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HPLC-ESI-MS法测定大鼠脑组织中神经递质L/D-丝氨酸的含量

, PP. 165-170

Keywords: 高效液相色谱—电喷雾离子化—质谱,脑组织,神经递质,D-丝氨酸,L-丝氨酸

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Abstract:

建立一种检测大鼠脑组织中L-丝氨酸和D-丝氨酸的高效液相色谱—电喷雾离子化—质谱联用技术(HPLC-ESI-MS)的测定方法.大鼠脑组织样品匀浆及离心后,取上清液并加入Marfey试剂对样品中L/D-丝氨酸进行衍生化.HPLC流动相为甲酸铵和乙腈,梯度洗脱;质谱采用电喷雾离子源,在负离子模式下进行定性定量分析.结果显示,L-丝氨酸和D-丝氨酸在22min内得以分离,在2.5~500ng/mg和1.2~250ng/mg浓度范围内分别与各自的色谱峰面积呈良好线性关系;L/D-丝氨酸的最低定量限分别为2.5ng/mg和1.2ng/mg,方法回收率均在81%~89%范围内.本法具有省时快速,灵敏度和选择性高等优点.将此法运用于成年和老年大鼠海马及皮层中L-丝氨酸和D-丝氨酸含量测定,结果表明,与成年鼠相比,老年鼠海马和皮层中L-丝氨酸的含量基本不变,而D-丝氨酸含量则明显减少,提示D-丝氨酸的变化与正常的衰老过程密切相关.

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