Main content area

Development of an UPLC–MS/MS method for simultaneous quantitation of 11 d-amino acids in different regions of rat brain: Application to a study on the associations of d-amino acid concentration changes and Alzheimer’s disease B Analytical technologies in the biomedical and life sciences

Li, Zhe, Xing, Yuping, Guo, Xingjie, Cui, Yan
Journal of chromatography 2017 v.1058 pp. 40-46
Alzheimer disease, acetonitrile, cerebellum, cerebral cortex, hippocampus, liquid chromatography, monitoring, olfactory bulb, patients, rats, serine, tandem mass spectrometry
There are significant differences in d-amino acid concentrations between healthy people and Alzheimer’s disease patients. In order to investigate the potential correlation between d-amino acids and Alzheimer’s disease, a simple and sensitive ultra high performance liquid chromatography-tandem mass spectrometry (UPLC–MS/MS) method has been developed. The method was applied to simultaneous determination of 11 d-amino acids in different regions of rat brain. Rat brain homogenates were firstly pretreated with protein precipitation procedure and then derivatized with (S)-N-(4-nitrophenoxycarbonyl) phenylalanine methoxyethyl ester [(S)-NIFE]. Baseline separation of the derivatives was achieved on an ACQUITY UPLC BEH C18 column (2.1 mm×50mm, 1.7μm). The mobile phase consisted of acetonitrile and water (containing 8mM ammonium hydrogen carbonate) and the flow rate was 0.6mLmin⁻¹. The derived analytes were sensitively detected by multiple reaction monitoring in the positive ion mode. The lower limits of quantitation ranged from 0.06 to 10ngmL⁻¹ with excellent linearity (r≥0.9909). The intra- and inter-day RSD were in the range of 3.6–12% and 5.7–12%, respectively. The recovery rate was 82.5%–95.3%. With this UPLC–MS/MS method, the 11 d-amino acids in hippocampus, cerebral cortex, olfactory bulb and cerebellum from Alzheimer’s disease rats and age-matched controls could be simultaneously determined. Compared with the normal controls, the concentrations of d-serine, d-alanine, d-leucine, and d-proline in hippocampus and cerebral cortex of Alzheimer’s disease rat brain were significantly decreased, while no differences in olfactory bulb and cerebellum of all the d-amino acids were observed. The different amounts and distribution of d-amino acids in brain between the two groups, which regulated by particular pathological changes of Alzheimer’s disease, would give new insights into further study in neuropathogenesis and provide novel therapeutic targets of Alzheimer’s disease.