HPLC Method for Analysis of Heparin on BIST™ A Column

Separation type: Bridge Ion Separation Technology, or BIST™ by SIELC Technologies

HPLC Method for Analysis of Heparin on BIST™ A Column by SIELC Technologies
High Performance Liquid Chromatography (HPLC) Method for Analysis of Heparin

Heparin is a WHO-recognized essential medicine known for its anticoagulant properties and used to treat heart attacks and angina. Due to its very polar and polymeric nature, separating and quantifying Heparin using traditional HPLC methods can be quite difficult. Using SIELC’s newly introduced BIST™ method, however, Heparin and its salts can be separated easily 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 anion 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. Other positively-charged buffers that can generate BIST™ include TMDAP, Calcium acetate, and Magnesium acetate. Using this new and unique analysis method, Heparin and its salts can be separated and retained with high selectivity and great peak shape. This method can be detected and is compatible with ELSD, CAD, and Mass Spectrometry (LC-MS).


ColumnBIST™ A, 4.6×50 mm, 5 µm, 100A
Mobile PhaseGradient MeCN
BufferN,N’-Dimethylpiperazine acetate pH 4.0
Flow Rate1.0 ml/min
DetectionELSD, 70C


Class of CompoundsDrug, Glycosaminoglycan
Analyzing CompoundsHeparin