期刊
FREE RADICAL BIOLOGY AND MEDICINE
卷 51, 期 9, 页码 1823-1829出版社
ELSEVIER SCIENCE INC
DOI: 10.1016/j.freeradbiomed.2011.08.012
关键词
Lipid peroxidation; Oxidative stress; LC-MS/MS; Malondialdehyde; Free radicals
资金
- National Science Council, Taiwan [NSC 97-2314-B-040-015-MY3, NSC 99-2314-B-040-018-MY3]
A highly sensitive quantitative LC-MS/MS method was developed for measuring urinary malondialdehyde (MDA). With the use of an isotope internal standard and online solid-phase extraction, urine samples can be directly analyzed within 10 min after 2,4-dinitrophenylhydrazine (DNPH) derivatization. The detection limit was estimated as 0.08 pmol. This method was further applied to assess the optimal addition of DNPH for derivatization and to measure urinary MDA in 80 coke oven emission (COE)-exposed and 67 nonexposed workers. Derivatization optimization revealed that to achieve complete derivatization reaction, an excess of DNPH is required (DNPH/MDA molar ratio: 893-8929) for urine samples that is about 100 times higher than that of MDA standard solutions (molar ratio: 10-80). Meanwhile, the mean urinary concentrations of MDA in COE-exposed workers were significantly higher than those in nonexposed workers (0.23 +/- 0.17 vs 0.14 +/- 0.05 mu mol/mmol creatinine, P<0.005). Urinary MDA concentrations were also significantly associated with the CUE (P<0.005) and smoking exposure (P<0.05). Taken together, this method is capable of routine high-throughput analysis and accurate quantification of MDA and would be useful for assessing the whole-body burden of oxidative stress. Our findings, however, raise the issue that derivatization optimization should be performed before it is put into routine biological analysis. (C) 2011 Elsevier Inc. All rights reserved.
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