Solute complexation degree with human serum albumin: biochromatographic approach.
A mathematical model was developed for the study of the D,L-dansylamino acid retention mechanism in reversed-phase liquid chromatography using a C18 column as a stationary phase and human serum albumin (HSA) as an eluent modifier. The solute retention factor is dependent on the HSA concentration in the eluent as well as the binding constant of the guest-HSA complex. A determination of the degree of complexation n(c) (the percent of the complexed guest) could be carried out. Different Van 't Hoff plot shapes of the degree of complexation were observed with different eluent pH, confirming a change in the solute complexation mechanism for physiological pH (between 7-7.5). Enthalpy-entropy compensation was also analysed in relation to this mathematical model to confirm the solute complexation behavior with HSA. These results finally confirmed that at physiological pH and temperature (approximately 35 degrees C) values the HSA was in a favorable structural conformation for its binding with a great majority of drugs.
Guillaum
Y C
YC
Laboratoire de Chimie Analytique, Faculté de Médecine et de Pharmacie, Place Saint-Jacques, Besançon, France. yves.guillaume@univ-fcomte.fr
Millet
J
J
Nicod
L
L
Truong-Than
T
T
Guinchard
C
C
Xicluna
A
A
Thomassin
M
M
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015203:Reproducibility of Results; D012709:Serum Albumin / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet; D013696:Temperature; D013816:Thermodynamics
D012709:Serum Albumin