柱前衍生化HPLC测定大鼠血浆和脑组织中与抑郁症相关的12种氨基酸

Determination of 12 depression-related amino acids in rat plasma and brain tissue by HPLC with pre-column derivatization

建立一种使用9,10-蒽醌-2-磺酰氯作为衍生化试剂的柱前衍生化HPLC法,用于测定大鼠血浆和脑组织中与抑郁症相关的12种氨基酸。血浆和脑组织样本经乙腈沉淀蛋白后,加入Kolthoff缓冲液(pH 8.8)和衍生化试剂,室温下反应5 min后进行色谱分析。采用Lichrospher C18柱(250 mm×4.6 mm,5μm);流动相为乙腈-20 mmol/L醋酸铵缓冲液(pH 7.5),梯度洗脱;流速:1.0 mL/min;柱温:35℃;检测波长:254 nm。12种氨基酸在各自的浓度范围内线性良好。本方法灵敏准确、专属性高,可用于大鼠血浆和脑组织中与抑郁症相关的氨基酸的测定。

A pre-column derivatization HPLC method for the analysis of 12 depression-related amino acids in rat plasma and brain tissue using 9, 10-anthraquinone-2-sulfonyl chloride as derivatization reagent was developed. Rat plasma and brain tissue were deproteinated by acetonitrile and derivatized for only 5 min at room temperature by addition of Kohhoff buffer（ pH 8.8） and derivatization reagent. The derivatives were ,separated on a C18 column at 35 ℃ and detected at 254 nm. The mobile phase was acetonitrile and 20 mmol/L ammonium acetate buffer （ pH 7.5） with a gradient program at 1.0 mL/min. Good linearity was achieved over the concentration range for each amino acid. The method has good sensitivity, accuracy and specificity, and can be used to analyze amino acids related with the depression in rat plasma and brain tissue.