Chromatographia

, Volume 76, Issue 17–18, pp 1125–1130 | Cite as

Simultaneous Determination of Key Osmoregulants in Halophytes Using HPLC–ELSD

  • Umme Kalsoom
  • Mary C. Boyce
  • Ian J. Bennett
  • Varaporn Veraplakorn
Original

Abstract

Osmoregulants are the substances produced by plants that assist in tolerating environmental stresses. Three commonly analysed osomoregulants include mannitol, betaine and proline. A simple, sensitive and rapid HPLC–ELSD method has been developed for the simultaneous analysis of these common osmoregulants in plant extracts. Osmoregulants were extracted using 80 % ethanol and separated on an NH2 column using 0.1 % formic acid and acetonitrile as the mobile phase. Retention time repeatability was 0.85, 1.50, and 0.93 % for mannitol, betaine and proline, respectively. The limit of detection (μmol) was 1.43 × 10−4, 7.81 × 10−5 and 1.08 × 10−4 for mannitol, betaine and proline, respectively. The developed method was applied to three different plant extracts, Stylosanthes guianensis, Atriplex cinerea and Rhagodia baccata. A second method using a C18 column with 0.1 % heptafluorobutyric acid and acetonitrile as the mobile phase proved to be a useful complementary method for verifying tentative peak identifications.

Keywords

Column liquid chromatography ELSD Betaine Mannitol Proline Osmoregulators 

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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Umme Kalsoom
    • 1
  • Mary C. Boyce
    • 1
  • Ian J. Bennett
    • 1
  • Varaporn Veraplakorn
    • 2
  1. 1.School of Natural SciencesEdith Cowan UniversityPerthAustralia
  2. 2.Department of Biotechnology, Faculty of ScienceRamkhamhaeng UniversityBangkokThailand

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