LC-MS/MS Determination of 5-Hydroxytryptamine and 5-Hydroxyindole Acetic Acid in Serum and Hippocampus and Its Ap-plication in the Evaluation of Antidepressant Efficacy of Paeoniae Radix Alba
10.16466/j.issn1005-5509.2024.07.018
- VernacularTitle:血清与海马内5-羟色胺和5-羟基吲哚乙酸的LC-MS/MS测定及其在白芍抗抑郁功效评价中的应用
- Author:
Yuan SU
1
,
2
;
Siyu CAI
;
Mengtian SHI
Author Information
1. 浙江中医药大学药学院 杭州 310053
2. 中国科学院大学杭州高等研究院 生命与健康科学学院 浙江省系统健康科学重点实验室
- Keywords:
LC-MS/MS;
5-HT;
5-HIAA;
mice serum;
hippocampus;
Paeoniae Radix Alba;
depression;
efficacy evaluation
- From:
Journal of Zhejiang Chinese Medical University
2024;48(7):862-871
- CountryChina
- Language:Chinese
-
Abstract:
[Objective]To establish a liquid chromatography-tandem mass spectrometry(LC-MS/MS)method for the determination of 5-hydroxytryptamine(5-HT)and 5-hydroxyindole acetic acid(5-HIAA)in mouse serum and hippocampus,applied to the evaluation of the antidepressant efficacy of traditional Chinese medicine.[Methods]The serum and hippocampus were taken and the concentrations of 5-HT and 5-HIAA were determined by LC-MS/MS after pretreatment of protein precipitation.Chromatographic separation was performed on Hypersil ODS(250 mm×4.6 mm,5 μm)at column temperature of 40℃.The mobile phase was 0.1%formic acid-water solution with 2 mmol·L-1 ammonium acetate and acetonitrile whereas flow rate was set at 1 mL·min-1.The mass spectrometry detection was performed using multiple reaction monitoring(MRM).One hundred and ninety-eight mice were randomly divided into normal control group,model group,fluoxetine hydrochloride group and 30 groups of Paeoniae Radix Alba with 6 rats in each group.Tail suspension test and forced swimming test were carried out.[Results]5-HT and 5-HIAA had good linear relationship within the concentration range of 0.1～1 000 ng·mL-1 and 1～500 ng·mL-1,respectively(r>0.999).Precision and recovery of this method were in line with the requirements of biological sample analysis.The degree of change in 5-HIAA/5-HT values in the Paeoniae Radix Alba group relative to the model group was correlated with the degree of change in indicators of the behavioral experiment,and the degree of change was significantly higher than that of the behavioral experiment,with a much smaller within-group relative standard deviation(RSD).[Conclusion]This LC-MS/MS method is simple and fast for operation,with accurate results,high sensitivity,good reproducibility and strong specificity which indicated that was suitable for the simultaneous determination of 5-HT and 5-HIAA in serum and hippocampus.It could be used for research and efficacy evaluation of neuroprotective drugs like Paeoniae Radix Alba.
