Analysis of heparins by size-exclusion and reversed-phase high-performance liquid chromatography with photodiode-array detection.

High-performance liquid chromatography (HPLC) of heparins was carried out with on-line photodiode-array detection. Average molecular weights and molecular weight distribution were calculated using computer data acquisition and handling; size-exclusion chromatography was performed using different selective microparticulate columns and narrow molecular weight distribution heparin standards for calibration; heparin peak purity was investigated using several methods for evaluation. Additional UV-absorbing compounds present in heparin preparations were characterized and quantitated by reversed-phase HPLC. These methods are useful for the analysis of the molecular weight distribution and peak purity of heparins and for the determination of additional drugs, additives or impurities.