Lilian Regina Barros Mariutti

Development of a novel micro-assay for evaluation of peroxyl radical scavenger capacity: Application to carotenoids and structure–activity relationship

A micro-assay was developed and validated, using a microplate reader in 96-well format, C(11)-BODIPY(581/591) as fluorescent probe and AIBN as ROO() generator. The structure-activity relationship was established for 15 carotenoid standards, indicating that the opening of the β-ionone ring and the increase of chromophore extension in the carotenoid structure were the major factors leading to the increase of ROO() scavenging capacity. The values for ROO() scavenging capacity were calculated using α-tocopherol as reference compound. Among the studied carotenoids, all-trans-lycopene was the most efficient ROO() scavenger (8.67±0.74) followed by all-trans-astaxanthin (6.50±0.62). All the carotenoids showed to be more effective ROO() scavengers than α-tocopherol and some hydrophilic compounds. Finally, the method was successfully applied to assay the ROO() scavenging capacity of carotenoid extracts from two Amazonian fruits, peach palm (7.83±0.21) and mamey (6.90±0.44).

Carotenoids and Phenolic Compounds from Solanum sessiliflorum, an Unexploited Amazonian Fruit, and Their Scavenging Capacities against Reactive Oxygen and Nitrogen Species

The composition of carotenoids and phenolic compounds from mana-cubiu (Solanum sessiliflorum), a fruit native to Amazonia, was determined by high-performance liquid chromatography coupled to diode array and mass spectrometry detectors (HPLC-DAD-MS(n)). The antioxidant capacities of the hydrophilic and carotenoid extracts against some reactive oxygen (ROO(•), H(2)O(2), HOCl, and HO(•)) and nitrogen (ONOO(-)) species were also determined. Seventeen carotenoids and three phenolic compounds were found in mana-cubiu. The major carotenoids were (all-E)-β-carotene (7.15 μg/g of dry weight) and (all-E)-lutein (2.41 μg/g of dry weight). The 5-caffeoylquinic acid (1351 μg/g of dry weight) was the major phenolic compound, representing more than 78% (w/w) of the total phenolic compounds. Moreover, two dihydrocaffeoyl spermidines were found in the hydrophilic extract. Both mana-cubiu extracts were able to scavenge all the tested reactive species. The carotenoid extract was shown to be a potent scavenger of peroxyl radical, while the hydrophilic extract was a potent hydrogen peroxide and hypochlorous acid scavenger.

Lipid and Cholesterol Oxidation in Chicken Meat Are Inhibited by Sage but Not by Garlic

UNLABELLED The effects of the addition of sage and garlic in chicken meat on lipid and cholesterol oxidation, having as prooxidant factors the addition of salt, thermal treatment, and frozen storage, were evaluated. The content of unsaturated fatty acids did not change in the presence of sage; on the contrary, with garlic, the content of these fatty acids decreased after cooking and storage. Hexanal and pentanal contents were lower in patties containing sage, and higher in those with garlic. The 7-ketocholesterol was the cholesterol oxide found in higher amount in raw chicken on day 0, while the formation of 7β- and 7α-hydroxycholesterol was verified only from day 30 on. Cooking and storage resulted in increase of total cholesterol oxides and decrease of α- and γ-tocopherol. Sage was effective in controlling lipid and cholesterol oxidation, minimizing the prooxidant effects of salt, cooking, and storage. However, garlic presented no effect as antioxidant and accelerated lipid oxidation. PRACTICAL APPLICATION The addition of sage to chicken meat (0.1 g/100 g) is a good alternative to prevent and delay the formation of compounds derived from lipid oxidation that are responsible for off-flavors and loss of nutritional quality during long-term frozen storage. Care must be taken when using garlic to seasoning chicken meat products, such as hamburgers and meatballs, especially cooked or precooked due to its potential to promote lipid oxidation and consequently raising the risk of having the product rejected by the consumer.

Effect of Annatto Seed and Coriander Leaves as Natural Antioxidants in Fish Meatballs during Frozen Storage

UNLABELLED The effects of annatto (0.1 g/100g) and coriander (0.5 g/100g) were assessed against lipid oxidation in white hake meatballs cooked in boiling water (95 ± 1 °C) for 30 min and stored at -18 °C for 120 d. The fatty acids (FA) and the nutritional quality, cholesterol, cholesterol oxides, thiobarbituric acid reactive substances (TBARS) values, and conjugated dienes were analyzed to follow the course of oxidation. Annatto and coriander were efficient in the control of lipid oxidation, also preserving the essential FA. At 120 d of storage, the eicosapentaenoic acid (EPA) concentration decreased respectively by 43%, 32%, 12%, and 9% in the control, coriander, annatto, and annatto + coriander patties. For docosahexaenoic acid (DHA), these concentrations decreased, respectively, 44%, 30%, 11%, and 7%, revealing a probable synergistic effect among the antioxidant compounds present in both spices. On the other hand, annatto and coriander were not able to act protecting the meatballs against lipid oxidation when they were cooked, also not exerted any effect in the cholesterol oxidation. PRACTICAL APPLICATION Spices, especially coriander and annatto, can be an alternative to substitute synthetic additives with antioxidants to prevent loss of important unsaturated fatty acids (UFA) in fish meatballs during frozen storage for 120 d. The maximum effect was observed when 0.5% coriander and 0.1% annatto were used in combination. Cooking did not induce the formation of cholesterol oxides, compounds that can cause more health damages than cholesterol itself; however, during storage the cholesterol oxides levels presented a little increase regardless of spice addition.

Optimization and validation of analytical conditions for cholesterol and cholesterol oxides extraction in chicken meat using response surface methodology.

The analytical conditions for the extraction of cholesterol and cholesterol oxides in chicken meat were optimized by means of response surface methodology. The separation and identification were performed by normal phase HPLC using UV and refractive index (RI) detectors, and the confirmation of the 11 cholesterol oxides identities in the samples was verified by HPLC-APCI-MS. The developed methodology showed good analytical performance, presenting recovery levels from 84 to 103% and detection limits varying from 0.01 to 0.06 microg/g for UV detection and from 1.98 to 2.12 microg/g for RI detection. The present study demonstrated the presence of 22 R-hydroxycholesterol, 24 S-hydroxycholesterol, and 22 S-hydroxycholesterol for the first time in chicken meat.

Further Insights on the Carotenoid Profile of the Echinoderm Marthasterias glacialis L

In this study, the carotenoid profile of the echinoderm Marthasterias glacialis L. was established using HPLC-DAD-APCI-MS/MS equipped with a C30 column. This approach rendered the identification of 20 compounds, eight of them reported for the first time in this marine organism. Differentiation of carotenoid isomers was also achieved.

Influence of salt on lipid oxidation in meat and seafood products: A review

Sodium chloride, commonly known as salt, is a widely used additive in food industry due to its preservation and antimicrobial properties provided by its ability to reduce water activity. Moreover, the addition of salt to meat and seafood aims at improving water retention capacity and enhancing flavor due to its influence on the activity of some enzymes responsible for flavor development. On the other hand, salt added in meat and seafood can favor lipid oxidation, which is one of the main responsibles for quality losses in the food industry. In this review, the main mechanisms of fatty acids and cholesterol oxidation are described as well as the influence of salt on lipid oxidation in meat and seafood. Besides, the possible mechanisms of the pro-oxidant action of sodium chloride are presented and potential solutions to inhibit the salt action in lipid oxidation and decrease the salt content in food are discussed.

Carotenoid esters in foods - A review and practical directions on analysis and occurrence

Carotenoids are naturally found in both free form and esterified with fatty acids in most fruits and some vegetables; however, up to now the great majority of studies presents data on carotenoid composition only after saponification. The reasons for this approach are that a single xanthophyll can be esterified with several different fatty acids, generating a great number of different compounds with similar chemical and structural characteristics, thus, increasing the complexity of analysis compared to the respective saponified extract. This means that since UV/Vis spectrum does not change due to esterification, differentiation between free and acylated xanthophylls is dependent at least on elution order and mass spectrometry (MS) features. The presence of interfering compounds, especially triacylglycerides (TAGs), in the non-saponified extract of carotenoids can also impair carotenoid ester analyses by MS due to high background noise and ionization suppression since TAGs can be present in much higher concentrations than the carotenoid esters. This leads to the need of development of new and effective clean-up procedures to remove the potential interferents. In addition, only few standards of xanthophyll esters are commercially available, making identification and quantification of such compounds even more difficult. Xanthophyll esterification may also alter some properties of these compounds, including solubility, thermostability and bioavailability. Considering that commonly consumed foods are dietary sources of xanthophyll esters and that it is the actual form of ingestion of such compounds, an increasing interest on the native carotenoid composition of foods is observed nowadays. This review presents a compilation of the current available information about xanthophyll ester analyses and occurrence and a practical guide for extraction, pre-chromatographic procedures, separation and identification by liquid chromatography-tandem mass spectrometry (LC-MS/MS).

An in vitro digestion method adapted for carotenoids and carotenoid esters: moving forward towards standardization

In vitro digestion methods are a useful approach to predict the bioaccessibility of food components and overcome some limitations or disadvantages associated with in vivo methodologies. Recently, the INFOGEST network published a static method of in vitro digestion with a proposal for assay standardization. The INFOGEST method is not specific for any food component; therefore, we aimed to adapt this method to assess the in vitro bioaccessibility of carotenoids and carotenoid esters in a model fruit (Byrsonima crassifolia). Two additional steps were coupled to the in vitro digestion procedure, centrifugation at 20 000g for the separation of the aqueous phase containing mixed micelles and exhaustive carotenoid extraction with an organic solvent. The effect of electrolytes, enzymes and bile acids on carotenoid micellarization and stability was also tested. The results were compared with those found with a simpler method that has already been used for carotenoid bioaccessibility analysis. These values were in the expected range for free carotenoids (5-29%), monoesters (9-26%) and diesters (4-28%). In general, the in vitro bioaccessibility of carotenoids assessed by the adapted INFOGEST method was significantly higher (p < 0.05) than those assessed by the simplest protocol, with or without the addition of simulated fluids. Although no trend was observed, differences in bioaccessibility values depended on the carotenoid form (free, monoester or diester), isomerization (Z/E) and the in vitro digestion protocol. To the best of our knowledge, it was the first time that a systematic identification of carotenoid esters by HPLC-DAD-MS/MS after in vitro digestion using the INFOGEST protocol was carried out.

The Amazonian fruit Byrsonima crassifolia effectively scavenges reactive oxygen and nitrogen species and protects human erythrocytes against oxidative damage

A hydrophilic extract of murici (Byrsonima crassifolia), a fruit native to the North and Northeast regions of Brazil, was evaluated in relation to its phenolic composition and in vitro antioxidant potential against some physiologically relevant reactive oxygen and nitrogen species. Additionally, the protective effect of murici extract against peroxyl radical (ROO)-induced toxicity to human erythrocytes was also determined. The major phenolic compound, determined by HPLC-DAD-MSn, was quercetin (2.72±0.35μg/mL). The extract was able to scavenge ROO (0.30±0.04μmoltroloxequivalent/mg), hypochlorous acid (IC50=10.0±0.1μg/mL), hydroxyl radical (IC50=7±1μg/mL) and peroxynitrite anion (IC50=21.0±0.6μg/mL and 17.0±1.6μg/mL, respectively, in absence and presence of NaHCO3). Human erythrocytes were subjected to oxidative damage, but murici extract was not able to inhibit hemolysis, even at the highest tested concentration. On the other hand, the extract inhibited hemoglobin oxidation (IC50=271±44μg/mL), lipid peroxidation (1000μg/mL) by 48±5%, depletion of glutathione (100μg/mL) by 49±2% and formation of its oxidized form (100μg/mL) by 96±4%.