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Simultaneous Determination of D-amino Acids in Rat Urine by Highperformance Liquid Chromatography-tandem Mass Spectrometry Method: Application to Investigate the Clinical Value of D-amino Acids in the Early Diagnosis of Alzheimer’s Disease

[ Vol. 17 , Issue. 8 ]

Author(s):

Min Zhang, Shuting Zhang, Weichao Yu, Xiaoyan Li, Ning Ma and Yan Cui*   Pages 1026 - 1035 ( 10 )

Abstract:


Background: D-amino acids are closely related to the development and progression of Alzheimer's disease (AD) and are expected as the novel biomarkers for AD diagnosis.

Objective: The aim was to investigate the potential clinical value of D-amino acids for Alzheimer's disease.

Methods: A simple and sensitive HPLC/MS-MS method was developed for the simultaneous determination of D-alanine, D-glutamine, D-proline and D-serine in rat urine. The samples were firstly pretreated by methanol, then derivatized by 7-chloro-4-nitrobenzoxadiazole with Fudosteine as internal standard, enantioseparated on Sumichiral OA-2500S column, using a mobile phase composed of acetonitrile- methanol (50:50, v/v) containing 0.5% formic acid, and detected with 4000 Qtrap MS/MS in electrospray-ionization source by negative ion mode.

Results: The established method was successfully applied to determine the D-amino acid levels in rat urine from 20 Alzheimer's disease rats and 20 age-matched normal controls. The mean levels of Damino acids in the urine of Alzheimer's disease rats were all significantly lower than those in normal controls. Based on the contents of D-amino acids, the distinction model between Alzheimer's disease rats and normal controls was established by the Bayesian discriminant analysis.

Conclusion: The relationship between Alzheimer's disease and D-amino acids revealed that D-amino acids would be potential biomarkers for Alzheimer’s disease.

Keywords:

D-amino acids, Alzheimer’s disease, biomarkers, HPLC/MS-MS, Simultaneous determination, enantioseparation.

Affiliation:

School of Functional Food and Wine, Shenyang Pharmaceutical University, 103 Wenhua Road, Shenyang 110016, School of Functional Food and Wine, Shenyang Pharmaceutical University, 103 Wenhua Road, Shenyang 110016, School of Pharmacy, Shenyang Pharmaceutical University, 103 Wenhua Road, Shenyang 110016, PAREXEL International Unit, 286 Qingnian Street, Shenyang 110004, The Second Affiliated Hospital of Liaoning University of Traditional Chinese Medicine, 60 North Yellow River Street, Shenyang,110034, School of Pharmacy, Shenyang Pharmaceutical University, 103 Wenhua Road, Shenyang 110016



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