Lipid peroxidation and its measurement
Roger L. McMullen in Antioxidants and the Skin, 2018
The formation of secondary lipid oxidation products, such as ketones and aldehydes, can be monitored by following their reaction with 2,4-dinitrophenylhydrazine to form 2,4-dinitrophenylhydrazones (Figure 5.13). This test for lipid oxidation decomposition products, like many of the assays for bulk lipids, has been in use for more than 100 years.40,141,146,147 The formation of 2,4-dinitrophenylhydrazones is normally followed with UV-visible spectrophotometry.148
Maple syrup urine disease (branched-chain oxoaciduria)
William L. Nyhan, Georg F. Hoffmann, Aida I. Al-Aqeel, Bruce A. Barshop in Atlas of Inherited Metabolic Diseases, 2020
Screening for the disease has been carried out by the addition of 2, 4-dinitrophenylhydrazine, which produces a yellow precipitate of dinitrophenylhydrazones [81] in the presence of oxoacids. The individual oxoacids have been distinguished by gas chromatography-mass spectrometry (GCMS) of the oximes [82]. The ferric chloride test on the urine may yield a greenish-gray color.
Ultraviolet and Light Absorption Spectrometry
Adorjan Aszalos in Modern Analysis of Antibiotics, 2020
Some colorimetric assay procedures have been also described [338—340] for griseofulvin. Amer et al. [339] applied 2,4-dinitrophenylhydrazine and measured the light absorption of the colored phenylhydrazone derivative. Additional methods [336,341,342] are generally based on the reactions of the α, β-unsaturated carbonyl group.
Omega-3 fatty acid attenuates oxidative stress in cerebral cortex, cerebellum, and hippocampus tissue and improves neurobehavioral activity in chronic lead-induced neurotoxicity
Published in Nutritional Neuroscience, 2019
Pramod Kumar Singh, Manish Kumar Singh, Rajesh Singh Yadav, Rajendra Nath, Anju Mehrotra, Akash Rawat, Rakesh Kumar Dixit
Protein carbonyl content (PCC) in brain regions was measured by the method of Levine et al.,44 using 2,4 – dinitrophenylhydrazine (DNPH) as a substrate. Brain regions individually were homogenized in phosphate buffer (50 mM, pH 7.4, 10% w/v) and centrifuged at 11 000×g for 15 min. The resulting supernatant was used for reaction with DNPH. The supernatant was divided into two equal volumes for each sample. Following this, four volumes of 10 mM DNPH dissolved in 2 M HCl were added to one of the sample pairs. Four volumes of 2 M HCl were also added to the other tube that served as a reagent blank assay. Following this, samples were incubated for 1 h at room temperature in the dark with continuous stirring and precipitated with an equal volume of 20% TCA. The tubes were kept for 10 min on ice and samples were centrifuged at 3000×g for 5 min. The supernatant was discarded and protein pellet washed in 10% TCA once and in ethanol/ethyl acetate (1:1) three times to remove free DNPH and additional lipid contaminants. Final protein precipitate was dissolved in 6 M guanidine hydrochloride solution. The difference in absorbance between the DNPH-treated and the HCl-treated samples was determined on spectrophotometer at 375 nm and the amount of carbonyl content (C) was calculated using a molar extinction coefficient (ε) of 22.0 mM−1 cm−1 for aliphatic hydrazones.
Effects of chronic treatment with gold nanoparticles on inflammatory responses and oxidative stress in Mdx mice
Published in Journal of Drug Targeting, 2020
Daniela Pacheco dos Santos Haupenthal, Jonathann Corrêa Possato, Rubya Pereira Zaccaron, Carolini Mendes, Matheus Scarpatto Rodrigues, Renata Tiscoski Nesi, Ricardo Aurino Pinho, Paulo Emilio Feuser, Ricardo Andrez Machado-de-Ávila, Clarissa M. Comim, Paulo Cesar Lock Silveira
Protein carbonyl content was measured using labelled protein-hydrazone derivatives with 2,4-dinitrophenylhydrazine. These derivatives were extracted with trichloroacetic acid followed by treatment with ethanol/ethyl acetate and dissolved in urea hydrochloride. Concentrations of incorporated 2,4-dinitrophenylhydrazine were measured by the absorbance at 370 nm and are shown as per milligram of protein [22]. Total thiol content was determined using 5,5′-dithiobis(2-nitrobenzoic acid) and quantified according to the absorbance at 412 nm using a spectrophotometer [23]. Formation of malondialdehyde (MDA) was measured using high-performance liquid chromatography (Prominence Shimadzu) on an Ascentis® C18 column (250 × 2.1 mm, 5 μm, Supelco Sigma-Aldrich, Bellefonte, PA), according to the protocol of Grotto et al. [24].
Antifouling activity of portimine, select semisynthetic analogues, and other microalga-derived spirocyclic imines
Published in Biofouling, 2018
Darby G. Brooke, Gunnar Cervin, Olivier Champeau, D. Tim Harwood, Henrik Pavia, Andrew I. Selwood, Johan Svenson, Louis A. Tremblay, Patrick L. Cahill
Detailed procedures for producing the semisynthetic derivatives are provided in Supplemental material. To access 5, the cyclic imine of 1 was reduced using NaBH4, as 1 proved unexpectedly impervious to reduction with NaCNBH3 in preliminary trials, akin to the non-susceptibility of the cyclic imine of pinnatoxin E to reduction with this reagent (Selwood et al. 2010). A mixture of 6 and 7 was produced by exposing 1 to acetylation conditions, whilst the reducing agent PtO2 was used to produce a mixture of alkenes and alkanes 8–10. In addition to the imine group, 1 contains a ketone group at the C-14 position that would be expected to be susceptible to hydrogenation. To confirm this group was intact in 5 and 8–10, solutions of the derivates were spotted on SiO2 TLC plates and treated with acidic 2,4-dinitrophenylhydrazine. If the spot became orange or yellow, this confirmed that the molecule (5, and the mixture of 8–10) had contained a ketone group, which had been transformed to its corresponding, coloured 2,4-dinitrophenylhydrazone by exposure to acidic 2,4-dinitrophenylhydrazine (Oscar 1926).
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