Speciation of nickel by HPLC-UV/MS in pea nodules

A new and sensitive methodology based on normal phase HPLC has been developed for the speciation of nickel in low-complexity plant extracts. The method combines a silica stationary phase column, a 9:1 (v/v) hexane:ethanol mixture as mobile phase, and the detection of nickel complexes by either UV or MS. The developed methodology was applied to the speciation of nickel complexes in the cytoplasm of pea root nodules. Results obtained indicate that nickel citrate and nickel malate accounts for 99% of nickel present in pea nodule cytoplasm fraction. The low detection limit of the method (<0.2 nM) enables nickel speciation in non-hyperaccumulator plants.