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Chromatographia

, Volume 30, Issue 5–6, pp 301–308 | Cite as

Retention mechanisms and selectivity in internal-surface reversed-phase liquid chromatography. Studies with cyanobacterial peptide toxins

  • J. A. O. Meriluoto
  • K. Isaksson
  • H. Soini
  • S. E. Nygård
  • J. E. Eriksson
Originals

Summary

Microcystins-LA,-LR,-RR,-YR and nodularin, cyanobacterial peptide toxins, were separated by internal-surface reversed-phase (ISRP), high-performance liquid chromatography. The capacity factors of the toxins were measured in the range pH 2–8 using acetonitrile, isopropanol or tetrahydrofuran in potassium dihydrogenphosphate mobile phase. The main retention mechanism of the ISRP column was reversed-phase interaction but cation-exchange offered additional selectivity at neutral and slightly acidic pH. At neutral pH (10% modifier, 0.1 M buffer) the elution order was microcystin-LA (two nonpolar residues leucine and alanine as the variable amino acids), nodularin, microcystin-LR,-YR and-RR (two basic arginines as the variable amino acids). The retention times of all toxins except microcystin-RR were substantially longer at acidic pH. At pH 2 (10% modifier, 0.1 M buffer) where the cation-exchange mechanism was inoperative the elution order was changed to microcystin-RR, nodularin, microcystin-LR,-YR and-LA. The best separation was achieved at pH 2 where even two desmethylated microcystin-RR analogs could be separated from microcystin-RR.

Key Words

Column liquid chromatography Internal-surface reversed-phases Cyanobacterial peptide toxins Microcystin Nodularin 

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Copyright information

© Friedr. Vieweg & Sohn Verlagsgesellschaft mbH 1990

Authors and Affiliations

  • J. A. O. Meriluoto
    • 1
  • K. Isaksson
    • 1
  • H. Soini
    • 1
  • S. E. Nygård
    • 1
  • J. E. Eriksson
    • 2
  1. 1.Department of Biochemistry and PharmacyÅbo Akademi UniversityTurkuFinland
  2. 2.Department of BiologyÅbo Akademi UniversityTurkuFinland

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