FEASIBILITY STUDY OF HEPARIN MASS CALIBRATOR AS A GPC CALIBRATOR FOR HEPARINS AND LOW-MOLECULAR-WEIGHT HEPARINS

The proposed European Pharmacopoeial (EP) method for molecular weight determination of low molecular weight heparins (LMWHs) has been shown to have a range which is too narrow to allow for the accurate molecular weight determinations of all LMWHs. We have recently shown that a chemically degraded benzyl ester of unfractionated heparin, ITH-3, is a better calibrator using this methodology. Data is presented here which indicates that this calibrator cannot only be used to determine the molecular weights of LMWHs but also of unfractionated heparins. Therefore, it is now renamed heparin mass calibrator or HMC because of this extension of range. Weight average molecular weight, number average molecular weight, peak molecular weight, and dispersity values were calculated using the HMC calibration and a reference narrow-range 19-calibrator method for various LMWHs and unfractionated heparins. Values for the parameters calculated using the HMC calibration were found not to be significantly different from those of the reference method until the molecular weight exceeded 15.0 kDa. In contrast, the molecular weight profile obtained with the proposed EP method was significantly different from the reference method for samples >8.0 kDa. The range exhibited by the HMC calibrator should allow it to be used for both LMWHs and unfractionated heparins.