CHARACTERIZATION OF THE CHEMICAL AND FUNCTIONAL STABILITY OF DEAE SEPHAROSE (R) FAST FLOW

The release of amines from the ion-exchange groups in DEAE Sepharose(R) Fast Flow has been studied under static and column conditions. The leakage compounds have been identified and quantified by gas chromatography-mass spectrometry. It was shown that the main leakage product under acidic (pH 1) and basic conditions (pH 14) was N,N,N',N'-tetraethylethylenediamine. Three other amines were also identified, namely N,N,N'-triethylethylenediamine, diethylaminoethanol and diethylamine. The leakage of amines from DEAE Sepharose Fast Flow treated at pH 1 or 14 for 672 h at 40-degrees-C corresponds to a reduction of only 1 % of the total ion-exchange capacity. The functional stability of DEAE Sepharose Fast Flow was also studied by separation of a protein mixture during repeated cleaning-in-place treatments with 0.10 M HCl or 1.0 M NaOH. The separation behaviour was unaltered after the gel had been treated for a total contact time of 672 h with 0.10 M HCl or 1.0 M NaOH. The clearance of ethanol from a DEAE Sepharose Fast Flow column stored in a 20 % ethanol aqueous solution was also studied.