SPDS 09052014

8/22/2014

Enzymatic HPLC Method

– Modifications to method that may improve  performance/reproducibility between labs I i i f Δdi 0S f id k • ncreas ng retent on o  ‐ away rom vo pea  (several labs did not achieve resolution) – USP method uses anion exchange (SAX) column • Performing reaction in 2‐ mL or 5‐ mL conical  bottomed reaction vials to eliminate transfer step • For raw materials, dry sample in oven prior to  weighing, and place desiccant in balance chamber

Concerns Surrounding Enzymatic HPLC Method – Lack of availability and purity issues associated with  individual disaccharide reference standards • Adopting USP’s method of using a USP Chondroitin – Limited supply of enzyme • IBEX was only supplier for the past couple of years but  recently Sigma started offering Chondroitinase AC again  so availability is no longer an issue – Expensive to run – Technically Challenging Sulfate RM (or other well characterized/qualified  Chondroitin RM) verses the individual disaccharides  would address this issue

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