The HPLC protocol has been well established and is routinely used in our laboratory (Jiang et al., 2008; Shen et al., 2017). Samples of the striatum were weighed and then homogenized in 120 μL of solution A (0.4 M perchloric acid). After initial centrifugation (12,000 rpm for 20 min at 4°C) (Eppendorf 5810R, Germany), 80 μL of the supernatant was transferred into Eppendorf tubes, and 40 μL of solution B [containing 20 mM citromalic acid potassium, 300 mM dipotassium phosphate, 2 mM ethylenediamine tetraacetic acid (EDTA)⋅2Na] was added. After additional centrifugation (12,000 rpm for 20 min at 4°C), 100 μL of the supernatant was assayed for dopamine (DA) and its metabolites homovanillic acid (HVA) and dihydroxyphenylacetic acid (DOPAC) by HPLC. Separation was achieved on an Agilent C18 reverse-phase column (4.6 mm × 150 mm × 5 μm) (Agilent, CA, United States). The mobile phase consisted of 20 mM citromalic acid, 50 mM sodium caproate, 0.134 mM EDTA⋅2Na, 3.75 mM sodium octane sulfonic acid, and 1 mM di-sec-butylamine in 5% (v/v) methanol; the flow rate was 1 mL/min. A 2,465 electrochemical detector (Waters, United States) was operated in screen mode. The results are expressed as ng/mg wet weight of brain tissue.

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