A. Ahsan et al., Isoamyl nitrite depolymerized heparin as a universal calibrator for heparins and low molecular weight heparins, CL APPL T-H, 6(3), 2000, pp. 169-174
Because the subcutaneous absorption of heparin depends on its molecular wei
ght, high performance size exclusion chromatography methods, among others,
have been developed for its determination. Because heparin consists of poly
mer chains of varying lengths, a large number of calibrators, 19 in our wor
k, are required for these molecular weight analyses. Because the preparatio
n of that many calibrants is both time-consuming and expensive, other metho
ds requiring fewer calibrants are being sought. In pursuit of these aims, a
heparinase-degraded heparin has been accepted as the first international r
eference preparation low molecular weight heparin for molecular weight cali
bration. We have previously proposed another calibrant, the Heparin Mass Ca
librator. In this article, we introduce another calibrant, called the Hepar
in Molecular Mass Calibrant. It offers the advantage of a much wider availa
bility because it is a commercial product. In addition to having a superior
refractive index chromatography run profile, when used in combination with
two Narrow Range Calibrators with molecular weights of 34.4 and 18.4 KDa,
this new calibrant covers both the higher and the lower molecular weight ra
nges. Besides, ultraviolet detection is not needed fur molecular weight cal
culations. Therefore, it is proposed that its feasibility as a universal re
ference calibrator for both heparins and low molecular weight heparins be c
onsidered.