An ‘off-line’ high performance liquid chromatography–graphite furnace atomic absorption spectrometry (HPLC–GF-AAS) method using a size exclusion chromatography (SEC) column was developed to investigate heavy metal ions in xylem sap samples of cucumber plants grown in hydroponics containing iron as Fe(III)-ethylenediaminetetraacetate (Fe(III) EDTA), Fe(III) citrate or FeCl 3 and exposed to lead, nickel or vanadium contamination. The SEC chromatogram of the samples contained the peak of nitrate ions (in significant concentration ≈1400 μg/ml) and some small, unidentified compounds with molecular weight lower than 700 Da. The results indicate that Cu and Mn — which were added to the hydroponics as nutrient elements — determined in the collected fractions during the chromatographic runs are transported in the xylem vessels together with small inorganic ions like nitrate ions. In case of nickel other low-molecular weight compounds eluting earlier than the nitrate ions may take part in its transport toward the shoots. Lead could not be detected in the above mentioned fractions. Determination of vanadium in the fractions was not expected since it could not be detected in the sap samples.