in this work. Structure–activity relationships are discussed in order to determine the influence of ancillary ligands and the number and type of metal atoms (silver or gold). Compounds 4 and 8 showed not only maximal potency on human cells but also some tumour selectivity. Remarkably, compound 13, with both gold and silver atoms, showed outstanding activity against both Gram-positive and Gram-negative strains (nanomolar range), thus having a cooperative effect between gold and silver, with MIC values which are similar or lower than those of gentamicine, ciprofloxacin and rifampicine. The broad spectrum antimicrobial efficacy of all these metallophosphazenes and particularly of heterometallic compound 13 could be very useful to obtain materials for surfaces with antimicrobial properties that are increasingly in demand.
  • Publication
    Characterisation and evaluation of PLA films containing an extract of Allium spp. to be used in the packaging of ready-to-eat salads under controlled atmospheres
    (Elsevier, 2015-07-22) Llana Ruíz-Cabello, María; Pichardo, Silvia; Baños, Alberto; Nuñez, Cristina; Bermúdez, José María; Guillamón, Enrique; Aucejo, Susana; Cameán, Ana María
    New food packaging systems are been developed by food industry. The present work proposes the use of polylactic acid (PLA) containing different percentages of an Allium spp. extract (2%, 5%, and 6.5% Proallium ®) to be used in the packaging of ready-to-eat salads. Mechanical and optical properties of PLA films studied did not show remarkable changes when the active substance was incorporated. Additionally, no significant antioxidant activity was observed, although remarkable antimicrobial activity was recorded, mainly in films containing 5% and 6.5% of the Allium spp. extract. Moreover, the antimicrobial effect in vivo was observed in lettuce. All developed films decreased enterobacterial growth in a concentrationdependent manner. Regarding aerobic bacteria, film with the highest concentration of active agent (6.5%) was found to be effective up to 5 days of storage, and even 7 days for moulds. Therefore, the PLAProallium ® film could be a promising antimicrobial active material, especially for ready-to-eat salads.
  • Publication
    Development of PLA films containing oregano essential oil (Origanum vulgare L. virens) intended for use in food packaging
    (Taylor and Francis, 2016-07-25) Llana Ruíz-Cabello, María; Pichardo, Silvia; Bermúdez, José María; Baños, Alberto; Nuñez, Cristina; Guillamón, Enrique; Aucejo, Susana; Cameán, Ana María
    Consumers’ concerns about the environment and health have led to the development of new food packaging materials avoiding petroleum-based matrices and synthetic additives. The present study has developed polylactic acid (PLA) films containing different concentrations of essential oil from Origanum vulgare L. virens (OEO). The effectiveness of this new active packaging was checked for use in ready-to-eat salads. A plasticising effect was observed when OEO was incorporated in PLA films. The rest of the mechanical and physical properties of developed films did not show much change when OEO was included in the film. An antioxidant effect was recorded only for films containing the highest percentages of the active agent (5% and 10%). In addition, films exhibited in vitro antibacterial activity against Staphylococcus aureus, Yersinia enterocolitica, Listeria monocytogenes, Enterococcus faecalis and Staphylococcus carnosus. Moreover, in ready-to-eat salads, antimicrobial activity was only observed against yeast and moulds, where 5% and 10% of OEO was the most effective.
  • Publication
    Use of micronucleus and comet assay to evaluate evaluate the genotoxicity of oregano essential oil (Origanum vulgare l. Virens) in rats orally exposed for 90 days
    (Taylor and Francis, 2018-03-14) Llana Ruíz-Cabello, María; Puerto, María; Maisanaba, Sara; Pichardo, Silvia; Cameán, Ana María
    Essential oils from Origanum spp. exhibit antioxidant and antimicrobial activities making them suitable for use as food additives. The incorporation of oregano essential oil in active food packaging is under study; however, it has been not authorized for this purpose thus far. In order to fulfill the requirements of the European Food Safety Authority (EFSA), the aim of the present study was to determine the genotoxic potential of oregano essential oil using both the micronucleus (MN) test and comet (standard and enzyme-modified) assays in Wistar rats treated with 50, 100, or 200 mg/kg body weight administered daily for 90 days. MN was performed in cells from the bone marrow and standard and enzyme-modified comet assays were conducted in stomach, liver and blood cells. The major compound detected in the analytical study of oregano essential oil from Origanum vulgare L. virens, was carvacrol (55.82%) followed by thymol (5.14%), as well as their precursors, γ-terpinene (16.39%), and ρ-cimne (4.71%). The results obtained in the genotoxicity assays indicated lack of effect in MN and standard comet assay under the conditions tested. Furthermore, no apparent oxidative damage was observed in the enzyme-modified comet assay in any of the tissues examined of rats exposed to oregano essential oil for 90 days. Therefore, this oregano essential oil appears to be safe in Wistar rats and might be considered as a potential active material in food packaging industry.
  • Publication
    Analysis of the use of cylindrospermopsin and/or microcystin-contaminatedwater in the growth, mineral content, and contamination of spinacia oleracea and lactuca sativa
    (MDPI, 2019-08-22) Llana Ruíz-Cabello, María; Jos, Angeles; Cameán, Ana María; Oliveira, Flavio; Barreiro, Aldo; Machado, Joana; Azevedo, Joana; Pinto, Edgar; Almeida, Agostino; Campos, Alexandre; Vasconcelos, Vitor; Freitas, Marisa
    Cyanobacteria and cyanotoxins constitute a serious environmental and human health problem. Moreover, concerns are raised with the use of contaminated water in agriculture and vegetable production as this can lead to food contamination and human exposure to toxins as well as impairment in crop development and productivity. The objective of this work was to assess the susceptibility of two green vegetables, spinach and lettuce, to the cyanotoxins microcystin (MC) and cylindrospermopsin (CYN), individually and in mixture. The study consisted of growing both vegetables in hydroponics, under controlled conditions, for 21 days in nutrient medium doped with MC or CYN at 10 g/L and 50 g/L, or CYN/MC mixture at 5 + 5 g/L and 25 + 25 g/L. Extracts from M. aeruginosa and C. ovalisporum were used as sources of toxins. The study revealed growth inhibition of the aerial part (Leaves) in both species when treated with 50 g/L of MC, CYN and CYN/MC mixture. MC showed to be more harmful to plant growth than CYN. Moreover spinach leaves growth was inhibited by both 5 + 5 and 25 + 25 g/L CYN/MC mixtures, whereas lettuce leaves growth was inhibited only by 25 + 25 g/L CYN/MC mixture. Overall, growth data evidence increased sensitivity of spinach to cyanotoxins in comparison to lettuce. On the other hand, plants exposed to CYN/MC mixture showed di erential accumulation of CYN and MC. In addition, CYN, but not MC, was translocated from the roots to the leaves. CYN and MC a ected the levels of minerals particularly in plant roots. The elements most a ected were Ca, K and Mg. However, in leaves K was the mineral that was a ected by exposure to cyanotoxins.
  • Publication
    Characterisation and antimicrobial activity of active polypropylene films containing oregano essential oil and Allium extract to be used in packaging for meat products.
    (Taylor and Francis, 2018-01-18) Llana Ruíz-Cabello, María; Pichardo, Silvia; Bermudez, José María; Baños, Alberto; Ariza, Juan José; Guillamón, Enrique; Aucejo, Susana; Cameán, Ana María
    Cooked ham is more prone to spoilage than other meat products, making preservation a key step in its commercialisation. One of the most promising preservation strategies is the use of active packaging. Oregano essential oil (OEO) and Proallium® (an Allium extract) have previously been shown to be useful in polylactic acid (PLA)-active films for ready-to-eat salads. The present work aims to study the suitability of polypropylene (PP) films containing OEO and Proallium® in the preservation of cooked ham. Concerning the technological features of the studied material, no significant changes in the mechanical or optical properties of PP films containing the active substances were recorded in comparison to the PP film without extracts. However, films containing both active substances were more flexible than the control film and less strong, highlighting the plasticisation effect of the natural extracts. Moreover, physical properties changed when active substances were added to the film. Incorporation of 4% Proallium® affected the transparency of the film to a higher extent compared to 8% OEO, undergoing decreases in transparency of 40% and 45%, respectively. Moreover, only the film containing the highest amount of OEO (8%) significantly decreased the thickness. Both active substances showed antibacterial properties; however, Proallium®-active films seemed to be more effective against Brochothrix thermosphacta than PP films containing OEO, with all percentages of Proallium® killing the bacterial population present in the ham after 60 days. In addition, materials containing the lowest Proallium® content exhibited higher acceptability by consumers in the sensory analyses with 63–100% willing to purchase, better even than the control package (56–89%). In fact, 2% of Proallium® obtained the best results in the odour study performed by the panellists.
  • Publication
    Cytotoxicity and morphological effects induced by carvacrol and thymol on the human cell line Caco-2
    (Elsevier, 2013-12-08) Llana Ruíz-Cabello, María; Gutiérrez-Praena, Daniel; Pichardo, Silvia; Moreno, Francisco Javier; Bermúdez, José María; Aucejo, Susana; Cameán, Ana María
    Essential oils used as additives in the food industry due to its flavour, antimicrobial and antioxidant properties. Therefore, human can be exposed orally to these compounds through the ingestion of foods. In this sense, the present work aims to assess toxicological effects of oregano essential oil on the digestive tract. In concrete, the cytotoxic effects of two components of the oregano essential oils, carvacrol and thymol, and their mixture, on the intestinal cells line Caco-2 after 24 and 48 h of exposure are studied. The basal cytotoxicity endpoints assayed (total protein content, neutral red uptake and the tetrazolium salt reduction) and the annexin/propidium iodide staining indicated that carvacrol and the mixture carvacrol/thymol induced toxic effects. Moreover, a morphological study was performed in order to determine the ultrastructural cellular damages caused by these substances. The main morphological alterations were vacuolated cytoplasm, altered organelles and finally cell death. In addition, although no cytotoxic effects were recorded for thymol at any concentration and time of exposure, ultrastructural changes evidenced cellular damage such as lipid degeneration, mitochondrial damage, nucleolar segregation and apoptosis.