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Phytochemical Analysis

Capillary electrophoresis of the major anions and cations in leaf extracts of woody species.

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Charles R Warren
Mark A Adams

Keywords

Abstract

Capillary electrophoresis methods are described for the analysis of the major inorganic anions (nitrite, nitrate, chloride, sulphate, phosphate), organic acids (oxalate, malate, citrate, succinate) and inorganic cations (ammonium, potassium, sodium, calcium, magnesium) in leaf extracts. Analytical performance was validated for extracts from leaves of four sclerophyllous species: Eucalyptus globulus, E. cladocalyx, E. nitens and Pinus radiata. Inorganic anions and organic acids were analysed in a single run within 5 min using a background electrolyte of 2,6-pyridinedicarboxylic acid (20 mM) and cetyltrimethylammonium bromide (0.5 mM). Cations were analysed in a separate run also within 5 min using imidazole (10 mM) and 18-crown-6 (2 mM) as background electrolyte. Replicate injections were highly repeatable when the capillary was rinsed between runs with hydrochloric acid (0.25 M) and background electrolyte, but not when the acid rinse was omitted or replaced by a rinse with sodium hydroxide (0.25 M). Standard curves for all analytes were linear over the range of 0.05-1 mm. Standard curves constructed by serial dilution of a leaf extract were also highly linear, and this, combined with the excellent recovery of added solutes in a spike and recovery test, suggests quantification was unaffected by the complex matrix that is present in un-purified, hot water extracts of leaves. There were significant differences in concentrations of the major anions and cations between the species studied.

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