Spectrophotometric Determination of Nitrate in Vegetables Using Phenol

A rapid and sensitive spectrophotometric method for the determination of nitrate in vegetables is described. The method is based on the measurement of the absorbance of yellow sodium nitrophenoxide formed via the reaction of phenol with the vegetable-based nitrate in presence of sulphuric acid. The analytical concentration of the acid has marked effect on the nitrate determined. The colour development was rapid and remained stable overnight. Analysis of six vegetable samples containing nitrate gave satisfactory mean recoveries of 76 to 123% in 18 determinations. The proposed method is reproducible and sensitive to lower level concentrations. @JASEM. The high concentration of nitrogen in soil may lead to the high nitrate levels in edible vegetables and toxic levels of nitrite may be produced by microbial activity in the gastrointestinal tract of the consumer of such vegetables (Tanaka et al., 1982). Nitrate containing vegetables are potential causes of methemoglobinemia, a disease which may occur in children older than 6 month (Sanchez-Echaniz et al., 2001). Since nitrate-nitrogen is lost during digestion (Rowell, 1994), the determination of nitrate as nitrate-nitrogen via digestion becomes difficult. Pfeiffer and Smith (1975) determined nitrate in baby food using ion selective electrode which showed a good correlation with the AOAC xylenol method. Some of the methods employed are based on the nitration of bi-substituted phenol such as 2,4-xylenol, 3,4-xylenol and 2,6-xylenol (Tanaka et al., 1982). Due to the involvement of distillation stage the 2,4xylenol method is time consuming (Tanaka et al., 1982) and therefore unsuitable for routine food analysis. The 2,4-xylenol method was modified by Toyoda et al. (1978) and a gas-liquid chromatographic method was developed. Basset et al. (1978) reported a titrimetric method for the nitrate determination which could be erroneous if applied to vegetables. Tanaka et al. (1982) reported a sensitive and direct spectrophotometric method for the determination of nitrate in vegetables using 2-secbutyl phenol. The basis for the method is that 2-secbutylphenol reacts quantitatively with nitrate in acidic solution. Beijaars et al. (1994) reported a determination of nitrate in vegetables by continuous flow procedure and subsequent reaction of the nitrate extracted with sulfanilamide and N-1naphthylethylenediamine to form a reddish-purple azo dye which was measured colorimetrically at 530nm. This research aimed at developing a method for the determination of nitrate in vegetable products by using phenol as the active reagent. The principle is based on the nitration of phenol and the formation of the corresponding sodium salt. The elementary processes involved are NO3 + H2SO4 → HSO4 + HNO3 .............. (1 HNO3 + H2SO4 → NO2 + OSO3H + H2O ......... (2