Following a published methodology, a 1-liter subsample press emission gas sample was passed through a DNPH
cartridge to absorb aldehydes (Miyamoto et al.
Two grams of the derivatizing agent DNPH
was dissolved in 1 liter of acetonitrile: methanol 50:50; the pH of the mixture was adjusted by 10% phosphoric acid to the range of 5-6.
Di-nitrophenyl hydrazine (DNPH
; Waters Sep-Pak[R]) cartridges were used to sample aldehydes and ketones, and pre-evacuated canisters (Entech) were used to sample [C.sub.5]-[C.sub.12] gases.
Total histones were then incubated with a solution of 0.5% 2,4-dinitrophenyl hydrazine (DNPH
) (w/v) in 10% trifluoroacetic acid (v/v).
Thiol group of glycated BSA samples was assayed according to the Ellman's method using 2, 4-dinitrophenylhydrazine (DNPH
This reagent will be dyed with a DNPH
into an orange colored sample that can be analyzed with an UV-Spectrophotometer.
Phenylmethylsulfonyl fluoride (PMSF), ethylenediaminetetraacetic acid (EDTA), 2,4-dinitrophenylhydrazine (DNPH
), 5,5'-dithiobis-2-nitrobenzoic acid (DTNB), 2,3,5-triphenyl tetrazolium chloride (TTC), oxidized glutathione (GSSG), NADPH, FeS[O.sub.4] x 7[H.sub.2]O, and menadione were obtained from Sigma-Aldrich Corporation (USA); diagnostic kits for determination of glucose were from Cormay (tomianki, Poland); N,N,N',N'-tetramethyl ethylenediamine (TEMED) and quercetin were from Reanal (Hungary); peptone and yeast extract were from Fluka (Germany).
Phenylmethylsulfonyl fluoride (PMSF), ethylenediamine-tetraacetic acid (EDTA), 2,4-dinitrophenylhydrazine (DNPH
), 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB), 2,3,5-triphenyltetrazolium chloride (TTC), Girard's reagent T, amyloglucosidase (#10115), trehalase (#T8778), peptone, and yeast extract were obtained from Sigma-Aldrich Corporation (USA); N,N,N',N'-tetramethylethylenediamine (TEMED) and quercetin were from Reanal (Hungary); disodium salt of alpha-ketoglutaric acid (99% purity) was from Protista AB (Sweden); all reagents for determination of glucose and triacylglycerides (TAG), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) activities were from PZ Cormay S.A.
Bovine serum albumin, acrylamide, N,N'-methylene-bisacrylamide, trichloroacetic acid (TCA), FeS[O.sub.4], sulfuric acid, cumene hydroperoxide (CHP), butylhydroxy-toluene, methanol, xylenol orange, di-nitrophenylhydrazine (DNPH
), guanidine, ethanol, ethyl acetate, hydrochloric acid, Coomassie[R] Brilliant Blue R-250, thioglycolic acid, NBD-Cl, and o-phthalaldehyde (OPA) were purchased from Sigma-Aldrich (St.
Briefly explained, 10 [micro]g of proteins was denatured and derivatized using 10 mM DNPH
in acid solution.
Protein carbonyls were determined in plasma samples using the 2,4-dinitrophenolhydrazine (DNPH
) spectrophotometric method described by Levine et al.
Active air samples were collected at both upstream and downstream of air cleaning systems simultaneously using adsorbent tubes (AirToxic) for VOC analysis by an automated thermal desorber combined with GC/MS, and DNPH
cartridges with KI ozone scrubbers for the analysis of ketones and aldehydes by HPLC.