Rapid and sensitive determination of nitrate in plant tissue using flow injection analysis

Abstract

The determination of nitrate in waters and soil extracts by the reduction of nitrate to nitrite by metallic or liquid reductants followed by the colorimetric determination of NO₂ ^‐using the Griess‐Ilosvay reaction has been automated for use with air‐segmented auto‐analyzers or flow injection techniques. However, this technique is not applicable to plant extracts as organic species in the extracts inactivate the reduction columns. The objective of this study was to develop an automated procedure that would allow the determination of NO₃ ^‐in plant extracts without the necessity of prior manual treatment. A flow injection technique was developed that successively traps and releases NO₃in an anion exchange column thereby removing pigments and other non‐ionic and cationic species that otherwise interfere with conversion of NO₃ ^‐ to NO₂ ^‐on a copperized cadmium column. This reduction step is subsequently followed by standard Griess‐Ilosvay colorimetric detection of this ion at a wavelength of 530 nm. The technique uses relatively simple and inexpensive equipment, principally a spectrophotometer equipped with a flow‐through cell and a pen recorder output, a 6‐channel peristaltic pump with accompanying tubing and a Perspex injector/commutator valve made in a laboratory workshop. The technique was found to avoid any significant interference of pigments or other organic compounds in the plant extracts, and the results compared favorably with those obtained using the manual transnitration technique. Analysis time was approximately 1.5 min per sample and could detect NO₃’ concentration as low as 0.1 ug NO₃’‐N ml/¹ in plant extracts (10 ug NO₃’‐N g‘¹of plant material).