title: RP-HPLC determination of dissociation constant using solely aqueous mobile phase.
abstract: The proposed HPLC method using solely or nearly 100% aqueous mobile buffer as mobile phase offers fast determination of dissociation constant for compounds in relatively wide range of lipophilicity (log P from -2.26 to 2.26). The dissociation constant value for simpler chemical compounds can be determined via only 8 chromatographic runs. The number of needed chromatographic separations depends on the structural complexity of the tested compound. Moreover, the proposed method does not require a measurement of Yasuda-Shedlovsky extrapolation that includes several pKadeterminations in solutions with different methanol content which speeds up considerably the procedure. The methodology is suitable for evaluation of large series of drug candidates, which can be present as complex mixtures and in small amounts.
authors.affiliation: Department of Organic Chemistry, Faculty of Science, Palacký University in Olomouc, 17. listopadu 12, 771 46 Olomouc, Czech Republic.
authors.forename: Tereza
authors.initials: T
authors.lastname: Volná
authors.affiliation: Department of Organic Chemistry, Faculty of Science, Palacký University in Olomouc, 17. listopadu 12, 771 46 Olomouc, Czech Republic. Electronic address: kamil.motyka@upol.cz.
authors.forename: Kamil
authors.initials: K
authors.lastname: Motyka
authors.affiliation: Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacký University in Olomouc, Hněvotínská 5, 779 00 Olomouc, Czech Republic.
authors.forename: Jan
authors.initials: J
authors.lastname: Hlaváč
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D014867:Water / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*
chemical_list: D014867:Water
keywords: Dissociation constant; HPLC; High-throughput measurement; Reversed phase