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Sphingolipids Analysis Service

Creative Proteomics offers high-resolution sphingolipid analysis services using LC-MS/MS and Orbitrap platforms to deliver accurate quantification of over 150 sphingolipid species, including ceramides, sphingomyelins, glycosphingolipids, and signaling lipids like S1P. Our service enables researchers to decode lipid pathway disruptions, identify disease biomarkers, and support drug development with precise, reproducible lipidomic data.

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List of Sphingolipids
Advantages
Workflow
Methods
Results and Data Analysis
Sample Requirements
FAQ

What are Sphingolipids?

Sphingolipids are a complex class of bioactive lipids involved in membrane structure, cell signaling, apoptosis, inflammation, and immune response. They include ceramides, sphingomyelins, glycosphingolipids, and sphingosine-1-phosphate, each playing distinct roles in physiological and pathological processes.

Quantitative sphingolipid profiling is essential for understanding lipid metabolism, identifying disease-specific lipid alterations, and discovering biomarkers in cancer, neurodegeneration, metabolic syndromes, and inflammatory conditions. Accurate analysis supports mechanistic studies, therapeutic targeting, and translational research.

Sphingolipids Analysis in Creative Proteomics

Targeted Sphingolipid Quantification

Absolute quantification of individual sphingolipids using class-specific internal standards and LC-MS/MS methods; ideal for biomarker validation.

Untargeted Sphingolipidomics Profiling

Global screening and relative quantification of known and novel sphingolipids using high-resolution UHPLC-QTOF-MS or Orbitrap platforms.

Sphingolipid Class Profiling

Focused analysis of specific lipid classes such as ceramides, sphingomyelins, or glycosphingolipids to investigate class-specific dysregulation.

Sphingolipid Metabolic Pathway Mapping

Analysis of key metabolites and intermediates in sphingolipid biosynthesis and degradation pathways (e.g., de novo pathway, salvage pathway, S1P pathway).

Isotope Tracer-Based Flux Analysis

Dynamic profiling of sphingolipid metabolic turnover using stable isotope-labeled precursors (e.g., 13C-palmitate, 15N-serine).

Comparative Lipidomics (Case-Control or Dose-Response)

Statistical comparison of sphingolipid profiles across experimental groups, drug treatments, or disease models.

Tissue-Specific Sphingolipid Distribution

Subcellular or tissue-specific profiling to reveal spatial heterogeneity (e.g., brain, liver, tumor, plasma, CSF).

Biomarker Discovery Services

Identification of differential sphingolipid signatures associated with pathological states using multivariate statistics and machine learning.

Custom Sphingolipid Panels

Tailor-made panels based on disease relevance, organism model, or analytical goals; ideal for translational research and companion diagnostics.

List of Detected Sphingolipids

Sphingolipid Class	Representative Detected Species	Associated Metabolites	Relevant Metabolic Pathways
Ceramides (Cer)	Cer(d18:1/16:0), Cer(d18:1/18:0), Cer(d18:1/20:0), Cer(d18:1/24:1), Cer(d18:1/24:0)	Dihydroceramides, Ceramide-1-phosphate, Inositolphosphoryl-ceramides	De novo synthesis, Ceramide salvage, Inflammatory signaling
Dihydroceramides (dhCer)	dhCer(d18:0/16:0), dhCer(d18:0/18:0), dhCer(d18:0/24:1)	Sphinganine	De novo ceramide biosynthesis
Sphingomyelins (SM)	SM(d18:1/16:0), SM(d18:1/18:1), SM(d18:1/24:0), SM(d18:1/24:1), SM(d18:1/20:0)	Ceramides, Choline	Sphingomyelin cycle, Membrane dynamics
Sphingosines & Derivatives	Sphingosine Base: Sphingosine (d18:1), Sphinganine (d18:0); Sphingosine-1-phosphate (S1P), Sphinganine-1-phosphate (Sa1P)	Phosphoethanolamine, NADPH	S1P signaling pathway, Ceramide breakdown
Ceramide-1-phosphate (C1P)	C1P(16:0), C1P(18:0), C1P(24:1)	Ceramides	Inflammation regulation, Cell survival
Glycosphingolipids (GSLs)	Cerebrosides: Glucosylceramide (GlcCer), Galactosylceramide (GalCer); Dihexosylceramides (CerG2): Lactosylceramide (LacCer); Gb3, Gb4	UDP-Glucose, UDP-Galactose	Glycosphingolipid biosynthesis, Lysosomal storage pathways
Globosides	Gb3, Gb4	UDP-Galactose, UDP-N-acetylglucosamine	Blood group antigen synthesis, Glycosphingolipid signaling
Hexosylceramides (HexCer)	HexCer(d18:1/16:0), HexCer(d18:1/22:0), HexCer(d18:1/24:1)	Ceramides, Monosaccharides	Glycosylation pathway
Sulfatides (ST)	Sulfo-GalCer, ST(d18:1/24:1)	GalCer, PAPS (3'-phosphoadenosine-5'-phosphosulfate)	Myelin maintenance, Neurodegeneration pathways
Gangliosides	GM3, GM2, GM1, GD1a, GD3	CMP-Neu5Ac, Lactosylceramide	Neuronal development, Ganglioside synthesis
Phytoceramides	PhytoCer(d18:0/24:0), PhytoCer(d18:0/20:0)	Phytosphingosine	Fungal sphingolipid metabolism (also relevant to plant models)
Sphingolipid Intermediates	3-Ketosphinganine, Palmitoyl-CoA, Serine	-	De novo biosynthesis pathway initiation

Why Choose Our Sphingolipids Assay?

Detection sensitivity as low as 1–5 ng/mL for key sphingolipids such as S1P and ceramides.
Quantification linear range spans four orders of magnitude, enabling both low- and high-abundance lipid detection.
Over 90% of detected sphingolipid species show RSD <10%, ensuring high analytical reproducibility.
Capable of profiling 150+ sphingolipid species, covering all major subclasses including rare glycosphingolipids.
Detection includes low-abundance species such as inositol-phosphoryl ceramides and sulfatides.

Analysis performed on Thermo Orbitrap Exploris 480 or SCIEX Triple Quad 6500+ mass spectrometers.
Mass accuracy maintained within ≤2 ppm and resolution up to 240,000 FWHM at m/z 200.
Quantitative MRM/PRM workflows ensure targeted accuracy for specific lipid panels.
Lipid extraction protocols achieve >90% recovery efficiency from plasma, tissue, and cell samples.
Internal standards for each lipid class used to normalize matrix effects and enhance quantitation precision.

How Creative Proteomics Provides Sphingolipid Assay?

Workflow for Sphingolipid Analysis

What Methods are Used for Our Sphingolipids Analysis?

Triple Quadrupole LC-MS/MS

Targeted sphingolipid quantification with femtomolar sensitivity

SCIEX Triple Quad™ 6500+ (Figure from Sciex)

Orbitrap MS

High-resolution profiling, isotope tracing

Thermo Fisher Q Exactive (Figure from Thermo Fisher)

UHPLC-QTOF-MS

Untargeted lipidomics and structural elucidation

Agilent 6545 Q-TOF(Figure from Agilent)

Sphingolipids Analysis Service: Results and Data Analysis

Standard Deliverables

Quantitative result tables including absolute or relative concentrations (e.g., ng/mL, pmol/µg protein) of all detected sphingolipid species.
Lipid class summary reports showing total abundance of each sphingolipid subclass (e.g., Cer, SM, HexCer) across all samples.
Normalized data matrices corrected by internal standards and sample loading amounts, compatible with statistical software and pathway tools.
Full raw data files (Thermo .raw or SCIEX .wiff/.mzML formats) for in-house reanalysis or publication.
Quality control reports, including technical replicates, RSD%, recovery rates, and instrument calibration curves.

Sphingolipid chromatographic traces by LC–MS/MS (Burrello, J., et al., 2020).

LC ESI-MS/MS elution profiles for the sphingolipids on reverse phase (A, B) and normal phase (C, D) chromatography (Shaner, Rebecca L., et al., 2009).

Advanced Data Analysis (Optional)

Multivariate statistical analysis: PCA, PLS-DA, hierarchical clustering for sample classification and biomarker discovery.
Differential lipid expression analysis: Fold-change, p-value, FDR-adjusted statistics between experimental groups.
Pathway enrichment and lipid ontology analysis: Mapping sphingolipids to KEGG, Reactome, or LIPID MAPS pathways to identify altered metabolic routes.
Lipid structural annotation: Fragment ion-based confirmation of isomeric or unusual species (e.g., odd-chain ceramides, ether-linked sphingolipids).

Delivery Formats

Excel and CSV tables for ease of integration into downstream workflows.
Graphical summary reports (PDF) including volcano plots, heatmaps, lipid class distributions, and pathway diagrams.
Technical summary and interpretation notes from our lipidomics experts, highlighting key findings and potential biological implications.

Explore our Lipidomics Solutions brochure to learn more about our comprehensive lipidomics analysis platform.

Download Brochure

What Our Sphingolipids Analysis Used For

Metabolism and Obesity

Profiling lipid changes in metabolic disorders like obesity, diabetes, and insulin resistance.

Neurodegenerative Diseases

Analyzing serum lipids in Alzheimer's, Parkinson's, and other neurological disorders.

Inflammation and Immunity

Investigating lipid profiles in inflammation and immune response, relevant to autoimmune diseases.

Aging and Age-Related Diseases

Exploring lipidomic shifts linked to aging and related pathologies.

Toxicology and Environmental Exposure

Examining lipid metabolism changes due to environmental toxins and pollutants.

Nutritional Impact

Assessing how diet, fasting, and supplements affect lipid metabolism

Sample Requirements for Sphingolipids Analysis Solutions

Sample Type	Recommended Amount
Plasma / Serum	≥ 200 µL
Whole Blood	≥ 500 µL
Tissue	≥ 50 mg (wet weight)
Cells	≥ 1 × 10⁷ cells or equivalent pellet
Cerebrospinal Fluid	≥ 500 µL
Urine	≥ 1 mL (preferably concentrated)
Feces	≥ 100 mg (fresh or lyophilized)
Liver Microsomes	≥ 200 µg protein
Lipid Extracts	≥ 50 µg total lipids
Plant Tissues	≥ 100 mg (fresh or flash-frozen)
Model Organisms (e.g. Drosophila, C. elegans)	≥ 50 mg pooled sample

FAQs for Sphingolipids Analysis Service

How should I prepare biological samples before submission?

For plasma/serum: Centrifuge at 1,500 × g for 10 min to remove cellular debris. Avoid repeated freeze-thaw cycles.
For tissues: Flash-freeze in liquid nitrogen and homogenize to a fine powder before lipid extraction.
For cells: Wash 3× with PBS to remove culture media contaminants.

Can you differentiate between structural isomers (e.g., C16:0 vs. C18:0 ceramides)?

Yes. Our UPLC-MS/MS platform achieves baseline separation of isomers using optimized gradients (e.g., 0.1% formic acid in water/acetonitrile) with retention time shifts <0.1 min.

Do you support time-course or dose-response study designs?

Absolutely. We design batch analyses to minimize inter-run variability (CV <8%) and include QC samples at every 10th injection to ensure longitudinal data consistency.

How are low-abundance sphingolipids (e.g., S1P) stabilized during processing?

We add 10 mM EDTA and 0.1% butylated hydroxytoluene (BHT) during extraction to inhibit enzymatic degradation and oxidation.

Can I request analysis of oxidized or modified sphingolipids?

Yes. Our methods detect oxidized species (e.g., hydroxylated ceramides) and sulfatides using precursor ion scanning (PIS) and neutral loss modes.

How do you handle lipid identification for novel/uncharacterized sphingolipids?

Untargeted profiling via high-resolution MS (Orbitrap Exploris 480) with MS/MS spectral matching to databases (e.g., LIPID MAPS, HMDB). Putative structures are reported with m/z accuracy <3 ppm.

What if my samples have high background interference (e.g., serum with high triglycerides)?

We apply a two-phase liquid-liquid extraction (chloroform/methanol/water) to remove neutral lipids, achieving >95% purity for sphingolipid fractions.

How do you handle samples with high lipid diversity (e.g., plant or microbial sphingolipids)?

Our methods are optimized for cross-kingdom compatibility, including plant-specific sphingolipids (e.g., phytoceramides) and microbial species (e.g., inositol phosphorylceramide). We use high-resolution mass spectrometry (Orbitrap Exploris 480) with customized spectral libraries to ensure accurate identification .

Can I analyze sphingolipids in cell culture media or extracellular vesicles?

Yes. We provide protocols for extracellular vesicle (EV) isolation and analysis, ensuring minimal contamination from culture media lipids. Sensitivity is maintained at 0.5 pmol/mL for low-abundance species like S1P in EVs .

What bioinformatics tools do you use for pathway analysis?

We employ PathVisio and KEGG pathway mapping to visualize sphingolipid metabolism and integrate data with transcriptomic/proteomic datasets. Custom pathway overlays are included in reports .

Are your methods compatible with stable isotope tracing for flux analysis?

Yes. We support 13C- or 2H-labeled tracer studies to monitor sphingolipid turnover rates. Detection limits for labeled species are <1% natural abundance, enabling precise metabolic flux modeling .

Can you analyze sphingolipids in formalin-fixed paraffin-embedded (FFPE) tissues?

Yes. We apply lipid recovery protocols for FFPE samples, achieving >80% extraction efficiency for ceramides and sphingomyelins compared to fresh-frozen tissues .

How are data reported for untargeted sphingolipid discovery?

Untargeted results include m/z values, MS/MS spectra, and putative identifications (matched to LIPID MAPS/HMDB databases). Annotated spectra are provided with mass accuracy <3 ppm .

What is the impact of freeze-thaw cycles on sphingolipid stability?

Sphingolipids like S1P are sensitive to freeze-thaw cycles. We recommend ≤2 cycles for plasma/serum and use stabilization buffers (e.g., 10 mM EDTA) to minimize degradation .

Publications

White matter lipid alterations during aging in the rhesus monkey brain. GeroScience, 2024. https://doi.org/10.1007/s11357-024-01353-3
Characterization of Dnajc12 knockout mice, a model of hypodopaminergia. bioRxiv, 2024. https://doi.org/10.1101/2024.07.06.602343
Annexin A2 modulates phospholipid membrane composition upstream of Arp2 to control angiogenic sprout initiation. The FASEB Journal, 2023. https://doi.org/10.1096/fj.202201088R
Lipid Membrane Engineering for Biotechnology (Doctoral dissertation, Aston University). Lipid Membrane Engineering for Biotechnology, 2023. https://doi.org/10.48780/publications.aston.ac.uk.00046663
Multi‐omics identify xanthine as a pro‐survival metabolite for nematodes with mitochondrial dysfunction. EMBO Journal, 2019. https://doi.org/10.15252/embj.201899558
Evidence for phosphate-dependent control of symbiont cell division in the model anemone Exaiptasia diaphana. mBio, 2024. https://doi.org/10.1128/mbio.01059-24
The olfactory receptor Olfr78 promotes differentiation of enterochromaffin cells in the mouse colon. EMBO reports, 2024. https://doi.org/10.1038/s44319-023-00013-5
Metabolomic Studies in Girls With Central and Peripheral Precocious Puberty. Fabad Journal of Pharmaceutical Sciences, 2023. https://doi.org/10.55262/fabadeczacacilik.1344851
Prospective randomized, double-blind, placebo-controlled study of a standardized oral pomegranate extract on the gut microbiome and short-chain fatty acids. Foods, 2023. https://doi.org/10.3390/foods13010015

References

Burrello, J., et al. "Sphingolipid composition of circulating extracellular vesicles after myocardial ischemia." Scientific reports 10.1 (2020): 16182. https://doi.org/10.1038/s41598-020-73411-7
Shaner, Rebecca L., et al. "Quantitative analysis of sphingolipids for lipidomics using triple quadrupole and quadrupole linear ion trap mass spectrometers [S]." Journal of lipid research 50.8 (2009): 1692-1707. https://doi.org/10.1194/jlr.D800051-JLR200

* Our services can only be used for research purposes and Not for clinical use.

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