Simultaneous determination of hydrophobicity and dissociation constant for a large set of compounds by gradient reverse phase high performance liquid chromatography-mass spectrometry technique

J Chromatogr A. 2015 Oct 16:1416:31-7. doi: 10.1016/j.chroma.2015.09.003. Epub 2015 Sep 6.

Abstract

Fast and reliable methods for the determination of hydrophobicity and acidity are desired in pre-clinical drug development phases to eliminate compounds with poor pharmacokinetic properties. Reversed-phase high-performance liquid chromatography (RP HPLC) coupled with time-of-flight mass spectrometry (RP HPLC-ESI-TOF-MS) is a convenient technique for that purpose. In this work we determined the chromatographic measure of hydrophobicity (logkw) and dissociation constant (pKa) simultaneously for a large and diverse group of 161 drugs. Retention times were determined by means of RP HPLC-ESI-TOF-MS for a series of pH and organic modifier gradients. We were able to measure retention times for 140 out of 161 (87%) compounds. For those analytes logkw and pKa parameters were calculated and compared with literature and ACD Labs-calculated data. The determined chromatographic measure of hydrophobicity and dissociation constant was closely related to literature and theoretically calculated values. Applied methodology achieved the medium-throughput screening rate of 100 compounds per day and proved to be a simple, fast and reliable approach of assessing important physicochemical properties of drugs. This technique has certain limitations as it is not applicable for very hydrophilic analytes (logP<0.5) and compounds with identical molar masses.

Keywords: Acidity; Gradient RP HPLC; Hydrophobicity; Throughput analysis; pH/organic modifier.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, High Pressure Liquid / methods*
  • Chromatography, Reverse-Phase / methods*
  • Hydrogen-Ion Concentration
  • Hydrophobic and Hydrophilic Interactions*
  • Pharmaceutical Preparations / analysis*
  • Pharmaceutical Preparations / chemistry*
  • Spectrometry, Mass, Electrospray Ionization / methods*

Substances

  • Pharmaceutical Preparations