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Separation, detection, and quantification of imine-linked opines by high-performance liquid chromatography

Authors
Journal
Journal of Chromatography A
0021-9673
Publisher
Elsevier
Publication Date
Volume
813
Issue
2
Identifiers
DOI: 10.1016/s0021-9673(98)00346-x
Keywords
  • Opines
  • Mannopine

Abstract

Abstract Protocols for reversed-phase high-performance liquid chromatography (RP-HPLC) with fluorescence detection were developed for the separation, detection and quantification of imine-linked opines, following 4-fluoro-7-nitrobenzoxadiazole (NBD-F) derivatization. Both natural opines (mannopine, mannopinic acid, cucumopine, octopine, octopinic acid, and nopaline) and opine analogs (nor-mannopine, galactopine, glucopine) were included in the study. The on-column detection limits for NBD-opines varied from 0.1 pmol to 5 pmol. These methods were applied to quantify mannopine on the leaf surfaces of transgenic tobacco ( Nicotiana tabacum) plants expressing mannityl opine synthesis ( mas) genes from Agrobacterium tumefaciens.

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