Identification and quantification of cholesterol oxides in foods

The types and levels of nine major cholesterol oxidation products was determined in a variety of foods. An estimate was made of the possible levels of cholesterol oxides in the Australian diet. These levels have been shown to cause endothelial damage in the coronary arteries of laboratory animals in previous studies. The health implications of these observations to humans is unknown.

Increase of Cholesterol Oxidation and Decrease of PUFA as a Result of Thermal Processing and Storage in Eggs Enriched with n-3 Fatty Acids

In this work, cholesterol oxide formation and alteration of fatty acid composition were analyzed in n-3 enriched eggs under different storage periods and two temperatures. The eggs enriched with n-3 fatty acids were stored at 5 or 25 degrees C for 45 days and subsequently boiled or fried. For each treatment, 12 yolks were analyzed every 15 days including time zero. The concentrations of the cholesterol oxides 7-ketocholesterol, 7 beta-hydroxycholesterol, and 7 alpha-hydroxycholesterol increased during the storage period and were higher in fried eggs. Only the 7-ketocholesterol was affected by the storage temperature, and its concentration was highest in eggs stored at 25 degrees C. There was no significant difference in the contents of cholesterol and vitamin E at the different storage periods; however, the concentration of vitamin E decreased with thermal treatment. In addition, the n-3 polyunsaturated fatty acids, especially 18:3, 20:5, and 22:6, were reduced throughout the storage at 5 and 25 degrees C.

HPLC analysis of plant sterol oxidation products

Increased interest in the cholesterol-lowering effect of plant sterols has led to development of plant sterol-enriched foods. When products are enriched, the safety of the added components must be evaluated. In the case of plant sterols, oxidation is the reaction of main concern. In vitro studies have indicated that cholesterol oxides may have harmful effects. Due their structural similarity, plant sterol oxidation products may have similar health implications. This study concentrated on developing high-performance liquid chromatography (HPLC) methods that enable the investigation of formation of both primary and secondary oxidation products and thus can be used for oxidation mechanism studies of plant sterols. The applicability of the methods for following the oxidation reactions of plant sterols was evaluated by using oxidized stigmasterol and sterol mixture as model samples. An HPLC method with ultraviolet and fluorescence detection (HPLC-UV-FL) was developed. It allowed the specific detection of hydroperoxides with FL detection after post-column reagent addition. The formation of primary and secondary oxidation products and amount of unoxidized sterol could be followed by using UV detection. With the HPLC-UV-FL method, separation between oxides was essential and oxides of only one plant sterol could be quantified in one run. Quantification with UV can lead to inaccuracy of the results since the number of double bonds had effect on the UV absorbance. In the case of liquid chromatography-mass spectrometry (LC-MS), separation of oxides with different functionalities was important because some oxides of the same sterol have similar molecular weight and moreover epimers have similar fragmentation behaviour. On the other hand, coelution of different plant sterol oxides with the same functional group was acceptable since they differ in molecular weights. Results revealed that all studied plant sterols and cholesterol seem to have similar fragmentation behaviour, with only relative ion abundances being slightly different. The major advantage of MS detection coupled with LC separation is the capability to analyse totally or partly coeluting analytes if these have different molecular weights. The HPLC-UV-FL and LC-MS methods were demonstrated to be suitable for studying the photo-oxidation and thermo-oxidation reactions of plant sterols. The HPLC-UV-FL method was able to show different formation rates of hydroperoxides during photo-oxidation. The method also confirmed that plant sterols have similar photo-oxidation behaviour to cholesterol. When thermo-oxidation of plant sterols was investigated by HPLC-UV-FL and LC-MS, the results revealed that the formation and decomposition of individual hydroperoxides and secondary oxidation products could be studied. The methods used revealed that all of the plant sterols had similar thermo-oxidation behaviour when compared with each other, and the predominant reactions and oxidation rates were temperature dependent. Overall, these findings showed that with these LC methods the oxidation mechanisms of plant sterols can be examined in detail, including the formation and degradation of individual hydroperoxides and secondary oxidation products, with less sample pretreatment and without derivatization.

Modificaciones tecnológicas para mejorar la seguridad y calidad del jamón curado

Se introdujeron varias modificaciones tecnológicas en la elaboración habitual del jamón curado español de cerdo blanco para mejorar su seguridad y calidad, así como para investigar la contribución relativa de los diversos procesos implicados en la calidad sensorial. Las modificaciones introducidas en cada uno de 3 experimentos (a, b, c) fueron: a) inoculación de un cultivo iniciador (CI) en la superficie del producto y el envasado del jamón al vacío durante la etapa de reposo (EV); b) aplicación de una atmósfera modificada con un contenido reducido de oxígeno (AMCRO) (durante la totalidad o la última parte del procesado) en dos procesos que diferían en las humedades relativas aplicadas; c) realización de un estufaje de 4 días a 35ºC y la aplicación repetida de pequeñas cantidades de grasa dorsal líquida sobre la superficie del jamón. En cada experimento, se siguió un diseño experimental de bloques incompletos para bloquear y evaluar el efecto de la materia prima en cada parámetro. La aplicación del CI evitó el crecimiento superficial de hongos, pero modificó el flavor del producto, dando lugar a la aparición de flavores impropios del jamón tradicional, al aumento de la incidencia de la coquera y a la reducción de la intensidad de notas características del mismo como el flavor añejo. Estos efectos fueron debidos a la acción directa del CI pero probablemente también a los cambios que provocó en la superficie del jamón, como la atenuación del "sudado" del jamón. El EV trajo consigo una reducción del crecimiento superficial de mohos; un mayor gradiente de humedad entre el interior y el exterior del jamón; una disminución de la pérdida de peso; un aumento del nitrógeno no proteico y cambios negativos en la textura, aspecto y flavor, como el aumento de la intensidad del velo blanco y del flavor a pienso, el aumento de la incidencia de la coquera y la atenuación del flavor añejo. Estos efectos fueron consecuencia del mayor contenido de humedad a que dio lugar dicha modificación tecnológica, de la potenciación de los efectos negativos del uso del CI, así como a los cambios que provocó en la superficie del jamón. El uso de una ACRO durante todo el proceso provocó un aumento del nitrógeno no proteico, una disminución de la concentración de óxidos de colesterol, un aumento de la intensidad del velo blanco y, en combinación con el uso de humedades relativas bajas, causó una disminución del crecimiento bacteriano y evitó el crecimiento de hongos, levaduras y ácaros en el interior y exterior del jamón y el desarrollo de la coquera. Asímismo, dio lugar a una drástica reducción de la intensidad del flavor del jamón debido a la disminución de la intensidad de la oxidación lipídica. Cuando esta ACRO se aplicó únicamente al final del proceso, se consiguió la eliminación de las formas móviles de los ácaros y la disminución de la intensidad de la coquera y el producto resultante poseía un flavor algo más intenso que aquél sometido a una ACRO durante todo el proceso. El aumento de la temperatura de 25-27 ºC a 35 ºC durante 4 días no tuvo ningún efecto sobre los parámetros estudiados. La aplicación de la grasa líquida en la superficie del jamón evitó el secado excesivo en superficie, previno el desarrollo de la coquera y causó un aumento de la intensidad del flavor añejo y una reducción de la incidencia de notas negativas como el tostado, hechos que indican que la liberación de grasa líquida en el jamón ("sudado") constituye un fenómeno determinante en su calidad sensorial. La materia prima fue el factor que afectó a un mayor número de parámetros.

Effect of natural antioxidant combinations on lipid oxidation in cooked chicken meat during refrigerated storage

The effect of combinations of sage, oregano and honey on lipid oxidation in cooked chicken meat during refrigeration at 4 degrees C for 96 h was determined. Chicken samples (thigh and breast) were then separated into five groups; control; butylated hydroxytoluene; oregano + sage; oregano + sage + 5%honey and oregano + sage + 10%honey. Quantitative measurements of thiobarbituric acid reactive substances, conjugated dienes, hexanal, fatty acids, cholesterol and cholesterol oxides were used as indicators of lipid oxidation. Acceptability and preference were also evaluated. The effectiveness of the natural antioxidants for reducing the velocity of lipid oxidation in cooked chicken thigh and breast was demonstrated after 48 and 96 h of refrigeration at 4 degrees C. The treatments that presented the lowest hexanal values after 96 h of refrigeration were oregano + sage + 5%honey and oregano + sage + 10%honey. Only traces of free cholesterol oxides were found (25-OH, 7-k, 7 alpha-OH and 7 beta-OH). The natural antioxidants protected cooked chicken meat from oxidation processes and resulted in great acceptability. (C) 2012 Elsevier Ltd. All rights reserved.

Perfil lipídico de hambúrgueres produzidos a partir de carne de bovinos alimentados com fontes de óleo na dieta

Objetivou-se nesse trabalho avaliar os efeitos de fontes de óleo vegetal na dieta de bovinos Nelore em fase de terminação sobre a aceitação sensorial, teores de colesterol e óxidos de colesterol, composição de ácidos graxos e estabilidade oxidativa de hambúrgueres bovinos, avaliados após serem congelados e acondicionados a -18°C por 90 dias, então avaliados crus e grelhados. Foram confinados 96 bovinos, machos, castrados, Nelore, com aproximadamente 380 kg de peso inicial e idade média de 24 meses, alimentados com óleos de soja, girassol, linhaça e tratamento controle sem adição de óleo por 82 dias. A partir do músculo Longissimus e da gordura subcutânea dos animais foram produzidos hambúrgueres Os hambúrgueres produzidos com carne de bovinos que receberam óleo de soja tiveram maior estabilidade lipídica, tanto para zero quanto para 90 dias de armazenamento quando comparado às outras dietas. O método cru de preparo também resultou em menores valores tanto para o tempo zero quanto para o tempo 90 dias de armazenamento, em relação ao método grelhado. Os hambúrgueres dos animais alimentados com o óleo linhaça resultaram em menores valores para o atributo sabor. O tempo zero dias de armazenamento dos hambúrgueres resultou em maiores notas dos consumidores para a textura e aceitabilidade global, em relação às amostras armazenadas por 90 dias. A porcentagem de ácidos graxos polinsaturados foi reduzida, independente dos tratamentos, após 90 dias de armazenamento. O CLA c9 t11 foi encontrado em maiores quantidades nas amostras dos animais alimentados com os óleos de linhaça e soja, para zero e 90 dias de armazenamento. O teor de colesterol aumentou após as amostras de hambúrgueres serem grelhadas, enquanto que não foi observado efeito para as fontes de óleos e os tempos de armazenamento. Foi observada a presença de óxidos de colesterol após as amostras serem armazenadas por 90 dias, independente das fontes de óleos utilizadas. O fornecimento de óleos insaturados para bovinos confinados aumenta a porcentagem de ácidos graxos polinsaturados em hambúrgueres produzidos com carne e gordura desses. A inclusão de óleos vegetais para bovinos aumenta a oxidação lipídica de hambúrgueres produzidos a partir de carne e gordura provenientes desses animais quando comparados a um tratamento sem inclusão de óleos vegetais. Hambúrgueres produzidos com carne e gordura de animais alimentados com óleos vegetais na dieta possuem maior oxidação lipídica após 90 dias de armazenamento quando comparados com os hambúrgueres do tempo zero e dos hambúrgueres do tratamento controle enquanto que o processamento térmico não foi responsável pela maior oxidação lipídica.

Efeito de fontes de óleos vegetais sobre o desempenho, características de carcaça e qualidade da carne de bovinos Nelore

A inclusão de óleos vegetais na dieta de bovinos tem sido utilizada para aumentar a densidade energética da dieta, melhorar a eficiência alimentar, além de produzir carnes com a composição de ácidos graxos mais favorável à saúde humana. Objetivou-se estudar os efeitos da inclusão de óleos de soja, girassol e linhaça na dieta de bovinos, sobre o desempenho, características quantitativas de carcaça e qualitativas da carne, perfil de ácidos graxos, oxidação lipídica e formação de compostos oxidados do colesterol. Foram confinados 96 bovinos Nelore, castrados, com aproximadamente 380 kg ± 34 kg de peso inicial e idade média de 20 meses. As dietas foram compostas de 79% de concentrado e 21% de volumoso (silagem de milho) e incluídos os óleos de soja, girassol e linhaça. Os animais foram pesados e avaliadas as características de carcaça por ultrassonografia nos dias 0, 28, 56 e 81 de confinamento. Nos dias zero e 81 dias de confinamento foi coletado sangue dos bovinos para avaliação do LDL-colesterol, HDL-colesterol, VLDL-colesterol e triacilgliceróis. Ao final de 81 dias de confinamento, os animais foram abatidos e foi avaliado o pH (uma e 48 horas após o abate) e foram retiradas amostras do músculo longissimus. Foi avaliado a cor, força de cisalhamento (FC) e perdas por cocção (PPC) em carnes não maturadas e maturadas por 14 dias. Duas amostras do longissimus foram expostas por um e três dias em condições semelhantes ao varejo. Nas carnes expostas por um dia foi avaliado a cor e o pH. Nas amostras expostas por três dias foi avaliado além da cor e pH, o perfil de ácidos graxos, TBARS, colesterol e a presença do 7-cetocolesterol. Foi realizada ainda a análise sensorial e determinado a quantidade de lipídios totais das carnes. A estabilidade oxidativa dos óleos utilizados na dieta também foi avaliada. O experimento foi conduzido em delineamento de blocos casualizados, sendo o peso inicial o bloco. O desempenho e as características de carcaça avaliadas por ultrassonografia não foram influenciadas pelas fontes de óleo. Os tratamentos não influenciaram o peso de abate, o peso de carcaça quente, o pH da carcaça uma hora e 48 horas após o abate, o rendimento de carcaça, a cor, as PPC e a FC. O pH foi maior nas carnes maturadas por 14 dias (P=0,01), em relação àquelas sem maturação. As PPC e a FC foram menores (P=0,01) nas carnes maturadas por 14 dias. O óleo de linhaça apresentou menor estabilidade oxidativa seguidos do óleo de girassol e soja. As fontes de óleo não afetaram a concentração dos lipídios do plasma sanguíneo, no entanto, os níveis de VLDL, LDL, HDL, colesterol e triacilgliceróis foram maiores (P<0,01) no final do experimento em relação ao início. Não houve interação entre a espessura de gordura subcutânea (EGS) avaliada no abate e as dietas e efeito das fontes de óleo sobre os valores de TBARS, lipídios e colesterol. Não foi encontrado o 7-cetocolesterol nas carnes. O pH das carnes expostas por um e três dias sob condições de varejo, não foram influenciadas pela dieta nem pela interação da dieta e dos dias de exposição. No entanto, foi observado o efeito de tempo (P<0,01), as carnes expostas por três dias tiveram valores de pH maiores que as carnes expostas por um dia. A cor L*, a* e b* das carnes expostas em gôndola, sob condições de varejo não foi influenciado pela dieta, pelos dias de exposição e nem pela interação dos dias de exposição e dietas. Os ácidos graxos C18:1 n-9, C20:3 n-6 e C20:5 n-3 apresentaram interação entre a EGS e a dieta (P<0,05). O C18:1 cis 6 apresentou maiores concentrações (P<0,05) nas carnes provenientes dos animais alimentados com óleo de linhaça e soja, em comparação com as carnes provenientes da dieta controle. O C18:3 n-3 apresentou maiores concentrações nas carnes de animais alimentados com linhaça (P<0,05), em comparação com os demais tratamentos. O aroma e a textura da carne avaliados em análise sensorial realizada com consumidores não foram alterados pelos tratamentos. A carne dos animais alimentados com óleo de girassol resultou em maiores notas para o sabor (P<0,01), em relação à carne proveniente de animais alimentados com óleo de soja. As dietas controle e girassol resultaram em carnes mais suculentas e com maior aceitabilidade global (P<0,01), em relação ao tratamento soja. Independentemente do tipo de óleo utilizado na dieta dos animais, não houve influência no desempenho e nas características da carcaça. O óleo de linhaça proporcionou carnes com perfil de ácidos graxos mais favorável para a saúde humana, pois apresentou maiores proporções do ácido linolênico e relações ideais de n6:n3 (4,15). O uso de óleos vegetais na dieta, não prejudicou a aparência das carnes e não proporcionaram oxidação lipídica com a formação de compostos de colesterol oxidados nas carnes expostas sob condições de varejo.

Cholesterol-lowering management of stroke : inconsistencies of evidence and practice

Cholesterol-lowering treatment by statins is an important and costly issue; however, its role in stroke has not been well documented. The aim of the present study was to review literature and current practice regarding cholesterol-lowering treatment for stroke patients. A literature review was conducted on lipids in stroke and their management with both statins and diet, including the cost-effectiveness of medical nutrition therapy. Qualifying criteria and prescription procedures of the Pharmaceutical Benefits Scheme (PBS) were also reviewed. Data on lipid levels and statin prescriptions were analysed for 468 patients admitted to a stroke unit. The literature shows that management with both medication and diet can be effective, especially when combined; however, 60% of patients with an ischaemic event had fasting total cholesterol measures ≥4 mmol/L (n = 231), with only 52% prescribed statins on discharge (n = 120). Hypercholesterolaemia is an underdiagnosed and undertreated risk factor within the stroke population. It appears that the PBS has not kept pace with advances in the evidence in terms of statin use in the stroke population, and review is needed. The present review should address the qualifying criteria for the stroke population and recommendations on referral to dietitians for dietary advice. Cholesterol-lowering treatment for both stroke patients and the wider population is an area that needs awareness raising and review by the PBS, medical practitioners and dietitians. The role of dietary and pharmacological treatments needs to be clearly defined, including adjunct therapy, and the cost-effectiveness of medical nutrition therapy realised.

meso-Porphyrinylphosphine Oxides: Mono- and Bidentate Ligands for Supramolecular Chemistry and the Crystal Structures of Monomeric [10,20-Diphenylporphyrinatonickel(II)-5,15-diyl]-bis-[P(O)Ph2] and Polymeric Self-Coordinated ([10,20-Diphenylporphyrinatozinc(II)-5,15-diyl]-bis-[P(O)Ph2])

A series of porphyrins substituted in one or two meso-positions by diphenylphosphine oxide groups has been prepared by the palladium catalysed reaction of diphenylphosphine or its oxide with the corresponding bromoporphyrins. Compounds {MDPP-[P(O)Ph2]n} (M = H2, Ni, Zn; H2DPP = 5,15-diphenylporphyrin; n = 1, 2) were isolated in yields of 60-95%. The reaction is believed to proceed via the conventional oxidative addition, phosphination and reductive elimination steps, as the stoichiometric reaction of η1-palladio(II) porphyrin [PdBr(H2DPP)(dppe)] (H2DPP = 5,15-diphenylporphyrin; dppe = 1,2-bis(diphenylphosphino)ethane) with diphenylphosphine oxide also results in the desired mono-porphyrinylphosphine oxide [H2DPP-P(O)Ph2]. Attempts to isolate the tertiary phosphines failed due to their extreme air-sensitivity. Variable temperature 1H NMR studies of [H2DPP-P(O)Ph2] revealed an intrinsic lack of symmetry, while fluorescence spectroscopy showed that the phosphine oxide group does not behave as a "heavy atom" quencher. The electron withdrawing effect of the phosphine oxide group was confirmed by voltammetry. The ligands were characterised by multinuclear NMR and UV-visible spectroscopy as well as mass spectrometry. Single crystal X-ray crystallography showed that the bis(phosphine oxide) nickel(II) complex {[NiDPP-[P(O)Ph2]2} is monomeric in the solid state, with a ruffled porphyrin core and the two P=O fragments on the same side of the average plane of the molecule. On the other hand, the corresponding zinc(II) complex formed infinite chains through coordination of one Ph2PO substituent to the neighbouring zinc porphyrin through an almost linear P=O---Zn unit, leaving the other Ph2PO group facing into a parallel channel filled with disordered water molecules. These new phosphine oxides are attractive ligands for supramolecular porphyrin chemistry.

Nanostructured metal oxides as adsorbents and photocatalysts

This research underlines the extensive application of nanostructured metal oxides in environmental systems such as hazardous waste remediation and water purification. This study tries to forge a new understanding of the complexity of adsorption and photocatalysis in the process of water treatment. Sodium niobate doped with a different amount of tantalum, was prepared via a hydrothermal reaction and was observed to be able to adsorb highly hazardous bivalent radioactive isotopes such as Sr2+ and Ra2+ions. This study facilitates the preparation of Nb-based adsorbents for efficiently removing toxic radioactive ions from contaminated water and also identifies the importance of understanding the influence of heterovalent substitution in microporous frameworks. Clay adsorbents were prepared via a two-step method to remove anionic and non-ionic herbicides from water. Firstly, layered beidellite clay was treated with acid in a hydrothermal process; secondly, common silane coupling agents, 3-chloro-propyl trimethoxysilane or triethoxy silane, were grafted onto the acid treated samples to prepare the adsorption materials. In order to isolate the effect of the clay surface, we compared the adsorption property of clay adsorbents with ƒ×-Al2O3 nanofibres grafted with the same functional groups. Thin alumina (£^-Al2O3) nanofibres were modified by the grafting of two organosilane agents 3-chloropropyltriethoxysilane and octyl triethoxysilane onto the surface, for the adsorptive removal of alachlor and imazaquin herbicides from water. The formation of organic groups during the functionalisation process established super hydrophobic sites along the surfaces and those non-polar regions of the surfaces were able to make close contact with the organic pollutants. A new structure of anatase crystals linked to clay fragments was synthesised by the reaction of TiOSO4 with laponite clay for the degradation of pesticides. Based on the Ti/clay ratio, these new catalysts showed a high degradation rate when compared with P25. Moreover, immobilized TiO2 on laponite clay fragments could be readily separated out from a slurry system after the photocatalytic reaction. Using a series of partial phase transition methods, an effective catalyst with fibril morphology was prepared for the degradation of different types of phenols and trace amount of herbicides from water. Both H-titanate and TiO2-(B) fibres coated with anatase nanocrystal were studied. When compared with a laponite clay photocatalyst, it was found that anatase dotted TiO2-(B) fibres prepared by a 45 h hydrothermal treatment followed by calcination were not only superior in performance in photocatalysis but could also be readily separated from a slurry system after photocatalytic reactions. This study has laid the foundation for the development of the ability to fabricate highly efficient nanostructured solids for the removal of radioactive ions and organic pollutants from contaminated water. These results now seem set to contribute to the development of advanced water purification devices in the future. These modified nanostructured materials with unusual properties have broadened their application range beyond their traditional use as adsorbents, to also encompass the storage of nuclear waste after concentrating from contaminated water.

Cholesterol enhances phospholipid-dependent activated protein C anticoagulant activity

The influence of cholesterol on activated protein C (APC) anticoagulant activity in plasma and on factor Va inactivation was investigated. Anticoagulant and procoagulant activities of phosphatidylcholine/phosphatidylserine (PC/PS) vesicles containing cholesterol were assessed in the presence and absence of APC using factor Xa-1-stage clotting and factor Va inactivation assays. Cholesterol at approximate physiological membrane levels (30%) in PC/PS (60%/10% w/w) vesicles prolonged the factor Xa-1-stage clotting time dose-dependently in the presence of APC but not in the absence of APC. APC-mediated cleavage of purified recombinant factor Va variants that were modified at specific APC cleavage sites (Q306/Q679-factor Va; Q506/Q679-factor Va) was studied to define the effects of cholesterol on APC cleavage at R506 and R306. When compared to control PC/PS vesicles, cholesterol in PC/PS vesicles enhanced factor Va inactivation and the rate of APC cleavage at both R506 and R306. Cholesterol also enhanced APC cleavage rates at R306 in the presence of the APC cofactor, protein S. In summary, APC anticoagulant activity in plasma and factor Va inactivation as a result of cleavages at R506 and R306 by APC is markedly enhanced by cholesterol in phospholipid vesicles. These results suggest that cholesterol in a membrane surface may selectively enhance APC activities. © 2005 International Society on Thrombosis and Haemostasis.