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Determination of urinary malondialdehyde by isotope dilution LC-MS/MS with automated solid-phase extraction: A cautionary note on derivatization optimization
- Source :
-
Free Radical Biology & Medicine . Nov2011, Vol. 51 Issue 9, p1823-1829. 7p. - Publication Year :
- 2011
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Abstract
- Abstract: 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 10min after 2,4-dinitrophenylhydrazine (DNPH) derivatization. The detection limit was estimated as 0.08pmol. 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±0.05μmol/mmol creatinine, P <0.005). Urinary MDA concentrations were also significantly associated with the COE (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. [Copyright &y& Elsevier]
Details
- Language :
- English
- ISSN :
- 08915849
- Volume :
- 51
- Issue :
- 9
- Database :
- Academic Search Index
- Journal :
- Free Radical Biology & Medicine
- Publication Type :
- Academic Journal
- Accession number :
- 66229875
- Full Text :
- https://doi.org/10.1016/j.freeradbiomed.2011.08.012