Quantification of sulfatides in dried blood and urine spots from metachromatic leukodystrophy patients by liquid chromatography/electrospray tandem mass spectrometry

Barcenas, Mariana; Suhr, Teryn; Scott, C. Ronald; Tureček, František; Gelb, Michael H.

doi:10.1016/j.cca.2013.12.016

Cited by 28 publications

(29 citation statements)

References 11 publications

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“…Plasma samples from human MLD patients show relatively high levels of C16:0 and C16:0-OH sulfatides. A similar observation for the C16:0 species was recently reported for sulfatides in blood spots from MLD patients ( 24,25 ) In addition to the sulfatide determination, we describe the determination of lysosulfatide, the deacylated form of the primary storage product sulfatide, by LC/MS/MS for containing small amounts of sulfatides in the presence of large amounts of neutral and phopholipids like plasma. Overall, fractionation on SPE led to better performance on LC/MS/MS and allowed for a much more sensitive detection of sulfatides in tissues, cells (not shown), and body fl uids that contain low amounts of sulfatides.…”

Section: Measurement Of Different Molecular Sulfatide Species In Fl Usupporting

confidence: 72%

“…MLD mice models and human patients can easily be distinguished from controls. In contrast to recently published methods on the quantifi cation of sulfatides in dried blood spots and urine ( 24,25 ) from MLD patients and normal controls, our method could easily quantify sulfatides in normal plasma and urine with a signal to noise ratio of >10 for all peaks quantifi ed. Because of its sensitivity, only 0.5 ml of a representative urine sample suffi ces for the analysis of MLD and normal urines instead of the large volumes or even 24 h urine collections, which are needed when traditional methods like TLC or HPLC are used ( 14,19,32 ).…”

Section: Measurement Of Different Molecular Sulfatide Species In Fl Umentioning

confidence: 90%

“…Several methods have been used to quantify sulfatides, including TLC ( 19 ), HPLC ( 14,20 ), and, more recently, MS ( 4,13,15,(21)(22)(23)(24)(25). Here we describe a LC/MS/MS method to quantify sulfatides and their molecular species over a wide range of concentrations in normal and diseased tissues, plasma, and urine.…”

Section: Extraction Of Sulfatides From Plasma and Tissue Homogenatesmentioning

confidence: 99%

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Quantification of sulfatides and lysosulfatides in tissues and body fluids by liquid chromatography-tandem mass spectrometry

Mirzaian

Kramer²,

Poorthuis

2015

Journal of Lipid Research

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This article is available online at http://www.jlr.org high concentrations are found in selected organs, including kidney, colon, pancreas, and gall bladder. Increased concentrations of sulfatides have been reported in renal cell carcinoma, adenocarcinoma of colon and lung, and ovarian cancer ( 2, 4 ). Sulfatides have been proposed to play a role as ion barriers to osmotic stress and as counterions of ammonium in the renal adaptation to chronic metabolic acidosis ( 5,6 ) Sulfatides are synthesized from their precursor galactosylceramide in the Golgi apparatus by 3-O-sulfation of the galactose residue by the enzyme 3 ′ -phosphoadenosine-5 ′ -phosphosulfate: cerebroside sulfotransferase (CST) (EC2.8.2.11). Galactosylceramide is synthesized in the endoplasmic reticulum from ceramides and UDP-galactose by the enzyme UDP-galactose:ceramide galactosyltransferase (CGT) (EC 2.4.1.45) and is then transported to the Golgi apparatus prior to sulfation to sulfatides. Sulfatides consist of many molecular species, with structures differing in acyl chain length and hydroxylation and sphingoid base. These sulfatide molecular species are cell and tissue specifi c, which may be explained by the cell-and tissuespecifi c expression of ceramide synthases and fatty acid 2-hydroxylase ( 1, 7 ). It is diffi cult to assign specifi c functions to the different molecular species of sulfatides, but sulfatides with C16:0 fatty acids were reported to be involved in the regulation of insulin secretion in rat pancreatic ␤ -cells ( 8 ), and elevated levels of C18:0 sulfatides have been implicated as a cause of audiogenic seizers in mice overexpressing CGT ( 9 ). There is also heterogeneity of the sphingoid base composition of sulfatides. Most of the Abstract Sulfatides are found in brain as components of myelin, oligodendrocytes, and neurons but are also present in various visceral tissues. Metachromatic leukodystrophy (MLD) is an inherited lysosomal storage disorder caused by a defi ciency of arylsulfatase A, leading to severe white matter disease due to the accumulation of sulfatides and lysosulfatides. To study the physiological role of sulfatides, accessible and sensitive quantitative methods are required. We developed a sensitive LC/MS/MS method to quantify total sulfatide and lysosulfatide content as well as individual molecular species in urine and plasma from MLD patients and plasma and tissues from an MLD mouse model. Our results demonstrate that the method can quantify a wide range of sulfatide concentrations and can be used to quantify total sulfatide content and levels of individual molecular species of sulfatides in tissues, cells, and body fl uids. Even though plasma sulfatides and lysosulfatides would seem attractive candidate biomarkers that could possibly correlate with the severity of MLD and be of use to monitor the effects of therapeutic intervention, our results indicate that it is unlikely that the determination of these storage products in plasma will be useful in this respect. Sulfatides are anionic sulfoglycolipids main...

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Section: Measurement Of Different Molecular Sulfatide Species In Fl Usupporting

confidence: 72%

Section: Measurement Of Different Molecular Sulfatide Species In Fl Umentioning

confidence: 90%

Section: Extraction Of Sulfatides From Plasma and Tissue Homogenatesmentioning

confidence: 99%

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Quantification of sulfatides and lysosulfatides in tissues and body fluids by liquid chromatography-tandem mass spectrometry

Mirzaian

Kramer²,

Poorthuis

2015

Journal of Lipid Research

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“…Similar to many other groups, we used reversed-phase chromatography for the separation of ST species prior to MS detection (23,24,26,28,38). Using this approach, the separation of individual species was mainly based on the difference in fatty acid moieties attached to the sphingosine backbone.…”

Section: Discussionmentioning

confidence: 99%

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

Blomqvist

Borén

Zetterberg

et al. 2017

Journal of Lipid Research

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This article is available online at http://www.jlr.org ceramide (sphingoid base and a fatty acid) and a sulfated galactose. STs are, together with galactosylceramides, the most typical myelin lipids, where the long-chain fatty acid moieties C24:0 and C24:1 are expressed in particular (1, 2). In addition to myelin, STs have been shown to be expressed in subpopulations of neurons and astrocytes in the gray matter of the brain (3, 4). STs and galactosylceramides are required for the stability and maintenance of the myelin sheet (5), whereas the STs found in other brain regions suggest more diverse functions of these glycosphingolipids (6). Furthermore, STs have been suggested to participate in other cellular processes, such as protein trafficking, cell adhesion, and aggregation and immune responses, among others (7). STs are also present in many other organs, including kidney, liver, heart, intestine, muscle, and pancreas (8), although in minor amounts compared with brain, except in kidney.Findings indicate that the release of STs from myelin is associated with the development of CNS diseases and that STs in cerebrospinal fluid (CSF) might function as a marker of demyelination, myelin damage, and myelin turnover (9-13). Changes in CSF and brain ST levels have also been observed in other neurological disorders, such as early Alzheimer's disease (AD) (14-16). Furthermore, the absence of hydroxylated forms of ST in CSF could be used as a diagnostic tool for fatty acid 2-hydroxylase deficiency (17). These examples accentuate the need for a robust and sensitive method for quantification of STs in CSF.In previous studies, CSF ST measurements have usually been performed by TLC with monoclonal antibody 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 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.-Blomqvi...

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“…However, NBS programs typically use DBS, and dried urine samples (DUS) are usually not available. Recent studies have shown that sulfatides are increased in DBS from MLD patients (8, 9), but the discriminatory power of sulfatide concentrations in DBS between MLD patients and controls is less pronounced than in DUS, raising questions about the potential for NBS. The interest in screening for MLD is timely owing to new treatment options being investigated in the clinic (10).…”

mentioning

confidence: 99%

Sulfatide Analysis by Mass Spectrometry for Screening of Metachromatic Leukodystrophy in Dried Blood and Urine Samples

Spáčil¹,

Kumar²,

Liao³

et al. 2016

Clinical Chemistry

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BACKGROUND Metachromatic leukodystrophy (MLD) is an autosomal recessive disorder caused by deficiency in arylsulfatase A activity, leading to accumulation of sulfatide substrates. Diagnostic and monitoring procedures include demonstration of reduced arylsulfatase A activity in peripheral blood leukocytes or detection of sulfatides in urine. However, the development of a screening test is challenging because of instability of the enzyme in dried blood spots (DBS), the widespread occurrence of pseudodeficiency alleles, and the lack of available urine samples from newborn screening programs. METHODS We measured individual sulfatide profiles in DBS and dried urine spots (DUS) from MLD patients with LC-MS/MS to identify markers with the discriminatory power to differentiate affected individuals from controls. We also developed a method for converting all sulfatide molecular species into a single species, allowing quantification in positive-ion mode upon derivatization. RESULTS In DBS from MLD patients, we found up to 23.2-fold and 5.1-fold differences in total sulfatide concentrations for early- and late-onset MLD, respectively, compared with controls and pseudodeficiencies. Corresponding DUS revealed up to 164-fold and 78-fold differences for early- and late-onset MLD patient samples compared with controls. The use of sulfatides converted to a single species simplified the analysis and increased detection sensitivity in positive-ion mode, providing a second option for sulfatide analysis. CONCLUSIONS This study of sulfatides in DBS and DUS suggests the feasibility of the mass spectrometry method for newborn screening of MLD and sets the stage for a larger-scale newborn screening pilot study.

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Quantification of sulfatides in dried blood and urine spots from metachromatic leukodystrophy patients by liquid chromatography/electrospray tandem mass spectrometry

Cited by 28 publications

References 11 publications

Quantification of sulfatides and lysosulfatides in tissues and body fluids by liquid chromatography-tandem mass spectrometry

Quantification of sulfatides and lysosulfatides in tissues and body fluids by liquid chromatography-tandem mass spectrometry

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

Sulfatide Analysis by Mass Spectrometry for Screening of Metachromatic Leukodystrophy in Dried Blood and Urine Samples

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