with frontal analysis (FA), the d e -dadsorpb i s o t h m d insulin on the composition of mobile phase in reversed phase liquidchromatography(RPLC) has been inwdgated. ' Ibi9 is elso a good example to empIoy the stoirhiometric dkpkmed theory (SDT) for i n * dute adsorpbion in physical &*j . six ldnasofmobllephrse in RPLC Wem anployed to study the eat& 011 the elution curves and adsorption isdherns of insulin. llle key ~o f t h k p a p e r a r e : ( 1 ) ' I b e s t a b i l l t y o f~d u e t o d e l a y time after p-, the organic sdveut cwcentrstian, the kind and the concentration of ion-peiring agent in mobile phase wen found toaffectbotb elutioo QVM and adsorptionisdherm vergser i d y . (2) To obtain a valid and ComparaMe result. the Campositloo dthe mobile paaSe employed in FA must be as same as to that in usual RPLC of either analytical d e or prepdive purtate these obtained adforption isotherm. 'Ibe q r e s i o n for sdute adsorption from sdution ofthe SDT wa3 found to have a better elu-cidafkmtotheimulinadsorpti~frommobUe~inRpu3r. pose. (3) Lengmuir Equation and the SDT m employed to hni-Keywords phase liguid C -(RPLC), did-liquid system, stoichiometric displacement theory (SDT) , adsarption mechanism, adsorption isotherm, fmntal adysie (FA), insulin *
with the combination of the the stoichiometric displacement model for retention (SDM-R) in reversed phase liquid chromatography (RPLC) and the stoichiometric disptacement model for adsorption (SDM-A) in physical chemistry, the total number of mdes of the re-solvated methanol of stationary phase side, N , and that of solute side in the mobile phase, q , c o -* toonemdeofthe desorbiing solute, were separately detennined and referred as the characterization parameters of the contriions of the adsorption m e c h a n i s n a n d p a r t i t i o n~ ' to the solute retention. respectively. A chromatographic system of insulin, using mobile phase conristing of the pseud~homologue of alcohols (methanol, ethanol and Zpropanol)-water and tritluoroacetic acid was employed. The maximum number of the methand layers on the stationary phase d a c e was found to be 10.6, only 3 of which b e i i valid in usual RPLC, traditionally referred as a vdume proces~ in partition mechanisn. However, it still follows the SDM-R. Both of q and N of insulin were found not to be zero, indicating that the retention adsorption mecharusn ' .Whenmethawlisusedastheorganicmod-Xer, the ratio of q/nr was 1.l3, indicating the contriition to insulin retention due to partition mecharusn . being a bit greater than that due to adsorption mecharusn * . A linear relationship between q , or nr and the carbon number of the peudc+homologue in the mobile phase was also found. As a methoddogy for invgtigating the retention m retention and behaviors of biopolymers, a homologue of organic solvents as the organic modifier in mobile phase lw also been explored. mechanisnofinsulin is a mixed mode of partition rrxhumn * a n d Keywords nism, mixed mode, stoichiometric displacement, insulin reversed phase liquid chromatography, retention mecha-*
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