1210824-61-5 Purity
97%
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Specification
Diphenylamine (DPA) has been used as an antioxidant in lipid peroxidation studies. Using peroxidized red blood cell (RBC) membranes, it was found that DPA actually promoted lipid hydroperoxide (LOOH) formation and oxygen consumption, while significantly inhibiting the generation of thiobarbituric acid reactive substances (TBARS). DPA abolished the formation of cyclooxygenase-dependent arachidonic acid conversion products. DPA was nearly as effective as BHT in inhibiting peroxyl radical-mediated destruction of phycoerythrin fluorescence. Studies on DPA analogs suggest that the antioxidant effect of DPA lies in its secondary amine functionality. Presumably, this results in the intermediate formation of a nitrogen radical, which allows redox cycling of this aromatic amine to stimulate further peroxidation. This dramatically illustrates the danger of relying solely on TBARS measurements to assess peroxidation.
Replicate 1 using 10 volumes of buffer A (10mM HEPES, 10mM glucose, 4mM KCl, NaCl to 290mOsmol/L, pH 7.4). For treatment with 2 mM tert-butyl hydroperoxide (tBuOOH), erythrocytes were suspended in buffer A to a hematocrit of 10% and incubated at 37°C for 30 min. Peroxidation incubations were performed with or without the addition of DPA (0.01-0.1 mM) or butylated hydroxytoluene (BHT, 0.01-0.5 mM). In additional experiments, the effect of 0.1 mM DPA in this system was compared to that of an equimolar amount of a DPA analog. Since the analog compound is soluble in ethanol, an equal amount of ethanol (0.1%) was added to the control and DPA samples.
Apoptosis is a distinct morphological and biochemical entity leading to cell death that occurs in response to a variety of pathological and physiological stimuli. The apoptotic process results in cytoskeletal disruption, cell shrinkage, membrane blebbing, nuclear condensation, and internucleosomal DNA fragmentation. Multiple methods exist to determine apoptosis along this pathway. The diphenylamine assay allows for quantification of degraded DNA. The protocol is simple and allows manipulation of all types of cell cultures. The resulting colorimetric reaction is easily quantified, and the assay is highly reproducible.
Harvest monolayer cultures by scraping cells into the culture medium with a rubber spatula (or directly into a centrifuge tube for suspension cultures) and centrifuge (300g) at 4°C for 10 min to pellet the cells. Resuspend the cell pellet in 0.8 mL 10 mM PBS (pH 7.4) and 0.7 mL ice-cold lysis buffer. Transfer the cell lysate to a microcentrifuge tube and incubate on ice for 15 min. Centrifuge the lysate (13,000g) at 4°C for 15 minutes to separate fragmented DNA from high molecular weight DNA. Transfer the entire supernatant (approximately 1.5 mL containing fragmented DNA) to a 5 mL glass tube. Resuspend the pellet containing intact DNA in 1.5 mL TE and transfer again to another 5 mL glass tube. Add 1.5 mL 10% TCA to each tube and incubate at room temperature for 10 minutes. Centrifuge (500g) at 4°C for 15 minutes and discard the supernatant. Resuspend the 10% TCA pellet in 0.7 mL 5% TCA, boil (100°C) for 15 minutes, cool to room temperature, and centrifuge (300g) at 4°C for 15 minutes. Transfer 0.5 mL of supernatant to a new glass tube (do not disturb the pellet). Add 1 mL diphenylamine reagent and incubate overnight at 30°C.
This work simplified the Burton method by a one-step diphenylamine reagent pretreatment and validated the reliability and accuracy of the method by measuring four model microorganisms grown in normal medium or medium containing solid particles.
Measurement of growth by the diphenylamine colorimetric method
· To prepare the diphenylamine reagent for colorimetric assay, 1.5 g of reagent-grade diphenylamine was dissolved in 100 ml of glacial acetic acid with the addition of 1.5 ml of concentrated H2SO4.
Prior to use, 1.6% aqueous acetaldehyde was added to 100 ml of the diphenylamine reagent. It is important to note that the reagent should be colorless and stored in a dark environment.
· To quantify growth using the diphenylamine colorimetric reaction, cell pellets were collected from 1 ml cultures through centrifugation at 10,000×g for 10 minutes, followed by two washes with an appropriate buffer. Subsequently, the cell pellets were suspended in 2 ml of diphenylamine reagent and incubated at 60 °C for 1 hour.
· The supernatants from the diphenylamine reactions were then transferred to 96-well microtiter plates after centrifugation, and A595 was determined using a multifunctional microtiter plate reader. The readings obtained from this reaction system fall within the standard error range of the optical density measurements.
Diphenylamine is the monomer for the synthesis of polydiphenylamine. Studies have shown that polydiphenylamine can be an effective corrosion inhibitor for iron in 0.5 M H2SO4, with a maximum efficiency of 96% observed at a very low concentration of 10 ppm. In addition, polydiphenylamine was found to improve the passivation characteristics of iron in 0.5 M H2SO4.
Synthesis procedure of polydiphenylamine
· Reagent grade diphenylamine was purified through distillation with a small amount of zinc dust.
The sodium salt of dodecyl benzene sulphonic acid was used as a dopant and was neutralized with concentrated hydrochloric acid.
· A solution of 0.1 M freshly distilled diphenylamine in 0.1 M HCl was added to this neutralized solution, which had been pre-cooled.
· A freshly prepared solution of 0.1 M ammonium persulphate, kept at a temperature of 5-10 °C, was then slowly added to the reaction mixture with constant stirring for 2 hours.
· The polymerization efficiency was approximately 50%, and characterization was done through UV-visible and FTIR spectroscopy. The molecular weight was determined using the GPLC method.
The molecular formula of Diphenylamine is C12H11NC6H5NHC6H5.
The molecular weight of Diphenylamine is 169.22 g/mol.
Diphenylamine has been used as a fungicide for the treatment of superficial scald in apples and pears. It also has other roles such as a carotogenesis inhibitor, an antioxidant, and a radical scavenger.
No, Diphenylamine is no longer approved for use as a fungicide in the European Union.
Yes, Diphenylamine is a natural product found in Camellia sinensis, Allium cepa, and other organisms.
The IUPAC name of Diphenylamine is N-phenylaniline.
The InChI of Diphenylamine is InChI=1S/C12H11N/c1-3-7-11(8-4-1)13-12-9-5-2-6-10-12/h1-10,13H.
The InChIKey of Diphenylamine is DMBHHRLKUKUOEG-UHFFFAOYSA-N.
The Canonical SMILES of Diphenylamine is C1=CC=C(C=C1)NC2=CC=CC=C2.
The CAS number of Diphenylamine is 122-39-4.