Heparin sodium oligosaccharides

Molecular weight and polydispersity of prepared heparin products was determined using size exclusion chromatography (SEC) as described earlier.^{38 (link),39 (link)} TSK-GEL G3000PWxl size exclusion column (Tosoh Bioscience, Minato, Japan), maintained at 40°C with an Eppendorf column heater, was connected to a HPLC system consisting of a Shimadzu LC-10Ai pump, a Shimadzu CBM-20A controller and a Shimadzu RID-10A refractive index detector. The mobile phase consisted of 0.1 M NaNO₃. A sample injection volume of 20 μL and a flow rate of 0.6 mL min⁻¹ were used. The SEC chromatograms were recorded with the LCsolution Version 1.25 software and molecular weight properties determined using the “GPC Postrun” function. Heparin sodium oligosaccharides (2,687, 4,300, 5,375, 6,449, and 8,060 Da) (Iduron, Manchester, UK) were used as calibrants for heparin, HS and Lovenox. Hyaluronic acid (HA) (81, 130, and 44.3 kDa) was used as calibrant for heparosan. The molecular weight measurements for processed GAGs/starting materials was carried out in triplicates/duplicates.

Molecular weight and polydispersity of prepared heparin products was determined using size exclusion chromatography as described earlier (Zhang et al., 2011 (link)). TSK-GEL G3000PWxl size exclusion column (Tosoh Bioscience), maintained at 40 °C with an Eppendorf column heater, was connected to a HPLC system consisting of a Shimadzu LC-10Ai pump, a Shimadzu CBM-20A controller and a Shimadzu RID-10A refractive index detector. The mobile phase consisted of 0.1 M NaNO₃. A sample injection volume of 20 μL and a flow rate of 0.6 mL/min were used. The SEC chromatograms were recorded with the LCsolution Version 1.25 software and molecular weight properties determined using the “GPC Postrun” function. Heparin sodium oligosaccharides (Iduron, UK) of different molecular weights (2687 Da, 4300 Da, 5375 Da, 6449 Da and 8060 Da) were used as calibrants for the standard curve. The molecular weight measurements were carried out in duplicates.