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According to the results of our pilot experiments (data not shown), the HSA concentration, the ratio of GSH to HSA, the temperature of the reduction reaction, the ratio of ATRA to HSA, the ratio of ATO and ATRA, the injection speed of ATRA solution, and the volume ratio of ethanol and water were set to 4%, 38.5%, 40 , 3%, 12.7%, 1.5 mL min 1 , and 5:1, respectively

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The conditions, such as a 75 mM sodium phosphate buffer (pH 3.0) with 1.7 mM 1-octanesulfonic acid, 25 M EDTA, 10% methanol, and a flow rate of 1 mL/min, are optimized for precision and sensitivity, ensuring accurate neurotransmitter profiling at a column temperature of 25C

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