Development of a rapid high performance liquid chromatography assay for the determination of dopamine in retinal tissue utilizing a simple extraction procedure
The extraction method presented here demonstrates a safe, simple, and rapid method for catecholamine extraction. High performance liquid chromatography (HPLC) with electrochemical detection (ED) was employed for quantitative analysis of dopamine. Comparisons of various chromatographic conditions were investigated including the effects of varying the mobile phase pH and the mobile phase composition. A mobile phase was developed to optimize separation of analyte peaks, while minimizing sample run time and maintaining excellent peak resolution. This method has successfully been applied to the analysis of catecholamines contained in the retinal tissue of zebrafish. The feasibility of accomplishing detection with UV-Vis detection was also investigated, however, the sensitivity of the UV-Vis detector was found to be insufficient for retinal catecholamine quantification.