HPLC Method for Enoxaparin on BIST A by SIELC Technologies

High Performance Liquid Chromatography (HPLC) Method for Analysis of  Enoxaparin

Enoxaparin sodium is a popular anticoagulant with the chemical formula (C₂₆H₄₀N₂O₃₆S₅)n. It is primarily used to treat deep vein thrombosis, pulmonary embolisms, and heart attacks. It is on the World Health Organization’s List of Essential Medicines and is available as a generic medication. It works by binding to and activating antithrombin, forming a complex that inactivates clotting factor Xa.

Using SIELC’s newly introduced BIST™ method, Enoxaparin, which ionizes in water, can be retained on a negatively-charged, cation-exchange BIST A column. There are two keys to this retention method: 1) a multi-charged, positive buffer, such as N,N’-Dimethylpiperazine (DMP), which acts as a bridge, linking the negatively-charged Enoxaparin analytes to the negatively-charged column surface and 2) a mobile phase consisting mostly of organic solvent (such as MeCN) to minimize the formation of a solvation layer around the charged analytes. With a fairly simple 2-stage gradient, Enoxaparin and sodium can be separated and retained on a BIST™ A column. The gradient is necessary because of the extremely strong retention in BIST. Mobile Phase A, with its high-organic concentration, induces the initial Enoxaparin retention. But an isocratic MP with just MP A would result in extremely long retention times. By introducing a low-organic mobile phase, the solvation layer increases, reducing BIST interactions and the analyte is able to elute in an appropriate amount of time. This method is compatible with mass spectrometry (MS), evaporative light scattering detection (ELSD), charged aerosol detection (CAD).