Binding measurements of indolocarbazole derivatives to immobilised human serum albumin by high-performance liquid chromatography.

The binding properties of six indolocarbazole derivative have been measured using immobilised human serum albumin (HSA) in an HPLC column. The compounds showed very strong binding to HSA which necessitated the application of a 30 to 40% concentration of 2-propanol in the mobile phase. This represents a much higher concentration than is recommended by the column manufacturers. This HSA column had not changed its binding property when it was used again with 4% 2-propanol and 96% phosphate buffer. The binding parameters were estimated by extrapolation to 0% 2-propanol and were above 99% for each indolocarbazole derivative. The correlation analysis, including the calculated octanol/water partition coefficient (log P), pKa values as well as measured reversed-phase retention data of the compounds revealed that the extremely strong binding can be explained by the hydrophobic and acidic properties of the compounds.