摘要
目的：建立检测小鼠脑组织中色氨酸、5-羟基色氨酸、5-羟色胺和5-羟基吲哚乙酸含量的LC-MS/MS分析方法。方法：采用TSK Gel amide 80（2.0 mm×15 cm,3μm）色谱柱,柱温35℃,乙腈：甲酸铵水溶液（15 mmol·L-1,p H5.5）（40∶60,v/v）为流动相,流速0.4 m L·min-1,采用正离子多反应监测（MRM）模式进行扫描。结果：4种待测物均在5 min内完成测定,色氨酸、5-羟基色氨酸、5-羟色胺和5-羟基吲哚乙酸最低定量限分别为0.2、0.1、0.2、0.2 ng·m L-1,最低检测限分别为0.05、0.02、0.05、0.05 ng·m L-1,在1~500 ng·m L-1浓度范围内线性关系均良好,方法的精密度和加样回收率均符合生物样品的分析要求。将该方法应用于口服萱草花总黄酮的小鼠脑提取物检测后发现：脑内四种待测物浓度明显发生改变,且变化规律与阳性药物帕罗西汀相同。结论：本方法简便、准确、灵敏度高,专属性强,重复性好,适用于脑组织中色氨酸-五羟色胺代谢过程中的4种成分的测定,可应用于神经精神类药物的药理作用机制研究。
Objective： To establish a LC- MS / MS method for the determination of tryptophan（ TRP） 5- hydroxy-tryptophan（ 5- HTP）,5- hydroxytryptamine（ 5- HT） and 5- hydroxyindole acetic acid（ 5- HIAA） in mouse brain tissues.Method： Chromatographic separation was performed on a TSK Gel amide 80 column（ 2.0 mm × 15 cm,3 μm） at column temperature of 35 ℃. The mobile phase system was a mixture of acetonitrile- ammonium formate solution（ 15 mmol·L- 1,p H = 5.5）（ 40 ∶ 60） whereas the flow rate was set at 0.4 m L·min- 1. The instrument was operated under the multiple reaction monitoring（ MRM） mode using electrospray ionization（ ESI） in the positive ion mode.Results： Four analyses were separated within 5 minutes. The lowest limits of quantification in tryptophan（ TRP） 5-hydroxy-tryptophan（ 5-HTP）,5- hydroxytryptamine（ 5- HT） and 5- hydroxyindole acetic acid（ 5- HIAA）were 0.2,0.1,0.2 ng·m L- 1and 0.2 ng·m L- 1,respectively.The minimum detection limit were 0.05,0.02,0.05 ng·m L- 1and 0.05 ng·m L- 1,respectively.A good linear relationship was achieved within the concentration ranges from 1 to500 ng·m L- 1for the analyses.Precision and recovery of this method were in line with the requirements of biological sample analysis. The method was applied to determine the concentrations of four analyses in brain after oral administration of Hemerocallis flavonoid extract and results showed that four kinds of analyses were significantly changed which was similar with the positive drug of paroxetine. Conclusion： This method was simple,accurate,highly sensitive,strongly,specific and repeatable which indicated that was suitable for the determination of fourmetabolites of tryptophan-5- HT metabolic pathways in mice. This method could be used to the pharmacological mechanism of the neuropsychiatric drugs.
出处
《食品工业科技》
CAS
CSCD
北大核心
2016年第1期315-319,330,共6页
Science and Technology of Food Industry
基金
重大新药创制(航天应激所致认知功能损伤防治新型化合物DS的成药性研究
2012ZX09J12110-04C)
