title: Determination of galactosamine impurities in heparin sodium using fluorescent labeling and conventional high-performance liquid chromatography.
abstract: Heparin is a sulfated glycosaminoglycan (GAG), which contains N-acetylated or N-sulfated glucosamine (GlcN). Heparin, which is generally obtained from the healthy porcine intestines, is widely used as an anticoagulant during dialysis and treatments of thrombosis such as disseminated intravascular coagulation. Dermatan sulfate (DS) and chondroitin sulfate (CS), which are galactosamine (GalN)-containing GAGs, are major process-related impurities of heparin products. The varying DS and CS contents between heparin products can be responsible for the different anticoagulant activities of heparin. Therefore, a test to determine the concentrations of GalN-containing GAG is essential to ensure the quality and safety of heparin products. In this study, we developed a method for determination of relative content of GalN from GalN-containing GAG in heparin active pharmaceutical ingredients (APIs). The method validation and collaborative study with heparin manufacturers and suppliers showed that our method has enough specificity, sensitivity, linearity, repeatability, reproducibility, and recovery as the limiting test for GalN from GalN-containing GAGs. We believe that our method will be useful for ensuring quality, efficacy, and safety of pharmaceutical heparins. On July 30, 2010, the GalN limiting test based on our method was adopted in the heparin sodium monograph in the Japanese Pharmacopoeia.
authors.affiliation: Division of Biological Chemistry and Biologicals, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan.
authors.forename: Satsuki
authors.initials: S
authors.lastname: Itoh
authors.forename: Yoko
authors.initials: Y
authors.lastname: Hiruta
authors.forename: Noritaka
authors.initials: N
authors.lastname: Hashii
authors.forename: Naho
authors.initials: N
authors.lastname: Fujita
authors.forename: Toru
authors.initials: T
authors.lastname: Natsuga
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Hattori
authors.forename: Aya
authors.initials: A
authors.lastname: Bando
authors.forename: Yuko
authors.initials: Y
authors.lastname: Sekimoto
authors.forename: Kazuyoshi
authors.initials: K
authors.lastname: Miyata
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Namekawa
authors.forename: Kazunori
authors.initials: K
authors.lastname: Mabuchi
authors.forename: Toru
authors.initials: T
authors.lastname: Sakai
authors.forename: Hirotoshi
authors.initials: H
authors.lastname: Shimahashi
authors.forename: Kenzo
authors.initials: K
authors.lastname: Kawai
authors.forename: Hikaru
authors.initials: H
authors.lastname: Yoden
authors.forename: Sadatoshi
authors.initials: S
authors.lastname: Koyama
authors.forename: Susanne Odgaard
authors.initials: SO
authors.lastname: Herr
authors.forename: Shunji
authors.initials: S
authors.lastname: Natsuka
authors.forename: Teruhide
authors.initials: T
authors.lastname: Yamaguchi
authors.forename: Nana
authors.initials: N
authors.lastname: Kawasaki
mesh_terms: D002809:Chondroitin Sulfates / Q000032:analysis / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003871:Dermatan Sulfate / Q000032:analysis / Q000737:chemistry; D004340:Drug Contamination / Q000517:prevention & control*; D005456:Fluorescent Dyes / Q000737:chemistry; D005688:Galactosamine / Q000032:analysis*; D006493:Heparin / Q000032:analysis* / Q000737:chemistry; D006868:Hydrolysis; D008956:Models, Chemical; D015203:Reproducibility of Results; D062366:para-Aminobenzoates / Q000737:chemistry
chemical_list: D005456:Fluorescent Dyes; D062366:para-Aminobenzoates; D003871:Dermatan Sulfate; D005688:Galactosamine; D006493:Heparin; D002809:Chondroitin Sulfates
keywords: Dermatan sulfate; Fluorescent labeling; Galactosamine; Heparin sodium; Limiting test