Application of size-exclusion chromatography–inductively coupled plasma mass spectrometry for fractionation of element species in seeds of legumes

Fractionation of soluble species of P, Mn, Fe, Co, Ni, Cu, Zn, Se and Mo in pea and lentil seeds was made by on-line hyphenation of size-exclusion chromatography (SEC) and inductively coupled plasma mass spectrometry. Seed samples were extracted with 0.02 mol l −1 Tris–HCl buffer solution, pH 7.5. SEC was performed on Superdex 75 and Superdex Peptide columns (300×10 mm) with the same buffer solution as the mobile phase. Monitoring of oxide ion 47(PO) + was used for detection of phosphorus compounds. Other elements were detected as ions of 55Mn, 57Fe, 59Co, 62Ni, 65Cu, 66Zn, 82Se and 95Mo nuclides. Elements in individual elution zones were quantified using external calibration. Complete chromatographic recoveries of elements were found in cases of phosphorus, nickel and copper. Substantial parts of manganese and zinc, as well as traces of cobalt, selenium and molybdenum are retained on the column. Injection of EDTA solution removes these elements from the column. Chromatographic profiles of pea and lentil samples are very similar for all elements except Mo. Main element species in the high-molecular-mass region (approx. 190 000 rel. mol. mass unit) were detected in case of Fe. Low-molecular-mass species (