Rapid determination of the applicability of hydrophilic interaction chromatography utilizing ACD labs log D suite: a bioanalytical application.
Hydrophilic interaction chromatography (HILIC) is an effective technique for retaining and separating polar compounds. This approach offers several advantages for bioanalytical liquid chromatography/mass spectrometry, considering that a majority of active pharmaceutical ingredients are polar amines. HILIC employs high concentrations of relatively polar organic mobile phase components (i.e. acetonitrile), providing enhanced desolvation and electrospray ionization efficiency, as well as allowing direct injection of many protein precipitation, liquid/liquid, and solid phase extracts. A set of 30 probe compounds was evaluated to demonstrate a relationship between a compound's HILIC capacity factor (k'), and pH dependent distribution coefficient (D), using three sets of generic isocratic conditions. Plots of logk' versus logD(pH3.0) produced correlation coefficients of 0.751, 0.696, and 0.689 at acetonitrile mobile phase concentrations of 85%, 90%, and 95% (v/v), respectively. For bioanalytical applications a k'>2 is typically targeted to ensure adequate retention of a given analyte relative to extracted matrix components. Using k'> or =2 as a measure of HILIC applicability, the linear relationships for each of the three acetonitrile levels predicted whether or not HILIC was able to meet this criterion for at least 90% of the compounds tested. Overall, the relationship between k' and logD can serve as a valuable tool for identifying the applicability of HILIC and a starting point for the chromatographic conditions, prior to the initiation of any laboratory activities. Additionally, this relationship can assist with the selection of appropriate chemical analog internal standards.
Kadar
Eugene P
EP
Pharmacokinetics, Dynamics & Metabolism, Pfizer Global Research and Development, Pfizer Inc., Groton, CT 06340, USA. eugene.p.kadar@pfizer.com
Wujcik
Chad E
CE
Wolford
David P
DP
Kavetskaia
Olga
O
D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002845:Chromatography / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D012015:Reference Standards; D012995:Solubility; D053719:Tandem Mass Spectrometry; D014867:Water
D007202:Indicators and Reagents; D014867:Water