Nitrate/Nitrite Assay Kit

Product Description

REAGEN^™ Nitrate/Nitrite Assay Kit is a cuvette-based enzymatic assay for the determination of Nitrate/Nitrite across a variety of food samples. The kit uses a spectrophotometric, kinetic assay to detect the enzymatic degradation of Nitrate/Nitrite directly from food samples.

The unique features of the kit are:

l        High sensitivity and low detection limit (0.02 ppm)

l          High reproducibility

l          A rapid (45 minute), robust enzyme-based assay

Procedure Overview

REAGEN^™ Nitrate/Nitrite Assay Kit uses a coupled enzymatic reaction scheme to detect nitrate and/or nitrite in food samples: nitrate in the sample is converted by nitrate reductase enzyme to make nitrite. The nitrite then reacts with sulfanilamide and N-(1-naphthyl)-ethylene-diamine to form a red diazo dye. This dye, measured at 540 nm, is proportional to the level of nitrate and/or nitrite in the sample.

The absorbance of each sample at 540 nm is measured using a spectrophotometer. The concentration of nitrate /nitrite in each sample is then directly determined from the diazo dye absorbance at 540 nm within 45 minutes.

Kit Contents, Storage and Shelf Life

REAGEN^™ Nitrate/Nitrite Assay Kit has the capacity for 48 determinations or 24 determinations in duplicate. Store the kit at 4°C. The shelf life of the kit is 12 months, after receipt, when the kit is properly stored.

Sensitivity (Detection Limit)

The linear range of the assay is 0.02 –40 mg/L (ppm).

Required Materials Not Provided With the Kit

l          Spectrophotometer (578 nm)

l          Low volume (0.3 mL) UV cuvettes, such as Brand Scientific catalog #7592 00

l          Deionized or distilled water

l          100 mL volumetric flask

l          50 mL plastic conical tubes

l          Carrez buffer I and Carrez buffer II (recipes included in protocol)

Warnings and Precautions

l          Do not use the kit past the expiration date.

l          Try to maintain a laboratory temperature of 20°–25°C (68°–77°F). Avoid running assays under or near air vents, as this may cause excessive cooling, heating and/or evaporation. Also, do not run assays  in  direct sunlight,  as this may cause excessive  heat and evaporation. Cold  bench tops should be avoided by placing several layers of paper towel or some other insulate material under the assay plates during incubation.

l          Make sure you are using only distilled or deionized water since water quality is very important.