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Standard of formaldehyde (36% methanol stabilized solution) and 2,4-dinitrophenylhydrazine (99%) were purchased form Sigma Aldrich.
Kunugita, "Derivatization of carbonyl compounds with 2,4-dinitrophenylhydrazine and their subsequent determination by high-performance liquid chromatography," Journal of Chromatography B, vol.
These tests included 2,4-dinitrophenylhydrazine (carbonyl group), anthrone (carbohydrates), Baljet (sesquiterpene lactones), sodium bicarbonate (carboxyl group), ferric chloride (tannins), Dragendorff (alkaloids), Liebermann-Burchard (sterols and triterpenes), potassium permanganate (double bonds), Shinoda (flavonoids), sodium hydroxide (coumarins), Borntrager (quinones), and sodium bicarbonate (saponins).
Vitamin C was extracted with 0.5% oxalic acid, under stirring, and after filtration the determination in the extract was carried out, using 2,4-dinitrophenylhydrazine and using ascorbic acid as the standard.
2,4-Dinitrophenylhydrazine, the most popular derivatization reagent, has been used before determination of formaldehyde by HPLC [11, 15, 22].
The supernatant was incubated for 15min with 500[micro]L of 2,4-dinitrophenylhydrazine (10 mM DNFH prepared in 2M HCl).
Briefly, to detect LDH capacity, the extract was incubated with the substrate buffer and coenzyme I at 37[degrees]C for 15 min, and then was color-rendered using 2,4-dinitrophenylhydrazine [24].
Briefly, the membranes were equilibrated in 20% methanol in TBS, washed for 5min with 10% trifluoroacetic acid (TFA), derivatized with 5 mM 2,4-dinitrophenylhydrazine (DNPH, Sigma-Aldrich) diluted in 10% TFA for 10min (protected from light), washed with 10% TFA to remove the excess of DNPH, and finally washed with 50% methanol.
The pellet was resuspended in 100 [micro]L of 0.2 M NaOH and the samples were incubated for 1 hour at room temperature with 2 M HCl (1: 3) added to 10 mM 2,4-dinitrophenylhydrazine (DNPH).
Protein carbonyl proteins were assayed by 2,4-dinitrophenylhydrazine reaction.
Janiak, "Synthesis and spectral characterization of hydrazone schiff bases derived from 2,4-dinitrophenylhydrazine. Crystal structure of salicylaldehyde-2,4-dinitrophenylhydrazone," Zeitschrift fur Naturforschung--Section B Journal of Chemical Sciences, vol.
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