Journal of East China Normal University(Natural Sc ›› 2013, Vol. 2013 ›› Issue (6): 165-170.

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HPLC-ESI-MS determination of neurotransmitters L-serine/D-serine in rat brain tissues

WANG Wei-li, DUAN Ya-le, LIU Sha, ZHANG Yao, ZHAO Zheng   

  1. Laboratory of Neuropharmacology and Toxicology, Key Laborarory of Brain Functional Genomics, Ministry of Education, Shanghai Key Laboratory of Brain Functional Genomics, East China Normal University, Shanghai 200062, China
  • Received:2013-01-01 Revised:2013-04-01 Online:2013-11-25 Published:2014-01-13

Abstract: A highly sensitive and selective high performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) assay was developed for the determination of neurotransmitters L-serine/D-serine in rat brain tissues. The rat brain was removed, homogenized and centrifuged.L-serine/D-serine in the supernatant was derivatized with the Marfey’s reagent. The derivatives were separated by HPLC under gradient elution of mobile phase consisting of ammonium formate and acetonitrile and detected by ESI-MS in the negative ion mode. Reverse phase chromatographic resolution of the enantiomers was obtained within 22 min. Good linear correlations of the concentration to the peak area were obtained in the ranges of 2.5~500 ng/mg for L-serine and 1.2~250 ng/mg for D-serine. The extraction recoveries for both analysts were in the range of 81% to 89%. The method was then applied to determine the levels of the neurotransmitters in cerebral cortex and hippocampus of the adult and aged rats. The results showed that L-serine levels kept unchanged,while D-serine levels decreased significantly, in cortex and hippocampus of the aged rats compared with those of the adult rats, suggesting an involvement of D-serine in the normal aging process.

Key words: HPLC-ESI-MS, brain tissue, neurotransmitter, D-serine, L-serine

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