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DETERMINATION OF TOTAL NITROGEN
IN SOIL BY SEMI-MICRO KJELDAHL
METHOD
- Abdullah Al Noman Neion
Department of Soil Science , Bangladesh Agricultural University
Importance of Nitrogen in Soil
Essential for plant
growth and
development
Component of
chlorophyll and
organic matter
Regulates soil pH
levels
Vital for soil
microbe growth
Limits plant
growth if
deficient
Commonly used
in agriculture
Involves complex
nitrogen cycling
Improves soil
health and
sustainable
agriculture
Objectives of Determining
Total Nitrogen in Soil
• Assess soil fertility
• Determine nitrogen requirements
• Monitor soil nitrogen changes
• Evaluate environmental impact
• Understand nitrogen cycling
• Support sustainable agriculture
• Guide fertilizer and soil management
• Monitor soil conservation efforts
Sources of
Nitrogen in
Soil
Atmospheric deposition
Nitrogen-fixing plants and bacteria
Decomposition of organic matter
Commercial fertilizers
Manure and compost
Nitrification of ammonium compounds
Fixation of nitrogen gas in soil
Application of sewage sludge
Release from plant roots and root exudates
Functions of
Nitrogen in Plants
Symptoms of
Nitrogen Deficiency
• Very slow growth and look weak and
stunted.
• Older leaves have a light green
or yellow colour.
• As nitrogen is mobile in the plant
and moves to where it is needed,
these older and lower leaves are
always first to show symptoms of
deficiency.
Leaching Loss
• The nitrate-nitrogen is lost in
drainage or with percolating
water.
• The amount of nitrogen lost
depends upon the climate and
cultural conditions.
• In humid region or a water- logged
condition, losses of nitrate by
leaching are significant.
• In arid and semi-arid regions, such
losses are minimum
Volatilization
• The loss of nitrogen to the atmosphere as ammonia gas.
• The greatest losses occur from surface application of fertilizer
containing ammonium (urea or blended dry fertilizers), particularly on
a high pH soil and/or high temperature during hot, windy weather, or
on dense crop residue which inhibits fertilizer-soil contact.
Denitrification
• Denitrification occurs when oxygen levels
are low, anaerobic microorganisms strip
oxygen from nitrate which produces
nitrogen gas, nitric oxide or nitrous oxide.
• When this occurs, these gases may
diffuse out of the soil into the air.
• The following four conditions that
contribute to denitrification are: Wet or
poorly drained soil; compaction; warm
soil temperatures; or readily
decomposable organic matter.
Methodology
Two methods have gained general
acceptance for determination of
total Nitrogen:
1. The Kjeldahl method
2. The Dumas method
Kjeldahl Method
It is a method for the quantitative determination of nitrogen contained
in organic substances, the nitrogen contained in the inorganic
compounds ammonia and ammonium.
Dumas Method
This method is based upon the fact that nitrogenous compound when
heated with cupric oxide in an atmosphere of CO2 yields free nitrogen.
Reagents Required
CONC. H2SO4 NaOH 35% H3BO3 INDICATOR
SOLUTION
CATALYST
MIXTURE
Functions of
Reagents
Digestion
Distillation
Indicator
Raises temperature and oxidation
Principle
Major steps:
• Digestion: Organic N converted to NH4
+
• Distillation: NH4
+ converted to NH3 and trapped
by indicator
• Titration: Trapped NH3 determined
Digestion:
Organic N is converted to NH4
+ by digestion with conc. H2SO4 in
presence of substances (catalyst mixture) that promote this conversion.
Catalyst mixture:
K₂SO4, CuSO4.5H₂O and Se in the ratio of 100:10:1 or 10:1:0.1
Distillation:
The NH4
+-N in the digest is converted to NH3 by distillation with
strong alkali (35% NaOH) and this is trapped with H3BO3 indicator
solution.
Titration
The NH3 trapped in H3BO3 is determined by titration with 0.01N H2SO4.
NH3H3BO3+H2SO4 (NH4)2SO4+H3BO3
Green Pink
Procedure
01
Take 1 g soil
sample
02
03
04
1.1 g of K2SO4
catalyst
mixture
5 ml H2SO4
(conc.)
Heat up to
360°C
(colorless)
Allow to cool
& add 20 ml
H2O, then
Swirl the flask
Procedure (contd.)
05
Vol. up to 100
ml with DW &
Transfer 25 ml
in test tube
06
07
08
Add 20 ml of
H3BO3
indicator &
place into
distillation unit
Steam supply
for 4 minutes
(pumping
NaOH)
Volatilized
NH3 trapped
with H3BO3 &
Titration with
0.01 N H2SO4
Calculation
1L 1N H2SO4 ≡ 14g N
Or, 1000mL 1N H2SO4 ≡14g N
Or, 1mL 1N H2SO4 ≡ 0.014g N
Or, 1mL 0.01N H2SO4 ≡ 0.00014g N
% N = {(Vol. of 0.01N H2SO4 required in titration – Blank)*100} ÷
Weight of soil taken
=0.0078
Result
The percentage Nitrogen of
the supplied soil sample was
0.0078
Please
Conserve Soil :)

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Determination of Total Nitrogen in Soil by Semi Micro Kjeldahl Method

  • 1. DETERMINATION OF TOTAL NITROGEN IN SOIL BY SEMI-MICRO KJELDAHL METHOD - Abdullah Al Noman Neion Department of Soil Science , Bangladesh Agricultural University
  • 2. Importance of Nitrogen in Soil Essential for plant growth and development Component of chlorophyll and organic matter Regulates soil pH levels Vital for soil microbe growth Limits plant growth if deficient Commonly used in agriculture Involves complex nitrogen cycling Improves soil health and sustainable agriculture
  • 3. Objectives of Determining Total Nitrogen in Soil • Assess soil fertility • Determine nitrogen requirements • Monitor soil nitrogen changes • Evaluate environmental impact • Understand nitrogen cycling • Support sustainable agriculture • Guide fertilizer and soil management • Monitor soil conservation efforts
  • 4. Sources of Nitrogen in Soil Atmospheric deposition Nitrogen-fixing plants and bacteria Decomposition of organic matter Commercial fertilizers Manure and compost Nitrification of ammonium compounds Fixation of nitrogen gas in soil Application of sewage sludge Release from plant roots and root exudates
  • 6. Symptoms of Nitrogen Deficiency • Very slow growth and look weak and stunted. • Older leaves have a light green or yellow colour. • As nitrogen is mobile in the plant and moves to where it is needed, these older and lower leaves are always first to show symptoms of deficiency.
  • 7. Leaching Loss • The nitrate-nitrogen is lost in drainage or with percolating water. • The amount of nitrogen lost depends upon the climate and cultural conditions. • In humid region or a water- logged condition, losses of nitrate by leaching are significant. • In arid and semi-arid regions, such losses are minimum
  • 8. Volatilization • The loss of nitrogen to the atmosphere as ammonia gas. • The greatest losses occur from surface application of fertilizer containing ammonium (urea or blended dry fertilizers), particularly on a high pH soil and/or high temperature during hot, windy weather, or on dense crop residue which inhibits fertilizer-soil contact.
  • 9. Denitrification • Denitrification occurs when oxygen levels are low, anaerobic microorganisms strip oxygen from nitrate which produces nitrogen gas, nitric oxide or nitrous oxide. • When this occurs, these gases may diffuse out of the soil into the air. • The following four conditions that contribute to denitrification are: Wet or poorly drained soil; compaction; warm soil temperatures; or readily decomposable organic matter.
  • 10. Methodology Two methods have gained general acceptance for determination of total Nitrogen: 1. The Kjeldahl method 2. The Dumas method
  • 11. Kjeldahl Method It is a method for the quantitative determination of nitrogen contained in organic substances, the nitrogen contained in the inorganic compounds ammonia and ammonium.
  • 12. Dumas Method This method is based upon the fact that nitrogenous compound when heated with cupric oxide in an atmosphere of CO2 yields free nitrogen.
  • 13. Reagents Required CONC. H2SO4 NaOH 35% H3BO3 INDICATOR SOLUTION CATALYST MIXTURE
  • 15. Principle Major steps: • Digestion: Organic N converted to NH4 + • Distillation: NH4 + converted to NH3 and trapped by indicator • Titration: Trapped NH3 determined
  • 16. Digestion: Organic N is converted to NH4 + by digestion with conc. H2SO4 in presence of substances (catalyst mixture) that promote this conversion. Catalyst mixture: K₂SO4, CuSO4.5H₂O and Se in the ratio of 100:10:1 or 10:1:0.1
  • 17. Distillation: The NH4 +-N in the digest is converted to NH3 by distillation with strong alkali (35% NaOH) and this is trapped with H3BO3 indicator solution.
  • 18. Titration The NH3 trapped in H3BO3 is determined by titration with 0.01N H2SO4. NH3H3BO3+H2SO4 (NH4)2SO4+H3BO3 Green Pink
  • 19. Procedure 01 Take 1 g soil sample 02 03 04 1.1 g of K2SO4 catalyst mixture 5 ml H2SO4 (conc.) Heat up to 360°C (colorless) Allow to cool & add 20 ml H2O, then Swirl the flask
  • 20. Procedure (contd.) 05 Vol. up to 100 ml with DW & Transfer 25 ml in test tube 06 07 08 Add 20 ml of H3BO3 indicator & place into distillation unit Steam supply for 4 minutes (pumping NaOH) Volatilized NH3 trapped with H3BO3 & Titration with 0.01 N H2SO4
  • 21. Calculation 1L 1N H2SO4 ≡ 14g N Or, 1000mL 1N H2SO4 ≡14g N Or, 1mL 1N H2SO4 ≡ 0.014g N Or, 1mL 0.01N H2SO4 ≡ 0.00014g N % N = {(Vol. of 0.01N H2SO4 required in titration – Blank)*100} ÷ Weight of soil taken =0.0078
  • 22. Result The percentage Nitrogen of the supplied soil sample was 0.0078