Journal
PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS
Volume 38, Issue 12, Pages 1171-1177Publisher
CHINESE ACAD SCIENCES, INST BIOPHYSICS
DOI: 10.3724/SP.J.1206.2011.00407
Keywords
quantitative analysis; endogenous formaldehyde; high performance liquid chromatography; porcine brain; 2,4-dinitrophenylhydrazine; cognitive impairment
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Funding
- National Basic Research Program of China [2010CB912303, 2012CB911004]
- CAS [KSCX2-YW-R-256, CAS-KSCX2-YW-R-119]
- National Natural Science Foundation of China [30970695]
- Johnson & Johnson Corporate Office of Science Technology [GJHZ1131]
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Chronic impairments of brain caused by accumulation of endogenous formaldehyde as ageing is thought as one of the risk factors for sporadic age-related dementia. Thus, it is necessary to determine the endogenous concentration of formaldehyde in brain. Using 2, 4-dinitrophenylhydrazine (2, 4-DNPH) in the assay, the concentrations of formaldehyde in parietal lobe, frontal lobe, temporal lobe, occipital lobe, hippocampus, cerebellum and brainstem of porcine brain (n=5) were measured through UV-HPLC. Homogenizing brain tissues with 10% trichloroacetic acid (pH=1.0) not only avoided the disturbance from contaminated protein and other compounds, but also eliminated the step extraction of FA-DNPH derivative (2, 4-dinitrophenylhydrazone) and improve the methodological sensitivity (P < 0.05). The porcine brain formaldehyde was 75.5 similar to 83.4 mu mol/kg fresh brain tissues measured by this improved method. Recoveries of spiked formaldehyde at low and high level were 95.96%similar to 102.04% with relative standard deviations less than 10%. This suggests that the modified 2, 4-DNPH method is effective enough to measure the endogenous formaldehyde in brain tissues.
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