47 / 116
Question
Answer
General
AOAC Method Number:
CHON-004
Method Title:
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 Author(s):
David Ji, Mark Roman, Joseph Zhou, Jana Hildreth
Reviewer Name:
Phil Koerner
Summary of Method
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.
Method Scope / Applicability:
quantitative method for determination of individual
disaccharides and total Chondroitin Sulfate
General Comments About the
Method:
very thorough SLV documented
potential interferences considered and tested
multiple sources for enzyme?
reliable availability of reference standards?
Method Clarity
well documented protocol, appears to be easy to follow
as written but requires skilled analyst
Pros/Strengths
Uses standard HPLC instrument with UV detection
HPLC column is easily obtained
HPLC analysis time <20 minutes
Cons/Weaknesses
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
CHON-004