Soil hydroxylamine reductase (HR) test kit

Product Details:
Soil hydroxylamine reductase(HR) Test BoxMeasurement significance

Soil hydroxylamine reductase can reduce the intermediate product hydroxylamine formed during nitrogen metabolism in soil to ammonia. The reduced compounds in soil can act as hydrogen donors, and their strength affects the ammonia volatilization loss of nitrogen during soil nitrogen metabolism, indirectly affecting the utilization efficiency of nitrogen fertilizer.

Measurement principle

Ammonium iron sulfateFe3+can oxidize hydroxylamine to nitrogen gas and reduce itself to Fe2+. Fe2+forms an orange red complex with o-phenanthroline under weak acid conditions, with an absorption peak at 510nm. Hydroxylamine reductase acts on hydroxylamine, reducing the amount of complex formation. The decrease in absorbance at 510nm reflects the activity of hydroxylamine reductase.

Bring your own experimental supplies and instruments

Balance, room temperature centrifuge, visible spectrophotometer/Enzyme linked immunosorbent assay (ELISA) reader, trace quartz colorimetric dish/96 well plate, shaker, nitrogen blower.
Reagent composition and preparation:

Reagent 1: Liquid1 bottle, stored at 4 ℃.

Reagent 2: Liquid1 bottle, stored at 4 ℃.

Reagent 3: Powder1 bottle (brown), stored at 4 ℃ in the dark. Add 667 μ L anhydrous ethanol before use, cover tightly and mix well. Then add 9.333 mL distilled water and mix well. Store in the dark.

Reagent 4: Powder1 tube, stored in the dark at 4 ℃. Add 1 mL of distilled water before use and dissolve thoroughly.

Standard product: Powder1 bottle, add 10 mL of distilled water before use, fully dissolve. 1 μ mol/mL standard solution, stored at 4 ℃ in the dark.
Product attributes:

Measurement operation:

Preheat the spectrophotometer/ELISA reader for 30 minutes, adjust the wavelength to 570 nm, and zero the distilled water.

2. Blank tube: Take an EP tube and add 10 μ L of distilled water, 100 μ L of reagent 2, 100 μ L of reagent 3, and 10 μ L of reagent 4. Mix well and cover tightly with a bottle cap (to prevent water loss). Place the tube in a boiling water bath for 15 minutes. After cooling, invert the EP tube several times and measure the absorbance value at 570nm, which is recorded as an empty white tube A. After color development, it is necessary to complete the measurement within 30 minutes.

3. Standard tube: Take an EP tube and add 10 μ L of standard substance, 100 μ L of reagent 2, 100 μ L of reagent 3, and 10 μ L of reagent 4. Mix well and cover tightly with a bottle cap (to prevent moisture loss). Place the tube in a boiling water bath for 15 minutes. After cooling, invert the EP tube several times and measure the absorbance value at 570nm, which is recorded as the A standard tube. After color development, it is necessary to complete the measurement within 30 minutes.

4. Measurement tube: Take an EP tube, add 10 μ L of supernatant, 100 μ L of reagent 2, 100 μ L of reagent 3, and 10 μ L of reagent 4, mix well, and cover tightly with a bottle cap (to prevent water loss). Place in a boiling water bath for 15 minutes, cool down, and repeatedly invert the EP tube several times. Measure the absorbance value at 570nm and record it as the A measurement tube. After color development, it is necessary to complete the measurement within 30 minutes.

Note: Blank tubes and standard tubes only need to be measured once.

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