Skip to Content
Merck
  • cIEF for rapid pKa determination of small molecules: a proof of concept.

cIEF for rapid pKa determination of small molecules: a proof of concept.

European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences (2014-07-06)
Stéphanie Romand, Julie Schappler, Jean-Luc Veuthey, Pierre-Alain Carrupt, Sophie Martel
ABSTRACT

A capillary isoelectric focusing (cIEF) method was developed for the determination of the ionization constants (pKa) of small molecules. Two approaches used to decrease the electroosmotic flow (EOF) were compared: (i) a hydroxypropylcellulose (HPC) coated capillary in aqueous medium and (ii) the addition of glycerol to act as a viscosifying agent. The cIEF method with the glycerol medium was selected, and the ionization constants of 22 basic and 21 acidic compounds, including 15 pharmaceutical drugs, were determined, resulting in pKa values from 3.5 to 7.4 and 6.4 to 9.3, respectively. cIEF offers an interesting alternative to other techniques for pKa determination with low sample consumption, high throughput and low cost.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Methanol, NMR reference standard
Supelco
Nitrofurantoin, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Acetaminophen Related Compound F, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Trimethoprim, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Cimetidine, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Theophylline, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
4-Chloroaniline, PESTANAL®, analytical standard
Supelco
4-Nitrophenol, PESTANAL®, analytical standard
Theophylline, European Pharmacopoeia (EP) Reference Standard
Acetazolamide, European Pharmacopoeia (EP) Reference Standard
USP
Trimethoprim, United States Pharmacopeia (USP) Reference Standard
USP
Theophylline, United States Pharmacopeia (USP) Reference Standard
Prilocaine, European Pharmacopoeia (EP) Reference Standard
Supelco
2,4-Dichlorophenol, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
Supelco
Trimethoprim, VETRANAL®, analytical standard
Supelco
Aniline, analytical standard
Sigma-Aldrich
Glycerol solution, 83.5-89.5% (T)
Sigma-Aldrich
Trimethoprim, ≥99.0% (HPLC)
Supelco
Nitrofurantoin, VETRANAL®, analytical standard
Sigma-Aldrich
p-Anisidine, 99%
Supelco
Methanol, analytical standard
Sigma-Aldrich
4-Chloroaniline, 98%
Sigma-Aldrich
Aniline, ACS reagent, ≥99.5%
Sigma-Aldrich
m-Toluidine, 99%
Sigma-Aldrich
Methyl nicotinate, 99%
Sigma-Aldrich
4-Methylpyridine, 98%
Sigma-Aldrich
Aniline, ReagentPlus®, 99%
Sigma-Aldrich
2,4-Dimethylaniline, ≥99%
Sigma-Aldrich
4-Nitrophenol solution, 10 mM
Sigma-Aldrich
2,6-Dichlorophenol, 99%