Rapid Determination of Nitrate in Brain Regions and Cerebrospinal Fluid of Transient Bilateral Common Carotid Artery Occlusion Rat Model by HPLC–UV

The present study is designed to determine the nitrate in the cortex, cerebellum, hippocampi, and cerebrospinal fluid (CSF) of Charles Foster albino rats with global ischemia. The global cerebral ischemia is performed by occluding both common carotid arteries for 1 h followed by reperfusion of 1 h. The level of nitrate is determined after 1 h of reperfusion in the brain samples and CSF by HPLC coupled with a UV detector at 212 nm. The mobile phase constitutes of 0.3 M potassium phosphate monobasic, 20% acetonitrile, and 20% methanol (pH 3.0), and 1.5 ml/min flow rate was kept for the analysis. The retention time for nitrate and nitrite was observed at 1.9 and 5.4 min, respectively. The concentration of nitrate in the cortex, hippocampi, cerebellum, and CSF was 21, 11, 23, and 06 µg/ml, respectively, in the normal rats as well as 114.97, 84.29, 92.58 and 18.07 µg/ml, respectively, in ischemic rats. The present method is simple, rapid, cost-effective and reproducible as well as suitable to analyze the nitrate level in the brain tissues and CSF in biochemical and pharmaceutical research.