SPDS Set 1 ERP Book
CHON-004
Question
Answer
General
AOAC Method Number:
CHON-004
Determination of Chondroitin Sulfate Content in Raw Materials and Dietary Supplements by High-Performance Liquid Chromatography with Ultraviolet Detection After Enzymatic Hydrolysis: Single-Laboratory Validation
Method Title:
Method Author(s):
David Ji, Mark Roman, Joseph Zhou, Jana Hildreth
Reviewer Name:
Phil Koerner
Quantitative ion-pair LC/UV method for determination of chondroitin sulfate in raw materials and dietary supplements from about 5 to 100% (w/w) chondroitin sulfate. Following aqueous extraction, chondroitin sulfate is selectively hydrolyzed by chondroitinase ACII enzyme to form un-, mono-, di-, and trisulfated unsaturated disaccharides; the resulting disaccharides are then quantified by ion-pairing HPLC with UV detection @ 240nm. Total chondroitin sulfate is sum of the individual disaccharides determined. quantitative method for determination of individual disaccharides and total Chondroitin Sulfate
Summary of Method
Method Scope / Applicability:
very thorough SLV documented potential interferences considered and tested multiple sources for enzyme? reliable availability of reference standards?
General Comments About the Method:
well documented protocol, appears to be easy to follow as written but requires skilled analyst Uses standard HPLC instrument with UV detection HPLC column is easily obtained HPLC analysis time <20 minutes Sample prep (enzymatic hydrolysis) is long (>3 hours); total assay time (5 calibration standards plus samples) is long, could take a day to assay multiple samples Secure and reliable source for enzyme (currently only
Method Clarity
Pros/Strengths
Cons/Weaknesses
Made with FlippingBook