title: Characterization of currently marketed heparin products: analysis of molecular weight and heparinase-I digest patterns.
abstract: We evaluated polyacrylamide gel electrophoresis (PAGE) and size exclusion chromatography coupled with multi-angle laser light scattering (SEC-MALLS) approaches to determine weight-average molecular weight (M(w)) and polydispersity (PD) of heparins. A set of unfractionated heparin sodium (UFH) and low-molecular-weight heparin (LMWH) samples obtained from nine manufacturers which supply the US market were assessed. For SEC-MALLS, we measured values for water content, refractive index increment (dn/dc), and the second virial coefficient (A(2)) for each sample prior to molecular weight assessment. For UFH, a mean ± standard deviation value for M(w) of 16,773 ± 797 was observed with a range of 15,620 to 18,363 (n = 20, run in triplicate). For LMWHs by SEC-MALLS, we measured mean M(w) values for dalteparin, tinzaparin, and enoxaparin of 6,717 ± 71 (n = 4), 6,670 ± 417 (n = 3), and 3,959 ± 145 (n = 3), respectively. PAGE analysis of the same UFH, dalteparin, tinzaparin, and enoxaparin samples showed values of 16,135 ± 643 (n = 20), 5,845 ± 45 (n = 4), 6,049 ± 95 (n = 3), and 4,772 ± 69 (n = 3), respectively. These orthogonal measurements are the first M(w) results obtained with a large heparin sample set on product being marketed after the heparin crisis of 2008 changed the level of scrutiny of this drug class. In this study, we compare our new data set to samples analyzed over 10 years earlier. In addition, we found that the PAGE analysis of heparinase digested UFH and neat LMWH samples yield characteristic patterns that provide a facile approach for identification and assessment of drug quality and uniformity.
authors: Division of Pharmaceutical Analysis, CDER, Food and Drug Administration, 1114 Market St. Rm 1002, St. Louis, MO 63101, USA.
authors: Cynthia D
authors: CD
authors: Sommers
authors: Hongping
authors: H
authors: Ye
authors: Richard E
authors: RE
authors: Kolinski
authors: Moheb
authors: M
authors: Nasr
authors: Lucinda F
authors: LF
authors: Buhse
authors: Ali
authors: A
authors: Al-Hakim
authors: David A
authors: DA
authors: Keire
mesh_terms: D000925:Anticoagulants / Q000737:chemistry* / Q000378:metabolism; D002850:Chromatography, Gel / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006493:Heparin / Q000737:chemistry* / Q000378:metabolism; D019763:Heparin Lyase / Q000378:metabolism*; D006495:Heparin, Low-Molecular-Weight / Q000737:chemistry* / Q000378:metabolism; D008027:Light; D008970:Molecular Weight; D012031:Refractometry; D012542:Scattering, Radiation
chemical_list: D000925:Anticoagulants; D006495:Heparin, Low-Molecular-Weight; D006493:Heparin; D019763:Heparin Lyase