Determination of the ‘apparent pKa’ of selected food hydrocolloids using ortho-toluidine blue

Leo F.W. Vleugels, Stella Ricois, Ilja K. Voets, Remco Tuinier

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Charged food hydrocolloids provide structure and texture in manufactured foods by their organization and electrostatic interactions with other species and small ions. These electrostatic interactions depend on their actual charge density which cannot be predicted from pKa values of weakly charged monomers. In practice, this is circumvented by the use of an ‘apparent pKa’ (pKa (app)) which depends on solution ionic strength and type of counter-ions. pKa (app) values for food hydrocolloids are poorly documented, especially in low ionic strength solvents. Titration of a metachromatic alginate-ortho-toluidine blue complex revealed a sigmoidal, pH-dependent, dye metachromasy response with its centre located at pH = pKa (app). To test the suitability of this approach for the determination of pKa (app), five food hydrocolloids and a synthetic polyanion are analysed and the results are compared to classical methods such as potentiometric titration. Metachromasy is shown to occur when the polymer inter-charge distance is 4.5 Å; the complex stoichiometry depending on inter-charge distance. pKa (app) of stoichiometric complexes at low ionic strength (1 mM phosphate buffer) were successfully determined (alginate 4.5, sodium carboxymethylcellulose 4.6, κ-carrageenan 1.2, polyphosphate 2.1 and polyacrylic acid 5.8) and are comparable to values obtained by potentiometric titration. Further, it is shown that pKa (app) determination by metachromatic dye titration is feasible for stoichiometric polymer-dye complexes at low concentrations (20–200 μM) in low ionic strength (<10 mM) aqueous solutions. Unlike classical methods, the metachromatic dye titration approach is suitable for low polymer concentrations in aqueous solutions at low ionic strength.

Original languageEnglish
Pages (from-to)273-283
Number of pages11
JournalFood Hydrocolloids
Publication statusPublished - 1 Aug 2018


  • Apparent pK
  • Low ionic strength
  • Metachromasy
  • Ortho-toluidine blue
  • Polyelectrolyte behaviour
  • Polysaccharides


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