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MEASUREMENT OF OXIDATIVE STRESS PARAMETERS USING LIQUID CHROMATOGRAPHY - TANDEM MASS SPECTROSCOPY (LC-MS/MS)
Citation:
WINNIK, W. M. AND K. T. KITCHIN. MEASUREMENT OF OXIDATIVE STRESS PARAMETERS USING LIQUID CHROMATOGRAPHY - TANDEM MASS SPECTROSCOPY (LC-MS/MS). TOXICOLOGY AND APPLIED PHARMACOLOGY. Academic Press Incorporated, Orlando, FL, 223(1):100-106, (2008).
Impact/Purpose:
Although oxidative stress is frequently cited as a cause of various adverse biological effects and diseases, convincing evidence of oxidative stress and its causality of adverse health outcomes is frequently lacking. In this article the advantages of LC-MS/MS based methods in measuring multiple oxidative stress markers are presented. Oxidative stress is an interdisciplinary research area spanning chemistry and biology. This review is aimed mainly at the biochemists with the intention to describe this versatile and sensitive analytical chemistry technique and its new applications in oxidative stress research.
Description:
What is the study?
An invited review article.
Measurement of oxidative stress parameters using liquid chromatography-tandem mass spectroscopy (LC-MS/MS)
Why was it done?
Although oxidative stress is frequently cited as a cause of various adverse biological effects and diseases, convincing evidence of oxidative stress and its causality of adverse health outcomes is frequently lacking. In this article the advantages of LC-MS/MS based methods in measuring multiple oxidative stress markers are presented. Oxidative stress is an interdisciplinary research area spanning chemistry and biology. This review is aimed mainly at the biochemists with the intention to describe this versatile and sensitive analytical chemistry technique and its new applications in oxidative stress research.
What is the impact to the field and the agency?
This review article describes the measurement and characterization of oxidative stress markers that are relevant to several on-going EPA research projects investigating the role of oxidative stress processes induced by chemical exposure in human health risk assessment.
Abstract
There is increasingly intense scientific and clinical interest in oxidative stress and the many parameters used to quantify the degree of oxidative stress. However, there remain many analytical limitations to currently available assays for oxidative stress markers. Recent improvements in software, hardware, and instrumentation design have made liquid chromatography and tandem mass spectroscopy (LC-MS/MS) methods optimal choices for the determination of many oxidative stress markers. In particular, LC-MS/MS often provides the advantages of higher specificity, higher sensitivity, and the capacity to determine multiple analytes (e.g. 4-11 oxidative stress markers per LC run) when compared to other available methods, such as gas chromatography-MS, immunoassays, spectrophotometric, or flourometric assays. LC-MS/MS methods are also compatible with cleanup and sample preparation methods including prior solid phase extraction or automated two dimensional LC/LC chromatography followed by MS/MS. LC-MS/MS provides three analytical filtering functions. (1) The LC column provides initial separation as each analyte elutes from the column. (2) The first MS dimension isolates ions of a particular mass-to-charge (m/z) ratio. (3) The selected precursor ion is fragmented into product ions that provide structural information about the precursor ion. Quantitation is achieved based on the abundances of the product ions. The sensitivity limits for LC-MS/MS usually lie within the range of fg-pg of analyte per LC on-column injection. In this article, the present capabilities of LC-MS/MS are briefly presented and specific examples of the strengths of these LC-MS/MS assays are discussed. The selected examples include methods for isoprostanes, oxidized proteins and amino acids, and DNA biomarkers of oxidative stress.