Comparison of anion-exchange and hydroxyapatite displacement chromatography for the isolation of whey proteins

J Chromatogr A. 1997 Jan 24;760(1):125-37. doi: 10.1016/s0021-9673(96)00797-2.

Abstract

In displacement chromatography, several substances may be isolated and concomitantly concentrated, which makes this separation procedure attractive for the processing of diluted product streams containing a number of high value substances. Here, the suitability of anion-exchange and hydroxyapatite displacement chromatography for the processing of technical dairy whey is investigated. The pH and flow-rate of the carrier, the displacer chemistry and, in case of the apatite, the particle diameter of the stationary phase are considered. As a consequence of the pH sensitivity of the beta-lactoglobulin, one major whey component, apatite displacement chromatography is less then successful in whey separation. At a denaturing carrier pH (> 8.5) the beta-lactoglobulin zone is broad and stretches over the entire displacement train. At a lower carrier pH, previously successful polyanionic displacers do not bring about separation, while low-molecular-mass ones do, but they tend to overrun and thus contaminate the protein zones. In the case of anion-exchange displacement chromatography, polyanions, especially polyacrylic acid (PAA, M(r) 6000), constitute suitable displacers. Here too, a carrier pH of 8.0 is most suited to the separation of the whey proteins. The low-molecular mass-displacer iminodiacetic acid (IDA, M(r) 133.4), on the other hand, displaces only alpha-lactalbumin. The beta-lactoglobulin remains on the column. PAA is used as the displacer to process a dairy whey sample.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anion Exchange Resins / chemistry*
  • Cattle
  • Chromatography, High Pressure Liquid / methods*
  • Hydrogen-Ion Concentration
  • Hydroxyapatites / chemistry*
  • Lactalbumin / analysis
  • Lactalbumin / chemistry
  • Lactalbumin / isolation & purification
  • Lactoglobulins / analysis
  • Lactoglobulins / chemistry
  • Lactoglobulins / isolation & purification
  • Microspheres
  • Milk Proteins / analysis
  • Milk Proteins / chemistry
  • Milk Proteins / isolation & purification*
  • Particle Size
  • Proteins / analysis
  • Whey Proteins

Substances

  • Anion Exchange Resins
  • Hydroxyapatites
  • Lactoglobulins
  • Milk Proteins
  • Proteins
  • Whey Proteins
  • Lactalbumin