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建立了超高效液相色谱-四极杆飞行时间质谱法(UPLC-Q-TOF-MS)对小鼠大脑中的多巴胺(DA)、5-羟色氨(5-HT)、二羟基苯乙酸(DOPAC)和高香草酸(HVA)4种单胺类神经递质的含量进行同时测定。样品以30%乙醇溶液(甲酸调至p H 5.0)匀浆处理后,冷冻离心,过滤。在优化色谱-质谱条件下,采用ESI和APCI复合源,正/负离子扫描方式进行数据采集。4种单胺类神经递质在0.5~1 000μg/L范围内呈良好线性关系,相关系数(r)大于0.999 3,方法的检出限为0.5~5.0μg/L,平均回收率为93.3%~101.8%,相对标准偏差为1.1%~3.3%。该方法具有操作简便、高效快速、回收率良好、灵敏度高、重现性好和准确度高等优点,可用于小鼠脑组织内4种单胺类神经递质的含量测定。
A high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was developed for the determination of dopamine (DA), 5- hydroxytryptamine (5-HT), dihydroxyphenylacetic acid (DOPAC) and high vanillic acid (HVA) four monoamine neurotransmitters were measured simultaneously. Samples were homogenized in 30% ethanol solution (formic acid adjusted to pH 5.0), frozen and centrifuged. Under optimized chromatographic-mass spectrometry conditions, ESI and APCI composite sources, positive / negative ion scan mode data acquisition. The linearity of the four monoamine neurotransmitters was in the range of 0.5-1 000 μg / L with a correlation coefficient (r) greater than 0.999 3. The detection limit was 0.5-5.0 μg / L with an average recovery of 93.3% ~ 101.8%, the relative standard deviation of 1.1% ~ 3.3%. The method has the advantages of simple operation, high efficiency and fastness, good recovery, high sensitivity, good reproducibility and high accuracy and can be used for determining the content of four monoamine neurotransmitters in mouse brain tissue.