Assessment and quantification of Dopamine in rat urine by a validated HPLC- Photodiode Array method (HPLC-PDA)

Abstract

Dopamine is a neurotransmitter that functions as a chemical messenger in the nervous system, allowing certain neurons to communicate
with one another. For the determination of dopamine HCl in rat urine, an easy, precise, and precise HPLC approach was devised and
validated. The separation was carried out on a C8 column (250 x 4.6 mm, 5 µm) with a combination of 0.1% ortho-phosphoric acid:
acetonitrile (70:30) (pH 2.2) as mobile phase. The mobile phase flow rate was kept constant at 0.5 mL/min. UV detection at 279 nm was
used to obtain high sensitivity. During the development of analytical methods, parameters according to ICH guidelines (Q2A; Q2B) were
examined. Dopamine had a retention duration of 4.1 minutes. For dopamine 8-42 µg/mL range was selected , the approach has been validated
(r2 =0.9871). This approach has proven to be highly reproducible and recoverable within the specified range. For routine analysis, this
approach can be employed successfully for assessing and quantification in rat urine samples