Simultaneous Determination of Key Osmoregulants in Halophytes Using HPLC-ELSD

Document Type

Conference Proceeding


Faculty of Health, Engineering and Science


School of Natural Sciences / Centre for Ecosystem Management


This paper was originally published as Kalsoom, U., Boyce, M. C., Bennett, I. J., & Veraplakorn, V. (2013). Simultaneous determination of key osmoregulants in halophytes using HPLC-ELSD. Paper presented at the 40th International Symposium on High Performance Liquid Phase Separations and Related Techniques. Hobart, Australia. 17-21st November 2013.


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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