The molecular weight profiles of low molecular weight heparin samples have been measured by high-performance gel permeation chromatography using as calibrant the heparinase-degraded material (90/686) now established as the 1st International Reference Preparation (IRP) Low Molecular Weight Heparin for Molecular Weight Calibration. Use of the calibrant as a broad molecular weight standard is described and a calibration table provided based on data collected over several years in one laboratory.
In order to confirm the assignment of degree of polymerisation to resolved oligosaccharide peaks in the calibrant, molecular weights of oligosaccharides fractionated from the 1st IRP were independently determined by fast atom bombardment mass spectrometry (FAB MS).
The molecular weight distributions of commercial low molecular weight heparins have been characterized. Measurements of molecular weight parameters of heparin molecular weight standards from several sources provide comparisons between the molecular weight scales of this and other studies.
References
1
Rodriguez HJ.
Accurate and reproducible determination of molecular weight distribution of sodium heparin USP by HPLC. Analytical Letts 1976; 9: 497-506
3
Teng L-N,
Teller DC.
Molecular weight determination of heparin sodium fractions by high pressure liquid chromatography. In: Chemistry and Biology of Heparin.
Lundblad RI,
Brown WV,
Mann KG,
Roberts HR.
(eds). Amsterdam: Elsevier; 1981. pp 65-72
5
Hennink WE,
van denBerg JWA,
Feijen J.
Standardization of heparins by means of h.p.l.c. equipped with a low angle laser light scattering detector. Thromb Res 1987; 45: 463-475
6
de VriesJX.
Analysis of heparins by size-exclusion and reversed-phase high-performance liquid chromatography with photodiode arraydetection. J Chromatogr 1989; 465: 297-304
7
Kristensen HI,
Tromborg EM,
Nielsen JR,
Nielsen JI,
Johansen KB,
Oster-gaard PB.
Development and validation of a size exclusion chromatography method for determination of molecular masses and molecular mass distribution in low molecular weight heparin. Thromb Res 1991; 64: 131-141
8
Komatsu H,
Yoshii K,
Ishimitsu S,
Okada S.
Molecular mass determination of low-molecular-mass heparins. Application of wide collection angle measurements of light scattering using a high-performance gel permeation chromatographic system equipped with a low-angle laser light-scattering photometer. J Chromatogr 1993; 644: 17-24
12
Ahsan A,
Jeske W,
Mardiguian J,
Fareed J.
Feasibility study of heparin mass calibrator as a GPC calibrator for heparins and low molecular weight heparins. J Pharm Sci 1994; 83: 197-201
14
Barrowcliffe TW,
Mulloy B,
Johnson EA,
Thomas DP.
The anticoagulant activity of heparin; measurement and relationship to chemical structure. J Pharm Biomed Anal 1989; 07: 217-226
16
Reinhold VN,
Carr SA,
Green BN,
Petitou M,
Choay J,
Sinay P.
Structural characterisation of sulfated glycosaminoglycans by fast atom bombardment mass spectrometry: application to heparin fragments prepared by chemical synthesis. CarbohydrRes 1987; 161: 305-313
19
Volpi N,
Bolognani L.
Glycosaminoglycans and proteins: different behaviours in high-performance size-exclusion chromatography. J Chromatogr 1993; 630: 390-396
