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Published on: April 16, 2013
Analytical method to determine keratan sulfate in the serum using HPLC
Yasuhiro Kurahashi1, Hiroyuki Masuda, Kyosuke Miyazaki
1Seikagaku Corporation, Yokosuka.
Rinsho Byori. the Japanese Journal of Clinical Pathology
|June 13, 2008
Summary
This study developed a new high-performance liquid chromatography (HPLC) method to measure keratan sulfate (KS) in serum. This offers a simpler way to assess articular cartilage damage compared to using synovial fluid.
Area of Science:
- Biochemistry
- Biomarker Discovery
- Analytical Chemistry
Background:
- Articular cartilage damage assessment traditionally uses keratan sulfate (KS) measurement in synovial fluid via HPLC.
- Serum analysis is less invasive and potentially reflects cartilage degeneration and injury.
- A validated method for serum KS determination is needed.
Purpose of the Study:
- To develop and validate a novel HPLC method for quantifying keratan sulfate (KS) in human serum.
- To establish serum KS as a viable biomarker for articular cartilage health.
Main Methods:
- Human serum samples were spiked with varying concentrations of sulfated keratan sulfate (KS).
- Protease digestion, anion-exchange chromatography, and desalting were used for KS extraction.
- Enzymatic digestion with keratanase II followed by HPLC analysis determined KS levels.
Main Results:
- The developed HPLC method demonstrated high recovery rates (≥98%) for serum KS, independent of sulfation degree.
- The method showed good linearity across a wide concentration range (50–100,000 ng/ml).
- Inter-day assay variability was low (6.0% over 10 days).
Conclusions:
- A robust HPLC method for determining serum keratan sulfate (KS) concentration has been successfully developed.
- This method is reliable and unaffected by the degree of KS sulfation.
- Serum KS measurement offers a promising, less invasive alternative for monitoring articular cartilage status.
