title: Analysis of crude heparin by (1)H NMR, capillary electrophoresis, and strong-anion-exchange-HPLC for contamination by over sulfated chondroitin sulfate.
abstract: We previously published a strong-anion-exchange-high performance liquid chromatography (SAX-HPLC) method for the detection of the contaminant over sulfated chondroitin sulfate (OSCS) in heparin sodium active pharmaceutical ingredient (API). While APIs have been processed to remove impurities, crude heparins contain insoluble material, chondroitin sulfates, heparan sulfate, and proteins that may interfere with the recovery and measurement of OSCS. We examined 500MHz (1)H NMR, capillary electrophoresis (CE), and SAX-HPLC to quantify OSCS in crude heparin. Using our standard API protocol on OSCS spiked crude heparin samples; we observed a weight percent LOD and LOQ for the NMR approach of 0.1% and 0.3%, respectively, while the SAX-HPLC method gave values of 0.03% and 0.09%, respectively. CE data was not amenable to quantitative measurement of OSCS in crude heparin. We developed a modified HPLC sample preparation protocol using crude dissolved at the 100mg/mL level with a 2.5M NaCl solution. This SAX-HPLC approach gave a weight percent LOD of 0.02% and a LOQ of 0.07% and had better performance characteristics than that of the protocol used for APIs.
authors: Food and Drug Administration, Division of Pharmaceutical Analysis, 1114 Market St., Saint Louis, MO 63101, USA. david.keire@fda.hhs.gov
authors: David A
authors: DA
authors: Keire
authors: Michael L
authors: ML
authors: Trehy
authors: John C
authors: JC
authors: Reepmeyer
authors: Richard E
authors: RE
authors: Kolinski
authors: Wei
authors: W
authors: Ye
authors: Jamie
authors: J
authors: Dunn
authors: Benjamin J
authors: BJ
authors: Westenberger
authors: Lucinda F
authors: LF
authors: Buhse
mesh_terms: D000837:Anion Exchange Resins; D002809:Chondroitin Sulfates / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004340:Drug Contamination; D019075:Electrophoresis, Capillary; D017408:Guidelines as Topic; D006493:Heparin / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D015203:Reproducibility of Results; D014481:United States; D014486:United States Food and Drug Administration
chemical_list: D000837:Anion Exchange Resins; D006493:Heparin; D002809:Chondroitin Sulfates