Absolute molecular weight distribution of low-molecular-weight heparins by size-exclusion chromatography with multiangle laser light scattering detection.
The absolute molecular weight (M(r)) distribution of seven low-molecular-weight (LMW) heparin products was determined by size-exclusion chromatography (SEC) coupled with multiangle laser light scattering (MALLS) detection. The SEC/MALLS technique does not rely on relative M(r) standards for column calibration and yields absolute M(r) estimates directly from the angular dependence of scattered light intensity as a function of concentration, as formulated by light scattering theory. The SEC/MALLS method we describe is rapid, precise, and accurate. In 1 h it yields results from triplicate injections that agree well with the manufacturers' own independent analyses and that exhibit coefficients of variation of approximately 1%. By eliminating the requirement for finite quantities of highly purified, well-characterized M(r) standards derived from heparin, the present procedure represents a clear improvement over relative methods of M(r) determination. Thus, it is concluded that the SEC/MALLS method is ideally suited to routine quality control of commercial LMW-heparin products.
Knobloch
J E
JE
Scientific Protein Laboratories, Waunakee, Wisconsin 53597, USA.
Shaklee
P N
PN
D000925:Anticoagulants / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods*; D017985:Dalteparin / Q000737:chemistry; D003911:Dextrans / Q000737:chemistry; D006493:Heparin / Q000737:chemistry*; D007834:Lasers; D008956:Models, Chemical; D008970:Molecular Weight; D011786:Quality Control; D012542:Scattering, Radiation
D000925:Anticoagulants; D003911:Dextrans; D006493:Heparin; D017985:Dalteparin