Integration of Miniaturized Solid Phase Extraction and LC-MS/MS Detection of 3-Nitrotyrosine in Human Urine for Clinical Applications

The overall goal of this miniaturized solid phase extraction combined with LC-MS/MS assay is to accurately quantify free 3-nitrotyrosine in human urine for clinical applications. Free 3-nitrotyrosine help in yields and a biomarker for oxidative stress. However, it will remain a special challenge to accurately quantify 3-nitrotyrosine in biological emergencies for clinical applications due to insufficient sensitivity, selectivity, and cumbersome sample preparations.

This technique incorporating an effective solid phase extraction in a highly sensitive LC-MS/MS interaction allows rapid, specific, and accurate determination of low levels of endogenous 3-nitrotyrosine in human urine without the need of constitution and two-dimensional LC.Though this method can provide insight into oxidative stress investigation, it can also be applied to other pathological conditions such as complex inflammatory and neurodegenerative disorders. The implications of this technique could be extended toward monitoring the effectiveness of antioxidant treatment because the 3-nitrotyrosine level can be reduced by antioxidants. To begin this procedure, vortex previously prepared and thawed urine samples, standards, and QC samples.