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首页> 外文期刊>Journal of mass spectrometry: JMS >Rapid analysis of neurotransmitters in rat brain using ultra-fast liquid chromatography and tandem mass spectrometry: Application to a comparative study in normal and insomnic rats
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Rapid analysis of neurotransmitters in rat brain using ultra-fast liquid chromatography and tandem mass spectrometry: Application to a comparative study in normal and insomnic rats

机译：超快速液相色谱和串联质谱快速分析大鼠脑中神经递质：在正常和失眠大鼠的比较研究中的应用

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摘要

Neurotransmitters and their metabolites in central nervous system were known to play a significant role in sedation and hypnosis. A rapid and sensitive UFLC-MS/MS method for simultaneous determination of serotonin, 5-hydroxyindole acetic acid (5-HIAA), tryptophan (Try), dopamine (DA), norepinephrine (NE), γ-aminobutyric acid (GABA), glutamic acid (Glu) and acetylcholine (Ach) in rat brain without derivatization, ion-pairing reagent or pre-concentration was developed. Analytes and IS were separated on a Inertsil ODS-EP column (150 mm × 4.6 mm, 5 μm particles) and analyzed in a single chromatographic run in less than 9.0 min, using gradient elution with the mobile phase consisting of methanol and 0.01% acetic acid in water at a flow rate of 1.2 ml min ~(-1). The detection of the analytes was performed on 4000Q UFLC-MS/MS system with turbo ion spray source in positive ion and multiple reaction monitoring mode. The developed method provided excellent linear calibration curves for the assay of analytes (R~2 ≥ 0.9915). Limits of quantification were in the range of 1.0 ng ml~(-1) to 1.0 μg ml ~(-1) for the analytes in rat brain. Intra- and inter-day precision and accuracy of analytes were well within acceptance criteria (15%). Mean extraction recoveries of analytes and IS from rat brain were all more than 80.0%. Furthermore, the validated method was successfully applied to comparing profiles of analytes in normal and insomnic rat brains. Results indicated that there were statistically significant differences for serotonin, 5-HIAA, DA, NE, Glu and Ach, but no significant difference for Try and GABA between two groups.
机译：已知中枢神经系统中的神经递质及其代谢产物在镇静和催眠中起重要作用。一种快速灵敏的UFLC-MS / MS方法，可同时测定5-羟色胺，5-羟基吲哚乙酸（5-HIAA），色氨酸（Try），多巴胺（DA），去甲肾上腺素（NE），γ-氨基丁酸（GABA），开发了大鼠脑中的谷氨酸（Glu）和乙酰胆碱（Ach），没有进行衍生化，离子配对试剂或预浓缩。在Inertsil ODS-EP色谱柱（150 mm×4.6 mm，5μm颗粒）上分离分析物和IS，并在9.0分钟内通过单色谱运行进行分析，使用由甲醇和0.01％乙酸组成的流动相进行梯度洗脱水中的酸浓度为1.2 ml min〜（-1）。在具有正离子和多反应监测模式的涡轮离子喷雾源的4000Q UFLC-MS / MS系统上进行分析物的检测。所开发的方法为分析物的测定提供了出色的线性校准曲线（R〜2≥0.9915）。大鼠脑中分析物的定量限在1.0 ng ml〜（-1）至1.0μgml〜（-1）范围内。日间和日间分析物的准确性和准确性均在可接受的标准之内（15％）。从大鼠脑中提取的分析物和IS的平均回收率均超过80.0％。此外，已验证的方法已成功应用于比较正常和失眠大鼠脑中分析物的分布。结果表明，两组之间5-羟色胺，5-HIAA，DA，NE，Glu和Ach差异有统计学意义，但Try和GABA差异无统计学意义。

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