Rapid analysis of neurotransmitters in rat brain using ultra-fast liquid chromatography and tandem mass spectrometry: application to a comparative study in normal and insomnic rats
Article first published online: 18 JUL 2013
Copyright © 2013 John Wiley & Sons, Ltd.
Journal of Mass Spectrometry
Volume 48, Issue 8, pages 969–978, August 2013
How to Cite
He, B., Bi, K., Jia, Y., Wang, J., Lv, C., Liu, R., Zhao, L., Xu, H., Chen, X. and Li, Q. (2013), Rapid analysis of neurotransmitters in rat brain using ultra-fast liquid chromatography and tandem mass spectrometry: application to a comparative study in normal and insomnic rats. J. Mass Spectrom., 48: 969–978. doi: 10.1002/jms.3243
- Issue published online: 10 JUL 2013
- Article first published online: 18 JUL 2013
- Manuscript Accepted: 3 JUN 2013
- Manuscript Revised: 22 MAY 2013
- Manuscript Received: 16 APR 2013
- brain samples;
- ODS-EP column;
Neurotransmitters and their metabolites in central nervous system were known to play a significant role in sedation and hypnosis. A rapid and sensitive UFLC-MS/MS method for simultaneous determination of serotonin, 5-hydroxyindole acetic acid (5-HIAA), tryptophan (Try), dopamine (DA), norepinephrine (NE), γ-aminobutyric acid (GABA), glutamic acid (Glu) and acetylcholine (Ach) in rat brain without derivatization, ion-pairing reagent or pre-concentration was developed. Analytes and IS were separated on a Inertsil ODS-EP column (150 mm × 4.6 mm, 5 µm particles) and analyzed in a single chromatographic run in less than 9.0 min, using gradient elution with the mobile phase consisting of methanol and 0.01% acetic acid in water at a flow rate of 1.2 ml min−1. The detection of the analytes was performed on 4000Q UFLC-MS/MS system with turbo ion spray source in positive ion and multiple reaction monitoring mode. The developed method provided excellent linear calibration curves for the assay of analytes (R2 ≥ 0.9915). Limits of quantification were in the range of 1.0 ng ml−1 to 1.0 µg ml−1 for the analytes in rat brain. Intra- and inter-day precision and accuracy of analytes were well within acceptance criteria (15%). Mean extraction recoveries of analytes and IS from rat brain were all more than 80.0%. Furthermore, the validated method was successfully applied to comparing profiles of analytes in normal and insomnic rat brains. Results indicated that there were statistically significant differences for serotonin, 5-HIAA, DA, NE, Glu and Ach, but no significant difference for Try and GABA between two groups. Copyright © 2013 John Wiley & Sons, Ltd.