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Simultaneous Determination of Key Osmoregulants in Halophytes Using HPLC–ELSD

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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.

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Authors and Affiliations

  1. School of Natural Sciences, Edith Cowan University, Perth, WA, 6027, Australia

    Umme Kalsoom, Mary C. Boyce & Ian J. Bennett

  2. Department of Biotechnology, Faculty of Science, Ramkhamhaeng University, Bangkok, 10240, Thailand

    Varaporn Veraplakorn

Authors
  1. Umme Kalsoom
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  2. Mary C. Boyce
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  3. Ian J. Bennett
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  4. Varaporn Veraplakorn
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Correspondence to Mary C. Boyce.

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Kalsoom, U., Boyce, M.C., Bennett, I.J. et al. Simultaneous Determination of Key Osmoregulants in Halophytes Using HPLC–ELSD. Chromatographia 76, 1125–1130 (2013). https://doi.org/10.1007/s10337-013-2494-7

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  • DOI: https://doi.org/10.1007/s10337-013-2494-7

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