QUANTITATIVE DETERMINATION OF NOREPINEPHRINE BY HPLC IN RODENT URINE SAMPLE
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 an 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.
Norepinephrine, HPLC-UV, method validation, rodent urine etc.