Column Chromatography Analysis of Brain Tissue: An Advanced Laboratory Exercise for Neuroscience Majors

Neurochemical analysis of discrete brain structures in experimental animals provides important information on synthesis, release, and metabolism changes following behavioral or pharmacological experimental manipulations. Quantitation of neurotransmitters and their metabolites following unilateral drug injections can be carried out using standard chromatographic equipment typically found in most undergraduate analytical laboratories. This article describes an experiment done in a six session (four hours each) component of a neuroscience research methods course. The experiment provides advanced neuroscience students experience in brain structure dissection, sample preparation, and quantitation of catecholamines using high performance liquid chromatography (HPLC) and protein analysis using ultraviolet spectroscopic methods. The students are exposed to useful laboratory techniques such as standard solution preparation and calibration curve construction, centrifugation, quantitative pipetting, and data evaluation and graphical presentation. Typically, only students that participate in independent neuroscience research are familiar with these types of quantitative skills. The usefulness of this type of experimental design for understanding behavioral or pharmacological effects on neurotransmitter systems is emphasized through a final report requiring a comprehensive literature search.