title: Determination of impurities in heparin by capillary electrophoresis using high molarity phosphate buffers.
abstract: Oversulfated chondroitin sulfate (OSCS), an impurity found in some porcine intestinal heparin samples was separated from intact heparin by capillary electrophoresis (CE) using a 600mM phosphate buffer, pH 3.5 as the background electrolyte in a 56cm x 25microm i.d. capillary. This method was confirmed in two separate labs, was shown to be linear, reproducible, robust, easy to use and provided the highest resolution and superior limits of detection compared to other available CE methods. Glycosoaminoglycans such as dermatan sulfate and heparan sulfate were separated and quantified as well during a single run. The heparin peak area response correlated well to values obtained using the official assay for biological activity. A high speed, high resolution version of the method was developed using 600mM lithium phosphate, pH 2.8 in a 21.5cm x 25microm i.d. capillary which provided limits of detection for OSCS that were below 0.1%.
authors.affiliation: Baxter Healthcare Corporation, Round Lake, IL 60073, USA. todd_Wielgos@baxter.com
authors.forename: Todd
authors.initials: T
authors.lastname: Wielgos
authors.forename: Karalyn
authors.initials: K
authors.lastname: Havel
authors.forename: Nadia
authors.initials: N
authors.lastname: Ivanova
authors.forename: Robert
authors.initials: R
authors.lastname: Weinberger
mesh_terms: D000818:Animals; D000925:Anticoagulants / Q000032:analysis / Q000737:chemistry*; D002021:Buffers; D002138:Calibration; D002809:Chondroitin Sulfates / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification; D003871:Dermatan Sulfate / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification; D004340:Drug Contamination; D004573:Electrolytes / Q000737:chemistry; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods; D006493:Heparin / Q000032:analysis / Q000737:chemistry*; D006497:Heparan Sulfate / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D015394:Molecular Structure; D009994:Osmolar Concentration; D010710:Phosphates / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D034421:Sus scrofa; D013696:Temperature; D013997:Time Factors
chemical_list: D000925:Anticoagulants; D002021:Buffers; D002809:Chondroitin Sulfates; D003871:Dermatan Sulfate; D004573:Electrolytes; D006493:Heparin; D006497:Heparan Sulfate; D010710:Phosphates