超高效液相色谱-三重四极杆质谱法评价白芍-甘草对抑郁大鼠大脑皮层中神经递质含量的影响

Effect of the herb pair Paeoniae Radix Alba-Glycyrrhizae Radix et Rhizoma on the content of neurotransmitters in the cerebral cortex of depressed rats by UPLC-MS/MS

目的建立超高效液相色谱-三重四极杆质谱(UPLC-MS/MS)法同时测定5-羟色胺(5-HT)、去甲肾上腺素(NE)和多巴胺(DA)等3种神经递质,考察白芍-甘草药对对抑郁大鼠大脑皮层中神经递质含量的影响。方法将50只SD大鼠随机分为空白组、模型组、阳性对照组、白芍组和白芍-甘草组,每组10只。利用长期不可预见性温和应激建立慢性应激大鼠抑郁模型,行为学测试后取大鼠大脑皮层,采用UPLC-MS/MS法分别测定5-HT、NE和DA的含量,以乙腈-0.1%甲酸水溶液等度洗脱,多反应监测进行分析。结果3种神经递质在各自质量浓度范围内与测定值线性关系良好,相关系数均不低于0.999,检测限为0.02~0.03μg·g^(-1),平均回收率为83.7%~94.1%。与空白组比较,模型组大鼠大脑皮层中5-HT、NE和DA的含量降低(P<0.01);与模型组比较,白芍组和白芍-甘草组大脑皮层中5-HT、NE和DA的含量均有提高(P<0.05,P<0.01)。结论UPLC-MS/MS法可用于单胺类神经递质的定量分析。白芍-甘草药对能显著提高抑郁大鼠大脑皮层中神经递质含量水平。

Objective To develop an ultra performance liquid chromatography-triple quadrupole mass spectrometric(UPLC-MS/MS)method to simultaneously determine 5-hydroxytryptamine(5-HT),norepinephrine(NE)and dopamine(DA),and to evaluate the effect of the herb pair Paeoniae Radix Alba-Glycyrrhizae Radix et Rhizoma on the content of neurotransmitters in the cerebral cortex of depressed rats.Methods Fifty SD rats were randomly divided into a blank group,a model group,a positive control group,a Paeoniae Radix Alba group and a Paeoniae Radix Alba-Glycyrrhizae Radix et Rhizoma group(10 rats in each group).Chronic stress rat model of depression was established by using long-term unpredictable mild stress.The cerebral cortex of the depressed rats was obtained after the behavior tests,and the content of 5-HT,NE and DA was determined by UPLC-MS/MS,respectively.The isocratic elution was performed with acetonitrile-0.1% formic acid solution,and the multi-reaction monitoring was used for the analysis.Results All three neurochemicals showed good linearity in their respective concentration ranges,and the correlation coefficients(r)were no less than 0.999.The limits of detection(LODs)ranged from 0.02 to 0.03μg·g^(-1).The average recoveries of the neurochemicals ranged 83.7%~94.1%.The content of 5-HT,NE and DA in the model group was significantly lower than that in the blank group(P<0.01),and the content of neurochemicals in the Paeoniae Radix Alba group and the Paeoniae Radix Alba-Glycyrrhizae Radix et Rhizoma group was higher than in the model group(P<0.05,P<0.01).Conclusion The herb pair Paeoniae Radix Alba and Paeoniae Radix Alba-Glycyrrhizae Radix significantly improves the level of monoamine neurotransmitters determined by UPLC-MS/MS in the cerebral cortex of depressed rats.