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Simultaneous Determination of Four Monoamine Neurotransmitters and Seven Effective Components of Zaoren Anshen Prescription in Rat Tissue using Uplc-Ms/Ms

Author(s):

Rong Li, Lin Wang, Xiao Wang, Dian Zhang, Yafeng Zhang, Zhuo Li and Minfeng Fang*   Pages 1 - 14 ( 14 )

Abstract:


Background: Zaoren Anshen prescription (ZAP) is widely used as a classic Chinese Traditional Medicine (TCM) prescription for the treatment of palpitations and insomnia in China. Some studies have identified the main active components for its anti-insomnia effect, and observed changes of some endogenous components that are closely related to its anti-insomnia effect. However, simultaneous determination of four monoamine neurotransmitters and seven effective components of ZAP and the investigation of their distribution behaviours in tissues by using ultra-performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS) have not been reported.

Methods: An ultra-performance liquid chromatography with tandem mass spectrometry method was developed and validated for simultaneous quantification of four monoamine neurotransmitters (norepinephrine, dopamine, 5-hydroxy tryptamine and 5-hydroxyindoleacetic acid) and seven prescription components (danshensu, protocatechualdehyde, spinosin, 6’’’-feruylspinosin, salviaolic acid B, schisandrin and deoxyschisandrin) in rats’ tissues. Tissue samples were prepared by protein precipitation with acetonitrile. Chromatographic separation was carried out on a C18 column with a gradient mobile phase consisting of acetonitrile and 0.01% formic acid water. An electrospray ionization triple quadrupole concatenation mass spectrometer was set to switch between positive and negative modes in a single run time. All components were quantitated by multiple-reaction monitoring scanning.

Results: The lower limits of quantitation for all analytical components were 0.78 ng/mL-1.99 ng/mL in heart, liver, spleen, lung, kidney and brain. All calibration curves displayed good linearity (r > 0.99544). The precision was evaluated by intra-day and inter-day assays, and the relative standard deviation (RSD) values were all within 12.67%. The relative errors of the accuracy were all within ± 19.88%. The recovery ranged from 76.00% to 98.78% and the matrix effects of eleven components were found to be between 85.10% and 96.40%.

Conclusion: This method was successfully applied to study the distribution of seven components from ZAP and the concentration changes of four monoamine neurotransmitters after oral ZAP in six tissues.

Keywords:

Zaoren Anshen prescription, UPLC-MS/MS, Simultaneous determination, Monoamine neurotransmitters, Effective components, Tissue distribution

Affiliation:

Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, Northwest University, Xi`an 710069, Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, Northwest University, Xi`an 710069, Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, Northwest University, Xi`an 710069, Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, Northwest University, Xi`an 710069, Xi`an Institute for Food and Drug Control, Xi`an 710054, Xi`an Institute for Food and Drug Control, Xi`an 710054, Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, Northwest University, Xi`an 710069



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