Kinetics and mechanism of the reaction of nitrous acid with 2,4-dinitrophenylhydrazine

Bernheim, Pierre; Dobos, Agnes; Doherty, Anne M. M.; Haine, Neil; Stedman, Geoffrey

doi:10.1039/p29960000275

Cited by 13 publications

(4 citation statements)

References 5 publications

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“…The ~ 100% efficiency of the ntrDNPH assay in removing unreacted DNPH may be due to the following reasons: Wash of unreacted DNPH by EA:hexane is performed with the protein in solution, at protein denaturing conditions (by urea and SDS), and at pH 7.0. Here, DNPH is uncharged and more easily soluble in organic solvents than when positively charged (as DNP-N 2 H 5 [54] , [55] ) at the near zero pH used by the stdDNPH assay. This together with the hydrogen bonding potential of the DNP-N 2 H 5 nitro groups (acting as proton acceptors in hydrogen bonding [88] ) will promote DNPH non-specific binding to proteins, resulting in at least 30% overestimation of protein carbonyls by the stdDNPH assay.…”

Section: Methods Proceduresmentioning

confidence: 99%

“…This limitation is an important factor of the high variability in the data obtained by the stdDNPH assay [32] . Any interfering unreacted DNPH that is non-specifically associated with the TCA(pKa 0.7)-pelleted protein is expected to be present as hydrazinium cation [54] , [55] . Being at a such highly polarized state, DNPH’s complete solubilization in the EtOH:EA organic mixture used by the stdDNPH assay’s wash procedure is greatly reduced and its removal from the pelleted protein is inefficient.…”

Section: Introductionmentioning

confidence: 99%

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Protein and cell wall polysaccharide carbonyl determination by a neutral pH 2,4-dinitrophenylhydrazine-based photometric assay

et al. 2018

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A new 2,4-dinitrophenylhydrazine (DNPH)-based photometric assay is developed for the quantification of carbonyls in protein samples from any biological source by protein carbonyl-DNPH hydrazone formation at acidic pH in the presence of denaturing urea, and subsequent hydrazone solubilization in the presence of SDS and stabilization from acid hydrolysis at pH 7.0. At this neutral (ntr) pH, interfering unreacted DNPH is uncharged and its thus increased hydrophobicity permits its 100% effective removal from the solubilizate with ethyl acetate/hexane wash. The ntrDNPH assay is more reliable and sensitive than the standard (std) DNPH photometric assay because it eliminates its main limitations: (i) interfering unreacted DNPH (pKa 1.55) that is nonspecifically bound to the TCA (pKa 0.7)-protein pellet is not effectively removed after wash with EtOH: ethyl acetate because it is positively charged, (ii) acid (TCA-induced) hydrolysis of the protein carbonyl-DNPH hydrazone, (iii) sample protein concentration re-determination, (iv) loss of sample acid (TCA)-soluble proteins, (v) DNA interference, and (vi) requires high protein quantity samples (≥ 1 mg). Considering ntrDNPH assay’s very low protein limit (1 µg), its cumulative and functional sensitivities are 2600- and 2000-fold higher than those of the stdDNPH assay, respectively. The present study elucidates the DNA interference mechanism on the stdDNPH assay, and also develops a standardized protocol for sample protein treatment and fractionation (into cytoplasmic/aqueous, membrane/lipid-bound, and histone/DNA-bound proteins; see Supplement section V) in order to ensure reproducible carbonyl determination on defined cell protein fractions, and to eliminate assay interference from protein samples containing (i) Cys sulfenic acid groups (via their neutralization with dithiothreitol), and (ii) DNA (via its removal by streptomycin sulfate precipitation). Lastly, the ntrDNPH assay determines carbonyl groups on cell wall polysaccharides, thus paving the way on studies to investigate cell walls acting as antioxidant defense in plants, fungi, bacteria and lichens.

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Section: Methods Proceduresmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Protein and cell wall polysaccharide carbonyl determination by a neutral pH 2,4-dinitrophenylhydrazine-based photometric assay

et al. 2018

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“…Many factors affecting the yield and purity of Curtius rearrangement products have been reported, including reaction kinetics, − solvents, ,,,, temperature, ,,− calorimetry, , energy release, residence time, , and analytics . We found that the use of toluene provided optimal rearrangement conditions at 85 °C. ,, Because of the potentially explosive and hazardous nature of acyl azide intermediates formed in situ, appropriate concentration of acyl hydrazide, reactor volume and flow, and residence time should be carefully considered to control the energy release.…”

mentioning

confidence: 87%

Preparation of Mono- and Diisocyanates in Flow from Renewable Carboxylic Acids

Hai

Backer

Cosford

et al. 2020

Org. Process Res. Dev.

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Diisocyanates used in polyurethanes are commonly prepared by phosgenation of petroleum-sourced diamines. This involves highly toxic phosgene and produces corrosive HCl, limiting synthetic applications. In our search for a renewable source for diisocyanates, we have developed a practical methodology for the production of isocyanates from algae-biomass-derived fatty acids or other renewable sources. This technique utilizes flow chemistry to prepare and convert high-energy intermediates, thus mitigating safety concerns. By the use of continuous flow, acyl azides are prepared from hydrazides and subsequently heated to undergo Curtius rearrangement, affording isocyanates in one scalable process. The method is efficient, safe, and sustainable, offers an opportunity to prepare isocyanates and diisocyanates from renewable feedstocks, and is amenable to distributed manufacturing processes.

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“…In order to improve the detection procedure, other probe systems are required. In 1996, Bernheim and co‐workers [40] reported that arylhydrazines react effectively with nitrous acid, and in 1999 Büldt and Karst [41] applied the nonfluorescent hydrazine derivative N ‐methyl‐4‐hydrazino‐7‐nitrobenzofurazan (MNBDH) for the quantification of nitrite as low as 50 n m in acidic solution. The subsequently generated fluorescent dye N ‐methyl‐4‐amino‐7‐nitrobenzofurazan (MNBDA) has a sensitivity limit of 5 n m in nonaqueous solution.…”

Section: Quantitative Determination Of N‐nitrosotryptophan Derivativesmentioning

confidence: 99%

N‐Nitrosomelatonin: synthesis, chemical properties, potential prodrug

Kirsch

Groot

2009

Journal of Pineal Research

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: Melatonin is easily nitrosated via various mechanisms at the nitrogen atom of the indole ring to give N‐nitrosomelatonin (NOMela). This Mini‐review provides a comprehensive view of this N‐nitroso compound. With an improved procedure NOMela can now economically synthesized with low laboratory expenditure. The major chemical property of NOMela, i.e. the (formally) transfer of the NO+ function to its target nucleophile, is explained in detail and a variety of detection methods using this reaction are suggested. As the suspected carcinogenical potential of NOMela is clearly overruled it seems attractive to apply this nitroso compound for endogenous generation of S‐nitrosothiols that act as nitric oxide donors in vivo.

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Kinetics and mechanism of the reaction of nitrous acid with 2,4-dinitrophenylhydrazine

Cited by 13 publications

References 5 publications

Protein and cell wall polysaccharide carbonyl determination by a neutral pH 2,4-dinitrophenylhydrazine-based photometric assay

Protein and cell wall polysaccharide carbonyl determination by a neutral pH 2,4-dinitrophenylhydrazine-based photometric assay

Preparation of Mono- and Diisocyanates in Flow from Renewable Carboxylic Acids

N‐Nitrosomelatonin: synthesis, chemical properties, potential prodrug

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