Heparan Sulfate & Heparin LC-MS/MS Quantification Service

Why Analyze Heparan Sulfate and Heparin?

Heparan sulfate (HS) and heparin are structurally diverse glycosaminoglycans (GAGs) that play essential roles in biological signaling, extracellular matrix integrity, and disease progression. Their functions are not dictated by a single sequence but by subtle differences in disaccharide composition, sulfation motifs, and chain length distribution.

These complex features govern interactions with growth factors, cytokines, and enzymes—making HS/heparin pivotal in coagulation regulation, lysosomal storage disorders (MPS), tumor progression, and host–pathogen interactions. Yet, conventional glycan detection methods often fail to resolve these fine structural determinants, leaving gaps in interpretation and comparability across samples.

At Creative Proteomics, we address this analytical challenge by offering quantitative, disaccharide-resolved LC–MS/MS-based HS/heparin profiling. Our platform allows researchers to:

• Accurately characterize N-, 2-O-, 6-O-, and 3-O-sulfation patterns
• Quantify total HS/heparin levels in serum, tissue, urine, and bioprocess samples
• Compare structural fingerprints across models, conditions, or drug lots

Whether you're working on MPS biomarkers, heparin product QC, or ECM remodeling, our GAG-targeted workflows provide actionable molecular resolution—turning structural complexity into scientific clarity.

What We Measure: Quantitative HS/Heparin Profiling

Our service delivers quantitative profiling of heparan sulfate and heparin directly from biological samples including serum, plasma, urine, tissues, cells, and feces. Beyond measuring total content, we resolve key structural and compositional features that govern their biological functions.

Total HS/Heparin Levels

We determine the absolute concentration of HS and/or heparin in each sample (e.g., ng/mL or μg/mg tissue). This global metric is ideal for:

• Monitoring GAG accumulation in metabolic or inflammatory models
• Comparing total GAG burden across conditions or treatment groups
• Supporting research on ECM remodeling or lysosomal storage disorders (RUO)

Disaccharide Composition Analysis

Following enzymatic digestion, HS/heparin chains are converted to discrete disaccharide units, which we quantify using LC–MS/MS with isotope-labeled internal standards. This enables:

• Molar and mass-based quantification of individual disaccharides
• Detection of domain shifts (e.g., NS- vs. NA-regions)
• Generation of sample-specific structural fingerprints

Sulfation Pattern and Epimerization Metrics

Biological specificity resides in where and how GAG chains are sulfated. We calculate:

• N-sulfation vs. N-acetylation ratios
• 2-O-, 6-O-, and 3-O-sulfation motif distribution
• Indicators of C5-epimerase activity (GlcA ↔ IdoA)

These readouts are essential for understanding pathway regulation, enzyme activity, and structural consistency in therapeutic applications.

Chain Length Distribution and GAG Panel Integration

For advanced projects, we can provide:

• Chain-length and polydispersity analysis (via GPC–MS or CE–MS)
• Cross-panel profiling of other GAGs (e.g., CS, DS, KS) or key glycan precursors (e.g., UDP-GlcNAc, PAPS)

This expanded scope supports integrative GAG metabolism studies within your broader omics pipeline.

Analytes Covered in the Heparan Sulfate/Heparin Panel

*Note: Some analytes are optional or matrix-dependent. TriS and 3-O-sulfation markers may require enhanced sensitivity.

Advantages of Our HS/Heparin Analysis Service

• Absolute Quantification with Internal Standards
  Isotope-labeled calibrants ensure accurate disaccharide-level measurement (R² ≥ 0.995, CV ≤ 10%).
• Disaccharide-Level Resolution
  Profile ≥8 HS/heparin disaccharides, including mono-, di-, and tri-sulfated motifs.
• Detailed Sulfation Pattern Analysis
  Distinguish N-, 2-O-, 6-O-, and optional 3-O-sulfation for structural interpretation.
• Compatible with Real Biological Matrices
  Validated for serum, plasma, urine, tissue, and cell samples—no purification required.
• Optional Chain-Length Profiling
  Assess molecular weight and polydispersity via GPC–MS or CE–MS.
• Comprehensive QC and Publication-Ready Data
  Receive raw files, quantitative tables, calibration curves, and recovery reports.
• Multi-GAG Expansion Available
  Add CS, DS, KS disaccharides or precursors like UDP-GlcNAc, all in one project.

Analytical Platforms and Technical Specifications for HS/Heparin Analysis

Our HS/heparin panel runs on a validated targeted metabolomics workflow with high-sensitivity LC–MS/MS at its core. The parameters below outline the data quality and reproducibility you can expect.

Analytical Platforms

Quantitative Method Highlights

Quality Control

• Batch-specific blanks, calibration standards, and QC samples included
• Continuous monitoring of internal standard recovery and retention time stability
• Review of peak shape and integration for each reported disaccharide

Q Exactive HF-X MS Figure from Thermo)

Triple Quad™ 6500+ (Figure from Sciex)

Step-by-Step Workflow for HS/Heparin LC–MS/MS Analysis

1

Project Scoping

Define research goals, sample types, and required depth (total HS only vs full disaccharide/sulfation ± size distribution).

2

Sample Receipt & Check

Log samples, verify volume/matrix/storage, and ensure conditions are suitable for HS/heparin stability.

3

Extraction & Enzymatic Digestion

Enrich HS/heparin from the matrix and digest into defined disaccharides, adding isotope-labeled standards to track recovery.

4

Instrumental Analysis

Perform LC–MS/MS for quantitative disaccharide profiling; optionally add GPC–MS or CE–MS for chain length and isomer resolution.

5

Data Processing & QC

Process spectra to obtain total HS/heparin, disaccharide levels, and structural indices, releasing only QC-approved results.

6

Reporting & Follow-Up

Deliver raw data, result tables, QC summary, and key plots, with optional consultation on data interpretation in your metabolomics context.

How to Submit Samples for HS/Heparin LC–MS/MS Analysis

General Handling Guidelines

• Use pre-chilled tubes where possible and minimize time at room temperature.
• Avoid repeated freeze–thaw cycles; prepare multiple small aliquots instead of one large vial.
• Clearly record matrix, species, treatment groups, collection time points, and any stabilizers or anticoagulants used.
• Ship samples on dry ice with sufficient coolant to maintain frozen conditions until arrival.

What You Receive: HS/Heparin Analysis Deliverables

Raw data files: LC–MS/MS instrument files (and GPC–MS/CE–MS, if included in the project), provided in native or agreed converted formats.

Quantitative result tables: Spreadsheet with per-sample results, including total HS/heparin levels, quantified disaccharides, and any agreed structural indices (e.g., NS/NA ratio, sulfation pattern metrics).

QC summary: Bief overview of calibration performance (e.g., linearity), internal standard recovery, and repeatability (CV) to support data reliability assessment.

Method summary: Short description of sample preparation, enzymatic digestion, analytical platform, and data processing workflow, suitable for use in internal reports or manuscript Methods

LC–MS/MS separation and identification of HS/heparin disaccharides.

Calibration and sensitivity for a representative HS disaccharide.

Use Cases of Heparan Sulfate/Heparin Quantification

GAG accumulation studies in metabolic disorders

Measure HS disaccharides in plasma, urine, or DBS for understanding glycosaminoglycan buildup in genetic models.

Vascular integrity and endothelial biology

Track HS fragments to study glycocalyx shedding under oxidative stress, inflammation, or mechanical injury.

Cancer biology and microenvironment research

Investigate HS structural patterns linked to ECM remodeling, angiogenic signaling, and cell–matrix interactions.

Growth factor signaling and regeneration pathways

Profile sulfation motifs that regulate FGF, VEGF, and Wnt activity during tissue repair or stem cell culture.

Neurobiology and glycan clearance mechanisms

Explore HS degradation and turnover in CNS tissue models of aging or neurodevelopment.

GAG product research and structural comparability

Compare the disaccharide composition and sulfation domains in natural vs. synthetic HS/heparin materials.

HS mimetic and GAG-targeted agent development

Support studies evaluating synthetic HS analogs used in bioengineering or ligand-binding modulation.

Embryonic development and morphogen modulation

Analyze HS profiles affecting morphogen gradients such as Hedgehog, BMP, and Wnt in model organisms.