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Soil Nitrate Nitrogen
KCl Extraction / Cd-Reduction Method

Robert Miller and Leticia Sonon

Application and Principle

This method involves the quantitative extraction of nitrate nitrogen (NO3-N) from soils using 2.0 M KCl. Nitrate is determined by reduction to nitrite (NO2-N) via a cadmium reactor, diazotized with sulfanilamide and is coupled to N-(1-Napthyl)-ethylenediamine dihydrochloride to form an azochromophore (red-purple in color) measured spectrophotometrically at 520 nm. The method is readily adapted to manual or automated techniques. The procedure follows that of Keeney and Nelson (1982) for determining nitrate nitrogen with a modification in which 25 mL of KCl and 5.0 g of soil are used instead of 100 mL and 10 g soil. Extending the shaking period to thirty minutes with 2.0 M KCl (Bremner and Keeney, 1965) permits the simultaneous extraction of ammonium and nitrate. Care must be taken to avoid contamination from filter paper and operator handling. Since cadmium is a hazardous material, follow manufacturer’s recommendations to properly handle this material. Soil NO3-N is available to plant uptake, therefore, it can be credited when determining crop nitrogen needs. The method detection limit is approximately 0.5 mg kg-1 (on a dry soil basis) and is generally reproducible ± 6%.

Equipment

1. Analytical balance, 100.0 g capacity, resolution ± 0.01 g

2. Repipette dispenser, calibrated to 25.0 ± 0.2 mL

3. Reciprocating horizontal mechanical shaker, capable of 180 oscillations per minute (opm)

4. Extraction vessels and associated filtration vessel

5. Whatman No. 42 or equivalent highly retentive filter paper

6. Spectrophotometer, autoanalyzer, or flow injection analyzer (FIA) instrument

Reagents

1. Deionized water, ASTM Type I grade

2. Potassium chloride extracting solution, 2.0 M KCl: Dissolve 150 g of reagent grade KCl in 500 mL deionized water and dilute to a 1000 mL (See Comment #1) 3. Standard calibration solutions of NO3-N. Prepare six calibration standards ranging from 0.1 to 20.0 mg L-1 in 2.0 M KCl extraction solution prepared from 1000 mg L-1 NO3-N standard solution.

Procedure

1. Weigh or scoop 5.0 ± 0.05 g of air-dried soil pulverized to pass 10 mesh sieve (< 2.0 mm) into extraction vessel. Add 25.0 mL of 2.0 M KCl extraction solution using repipette dispenser (See Comment #2). Include a method blank.

2. Place extraction vessel(s) on reciprocating mechanical shaker and shake for thirty (30) minutes.

3. Filter extract (See Comment #3), refilter if filtrate is cloudy (comment #4).

4. Nitrate-N content of the extract is determined using a spectrophotometer, automated flow analyzer (Technicon Method No. 329-74W/A) or FIA instrument. Calibrate using standard calibration solutions and operate instrument in accordance with manufacturer instructions. Determine nitrate concentration of KCl extract, method blank, unknown samples and record results as mg L-1 of nitrate in extract solution (See Comment #5).

Calculation

1. NO3-N mg kg-1 in soil = (NO3-N mg L-1 in filtrate - method blank) × 5

2. Report soil nitrate nitrogen concentration to the nearest 0.1 mg kg-1 (See Comment #6).

Precision and Accuracy

1. Soil NO3-N measurements can be made to the nearest 0.1 mg kg-1.

2. Typical measurements of inter-laboratory precision for soil NO3-N are shown below.

|Sample 1 |Mean |Standard deviation |
| |mg kg-1 |mg kg-1 |
|SRS - 1003 |4.6 |0.3 |
|SRS - 1006 |11.4 |0.6 |
|SRS - 1008 |24.6 |0.9 |
|SRS - 1010 |46.3 |1.2 |

1 Data from 2010 ALP Program, based on three replications.

Interferences

1. N nitrite (NO2-N) in soil will result in a positive interference. It is very unlikely that soil samples that have been thoroughly dried will contain NO2-N. To verify concentration repeat extraction and analyzed extract for NO2-N without the use of the cadmium reduction column.

Effects of Storage

1. Air-dried soils may be stored several months without affecting the soil nitrate provided they are stored in an moisture free environment.

Safety and disposal

1. The chemicals used in this procedure pose no safety risk and therefore can be stored and disposed of according to routine laboratory procedures.

Comments

1. Soils can be extracted with 1.0 M KCl (Kachurina et al., 200).

2. Check repipette dispensing volume calibration using an analytical balance.

3. Check filter paper supply for possible contamination of NO3-N. If contamination is greater than 0.2 mg L-1 on a solution basis, rinse filter paper with 2.0 M KCl.

4. Soil KCl extract may be stored up to three weeks if stored at 4oC and/or with 100 uL of toluene or thymol.

5. Samples having nitrate concentrations exceeding the highest standard will require dilution and reanalysis.

6. Nitrate-nitrogen (NO3-N) results can be expressed on a volume basis. Assuming the sample represents a 0-6 inch (0-15 cm) depth of the soil, then: NO3-N mg kg-1 × 2.0 ≅ NO3-N lbs ac-1

References

Bremmer, J. M. and D.R. Keeney. 1965. Determination and isotopic ratio analysis of different forms of nitrogen in soils: I. Apparatus and procedure for distillation for and determination of ammonium. Soil Sci. Soc. Am. Proc. 29:504-507.

Dahnke, W.C. 1990. Testing soils for available nitrogen. p. 120-140. In: R.L. Westerman (ed.) Soil testing and plant analysis. Soil Sci. Soc. Am. Book series 3 ASA Madison WI.

Dorich, R. A. and D. W. Nelson. 1984. Evaluation of manual cadmium reduction methods for determination of nitrate in potassium chloride extracts of soils. Soil Sci. Soc Am J. 48:72-75.

Gelderman, R.H. and P.E. Fixen. 1988. Recommended nitrate-N tests. p. 10-11. In: W.C. Dahnke (ed.) Recommended chemical soil test procedures for the North Central Region. North Dakota Agr. Expt. Sta. Bulletin No. 499 (revised).

Kachurina, O.M., H. Zhang, W.R. Raun, and E.G. Krenzer. 2000. Simultaneous determination of soil aluminum, ammonium- and nitrate-nitrogen using 1 M potassium chloride extraction. Commun. Soil Sci. and Pl. Anal. 31: 893-903.

Keeney, D.R. and D.W. Nelson. 1982. Nitrogen - inorganic forms. In A.L. Page (eds.) Methods of soil analysis, part 2. Agron. Monogr. 9, 2nd ed. ASA and SSSA, Madison, WI. p. 643-698.

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