A modified xylenol orange method to evaluate formation of lipid hydroperoxides during storage and processing of small pelagic fish

Eymard, Sylvie; Génot, Claude

doi:10.1002/ejlt.200300768

Cited by 55 publications

(37 citation statements)

References 11 publications

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“…Depending on the nature of the sample, different ratios of methanol/water in the FOX reagent have been used to enhance hydroperoxide solubility and avoid turbidity in the assay (19,20). The results (Fig.…”

Section: Resultsmentioning

confidence: 98%

Optimization of enzymatic assay for the measurement of lipoxygenase activity in organic solvent media

Vega

Karboune

Husson³

et al. 2005

J Americ Oil Chem Soc

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Lipoxygenases (LOX; EC 1.13.11.12) are an important class of non-heme iron enzymes that catalyze the di-oxidation of PUFA to hydroperoxy FA, which can be measured by the xylenol orange method (FOX). To determine the enzymatic production of these FA in organic solvent media, the FOX assay was optimized using the standard cumene hydroperoxide. An increase in the proportion of methanol from 0 to 75% in the FOX reagent resulted in a 93% increase in the molar absorption coefficients at 560 nm. In addition, the presence of linoleic acid in the cumene hydroperoxide sample enhanced the formation of the FOX complex, resulting in a 50% increase in the sensitivity of the method. Moreover, when perchloric acid was used, the source of ferrous ions and presence of denatured LOX had little effect on the sensitivity of the FOX assay whereas sensitivity decreased by 40-46% with sulfuric acid. The overall results demonstrated that the modified FOX assay may be used for the precise and accurate measurement of hydroperoxy FA obtained by LOX activity in organic solvent media.Paper no. J11120 in JAOCS 82, 817-823 (November 2005).KEY WORDS: FOX assay, lipid hydroperoxides, lipoxygenase, organic solvent media, xylenol orange.Lipoxygenases (LOX; EC 1.13.11.12) are an important class of non-heme iron enzymes that catalyze the regio-and stereospecific di-oxidation of PUFA containing a cis,cis-1,4-pentadiene moiety to hydroperoxy-FA, considered as precursors of flavor compounds. In addition, LOX are ubiquitously found in plants, microorganisms, and various animal tissues, where they are implicated in physiological activities (1). Most spectrophotometric methods reported for the determination of LOX activity were developed for an aqueous medium, with the most common being based on the absorption of hydroperoxy-FA containing a cis,cis-1,4-pentadiene moiety at 234 nm (2); however, this method remains limited by its inability to detect hydroperoxides that lack a conjugated diene chromophore (3). The ferrous oxidation assay, using xylenol orange (FOX) or ferrous thiocyanate, was reported as an alternative spectrophotometric method for the determination of lipid hydroperoxides, with the latter assay showing lower sensitivity compared with the FOX assay (4) and used for the quantification of high amounts of lipid hydroperoxides.The FOX assay is based on the oxidation of ferrous ions (Fe 2+ ) by hydroperoxides into their ferric counterparts (Fe 3+ ), which, in turn complex with the xylenol orange salt to form a chromophore that absorbs at 560 nm (5), with a molar absorption coefficient (ε 560 ) of 4.5 × 10 5 M −1 cm −1 for lipid hydroperoxides in a methanol-based reagent (6). To measure hydroperoxides in liposome or lipoprotein suspensions, Jiang et al. (7) also used methanol in the preparation of the FOX reagent. In addition, the specificity of the FOX assay to measure lipid hydroperoxides in an extract can be improved by the addition of triphenylphosphine (TPP) (8,9), and its sensitivity can be increased by the use of sucrose (10) or the replacem...

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Section: Resultsmentioning

confidence: 98%

Optimization of enzymatic assay for the measurement of lipoxygenase activity in organic solvent media

Vega

Karboune

Husson³

et al. 2005

J Americ Oil Chem Soc

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“…Brain lipid hydroperoxides were assayed using the previous method of Eymand and Genot (15), with ferrous ammonium sulfate and xylenol orange (3,3'-Bis[N,N-bis(carboxymethyl)-aminomethyl]-o-cresolsulfonephthalein). The color that developed was read at 560 nm.…”

Section: Animals and Dietsmentioning

confidence: 99%

A comparison between the impact of two types of dietary protein on brain glucose concentrations and oxidative stress in high fructose-induced metabolic syndrome rats

Madani

Malaisse²,

Ait-Yahia

2015

Biomedical Reports

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Abstract. The present study explored the potential of fish proteins to counteract high glucose levels and oxidative stress induced by fructose in the brain. A total of 24 male Wistar rats consumed sardine protein or casein with or without high fructose (64%). After 2 months, brain tissue was used for analyses. The fructose rats exhibited an increase in body mass index (BMI), body weight, absolute and relative brain weights and brain glucose; however, there was a decrease in food and water intake. Fructose disrupts membrane homeostasis, as evidenced by an increase in the brain hydroperoxides and a decrease in catalase (CAT) and glutathione peroxidase (GSH-Px) compared to the control. The exposure to the sardine protein reduced BMI, food intake, glucose and hydroperoxides, and increased CAT and GSH-Px in the brain. In conclusion, the metabolic dysfunctions associated with the fructose treatment were ameliorated by the presence of sardine protein in the diet by decreasing BMI, brain glucose and lipid peroxidation, and increasing CAT and GSH-Px activities.

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“…They determined the absolute values of lipid hydroperoxides in different experimental groups of chickens. Eymard et al [56] modified the FOX1 method used by Hermes-Lima et al [96] for the determination of lipid hydroperoxides in small pelagic fish. They used methanol extracts of ground tissues of the Atlantic horse mackerel (Trachurus trachurus Linnaeus).…”

Section: Measurement Of Lipid Peroxidation In Animal Tissuesmentioning

confidence: 99%

Automation of Methods for Determination of Lipid Peroxidation

Sochor¹,

Ruttkay-Nedecký²,

Babula³

et al. 2012

Lipid Peroxidation

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A modified xylenol orange method to evaluate formation of lipid hydroperoxides during storage and processing of small pelagic fish

Cited by 55 publications

References 11 publications

Optimization of enzymatic assay for the measurement of lipoxygenase activity in organic solvent media

Optimization of enzymatic assay for the measurement of lipoxygenase activity in organic solvent media

A comparison between the impact of two types of dietary protein on brain glucose concentrations and oxidative stress in high fructose-induced metabolic syndrome rats

Automation of Methods for Determination of Lipid Peroxidation

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