HPLC Determination of Dopamine and Its Metabolites in Mice

Stripped striatum is weighed, and 10 μl/mg PBS containing 10 mM EDTA is added. After being on ice for 15 min, the tissues were sonicated and centrifuged at 15000 rpm for 10 min at 4°C. Collected the supernatant and calculated the supernatant volume for each sample. Then the supernatant was added to an equal volume of ice-cold 0.4 M perchloric acid containing 10 mM EDTA and vortexed. After standing on ice in the dark for 15 min, the medium was centrifuged at 15000 rpm for 15 min at 4°C. The middle layer was used to detect the levels of dopamine (DA), dihydroxypheny lacetic acid (DOPAC), and HVA. The supernatant of the striatal tissue (20 μl) was injected directly into the column and detected as described in our previously described method (Beal et al., 1992).