A 10 mM stock solution of 2,4-DNPH (Sigma-Aldrich, St. Louis, MO, USA) in 100% ethanol was used to create a working solution of 0.2 mM 2,4-DNPH with 12 mL HCl (36%) per 100 mL ethanol. A working solution of 1 mM MG (Sigma) was prepared from a stock solution. The reaction consisted of 950 µL 0.2 mM 2,4-DNPH with different volumes of 1 mM MG, filled to 1 mL with distilled water for the blank, or 50 mL of media from cerebellar neuron culture. The samples and blanks were heated in an Eppendorf Thermomixer at 42 °C for 45 mins and 600 rpm, and were incubated at room temperature for 5 minutes. Spectrophotometer measurements were performed at 432 nm, according to absorbance of MG-bis-2,4-DNPH-hydrazone for calculating concentration of MG.
2 4 dnph
Manufactured by Merck Group
Sourced in United States
2,4-DNPH is a chemical compound commonly used as a reagent in analytical chemistry. It is a yellow crystalline solid that is utilized for the detection and quantification of carbonyl compounds, including aldehydes and ketones. The core function of 2,4-DNPH is to form hydrazones, which can be identified and measured using various analytical techniques.
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Lab products found in correlation
Thermomixer, by Eppendorf (1 mentions)
Acetonitrile, by Thermo Fisher Scientific (1 mentions)
Ammonium acetate, by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Acetone, by Merck Group (1 mentions)
Aluminium chloride, by Merck Group (1 mentions)
Isooctane, by Merck Group (1 mentions)
Sodium nitrite, by Merck Group (1 mentions)
Potassium iodide, by Merck Group (1 mentions)
Orthophosphoric acid, by Merck Group (1 mentions)
2 6 di tert butyl 4 methylphenol, by Merck Group (1 mentions)
Glacial acetic acid, by Merck Group (1 mentions)
Absolute ethanol, by Merck Group (1 mentions)
6 hydroxy 2 5 7 8 tetramethylchroman 2 carboxylic acid trolox, by Merck Group (1 mentions)
Anhydrous sodium sulphate, by Merck Group (1 mentions)
Soluble starch, by Merck Group (1 mentions)
Trichloroacetic acid, by Merck Group (1 mentions)
Linoleic acid, by Merck Group (1 mentions)
Sodium hydroxide (naoh), by Merck Group (1 mentions)
4 protocols using 2 4 dnph
1
Methylglyoxal Quantification Assay
2
Analysis of Carbonyl Compounds by HPLC
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Rezorian sorbent-style cartridges containing
350 mg of high-purity silica coated with 2,4-DNPH were purchased from
Sigma Aldrich (St. Louis, MO). A made-to-order calibration standard
was purchased from O2Si Smart Solutions (Charleston, SC) containing
target analytes in Acetonitrile at the following concentrations: formaldehyde
(5000 mg/L), acetaldehyde (5000 mg/L), acrolein (500 mg/L), and crotonaldehyde
(125 mg/L). An isotopically labeled analog solution was also purchased
from O2Si Smart Solutions with the following concentrations in Acetonitrile:
formaldehyde-d2-DNPH (1000 mg/L), acetaldehyde-d4-DNPH (1000 mg/L), acrolein-2,4-DNPH-3,5,6-d3 (100 mg/L),
and crotonaldehyde-2,4-DNPH-3,5,6-d3 (25 mg/L). Ammonium
acetate (molecular biology grade) was purchased from Sigma Aldrich.
Acetonitrile and water (HPLC grade) were purchased from Fisher Scientific
(Houston, TX).
350 mg of high-purity silica coated with 2,4-DNPH were purchased from
Sigma Aldrich (St. Louis, MO). A made-to-order calibration standard
was purchased from O2Si Smart Solutions (Charleston, SC) containing
target analytes in Acetonitrile at the following concentrations: formaldehyde
(5000 mg/L), acetaldehyde (5000 mg/L), acrolein (500 mg/L), and crotonaldehyde
(125 mg/L). An isotopically labeled analog solution was also purchased
from O2Si Smart Solutions with the following concentrations in Acetonitrile:
formaldehyde-d2-DNPH (1000 mg/L), acetaldehyde-d4-DNPH (1000 mg/L), acrolein-2,4-DNPH-3,5,6-d3 (100 mg/L),
and crotonaldehyde-2,4-DNPH-3,5,6-d3 (25 mg/L). Ammonium
acetate (molecular biology grade) was purchased from Sigma Aldrich.
Acetonitrile and water (HPLC grade) were purchased from Fisher Scientific
(Houston, TX).
3
Carbonyl Compound Analysis Protocol
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2,4-DNPH (Sigma-Aldrich, 97%), acetone (Merck, 99.8%), acetonitrile (Sigma-Aldrich, 99.8%), and methanol (Merck, 99.8%) were used in this study. All other chemicals were of analytical reagent grade and used without further purification.
4
Characterization of Antioxidant Compounds
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Methanol (analytical grade) was purchased from VWR Chemicals (Fontenay-sous-Bois, France). Methanol and sulfuric acid (HPLC grade), anhydrous sodium sulphate, isooctane, glacial acetic acid, trichloroacetic acid, orthophosphoric acid (all analytical grade), sodium carbonate, absolute ethanol, sodium hydroxide, chloroform, all were purchased from Merck (Darmstadt, Germany). 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,6-di-tert-butyl-4-methylphenol (BHT), 2-(1,1dimethylethyl)-1,4-benzenediol (TBHQ), (-)-epicatechin (purity ≥90%), (-)-epigallocatechin gallate (purity ≥80%), (-)-epigallocatechin gallate (purity ≥95%), 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox) (purity 97%), Tween ® 40, linoleic acid, βcarotene, Phenol reagent Folin & Ciocalteu`s, sodium nitrite, aluminium chloride, potassium iodide, soluble starch, p-anisidine, thiobarbituric acid, 2,4-DNPH, were all purchased from Sigma-Aldrich (Steinheim, Germany). n-hexane for spectroscopy was supplied from BDH Prolabo (Leuven, Belgium).
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