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World Bank & Government of The Netherlands funded
Training module # WQ - 38
How to measure Ammonia and
Organic Nitrogen: Kjeldahl
Method
New Delhi, March 2000
CSMRS Building, 4th Floor, Olof Palme Marg, Hauz Khas,
New Delhi – 11 00 16 India
Tel: 68 61 681 / 84 Fax: (+ 91 11) 68 61 685
E-Mail: dhvdelft@del2.vsnl.net.in
DHV Consultants BV & DELFT HYDRAULICS
with
HALCROW, TAHAL, CES, ORG & JPS
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 1
Table of contents
Page
1 Module context 2
2 Module profile 3
3 Session plan 4
4 Overhead/flipchart master 5
5 Evaluation sheets 16
6 Handout 18
7 Additional handout 22
8 Main text 24
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 2
1. Module context
This module deals with the significance of nitrogen to water quality and Kjeldahl method for
measuring ammonia and organic nitrogen. Modules in which prior training is required to
complete this module successfully and other available, related modules in this category are
listed in the table below.
While designing a training course, the relationship between this module and the others
would be maintained by keeping them close together in the syllabus and placing them in a
logical sequence. The actual selection of the topics and the depth of training would, of
course, depend on the training needs of the participants, i.e. their knowledge level and skills
performance upon the start of the course.
No. Module title Code Objectives
1. Basic water quality concepts WQ - 01 • Discuss the common water quality
parameters
• List important water quality issues
2. Basic chemistry concepts WQ - 02 • Convert units from one to another
• Discuss the basic concepts of
quantitative chemistry
• Report analytical results with the
correct number of significant digits.
3. Basic ecology concepts WQ - 26 • Explain how energy flows through an
aquatic ecosystem
• Explain how nutrients are cycled in the
environment
• Explain the causes and problems of
eutrophication
4. Surface water quality
planning concepts
WQ - 27 • Understand principles of WQ
monitoring and assessment
• Know of simple data analysis methods
5. Use of ion selective probes WQ - 33 • Precautions required in use of ion
selective electrodes
6. Absorption spectroscopy WQ - 34 • Understand theory and applications of
absorption spectroscopy
7. How to measure Oxidised
Nitrogen: Cd reduction
method and UV
Spectrophotometric methods
WQ - 37 • Measure oxidised nitrogen by Cd-
reduction and UV spectrophotometric
methods
• Appreciate limitations of the UV
method
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 3
2. Module profile
Title : How to measure Ammonia and Organic Nitrogen: Kjeldahl
Method
Target group : HIS function(s): Q2, Q3, Q5, Q6
Duration : 1 Theoretical session of 30 min., plus
1 Practical Laboratory session of 120 min, plus
1 Report writing session of 30 min.
Objectives : After the training the participants will be able to:
• Understand the relevance of nitrogen to water quality
• Know how to make analysis of ammonia and organic nitrogen
by Kjeldahl Method
Key concepts : • Kjeldahl titrimetric method
Training methods : Lecture, Laboratory Analytical Exercise, Report preparation
Training tools
required
: Board, flipchart, OHS, Complete Laboratory Facilities for Ammonia
and Organic Nitrogen Analysis
Handouts : As provided in this module,
Including SAP for Analysis of Nitrogen, Ammonia and Organic
Further reading
and references
: • Chemistry for environmental engineers - C. N. Sawyer, P. L.
McCarty & G. F. Parkin, McGraw - Hill, Inc., 1994
• Standard methods for the examination of water and
wastewaters, AWWA, 19th
edition, 1995
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 4
3. Session plan
No Activities Time Tools
1 Preparations
• Reagents and glassware for the test
• Collect samples A, B and C as described in the text
2 Introduction:
• Introduce the session and the subject of ammonia and
organic nitrogen
• Discuss the forms of ammonia found in water
• Discuss the problems of ammonia and organic nitrogen
in water and why it is a useful parameter
10 min OHS
3 Ammonia & Organic Nitrogen Methods
• Describe the basis of the Kjeldahl method in terms of its
chemistry.
• Refer to SAP and allow time to read it.
20 min OHS, SAP
4 Practical Session:
• Divide the class in working group of 3-4 persons.
• Allow participants to conduct analysis according to SAP.
120 min Laboratory
5 Report Writing and wrap up
• Allow participants to complete their reports
• Discuss results and discrepancies
30 min Board
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 5
4. Overhead/flipchart master
OHS format guidelines
Type of text Style Setting
Headings: OHS-Title Arial 30-36, with bottom border line (not:
underline)
Text: OHS-lev1
OHS-lev2
Arial 24-26, maximum two levels
Case: Sentence case. Avoid full text in UPPERCASE.
Italics: Use occasionally and in a consistent way
Listings: OHS-lev1
OHS-lev1-Numbered
Big bullets.
Numbers for definite series of steps. Avoid
roman numbers and letters.
Colours: None, as these get lost in photocopying and
some colours do not reproduce at all.
Formulas/Equat
ions
OHS-Equation Use of a table will ease horizontal alignment
over more lines (columns)
Use equation editor for advanced formatting
only
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 6
Ammonia and Organic Nitrogen
• Nitrogen cycle
- atmospheric fixation precesses, chemical and biochemical
- conversion of ammonia nitrogen into organic nitrogen
- wastes: organic and ammonia nitrogen
- nitrification and denitrification
• Ammonia and organic nitrogen in water bodies is an important
parameter of their quality.
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 7
Nitrogen cycle
• Fixation of atmospheric nitrogen, N2
- electrical discharge, chemical production, biological fixation
• NO3 plants animals death & wastes
• Waste org. N decomposers NH3
• NH3 nitrifiers NO2 & NO3
• NO2 & NO3 denitrifiers N2
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 8
Nitrogen and Water quality (1)
• Water which has been polluted by organic wastes contains:
- ammonia nitrogen (free gas NH3 or ionic specie NH4
+
)
- organic nitrogen (proteins, urea)
• Both forms influence water quality
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 9
Nitrogen and water quality (2)
• Ammonia in water can be problematic in two ways:
- The free (un-ionised) form (NH3) is toxic to fish
- Nitrification of ammonia removes dissolved oxygen from the
water:
NH3 + 2O2 HNO3 + H2O
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 10
Nitrogen and Water Quality (3)
• Organic nitrogen compounds are:
- Excreted by animals or released when animals and plants die
- Present in sewage effluents
• Bacterial decomposition of organic wastes produces ammonia
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 11
Kjeldahl Method (1)
• Ammonia by distillation followed by titration of distillate with
sulphuric acid
• Organic nitrogen, on the residue from the above, by
conversion to ammonia followed by determination as above
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 12
Kjeldahl Method (2)
• Ammonia
- conversion to gaseous from
NH4
+
+ OH-
NH3 + H2O, pH = 9.5
- distillation and absorption
NH3 + H3BO3 NH4
+
+ H2BO3
-
- titration
H2BO3
-
+ H+
H3BO3
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 13
Kjeldahl Method (3)
• Organic nitrogen
- Acid digestion
Organic nitrogen + H2SO4 (NH4)2SO4 + CO2 + H2O
- K2SO4 & CuSO4 added to raise digestion temperature to 370o
C
- Digestion is completed when digesting liquor clarifies with
release of fumes.
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 14
Experiment
• Aim
- To determine the concentration of ammonia and organic
nitrogen using Kjeldahl method
Sample Source Expected concentration, mg/L
NH3 – N Organic - N
A Surface water 0 – 5 1 – 10
B Polluted drain 10 – 20 10 – 50
C Distilled water 0 0
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 15
Report
• the aim of the investigation
• the results you have produced
• the results compared to original predictions and the reasons
for any differences
• possible reasons for presence of nitrogen in blank, if any
• what the ammonia and organic nitrogen results mean in terms
of water quality
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 16
5. Evaluation sheets
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 17
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 18
6. Handout
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 19
Ammonia and Organic Nitrogen
• Nitrogen cycle
- atmospheric fixation precesses, chemical and biochemical
- conversion of ammonia nitrogen into organic nitrogen
- wastes: organic and ammonia nitrogen
- nitrification and denitrification
• Ammonia and organic nitrogen in water bodies is an important parameter of their
quality.
Nitrogen cycle
• Fixation of atmospheric nitrogen, N2
- electrical discharge, chemical production, biological fixation
• NO3 plants animals death & wastes
• Waste org. N decomposers NH3
• NH3 nitrifiers NO2 & NO3
• NO2 & NO3 denitrifiers N2
Nitrogen and Water quality (1)
• Water which has been polluted by organic wastes contains:
- ammonia nitrogen (free gas NH3 or ionic specie NH4
+
)
- organic nitrogen (proteins, urea)
• Both forms influence water quality
Nitrogen and water quality (2)
• Ammonia in water can be problematic in two ways:
- The free (un-ionised) form (NH3) is toxic to fish
- Nitrification of ammonia removes dissolved oxygen from the water:
NH3 + 2O2 HNO3 + H2O
Nitrogen and Water Quality (3)
• Organic nitrogen compounds are:
- Excreted by animals or released when animals and plants die
- Present in sewage effluents
• Bacterial decomposition of organic wastes produces ammonia
Kjeldahl Method (1)
• Ammonia by distillation followed by titration of distillate with sulphuric acid
• Organic nitrogen, on the residue from the above, by conversion to ammonia
followed by determination as above
Kjeldahl Method (2)
• Ammonia
- conversion to gaseous from
NH4
+
+ OH-
NH3 + H2O, pH = 9.5
- distillation and absorption
NH3 + H3BO3 NH4
+
+ H2BO3
-
- titration
H2BO3
-
+ H+
H3BO3
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 20
Kjeldahl Method (3)
• Organic nitrogen
- Acid digestion
Organic nitrogen + H2SO4 (NH4)2SO4 + CO2 + H2O
- K2SO4 & CuSO4 added to raise digestion temperature to 370o
C
- Digestion is completed when digesting liquor clarifies with release of
fumes.
Experiment
• Aim
- To determine the concentration of ammonia and organic nitrogen using
Kjeldahl method
Sample Source Expected concentration, mg/L
NH3 – N Organic - N
A Surface water 0 – 5 1 – 10
B Polluted drain 10 – 20 10 – 50
C Distilled water 0 0
Report
• the aim of the investigation
• the results you have produced
• the results compared to original predictions and the reasons for any differences
• possible reasons for presence of nitrogen in blank, if any
• what the ammonia and organic nitrogen results mean in terms of water quality
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 21
Add copy of Main text in chapter 8, for all participants.
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 22
7. Additional handout
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 23
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 24
8. Main text
Contents
1. Introduction 1
2. Kjeldahl Method of analysis 2
3. Experiment 2
SAP for Nitrogen, Ammonia (1.14) 4
SAP for Nitrogen, Organic (1.15) 6
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 1
How to measure Ammonia and Organic Nitrogen: Kjeldahl Method
1. Introduction
An understanding of the chemistry and biochemistry of nitrogen is a very important factor in
the management of water quality. As an aid to this understanding, Figure 1 shows a
simplified representation of the nitrogen cycle in nature. Many of the interactions shown in
the diagram also apply in the aquatic environment as discussed below.
Simplified Representation of the Nitrogen (N) Cycle in Nature
Nitrate,
NO3
-
Ammonia,
NH3
Nitrite,
NO2
-
Organic N
(plants, micro-
organisms)
Organic N
(animals)
Sun
Animals consume plants
and micro-organisms
Bacterial Oxidation
(nitrification)
Bacterial Oxidation
(nitrification)
Bacterial decomposition
of waste products
Death and bacterial
decomposition
taken from: ‘Water Quality’, Tchnobanoglous G and Schroeder E D, Addison-Wesley Publishing Company, 1987, p 183.
Atmosphere,
N2
Electrical
Discharge
Nitrogen
Fixation
Bacterial Reduction
(denitrification)
Chemical
Production
Bacterial
Reduction
Figure 1: Simplified Representation of the Nitrogen Cycle in Nature
Water which has been subject to pollution from waste materials, including sewage, often
contains nitrogenous organic compounds and ammonia. Ammonia is also produced when
compounds, such as, proteins, urea, etc., are decomposed through microbial action.
Ammonia in water can be problematic in two ways. Firstly, the free (un-ionised) form of the
gas is toxic to fish in reasonably low concentrations (approximately 0.2mg/L can cause death
in some species). Secondly, as can be seen from Figure 1, ammonia in the aquatic
environment is normally oxidised by bacteria to nitrite (NO2
-
) and then nitrate (NO3
-
) in a
process known as ‘nitrification’. This process consumes dissolved oxygen in the water
which can lead to distress or death for aquatic life if sufficient oxygen is lost. In addition,
nitrate is an important factor in the eutrophication of surface waters which can lead to further
water quality problems such as explosive algal growth, low dissolved oxygen and fish
deaths. Further, when present in drinking water, nitrate has been associated with
methaemoglobinaemia (blue baby disease) in human infants.
This experiment deals with measurement of nitrogen existing as free or ionised ammonia
and that occurring in combination with organic compounds. Waters that contain mostly
organic and ammonia nitrogen are considered to have been recently polluted and therefore
potentially dangerous from public health view point.
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 2
2. Kjeldahl Method of analysis
This method, when used to determine ammonia and organic nitrogen, consists of two steps
as follows:
• distillation of the ammonia from the sample into a solution of boric acid and titration of
the ammonia against a standard sulphuric acid solution
• after the ammonia has been distilled from the sample, the residue is used to determine
organic nitrogen. Nitrogen contained in many organic species is converted to
ammonium sulphate when heated in the presence of sulphuric acid. Potassium sulphate
and copper sulphate catalysts are added to raise the boiling point of sulphuric acid to
370o
C. The organic matter is oxidised to carbon dioxide. Once the ammonia from
organic nitrogen has been released, it is distilled and determined as above.
In both cases, before distillation the pH of the sample or digested liquor is raised to pH of 9.5
to encourage formation of ammonia which can be distilled with steam:
NH4
+
+ OH-
NH3 + H2O
The distilled ammonia is absorbed in boric acid:
NH3 + H3BO3 NH4
+
+ H2BO3
-
The ammonia which reacts with Boric Acid can be determined by back titrating with a strong
standard acid
H2BO3
-
+ H+
H3BO3
3. Experiment
Aim
a. To determine the concentration of ammonia and organic nitrogen in water samples by
Kjeldahl method.
Method
a. Read the SAP for determination of ammonia nitrogen by distillation titrimetric method
and organic nitrogen by Kjeldahl method.
b. Collect samples for analysis. The source of samples and expected analyte
concentrations are given below.
Sample Source Expected concentration, mg/L
NH3 – N Organic - N
A Surface water 0 – 5 1 – 10
B Polluted drain 10 – 20 10 – 50
C Distilled water 0 0
c. Choose appropriate sample aliquots and start with the determination of ammonia
nitrogen.
d. After ammonia distillation is complete and the residue in the boiling (Kjeldahl) flask has
cooled down start digestion for the determination of organic nitrogen.
e. While the digestion is proceeding, titrate the distillate obtained in ‘d’ above.
f. Complete digestion and start distillation for organic nitrogen and titrate for organic
nitrogen.
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 3
Observations & calculations
a) Calculate the concentration of the two forms of nitrogen and fill in the following table.
Use the values obtained for the blank to correct the values for the samples A and B
Sample Nitrogen, mg/L
NH3 – N Organic - N
A
B
C
Report
When writing your report the following aspects should be addressed:
• the aim of the investigation
• the results that you have produced
• whether the results were as originally predicted and, if they were not, the reasons
why they deviated
• possible reasons for the presence of nitrogen in the blank, if any
• the ammonia and organic nitrogen concentration of the samples and what this
could mean in terms of water quality
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 4
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 5
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 6
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 7
HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 8

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Download-manuals-water quality-wq-training-38measurementofammoniaandorganicnitrogen

  • 1. World Bank & Government of The Netherlands funded Training module # WQ - 38 How to measure Ammonia and Organic Nitrogen: Kjeldahl Method New Delhi, March 2000 CSMRS Building, 4th Floor, Olof Palme Marg, Hauz Khas, New Delhi – 11 00 16 India Tel: 68 61 681 / 84 Fax: (+ 91 11) 68 61 685 E-Mail: dhvdelft@del2.vsnl.net.in DHV Consultants BV & DELFT HYDRAULICS with HALCROW, TAHAL, CES, ORG & JPS
  • 2. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 1 Table of contents Page 1 Module context 2 2 Module profile 3 3 Session plan 4 4 Overhead/flipchart master 5 5 Evaluation sheets 16 6 Handout 18 7 Additional handout 22 8 Main text 24
  • 3. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 2 1. Module context This module deals with the significance of nitrogen to water quality and Kjeldahl method for measuring ammonia and organic nitrogen. Modules in which prior training is required to complete this module successfully and other available, related modules in this category are listed in the table below. While designing a training course, the relationship between this module and the others would be maintained by keeping them close together in the syllabus and placing them in a logical sequence. The actual selection of the topics and the depth of training would, of course, depend on the training needs of the participants, i.e. their knowledge level and skills performance upon the start of the course. No. Module title Code Objectives 1. Basic water quality concepts WQ - 01 • Discuss the common water quality parameters • List important water quality issues 2. Basic chemistry concepts WQ - 02 • Convert units from one to another • Discuss the basic concepts of quantitative chemistry • Report analytical results with the correct number of significant digits. 3. Basic ecology concepts WQ - 26 • Explain how energy flows through an aquatic ecosystem • Explain how nutrients are cycled in the environment • Explain the causes and problems of eutrophication 4. Surface water quality planning concepts WQ - 27 • Understand principles of WQ monitoring and assessment • Know of simple data analysis methods 5. Use of ion selective probes WQ - 33 • Precautions required in use of ion selective electrodes 6. Absorption spectroscopy WQ - 34 • Understand theory and applications of absorption spectroscopy 7. How to measure Oxidised Nitrogen: Cd reduction method and UV Spectrophotometric methods WQ - 37 • Measure oxidised nitrogen by Cd- reduction and UV spectrophotometric methods • Appreciate limitations of the UV method
  • 4. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 3 2. Module profile Title : How to measure Ammonia and Organic Nitrogen: Kjeldahl Method Target group : HIS function(s): Q2, Q3, Q5, Q6 Duration : 1 Theoretical session of 30 min., plus 1 Practical Laboratory session of 120 min, plus 1 Report writing session of 30 min. Objectives : After the training the participants will be able to: • Understand the relevance of nitrogen to water quality • Know how to make analysis of ammonia and organic nitrogen by Kjeldahl Method Key concepts : • Kjeldahl titrimetric method Training methods : Lecture, Laboratory Analytical Exercise, Report preparation Training tools required : Board, flipchart, OHS, Complete Laboratory Facilities for Ammonia and Organic Nitrogen Analysis Handouts : As provided in this module, Including SAP for Analysis of Nitrogen, Ammonia and Organic Further reading and references : • Chemistry for environmental engineers - C. N. Sawyer, P. L. McCarty & G. F. Parkin, McGraw - Hill, Inc., 1994 • Standard methods for the examination of water and wastewaters, AWWA, 19th edition, 1995
  • 5. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 4 3. Session plan No Activities Time Tools 1 Preparations • Reagents and glassware for the test • Collect samples A, B and C as described in the text 2 Introduction: • Introduce the session and the subject of ammonia and organic nitrogen • Discuss the forms of ammonia found in water • Discuss the problems of ammonia and organic nitrogen in water and why it is a useful parameter 10 min OHS 3 Ammonia & Organic Nitrogen Methods • Describe the basis of the Kjeldahl method in terms of its chemistry. • Refer to SAP and allow time to read it. 20 min OHS, SAP 4 Practical Session: • Divide the class in working group of 3-4 persons. • Allow participants to conduct analysis according to SAP. 120 min Laboratory 5 Report Writing and wrap up • Allow participants to complete their reports • Discuss results and discrepancies 30 min Board
  • 6. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 5 4. Overhead/flipchart master OHS format guidelines Type of text Style Setting Headings: OHS-Title Arial 30-36, with bottom border line (not: underline) Text: OHS-lev1 OHS-lev2 Arial 24-26, maximum two levels Case: Sentence case. Avoid full text in UPPERCASE. Italics: Use occasionally and in a consistent way Listings: OHS-lev1 OHS-lev1-Numbered Big bullets. Numbers for definite series of steps. Avoid roman numbers and letters. Colours: None, as these get lost in photocopying and some colours do not reproduce at all. Formulas/Equat ions OHS-Equation Use of a table will ease horizontal alignment over more lines (columns) Use equation editor for advanced formatting only
  • 7. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 6 Ammonia and Organic Nitrogen • Nitrogen cycle - atmospheric fixation precesses, chemical and biochemical - conversion of ammonia nitrogen into organic nitrogen - wastes: organic and ammonia nitrogen - nitrification and denitrification • Ammonia and organic nitrogen in water bodies is an important parameter of their quality.
  • 8. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 7 Nitrogen cycle • Fixation of atmospheric nitrogen, N2 - electrical discharge, chemical production, biological fixation • NO3 plants animals death & wastes • Waste org. N decomposers NH3 • NH3 nitrifiers NO2 & NO3 • NO2 & NO3 denitrifiers N2
  • 9. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 8 Nitrogen and Water quality (1) • Water which has been polluted by organic wastes contains: - ammonia nitrogen (free gas NH3 or ionic specie NH4 + ) - organic nitrogen (proteins, urea) • Both forms influence water quality
  • 10. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 9 Nitrogen and water quality (2) • Ammonia in water can be problematic in two ways: - The free (un-ionised) form (NH3) is toxic to fish - Nitrification of ammonia removes dissolved oxygen from the water: NH3 + 2O2 HNO3 + H2O
  • 11. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 10 Nitrogen and Water Quality (3) • Organic nitrogen compounds are: - Excreted by animals or released when animals and plants die - Present in sewage effluents • Bacterial decomposition of organic wastes produces ammonia
  • 12. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 11 Kjeldahl Method (1) • Ammonia by distillation followed by titration of distillate with sulphuric acid • Organic nitrogen, on the residue from the above, by conversion to ammonia followed by determination as above
  • 13. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 12 Kjeldahl Method (2) • Ammonia - conversion to gaseous from NH4 + + OH- NH3 + H2O, pH = 9.5 - distillation and absorption NH3 + H3BO3 NH4 + + H2BO3 - - titration H2BO3 - + H+ H3BO3
  • 14. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 13 Kjeldahl Method (3) • Organic nitrogen - Acid digestion Organic nitrogen + H2SO4 (NH4)2SO4 + CO2 + H2O - K2SO4 & CuSO4 added to raise digestion temperature to 370o C - Digestion is completed when digesting liquor clarifies with release of fumes.
  • 15. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 14 Experiment • Aim - To determine the concentration of ammonia and organic nitrogen using Kjeldahl method Sample Source Expected concentration, mg/L NH3 – N Organic - N A Surface water 0 – 5 1 – 10 B Polluted drain 10 – 20 10 – 50 C Distilled water 0 0
  • 16. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 15 Report • the aim of the investigation • the results you have produced • the results compared to original predictions and the reasons for any differences • possible reasons for presence of nitrogen in blank, if any • what the ammonia and organic nitrogen results mean in terms of water quality
  • 17. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 16 5. Evaluation sheets
  • 18. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 17
  • 19. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 18 6. Handout
  • 20. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 19 Ammonia and Organic Nitrogen • Nitrogen cycle - atmospheric fixation precesses, chemical and biochemical - conversion of ammonia nitrogen into organic nitrogen - wastes: organic and ammonia nitrogen - nitrification and denitrification • Ammonia and organic nitrogen in water bodies is an important parameter of their quality. Nitrogen cycle • Fixation of atmospheric nitrogen, N2 - electrical discharge, chemical production, biological fixation • NO3 plants animals death & wastes • Waste org. N decomposers NH3 • NH3 nitrifiers NO2 & NO3 • NO2 & NO3 denitrifiers N2 Nitrogen and Water quality (1) • Water which has been polluted by organic wastes contains: - ammonia nitrogen (free gas NH3 or ionic specie NH4 + ) - organic nitrogen (proteins, urea) • Both forms influence water quality Nitrogen and water quality (2) • Ammonia in water can be problematic in two ways: - The free (un-ionised) form (NH3) is toxic to fish - Nitrification of ammonia removes dissolved oxygen from the water: NH3 + 2O2 HNO3 + H2O Nitrogen and Water Quality (3) • Organic nitrogen compounds are: - Excreted by animals or released when animals and plants die - Present in sewage effluents • Bacterial decomposition of organic wastes produces ammonia Kjeldahl Method (1) • Ammonia by distillation followed by titration of distillate with sulphuric acid • Organic nitrogen, on the residue from the above, by conversion to ammonia followed by determination as above Kjeldahl Method (2) • Ammonia - conversion to gaseous from NH4 + + OH- NH3 + H2O, pH = 9.5 - distillation and absorption NH3 + H3BO3 NH4 + + H2BO3 - - titration H2BO3 - + H+ H3BO3
  • 21. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 20 Kjeldahl Method (3) • Organic nitrogen - Acid digestion Organic nitrogen + H2SO4 (NH4)2SO4 + CO2 + H2O - K2SO4 & CuSO4 added to raise digestion temperature to 370o C - Digestion is completed when digesting liquor clarifies with release of fumes. Experiment • Aim - To determine the concentration of ammonia and organic nitrogen using Kjeldahl method Sample Source Expected concentration, mg/L NH3 – N Organic - N A Surface water 0 – 5 1 – 10 B Polluted drain 10 – 20 10 – 50 C Distilled water 0 0 Report • the aim of the investigation • the results you have produced • the results compared to original predictions and the reasons for any differences • possible reasons for presence of nitrogen in blank, if any • what the ammonia and organic nitrogen results mean in terms of water quality
  • 22. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 21 Add copy of Main text in chapter 8, for all participants.
  • 23. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 22 7. Additional handout
  • 24. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 23
  • 25. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 24 8. Main text Contents 1. Introduction 1 2. Kjeldahl Method of analysis 2 3. Experiment 2 SAP for Nitrogen, Ammonia (1.14) 4 SAP for Nitrogen, Organic (1.15) 6
  • 26. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 1 How to measure Ammonia and Organic Nitrogen: Kjeldahl Method 1. Introduction An understanding of the chemistry and biochemistry of nitrogen is a very important factor in the management of water quality. As an aid to this understanding, Figure 1 shows a simplified representation of the nitrogen cycle in nature. Many of the interactions shown in the diagram also apply in the aquatic environment as discussed below. Simplified Representation of the Nitrogen (N) Cycle in Nature Nitrate, NO3 - Ammonia, NH3 Nitrite, NO2 - Organic N (plants, micro- organisms) Organic N (animals) Sun Animals consume plants and micro-organisms Bacterial Oxidation (nitrification) Bacterial Oxidation (nitrification) Bacterial decomposition of waste products Death and bacterial decomposition taken from: ‘Water Quality’, Tchnobanoglous G and Schroeder E D, Addison-Wesley Publishing Company, 1987, p 183. Atmosphere, N2 Electrical Discharge Nitrogen Fixation Bacterial Reduction (denitrification) Chemical Production Bacterial Reduction Figure 1: Simplified Representation of the Nitrogen Cycle in Nature Water which has been subject to pollution from waste materials, including sewage, often contains nitrogenous organic compounds and ammonia. Ammonia is also produced when compounds, such as, proteins, urea, etc., are decomposed through microbial action. Ammonia in water can be problematic in two ways. Firstly, the free (un-ionised) form of the gas is toxic to fish in reasonably low concentrations (approximately 0.2mg/L can cause death in some species). Secondly, as can be seen from Figure 1, ammonia in the aquatic environment is normally oxidised by bacteria to nitrite (NO2 - ) and then nitrate (NO3 - ) in a process known as ‘nitrification’. This process consumes dissolved oxygen in the water which can lead to distress or death for aquatic life if sufficient oxygen is lost. In addition, nitrate is an important factor in the eutrophication of surface waters which can lead to further water quality problems such as explosive algal growth, low dissolved oxygen and fish deaths. Further, when present in drinking water, nitrate has been associated with methaemoglobinaemia (blue baby disease) in human infants. This experiment deals with measurement of nitrogen existing as free or ionised ammonia and that occurring in combination with organic compounds. Waters that contain mostly organic and ammonia nitrogen are considered to have been recently polluted and therefore potentially dangerous from public health view point.
  • 27. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 2 2. Kjeldahl Method of analysis This method, when used to determine ammonia and organic nitrogen, consists of two steps as follows: • distillation of the ammonia from the sample into a solution of boric acid and titration of the ammonia against a standard sulphuric acid solution • after the ammonia has been distilled from the sample, the residue is used to determine organic nitrogen. Nitrogen contained in many organic species is converted to ammonium sulphate when heated in the presence of sulphuric acid. Potassium sulphate and copper sulphate catalysts are added to raise the boiling point of sulphuric acid to 370o C. The organic matter is oxidised to carbon dioxide. Once the ammonia from organic nitrogen has been released, it is distilled and determined as above. In both cases, before distillation the pH of the sample or digested liquor is raised to pH of 9.5 to encourage formation of ammonia which can be distilled with steam: NH4 + + OH- NH3 + H2O The distilled ammonia is absorbed in boric acid: NH3 + H3BO3 NH4 + + H2BO3 - The ammonia which reacts with Boric Acid can be determined by back titrating with a strong standard acid H2BO3 - + H+ H3BO3 3. Experiment Aim a. To determine the concentration of ammonia and organic nitrogen in water samples by Kjeldahl method. Method a. Read the SAP for determination of ammonia nitrogen by distillation titrimetric method and organic nitrogen by Kjeldahl method. b. Collect samples for analysis. The source of samples and expected analyte concentrations are given below. Sample Source Expected concentration, mg/L NH3 – N Organic - N A Surface water 0 – 5 1 – 10 B Polluted drain 10 – 20 10 – 50 C Distilled water 0 0 c. Choose appropriate sample aliquots and start with the determination of ammonia nitrogen. d. After ammonia distillation is complete and the residue in the boiling (Kjeldahl) flask has cooled down start digestion for the determination of organic nitrogen. e. While the digestion is proceeding, titrate the distillate obtained in ‘d’ above. f. Complete digestion and start distillation for organic nitrogen and titrate for organic nitrogen.
  • 28. HP Training Module File: “ 38 Measurement of Ammonia and Organic Nitrogen.doc” Version March 2000 Page 3 Observations & calculations a) Calculate the concentration of the two forms of nitrogen and fill in the following table. Use the values obtained for the blank to correct the values for the samples A and B Sample Nitrogen, mg/L NH3 – N Organic - N A B C Report When writing your report the following aspects should be addressed: • the aim of the investigation • the results that you have produced • whether the results were as originally predicted and, if they were not, the reasons why they deviated • possible reasons for the presence of nitrogen in the blank, if any • the ammonia and organic nitrogen concentration of the samples and what this could mean in terms of water quality
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