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

• Published in: Journal of lipid research Vol. 58; no. 7; pp. 1482 - 1489
• Main Authors: Blomqvist, Maria, Borén, Jan, Zetterberg, Henrik, Blennow, Kaj, Månsson, Jan-Eric, Ståhlman, Marcus
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.07.2017; Journal of Lipid Research; The American Society for Biochemistry and Molecular Biology; Elsevier
• Subjects: Adult; Aged; Alzheimer's disease; alzheimers-disease; Automation; Biochemistry & Molecular Biology; central-nervous-system; Cerebrospinal fluid; Chemical Fractionation - methods; Chromatography, High Pressure Liquid - methods; Clinical Chemistry Tests - methods; Clinical Medicine; dried blood; Glycosphingolipids; Humans; Klinisk medicin; Liquid chromatography; Mass spectroscopy; metachromatic leukodystrophy; Methods; Middle Aged; Molecular Biology; Molekylärbiologi; multiple-sclerosis; Neurological diseases; neurological disorders; normal-pressure hydrocephalus; sulfatides; Sulfoglycosphingolipids - cerebrospinal fluid; Sulfoglycosphingolipids - isolation & purification; Tandem Mass Spectrometry - methods; tandem mass-spectrometry; total lipid extraction; ultra-performance liquid chromatography-tandem mass spectrometry; ultra-performance liquid chromatographytandem mass spectrometry; white-matter lesions; sulfatides; ultra-performance liquid chromatography-tandem mass spectrometry; Alzheimer's disease; Alzheimer’s disease
• ISSN: 0022-2275; 1539-7262; 1539-7262
• DOI: 10.1194/jlr.D076588

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 μl 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.