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High-throughput analysis of sulfatides in cerebrospinal fluid using automated extraction and UPLC-MS/MS

Journal article
Authors Maria K. Blomqvist
Jan Borén
Henrik Zetterberg
Kaj Blennow
Jan-Eric Månsson
Marcus Ståhlman
Published in Journal of Lipid Research
Volume 58
Issue 7
Pages 1482-1489
ISSN 0022-2275
Publication year 2017
Published at Institute of Biomedicine, Department of Clinical Chemistry and Transfusion Medicine
Institute of Neuroscience and Physiology, Department of Psychiatry and Neurochemistry
Institute of Medicine, Department of Molecular and Clinical Medicine
Pages 1482-1489
Language en
Links doi.org/10.1194/jlr.D076588
Keywords ultra-performance liquid chromatographytandem mass spectrometry, sulfatides, Alzheimer's disease, tandem mass-spectrometry, normal-pressure hydrocephalus, central-nervous-system, total lipid extraction, white-matter lesions, metachromatic leukodystrophy, alzheimers-disease, dried blood, neurological disorders, multiple-sclerosis, Biochemistry & Molecular Biology
Subject categories Biochemistry and Molecular Biology, Clinical Medicine

Abstract

Sulfatides (STs) are a group of glycosphingolipids that are highly expressed in brain. Due to their importance for normal brain function and their potential involvement in neurological diseases, development of accurate and sensitive methods for their determination is needed. Here we describe a high-throughput oriented and quantitative method for the determination of STs in cerebrospinal fluid (CSF). The STs were extracted using a fully automated liquid/liquid extraction method and quantified using ultra-performance liquid chromatography coupled to tandem mass spectrometry. With the high sensitivity of the developed method, quantification of 20 ST species from only 100 mu l of CSF was performed. Validation of the method showed that the STs were extracted with high recovery (90%) and could be determined with low inter-and intra-day variation. Our method was applied to a patient cohort of subjects with an Alzheimer's disease biomarker profile. Although the total ST levels were unaltered compared with an age-matched control group, we show that the ratio of hydroxylated/nonhydroxylated STs was increased in the patient cohort. In conclusion, we believe that the fast, sensitive, and accurate method described in this study is a powerful new tool for the determination of STs in clinical as well as preclinical settings.-Blomqvist, M., J. Boren, H. Zetterberg, K. Blennow, J-E. Mansson, and M. Stahlman. High-throughput analysis of sulfatides in cerebrospinal fluid using automated extraction and UPLC-MS/MS.

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