Nitrate-N determination in leafy vegetables: Study of the effects of cooking and freezing

Nitrate upon reduction to nitrite can cause methaemoglobinaemia or act as precursor in the endogenous formation of carcinogenic nitrosamines. The leafy vegetables are the major vehicle for the entry of nitrate into the human system. The present study was conducted to establish a flow injection analysis (FIA) technique to investigate the nitrate-N contents of four commonly consumed fresh leafy vegetables (Chinese cabbage, celery, lettuce and English cabbage) from market in Fiji. Two extraction techniques (activated carbon and alkaline extraction) were assessed to extract nitrate-N and the activated carbon extraction was preferred over alkaline extraction and applied. The recoveries of spiked nitrate-N in vegetable matrices ranged from 90.40% to 112.80% in activated carbon extraction with an average of 100.62%. The effects of cooking (boiling, baking and frying) and deep-freezing on the nitrate-N contents were also studied. Nitrate contents in selected leafy vegetables were determined by FIA coupled with Greiss protocol involving sulfanilamide and N-(1-naphthyl)ethylenediamine dihydrochloride as color reagents. Nitrate was determined in the linear range from 1.0 to 20.0 mg L⁻¹ with the method detection limit of 0.042 mg L⁻¹ (0.34 mg kg⁻¹). The results of the study show that nitrate contents in fresh leafy vegetables ranged from 1297 to 5658 mg kg⁻¹. Boiling reduces nitrate content by 47–56% whereas frying in Soya bean oil elevates nitrate content by as much as 159–307%. No significant change was observed in nitrate content after baking. The deep-freezing of the selected leafy vegetables shows that nitrate-N content fluctuates slightly from the original nitrate-N values over the seven day period. The FIA throughput was 38 samples h⁻¹.