| 摘要: |
| 目的 建立超高效液相色谱-串联质谱（UPLC-MS/MS）法，同时检测抑郁模型大鼠脑脊液中5-羟色胺及其代谢产物5-羟吲哚乙酸（5-HIAA）的水平，并观察氟西汀治疗后抑郁模型大鼠脑脊液中5-羟色胺和5-HIAA的含量变化。方法 将24只SD大鼠随机分为对照组、模型组和氟西汀治疗组，每组8只。造模第35天，待行为学测试后取大鼠脑脊液，用蛋白沉淀法进行前处理，然后采用UPLC-MS/MS测定5-羟色胺和5-HIAA的含量。色谱柱为ACE C18-PFP柱（4.6 mm×150 mm，3 μm），流动相为乙腈-0.2%甲酸水，梯度洗脱，流速为0.4 mL/min，柱温为25℃，质谱检测采用动态多反应监测（DMRM）模式。结果 大鼠脑脊液中5-羟色胺和5-HIAA分别在0.10~100.00 ng/mL和1.00~1 000.00 ng/mL浓度范围内线性关系良好（r>0.999），日内和日间精密度均<15.00%，提取回收率为86.86%~98.49%，基质效应为89.80%~95.25%，样品稳定性良好。与对照组比较，模型组大鼠脑脊液中5-羟色胺的含量降低（P<0.01），5-HIAA含量上升（P<0.01）；与模型组比较，氟西汀组大鼠脑脊液中5-羟色胺含量上升（P<0.01），5-HIAA含量降低（P<0.01）。结论 本实验所建立的方法操作简便，结果准确，灵敏度高，重现性好，适用于检测抑郁大鼠脑脊液中5-羟色胺和5-羟吲哚乙酸水平。 |
| 关键词: 超高效液相色谱法 串联质谱法 抑郁 脑脊液 5-羟色胺 5-羟吲哚乙酸 |
| DOI：10.16781/j.0258-879x.2018.12.1354 |
| 投稿时间：2018-06-05修订日期：2018-07-09 |
| 基金项目: |
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| Simultaneous determination of 5-hydroxytryptamine and 5-hydroxyindoleacetic acid in cerebrospinal fluid of depressive rats by ultra high performance liquid chromatography-tandem quadrupole mass spectrometry |
| CHEN Lei1,DU Yi2,LI Cheng-jian3,ZHOU Jin3,CAI Zhen1* |
(1. Department of Pharmacy, Changhai Hospital, Navy Medical University(Second Military Medical University), Shanghai 200433, China;
2. Department of Pharmacy, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China;
3. Department of Pharmacy, Eastern Hepatobiliary Surgery Hospital, Navy Medical University(Second Military Medical University), Shanghai 200438, China
*Corresponding author) |
| Abstract: |
| Objective To establish an ultra high performance liquid chromatography-tandem quadrupole mass spectrometry (UPLC-MS/MS) method for simultaneous determination of 5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) in cerebrospinal fluid of depressive rats, and to investigate the concentrations of 5-HT and 5-HIAA in cerebrospinal fluid of depression rats and fluoxetine treated rats. Methods Twenty-four SD rats were randomly divided into control group, model group and fluoxetine group, with 8 rats in each group. At 30 d of model building, the cerebrospinal fluids were taken after the test of behaviors, and the concentrations of 5-HT and 5-HIAA were determined by UPLC-MS/MS after pretreatment of protein precipitation. Chromatographic column was ACE C18-PFP column (4.6 mm×150 mm, 3 μm), mobile phase was acetonitrile and 0.2% formic acid-water solution, flow rate was 0.4 mL/min, and the column temperature was 25℃. The mass spectrometry detection was performed using dynamic multiple reaction monitoring (DMRM). Results 5-HT and 5-HIAA in cerebrospinal fluid of depression rats had good linear relationship within the concentration range of 0.10-100.00 ng/mL and 1.00-1 000.00 ng/mL, respectively (r>0.999). Intra-and inter-day precision was less than 15.00%; the recoveries ranged from 86.86% to 98.49%; and the matrix effects ranged from 89.80% to 95.25%. The samples remained stable during analysis. Compared with the control group, the concentration of 5-HT in cerebrospinal fluid of rats in the model group was significantly decreased (P<0.01), and the concentration of 5-HIAA was significantly increased (P<0.01). Compared with the model group, the concentration of 5-HT in cerebrospinal fluid of rats in the fluoxetine group was significantly increased (P<0.01), and the concentration of 5-HIAA was significantly decreased (P<0.01). Conclusion The UPLC-MS/MS method is simple, accurate, sensitive, reproducible and suitable for simultaneous determination of 5-HT and 5-HIAA in cerebrospinal fluid of depression rats. |
| Key words: ultra high performance liquid chromatography tandem quadrupole mass spectrometry depression cerebrospinal fluid 5-hydroxytryptamine 5-hydroxyindoleacetic acid |
