Determination of Total

EXPERIMENT 4 DETERMINATION OF Available Nitrogen in a

Soil Sample
TOTAL AVAILABLE
NITROGEN IN A SOIL
SAMPLE
Structure
4.1 Introduction
Objectives
4.2 Principle
4.3 Requirements
4.4 Solutions Provided
4.5 Procedure
4.6 Observations and Calculations
4.7 Results

4.1 INTRODUCTION
In the previous experiment you learnt how percent organic matter in the soil can be
determined by wet oxidation method. The present and next three experiments are
related to the analysis of soil for some of its nutrient elements. In Unit 3 of Block 1
you read about various macro and micronutrients present in soil and the dependence
of soil productivity on these. It is important to analyse nutrients present in a soil
sample so as to gauge its productivity level and recommend suitable remedial
measures in case of any imbalance of these nutrients. In this experiment you would
learn and perform the determination of nitrogen.

Nitrogen, an important plant nutrient is present in the soil in both organic as well as
inorganic forms. The organic component of the soil nitrogen is present primarily as
plant, animal and microbial remains and as such is unavailable to plants. It becomes
available through mineralisation and organic matter decomposition processes. In fact,
the organic form of nitrogen in the soil can be visualised as a reservoir of nitrogen
and reflects the nitrogen supplying capacity of the soil. Only a minute fraction say
about two percent gets mineralised due to microbial activity in the growing season.
The inorganic component of the soil nitrogen is present in the form of ammonium
(NH+4 ), nitrate (NO−3 ) and nitrite (NO−2 ) ions. Of these, nitrogen is taken up by
plants in (NH +4 ) - N and (NO −3 ) - N forms and reflects the extent of mineralisation.
The amount of nitrogen in these forms is an indicator of the nitrogen availability
for the current crop and in turn determines the immediate requirements of N-
fertilizer.

Determination of nitrogen in the soil is done in a number of ways depending on

whether the objective is to determine total nitrogen content or individual components
like, organic nitrogen or (NH +4 ) , - N, (NO 3− ) - N or (NO −2 ) - N or their combinations.
In this experiment you will learn to determine total available ( or mineralisable)
nitrogen from the organic matter using alkaline potassium permanganate method. As
mentioned above, it is a measure of nitrogen reservoir or the nitrogen supplying
capacity of the soil.

Objectives
After studying and performing this experiment, you should be able to: 29
Experiments • explain different types of nitrogen availability in the soil,
• explain the principle and procedure involved in the potassium permanganate
method of total available N determination, and
• interpret the nitrogen value in terms of nitrogen supplying capacity of the soil .

4.2 PRINCIPLE
The easily hydrolysable and oxidisable fractions of organic nitrogen present in a soil
sample can be extracted out using alkaline KMnO4 - a mild oxidising agent. For this
purpose a known amount of soil sample is boiled with a measured quantity of alkaline
KMnO4. It leads to evolution of ammonia which is absorbed in a known volume of
standard sulphuric acid. A part of the acid is neutralised by ammonia and the excess is
titrated with a standard solution of alkali using methyl red as indicator.
This method gives the amount of available nitrogen content as the percent of the soil.
This value is also expressed in terms of Kg/ha. For this purpose it is assumed that 1 ha
of furrow slice (0-15cm) of the soil weighs 2.24 million kg. If the amount of available
nitrogen is found to be less than 272 kg/ha. the sample is rated low, if it is 272-
544 kg/ha, it is rated medium and if the available nitrogen content is more than 544
kg/ha it is rated as high.

4.3 REQUIREMENTS
Apparatus Chemicals
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1. Kjeldahl flask (800 cm ) 1. Potassium permanganate
2. Distillation apparatus 2. Sodium hydroxide
3. Measuring cylinder 3. Sulphuric acid
4. Pipette 4. Methyl red indicator
5. Burette
6. Conical flas k

4.4 SOLUTIONS PROVIDED

You will be provided with these solutions. However, if required these can be prepared
by the following procedures:

1. 0.32% KMnO4 : Weigh 3.2 g KMnO4 and transfer it to a 1 dm3 or 1000 cm3
volumetric flask. Add some distilled water to the flask and dissolve it by gently
swirling the flask. Carefully make up the volume to 1 dm 3.

2. 2.5% NaOH : Weigh 25 g of NaOH pellets and transfer to a 1 dm3 graduated

beaker or measuring cylinder. Dissolve it in distilled water and carefully make up
the volume to 1 dm3.

3. 0.02 M NaOH: Dissolve about 1.0 g of NaOH pallets in water in a standard flask
or a measuring cylinder. Standardise by titrating against 0.01 M oxalic acid using
methyl red indicator. Adjust the strength to 0.02 M.

4. 0.01 M H2SO4 : Carefully add 0.6 cm3 of concentrated H2SO4 (sp. gr. 1.84) to one
dm3 of distilled water. Standardise by titrating against a standard alkali solution
using methyl red indicator. Adjust the strength to 0.01 M.

5. 0.15% methyl red indicator : Dissolve 0.15 g of methyl red powder in 50 cm3 of
ethyl alcohol and make up the volume to 100 cm3 with water.

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 Determination of Total
4.5 PROCEDURE Available Nitrogen in a
Soil Sample
Perform the following steps to determine total available ( mineralisable) nitrogen in a
soil sample.
1. Set up the distillation apparatus with the help of your counsellor.
2. Weigh 20 g of given soil sample and transfer it carefully into Kjeldahl distillation
flask.
3. Moisten the sample with about 10 cm3 of distilled water. Wash down the soil
adhering to the neck of the flask.
4. Add 100 cm3 of 0.32% KMnO4 solution and 100 cm3 of 2.5% NaOH solution and
a few glass beads or broken pieces of glass rod to avoid bumping to the above
sample and immediately stopper the flask.
5. Take 25 cm3 of 0.01 M H2SO 4 in a 150 cm3 conical flask and add 3-4 drops of
methyl red to it. Dip the end of the delivery tube of the distillation apparatus into
it.
6. Heat the distillation flask steadily to distill 100 cm3 of liquid ammonia in about 30
minutes time.
7. Titrate the excess of standard H2SO4 left in the conical flas k with 0.02 M NaOH
and note the volume used (X cm 3 ) in the observation table.

4.6 OBSERVATIONS AND CALCULATIONS

Observation Table

Weight of soil taken = 20 g

25 cm3
Volume of 0.01 M H2SO4 =

Volume of 0.02 M NaOH required for back ---------------- cm3

titration = X cm3

Volume of 0.01 M H2SO4 used by NH3 evolved ---------------- cm3

= (25 − X) cm3

Calculations

1 cm3 of 0.01 M H2SO4 = 0.00028 g of nitrogen

Amount of nitrogen in 20 g of soil = (25 −X) ×0.00028 g = Z g
% available nitrogen ( per 100 g of soil) = Z × 100/20 % = Z’ %
Available nitrogen in ppm = Z’ × 10,000
Available nitrogen in Kg/ha = Z ’ × 22400

4.7 RESULTS
The amount of available nitrogen in the given sample of soil = ……… Kg/ha

The soil sample on the basis of its available nitrogen value is rated as:

low / medium / high. ( Tick the appropriate option)

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