Analysis of heparin sodium by SAX/HPLC for contaminants and impurities.
A chromatographic method was developed for the detection and quantification of the contaminant oversulfated chondroitin sulfate (OSCS) and the impurity dermatan sulfate in heparin active pharmaceutical ingredient (API). The HPLC analysis of heparin is carried out using a polymer-based strong anion exchange (SAX) column with gradient elution from 0.125 M sodium chloride to 2.5M sodium chloride buffered mobile phase. The limit of detection (LOD) and limit of quantitation (LOQ) for the contaminant OSCS in heparin were determined to be 0.03% and 0.1%, respectively. The LOD and LOQ for dermatan sulfate, an impurity in heparin sulfate, were determined to be 0.1% and 0.8%, respectively. This manuscript is not a policy document and is not intended to replace either of the methods (capillary electrophoresis and NMR) currently required by the FDA.
Trehy
Michael L
ML
Food and Drug Administration, Division of Pharmaceutical Analysis, Saint Louis, MO 63101, USA. michael.trehy@fda.hhs.gov
Reepmeyer
John C
JC
Kolinski
Richard E
RE
Westenberger
Benjamin J
BJ
Buhse
Lucinda F
LF
D000925:Anticoagulants / Q000032:analysis*; D002809:Chondroitin Sulfates / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003871:Dermatan Sulfate / Q000032:analysis; D004340:Drug Contamination; D019075:Electrophoresis, Capillary; D006025:Glycosaminoglycans / Q000032:analysis; D006493:Heparin / Q000032:analysis*; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D012015:Reference Standards; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet
D000925:Anticoagulants; D006025:Glycosaminoglycans; D007202:Indicators and Reagents; D003871:Dermatan Sulfate; D006493:Heparin; D002809:Chondroitin Sulfates