| 高效液相色谱-荧光法测定小鼠脑组织中单胺递质及其相关物质 |
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| 引用本文： | 张璐璐, 冯芳, 叶小敏, 朱杰, 马冯飞. 高效液相色谱-荧光法测定小鼠脑组织中单胺递质及其相关物质[J]. 中国药科大学学报, 2010, 41(4): 367-371. |
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| 作者姓名： | 张璐璐 冯芳 叶小敏 朱杰 马冯飞 |
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| 作者单位： | 1.中国药科大学药物分析教研室;2.药物质量与安全预警教育部重点实验室 |
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| 基金项目： | 国家自然科学基金资助项目（No.30973858） |
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| 摘 要： | 建立高效液相色谱-荧光分析方法，研究小鼠脑皮层、海马、纹状体、丘脑等不同部位的单胺类神经递质及其相关物质。脑组织样本经高氯酸沉淀蛋白后进行色谱分析，采用Lichrospher C18柱(250 mm×4.6 mm，5 μm)，流动相为pH 3.06缓冲液(含1 mmol/L庚烷磺酸钠；30 mmol/L磷酸二氢钠-磷酸溶液)-甲醇(85∶15)；流速：1.0 mL/min；柱温：35 ℃。荧光检测：激发波长为280 nm，发射波长为315 nm。采用建立的方法，鉴别出小鼠脑中存在去甲肾上腺素(NE)、多巴胺(DA)、5-羟色胺(5-HT)、酪氨酸、色氨酸、3，4-二羟苯乙酸和5-羟吲哚乙酸等物质；对NE、DA和5-HT进行定量研究，确定它们的线性范围分别为2.4～471.0 ng/mL(r=0.999 6)，2.6～519.5 ng/mL(r=0.999 4)，2.4～469.3 ng/mL(r=0.999 1)，日内、日间RSD均小于9.0%，准确度达到限度要求，提取回收率大于90%。本法专属性好，准确度高，可以反映不同鼠脑区域单胺类神经递质及相关物质含量上的差异。
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| 关 键 词： | 高效液相色谱-荧光分析 单胺类神经递质 相关物质 脑组织 |
Determination of monoamine neurotransmitters and related substances in mouse brain tissue by HPLC-FLD |
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| Determination of monoamine neurotransmitters and related substances in mouse brain tissue by HPLC-FLD[J]. Journal of China Pharmaceutical University, 2010, 41(4): 367-371. |
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| Abstract: | To establish an HPLC-FLD method for the investigation of monoamine neurotransmitters and related substances in mouse frontal cortex，hippocampus，striatum，and hypothalamus.Mouse brain samples were deproteinized by perchloric acid and analyzed.The separation was carried out on a Lichrospher C18 column(250 mm×4.6 mm，5 μm).The mobile phase was composed of pH 3.06 buffer(including 1 mmol/L heptanesulfonic acid sodium salt，30 mmol/L NaH2PO4-H3PO4 solution) and methanol in the ratio of 85∶15；the flow rate was 1.0 mL/min and the column temperature was 35 °C.Fluorescence detection：the excitation and emission wavelengths were set at λex=280 nm，λem=315 nm.Under the optimized conditions，norepinephrine(NE)，dopamine(DA)，5-hydroxytryptamine (5-HT)，3，4-dihydroxy-phenyl acetic acid(DOPAC)，5-hydroxyindoleacetic acid(5-HIAA)，tyrosine(Tyr)，and tryptophan(Trp) in mouse brain were detected.NE，DA，and 5-HT were quantified.The linearity ranges of NE，DA， and 5-HT were 2.4-471.0 ng/mL(r=0.999 6)，2.6-519.5 ng/mL (r=0.999 4)，and 2.4-469.3 ng/mL(r=0.999 1)，respectively.The intra- and inter-day precision was always lower than 9.0%. The accuracy could attain the demands and the absolute recovery values were always higher than 90%. The proposed method has good specificity and accuracy.It could be used to analyze the content differences of monoamine neurotransmitters and related substances in different mouse brain areas. |
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| Keywords: | HPLC-FLD monoamine neurotransmitter related substances brain tissue |
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