Quantitative Determination of Norepinephrine by HPLC in Rodent Urine Sample

Abstract

Norepinephrine, a key neurotransmitter that has been linked to a variety of neuropsychiatric diseases. However, there is limited work on employing HPLC-UV to estimate monoamines in biological samples using HPLC-UV method. The present study explores the detection of norepinephrine in rat urine sample, developing a easy, precise validated HPLC method. The o-phosphoric acid (80%):acetonitrile (70:30) combination was used as the mobile phase, and a flow rate of 0.5 ml/min was used to produce the chromatographic separation on a C8 column (250 x 4.6 mm, 5 m). The detection was observed at λmax 275 nm with better sensitivity. According to the parameters indicated in the ICH guidelines (Q2A; Q2B), the procedure was verified. The linearity range was selected from 10-35 µg/mL, r² =0.966, LOD (1.17 µg/mL), LOQ (3.55 µg/mL), ret. time (4 min). The technique has demonstrated that it is repeatable and recoverable within the given range. Thus can be used for routine analysis of norepinephrine in urine samples.