Environmental Engineering and Microbiology Lab – II

M.Tech., YEAR: I Year SEM: I ACADEMIC YEAR: 2019-2020

FACULTY MANUAL
DEPARTMENT OF CIVIL ENGINEERING

Prepared by
P.Srinivasa Rao
 DHANEKULA INSTITUTE OF ENGINERING AND TECHNOLOGY
(An ISO 9001:2008 certified & AICTE approved institution affiliated to JNTUK), Ganguru-
521139, Andhra Pradesh

INDEX

Environmental Engineering and Microbiology Lab – II

Page No
S. No. NAME OF THE EXPERIMENT
From To
Ambient Air Quality Monitoring: Concentration of particulate
1 matter present in air (PM 10 & PM 2.5), SO2 and NOx by using
High Volume Air Sampler.
Stack Monitoring: Concentration of particulate matter present in
2 air (PM 10 & PM 2.5), SO2 and NOx and other parameters.
3 To determine the dissolved oxygen and BOD present in a given
sample
To determine the chemical oxygen demand present in waste water
4 sample
Type- II settling of particle sedimentation
5
Break point chlorination test
6
Media preparation , Inoculation and Plate count test
7

Most Probable Number (MPN) test

8
Membrane filtration techniques
9
Noise Isopleths in Institution or Industry
10

11 TCLP – Leachate from Landfills.

12 Micrometeorology –Wind Direction, Wind speed, Humidity

Temperature, Rainfall.
13 Automobile emission test.
 DHANEKULA INSTITUTE OF ENGINERING AND TECHNOLOGY
DEPARTMENT OF CIVIL ENGINEERING
Environmental Engineering and Microbiology Lab – II

JNTUK Syllabus
List of Experiments
1. Ambient Air Quality Monitoring: Concentration of particulate
matter present in air (PM 10 & PM 2.5), SO2 and NOx by using
High Volume Air Sampler.
2. Stack Monitoring: Concentration of particulate matter present in
air (PM 10 & PM 2.5), SO2 and NOx and other parameters.
3. To determine the dissolved oxygen and BOD present in a given
sample
4. To determine the chemical oxygen demand present in waste water
sample
5. Type- II settling of particle sedimentation

6. Break point chlorination test

7. Media preparation , Inoculation and Plate count test

8. Most Probable Number (MPN) test

9. Membrane filtration techniques

10. Noise Isopleths in Institution or Industry

11. TCLP – Leachate from Landfills.

12. Micrometeorology –Wind Direction, Wind speed, Humidity

Temperature, Rainfall.
13. Automobile emission test.
 DHANEKULA INSTITUTE OF ENGINERING AND TECHNOLOGY
DEPARTMENT OF CIVIL ENGINEERING
ENVIRONMENTAL ENGINEERING LABORATORY

List of conducted experiments

1. Ambient Air Quality Monitoring: Concentration of particulate
matter present in air (PM 10 & PM 2.5), SO2 and NOx by using
High Volume Air Sampler.
2. Stack Monitoring: Concentration of particulate matter present in
air (PM 10 & PM 2.5), SO2 and NOx and other parameters.
3. To determine the dissolved oxygen and BOD present in a given
sample
4. To determine the chemical oxygen demand present in waste water
sample
5. Type- II settling of particle sedimentation

6. Break point chlorination test

7. Media preparation , Inoculation and Plate count test

8. Most Probable Number (MPN) test

9. Membrane filtration techniques

10. Noise Isopleths in Institution or Industry

11. TCLP – Leachate from Landfills.

12. Micrometeorology –Wind Direction, Wind speed, Humidity

Temperature, Rainfall.
13. Automobile emission test.
Year/sem: I Year/I Sem Academic year: 2019-2020

Vision, Mission of the Institute & Department

Institute Vision
 Pioneering Professional Education through Quality

Institute Mission
 Providing Quality Education through state-of-art infrastructure, laboratories and
committed staff.
 Moulding Students as proficient, competent and socially responsible engineering
personnel with ingenious intellect.
 Involving faculty members and students in research and development works for
betterment of society.

Department Vision

 To become a pioneering center of learning and research in Civil Engineering

Department Mission

 Providing high quality education in an atmosphere of innovation and critical thinking.

 An integrated development of Civil Engineering Professionals Possessing Technical &
Managerial skills, Environmental, Ethical and Human values.
 Inculcating research and consultancy culture by involving faculty and students.
 Program Educational Objective (PEO)

PEO 1: Domain Knowledge

Graduates will have the fundamental knowledge of mathematics, science, economics and
computing and in-depth knowledge in Civil Engineering concepts through theoretical,
laboratory and project based experiences so as to design, develop and solve engineering
problems.

PEO 2: Employment & Higher Studies

Graduates will succeed in their chosen engineering careers with sustainable development in
their profession and have the ability to pursue advanced degrees in engineering and other
fields

PEO 3: Professional Citizenship

Graduates will have ability to organize and Communicate effectively in multidisciplinary

engineering projects through lifelong learning, practice ethics with a sense of social
responsibility.

Programme Outcomes (PO)

Engineering Graduates will be able to:
1. Engineering knowledge: Apply the knowledge of mathematics, science, engineering
fundamentals, and an engineering specialization to the solution of complex engineering
problems.

2. Problem analysis: Identify, formulate, review research literature, and analyze complex
engineering problems reaching substantiated conclusions using first principles of
mathematics, natural sciences, and engineering sciences.

3. Design/development of solutions: Design solutions for complex engineering problems

and design system components or processes that meet the specified needs with
appropriate consideration for the public health and safety, and the cultural, societal, and
environmental considerations.

4. Conduct investigations of complex problems: Use research-based knowledge and

research methods including design of experiments, analysis and interpretation of data,
 and synthesis of the information to provide valid conclusions.

5. Modern tool usage: Create, select, and apply appropriate techniques, resources, and
modern engineering and IT tools including prediction and modeling to complex
engineering activities with an understanding of the limitations.

6. The engineer and society: Apply reasoning informed by the contextual knowledge to
assess societal, health, safety, legal and cultural issues and the consequent
responsibilities relevant to the professional engineering practice.

7. Environment and sustainability: Understand the impact of the professional engineering

solutions in societal and environmental contexts, and demonstrate the knowledge of, and
need for sustainable development.

8. Ethics: Apply ethical principles and commit to professional ethics and responsibilities
and norm s of the engineering practice.
9. Individual and team work: Function effectively as an individual, and as a member or
leader in diverse teams, and in multidisciplinary settings.

10. Communication: Communicate effectively on complex engineering activities with the

engineering community and with society at large, such as, being able to comprehend and
write effective reports and design documentation, make effective presentations, and give
and receive clear instructions.
11. Project management and finance: Demonstrate knowledge and understanding of the
engineering and management principles and apply these to one’s own work, as a member
and leader in a team, to manage projects and in multidisciplinary environments.
12. Life-long learning: Recognize the need for, and have the preparation and ability to
engage in independent and life-long learning in the broadest context of technological
change.
Program Specific Outcomes (PSO)

1. PSO1: Knowledge of contemporary issues in the civil engineering industry to solve

societal issues.

2. PSO2: Qualify in competitive examinations for higher education and employment.

Course Objectives
The students will:

S.No Course Objectives (Cobs) COs

learn the laboratory practices to estimate the chemical
properties like pH, alkalinity, acidity, hardness, chlorides,
R16CObj 315.1 total, suspended and dissolved solids, D.O., B.O.D., COD and R16C315.1
residual chlorine present in the water and waste water
samples
learn the laboratory practices to estimate the physical
R16CObj 315.2 R16C315.2
properties like turbidity in the water and waste water samples

Course Outcomes

At the end of the Course/Subject, the students will be able to :

S.No Course Outcomes(COs) POS PSOs

Evaluate water quality based on chemical analysis of 1,2,7 1,2
R16C315.1
given samples
Evaluate water quality based on physical analysis of 1,2,7 1,2
R16C315.2
given samples

Course Outcomes vs POs Mapping

Course Outcome P01 P02 P03 P04 P05 P06 P07 P08 P09 P10 P11 P12

R16C315.1 2 - - 3 - 2 3 - - - - -
R16C315.2 2 - - 3 - 2 3 - - - - -
Total 4 - - 6 - 6 6 - - - - -
R16C315 2 - - 3 - 2 3 - - - - -

Course Outcomes vs PSOs Mapping

Courses Out Comes PSO1 PSO2

R16C315.1 2 3
R16C315.2 2 3
Total 4 6
R16C315 2 3
 DOs & DONTs

1. Conduct yourself in a responsible manner at all times in the laboratory.

2. Carefully follow all written and verbal instructions carefully.
3. If you do not understand a direction or part of a procedure, ASK YOUR TEACHER
BEFORE PROCEEDING WITH THE ACTIVITY.
4. Never work alone in the laboratory.
5. Do not touch any equipment, chemicals, or other materials in the laboratory area until
you are instructed to do so.
6. Do not eat food, drink beverages, or chew gum in the laboratory.
7. Do not use laboratory glassware as containers for food or beverages.
8. Read all procedures thoroughly before entering the laboratory.
9. Observe good housekeeping practices.
10. Work areas should be kept clean and tidy at all times.
11. Be alert and proceed with caution at all times in the laboratory.
12. Notify the instructor immediately of any unsafe conditions you observe.
13. Dispose of all chemical waste properly.
14. Never mix chemicals in sink drains. Labels and equipment instructions must be read
carefully before use.
15. Set up and use the equipment as directed by your laboratory instructor.
16. Keep hands away from face, eyes, mouth, and body while using chemicals or lab
equipment.
17. Experiments must be personally monitored at all times.
18. Know the locations and operating procedures of all safety equipment including: first aid
kit(s), and fire extinguisher.
19. Know where the fire alarm and the exits are located.
20. Do not panic in any situation.
21. Report any accident (spill, breakage, etc.) or injury (cut, burn, etc.) immediately, no
matter how trivial it seems.
22. If you or your lab partner is hurt, immediately (and loudly) yell out the teacher's name to
get the teacher's attention.
23. If a chemical should splash in your eye(s) or on your skin, immediately flush with
running water for at least 20 minutes.
24. All chemicals in the laboratory are to be considered dangerous.
25. Avoid handling chemicals with BARE HANDS.
26. When making an observation, keep at least 1 foot away from the specimen.
27. Do not taste, or smell any chemicals.
28. Check the label on all chemical bottles twice before removing any of the contents.
29. Take only as much chemical as you need. Never return unused chemicals to their original
container. Never remove label of any chemical bottle.
30. Never handle broken glass with your bare hands.
1. Use a brush and dustpan to clean up broken glass.
31. Place broken glass in the designated glass disposal container.
2. Examine glassware before each use. Never use chipped, cracked, or dirty glassware.
3. If you do not understand how to use a piece of equipment, ASK THE TEACHER FOR
HELP! Do not immerse hot glassware in cold water. The glassware may shatter.
32. Heated glassware remain very hot for a long time.
33. They should be set aside in a designated place to cool, and picked up with caution.
34. Use tongs or heat protective gloves if necessary.
35. Never look into a container that is being heated.
36. Do not place hot apparatus directly on the laboratory desk. Always use an insulated pad.
37. Allow plenty of time for hot apparatus to cool before touching it.
38. Any time chemicals, heat, or glassware are used, students will wear safety goggles.
39. NO EXCEPTIONS TO THIS RULE! Contact lenses may be not be worn in the
laboratory.
40. Dress properly during a laboratory activity.
41. Long hair, dangling jewellery, and loose or baggy clothing are a hazard in the laboratory.
42. Long hair must be tied back, and dangling jewellery and baggy clothing must be secured.
Shoes must completely cover the foot.
43. No sandals allowed on lab days. A lab coat or smock should be worn during laboratory
experiments

.
 JAWAHARLAL NEHRU TECHNOLOGICAL UNIVERSITY: KAKINADA KAKINADA – 533
003, Andhra Pradesh, India

ENVIRONMENTAL ENGINEERING AND MICROBIOLOGY LAB

EXP1
Objective
Ambient Air Quality Monitoring: Concentration of particulate matter present in air (PM 10 & PM
2.5), SO2 and NOx by using High Volume Air Sampler.

Particulate matter present in air (PM 10)

Aim: Sampling of Particulate matter (PM10) in ambient air and the determination of its
concentration.

Reference: IS 5182 (Part 23): 2006/CPCB Guidelines Volume-I

Method: - Gravimetric Method

Principle: Air with dust enters the sampler through circular inlet and passed through the impactor
where all particles size more than 10 microns are removed. Particles equal or less than 10 micron
passed through impactor. Design of impactor has been done for a cut off at 10 micron at flow rate 2.3
m3/hr. Particles greater than 10 microns hit on greased metallic surface and remains on the surface
while dust less than 10 microns follow trajectory and reach to pre weighted filter paper where it
accumulated. Accumulated dust on filter paper is obtained by weight difference while total volume of
air sampled is given by difference of final and initial DG reading. PM10 dust concentration calculated
simply by dividing weight of dust by volume of air passed through filters.

Instrument/Equipment: -
1. Analytical balance
2. Sampler : Combo PM10 and PM2.5 Sampler with size selective inlet for PM2.5 and
automatic volumetric flow control
3. Filter Jacket
4. Calibrated flow-measuring device to control the airflow at 16.67 l/min
5. High Volume sampler (HVS)
6. Whatman filter paper
7. Impingers

Absorbing Media: - Filter Media – A Glass fiber filters of 47 mm size.

Procedures:

1. Filter inspection: Inspect the filter for pin holes using a light table.
2. Loose particles should be removed with a soft brush. Apply the filter identification number or a
code to the filter if it is not a numbered.
3. Condition the filter in conditioning room maintained within 20-30° C and 40-50% relative
 humidity or in an airtight desiccator for 24 hours.
4. Take initial weight of the filter paper (Wi) before sampling.
5. Place the filter paper in the sampling system securely and tighten the screws of the bracket.
6. Note down the initial dry gas meter reading and time.
7. Start the sampler and adjust flow rate as per guidelines.
8. At the end stop the pump, note down final dry gas meter reading and time.
9. Remove filter cassette from the filter holder and store in filter carrier immediately and transfer
it to desiccators as early as possible.
10. Condition the filter after sampling in conditioning room maintained within 20 30° C and 40-
50% relative humidity or in an airtight desiccator for 24 hours.
11. Take final weight of the filter paper (Wf)

Calculation:

Concentration of PM10 in µg/m3 PM10 = (Wf-Wi) x 106/V

Where:
Wi: Initial mass of the conditioned filter before sample collection (gm)
Wf: Final mass of the conditioned filter after sample collection (gm)
106: Unit conversion factor for grams (gm) to micrograms (μg)
V : Volume of air sampled (m3)
[V= Final Dry Gas Meter Reading – Initial Dry Gas Meter Reading]
 Particulate matter present in air (PM 2.5)

Reference: CPCB Guidelines Volume-I

Method: - Gravimetric Method

Principle: Air with dust enters in the sampler through circular omni directional inlet reaches to
impactor where all particles having size more than 10 microns are removed and retained. Now air
having particles less than 10 microns further proceed and pass through WINS impactor which is well
shaped. A filter dipped in impaction oil is kept in the well where particles hit at a specific velocity
(maintained by top critical hole of WINS impactor). This results in separation of particles above 2.5
micron to 10 micron. Only particles having size 2.5 micron and below proceed further and
accumulated on PTFE membrane filter. Accumulated dust on filter is obtained by weight difference
while total volume of air sampled is given by the difference of final and initial DG readings. PM2.5
dust concentration can be calculated simply by dividing weight of dust by volume of air sampled. The
lower detection limit of PM2.5 dust is about 2μg/m3 which depends upon the accuracy of the digital
balance. A sensitive digital balance with minimum detection of 0.00001 g is basic need for
measurement of accumulated dust. If 6 digit balance (recommended by USEPA) is available one can
measure weight more accurately and thus PM2.5 values. Through filters

Instruments
1. Analytical balance
2. Sampler : Combo PM10 and PM2.5 Sampler with size selective inlet for PM2.5 and
automatic volumetric flow control
3. Filter Jacket
4. Calibrated flow-measuring device to control the airflow at 16.67 l/min
5. High Volume sampler (HVS)
6. Whatman filter paper
7. Impingers

Absorbing Media: - Filter Media – A Glass fibre filter of 47 mm size and supported filter paper 37 mm
size.

Procedures:

1. Filter inspection: Inspect the filter for pin holes using a light table.
2. Loose particles should be removed with a soft brush. Apply the filter identification number or a
code to the filter if it is not a numbered.
3. Condition the filter in conditioning room maintained within 20-30° C and 40-50% relative
humidity or in an airtight desiccator for 24 hours.
4. Take initial weight of the filter paper (Wi) before sampling.
5. Place the filter paper in the sampling system securely and tighten the screws of the bracket.
6. Note down the initial dry gas meter reading and time.
7. Start the sampler and adjust flow rate as per guidelines.
 8. At the end stop the pump, note down final dry gas meter reading and time.
9. Remove filter cassette from the filter holder and store in filter carrier immediately and transfer
it to desiccators as early as possible.
10. Condition the filter after sampling in conditioning room maintained within 20 30° C and 40-
50% relative humidity or in an airtight desiccator for 24 hours.
11. Take final weight of the filter paper (Wf)

Calculation:

Concentration of PM2.5 in µg/m3 PM2.5 = (Wf-Wi) x 106/V

Where:
Wi: Initial mass of the conditioned filter before sample collection (gm)
Wf: Final mass of the conditioned filter after sample collection (gm)
106: Unit conversion factor for grams (gm) to micrograms (μg)
V : Volume of air sampled (m3)
[V= Final Dry Gas Meter Reading – Initial Dry Gas Meter Reading]

SO2 (Sulphur Dioxide)

Aim: Measurement of Sulphur dioxide concentration in ambient air.

Reference: IS 5182 (Part 2): 2001/CPCB Guidelines Volume-I Method: -

Improved West and Gaeke method

Principle: Sulphur dioxide is absorbed from air in a solution of potassium tetra chloro mercurate
(TCM). A dichlorosulphitomercurate complex which resists oxidation by the oxygen in the air is
formed. This complex is stable to strong oxidants such as ozone and oxides of nitrogen and
therefore the absorber solution may be stored for some time prior to analysis. The complex is made
to react with pararosaliniline and methylsulphonic acid. The absorbance of the solution is
measured by means of a suitable spectrophotometer.

APPARATUS & GLASSWARES: -

 Gaseous Pollutant Sampler

 Glass Impingers
 Spectrophotometer
 Volumetric flasks
 Pipettes
 Burette

REAGENTS:

 Distilled Water
 Absorbing Reagent (Potassium Tetrachloromercurate–TCM-0.04M): Dissolve
10.86 gm mercuric chloride, 0.066 gm EDTA, and 6.0 gm potassium chloride in distilled water
and bring to the mark in a 1 liter volumetric flask. The pH of this reagent shall be
approximately 4.0 but it has been shown that there is no appreciable difference in collection
efficiency over the range of pH 5to 3. The absorbing reagent is normally stable for 6 months. If
a precipitate forms, discard the reagent after recovering the mercury.

 Sulphamic Acid (0.6%): Dissolve 0.6 gm of Sulphamic acid in 100 mL of distilled water. Prepare
fresh, when needed.
 Formaldehyde Solution (0.6%): Dilute 5 mL of formaldehyde solution (36 to 38 %) to 1 liter with
Distilled water. Prepare fresh, when needed.
 Stock Iodine Solution (0.1 N):Take 12.7 gm of iodine in a 250 mL beaker; add 40 gm of potassium
iodide and 25 mL of water. Stir to dissolve completely, then dilute to 1 liter with distilled water.

 Iodine Solution (0.01 N): Dilute 50 mL of stock solution to 500 mL with distilled water.
 Starch Indicator Solution: Triturate 0.4 gm of soluble starch and 0.002 gm of mercuric iodide
preservative with a little water and add the paste slowly to 200 mL boiling water. Continue boiling until
the solution clear, cool and transfer to a glass stoppered bottle.
 Stock Sodium Thiosulphate Solution (0.1 N):Take 25 gm of sodium thiosulphate pentahydrate in a
beaker, add 0.1 gm of sodium carbonate, and dissolve using boiled distilled water making the solution
up to final volume of 1 liter. Allow the solution to stand one day before standardizing

To standardize accurately weigh, to the nearest 0.1 mg, 1.5 gm of primary standard potassium
iodate dried at 180°C, dissolve and dilute to 500 mL in a volumetric flask. Take 50 mL of
iodate solution by pipette into a 500 mL iodine flask, add 2 g of potassium iodide and 10 mL
(1:10) hydrochloric acid and stopper the flask. After 5 min titrate with stock thiosulphate
solution to a pale yellow color. Add 5 mL starch indicator solution and continue the titration
until the blue color disappears.

 Sodium Thiosulphate Solution (0.01 N): Dilute 100 mL of the stock thiosulphate solution to 1 liter
with freshly boiled distilled water.
 Standardized sulphite Solution for preparation of working sulphate-TCM Solution: Dissolve 0.30
g of sodium meta bisulphite (NaHSO3) or 0.40 gm of sodium sulphite (Na2SO3) in 500 mL of freshly
boiled and cooled distilled water. Sulphite solution is unstable, it is therefore important to use water of
the highest purity to minimize this instability. This solution contains the equivalent of 320-400 μg/mL
of SO2.
The actual concentration of the sulphite solution is determined by adding excess iodine and
back titrating with standard sodium thiosulphate. To standardize pipette out 50 mL of the 0.01
N iodine solution into each of two 500 mL iodine flask A (blank) and B (sample). To flask A
add 25 mL of distilled water and into flask B measure 25 mL of sulphite solution. Stopper the
flask and allow reacting for 5 min. Titrate with standardized 0.01 N thiosulphate, to a pale
yellow color. Then add 5 mL of starch solution and continue the titration until the blue color
disappears.
 Working sulphite- TCM Solution: Take 2 mL of the standard sulphite solution into 100 mL
volumetric flask to dilute with 0.04 M TCM.
 Preparation of stock Para Rosaniline Solution: Dissolve 0.5 gm of para rosaniline chloride in 100
 mL distilled water. Keep it for 2 days and filter the solution. The filtrate solution is stable for 3 months,
if stored in refrigerator.
 Working Para Rosaniline Solution: Add 15 mL concentrated hydrochloric acid to 10 mL stock para
rosaniline solution and dilute to 250 mL with distilled water in a 250 mL volumetric flask. It may be
stored at room temperature in an amber colored bottle for one to two weeks, if stored in a refrigerator.

PROCEDURES:

Sampling:

 Assemble the sampling apparatus at the sampling site. Procedures are described for short-term (30
minutes, 1 hour, 4 hours long-term (24 hours) sampling.
 30 minutes, 1 hour, 4 hours Sampling: insert a midget impinger into the sampling system. Add 10 ml
of TCM solution to the impinger (30 ml TCM solution for 4 hours sampling)
 Collect sample at the rate of 1 l/min for 30 min, or 0.5 l/min for 1 hour and 4 hours using either a
rotameter.
 24 hours Sampling: Place 50 ml of TCM solution in a larger absorber and collect the sample at 0.2
l/min for 24 hour from midnight to midnight.
 Shield the absorbing reagent form direct sunlight during and after sampling by covering the impinger
with aluminium foil to prevent deterioration.
 Start sampling only after obtaining a flow rate. Ensure packing of ice around bubbler for low
temperature to improve absorption efficiency.
 Record the exact sampling time by recording initial time (ti) and final time (tf) of sampler.
 Measure and record the flow rate (fi) before the sampling and flow rate (ff) after the sampling.
 Recorded the atmospheric pressure and temperature.
 Remove the stopper the impinger. If the sample has to be stored for more then a day before analysis,
keep it at 5° C in a refrigerator; during hot weather, sampling is to not recommended unless it is
possible to refrigerate the samples.
 After sampling measure the volume of sample and transfer to a sample storage bottle.
 Mark identification number on sample storage bottle and transport to the laboratory for analysis.

Sample Preparation:

30-Minutes and 1-Hour Sample: Transfer the sample quantitavely to a 25 ml volumetric flask
using about 5 ml of distilled water for rinsing. Delay the analysis for 20 min to allow any ozone to
decompose.

24-Hour Samples: Dilute the entire sample to 50 ml with absorbing solution. Measure 5 ml of the
sample into a 25 ml volumetric flask by pipette for chemical analysis. Bring volume to 10 ml with
absorbing reagent. Delay analysis for 20 min to allow any ozone to decomposes.
Analysis:

 For each set of determinations prepare a reagent blank by adding 10 ml of

unexposed TCM solution to a 25 ml volumetric flask.
 Prepare a control solution by measuring 2 ml of working sulphite-TCM solution into
a 25 ml volumetric flask by pipette.
 To each flask containing sample or control solution or reagent blank, add 1 ml of
0.6% sulphamic acid and allow to react for 10 min to destroy the nitrite resulting
from oxides of nitrogen.
 Add 2 ml of 0.2% formaldehyde solution and 5 ml of pera rosaniline solution.
 Start a laboratory timer that has set for 30 min.
 Determine the absorbance of the sample, reagent blank, and control solution at 560
nm using cells with a 1 cm path length.

Calibration:

Preparation of Calibration Curve:

 Measure 0.5 ml, 1.0 ml, 1.5 ml, 2.0 ml, 2.5 ml, 3.0 ml, 3.5 ml and 4.0 ml of working
sulphite TCM solution in 25 ml volumetric flask.
 Add sufficient TCM solution to each flask to bring the volume to approximately 10
ml.
 Then add the remaining reagents as described in the procedure for analysis.
 A reagent blank with 10 ml absorbing solution is also prepared.
 Read the absorbance of each standard and reagent blank.

Standard Curve:

 Plot Plot a curve absorbance (Y axis) versus concentration (X axis). Draw a line ofbest
fit and determine the slope.
 The reciprocal of slope gives the calibration factor (CF).

CALCULATION:

SO2 Concentration in Air Sample: Calculate SO2 as µg per cubic meter of air as follows:
(A – Ao) × 103× B × D
SO2 (µg/m ) =
3

Vr

Where:
A : Sample absorbance, (µg)
Ao : Reagent blank absorbance, (µg)
103 : Conversion of factor to cubic meters
Vr : The sample corrected to 25 C and
760 mm Hg B : Calibration factor,
/absorbance unit
D : Dilution factor: for 30 min and 1 hr samples, D=1; for 24-hr samples, D=10

Conversion of µg/m3into ppm

SO2 (ppm) = (SO2µg/m3) ×3.82 × 10-4

 Nitrogen dioxide

Aim: Measurement of Nitrogen dioxide concentration in ambient air.

Reference: IS 5182 (Part 6): 2006/CPCB Guidelines Volume-I

Method: - Modified Jacob and Hochheiser Method

Principle: Ambient nitrogen dioxides are collected by bubbling air through a solution of
Sodium hydroxide and sodium arsenite. The concentration of nitrite ion (NO 2) produced
during sampling is determined colorimetrically by reacting the nitrite ion with
phosphoric acid, sulphanilamide and N-(1-naphtyl)-ethylenediamine di-hydrochloride
(NEDA) and measuring the absorbance of the highly colored azo-dye at 540 nm.

APPARATUS & GLASSWARES: -

 Gaseous Pollutant Sampler

 Glass Impingers
 Spectrophotometer
 Volumetric flasks
 Pipettes
 Burette

REAGENTS:

 Distilled Water
 Absorbin Reagents: Dissolve 4.0 g of sodium hydroxide in distilled water,
add 1.0 g of sodium Arsenite, and dilute to 1000 ml with distilled water.
 Hydrogen Peroxide, 30 percent
 Hydrogen Peroxide Solution: Dilute 0.2 ml of 30% hydrogen peroxide to 250
ml with distilled water. This solution may be used for one month, if,
refrigerated and protected from light.
 N-(1-Naphthyl)-ethylenediamine Di-hydrochloride (NEDA): A 1%aqueous
solution should have only one absorption peak at 320 nm over the range of
260-400 nm. NEDA showing more than one absorption peak over this range
is impure and should not be used
 NEDA Solution: Dissolve 0.5 g of NEDA in 500 ml of distilled water. This
solution is stable for one month, if refrigerated and protected from light.
 Phosphoric Acid, 85 percent
 Sodium Arsenite
 Sodium Hydroxide
 Sodium Nitrate: Assay of 97 percent NaNO2 or greater.
 Sodium Nitrate Stock Solution (1000 µg NO2/ml):Dissolve 1.5 gm of
desiccated sodium nitrate in distilled water and dilute to 1000 ml such that a
solution containing 1000 µg NO2/ml is obtained. The amount of NaNO2 to be
used if the assay percent is less than 100 percent , is calculated as follows:

G = 1.500/A

Where: G: Amount of NaNO2 in gm

1.500: Gravimetric Conversion factor

A: Assay percent (should be 97 or greater)

Note: This stock solution can be stored for six weeks, if

refrigerated.
 Sodium Nitrate Solution (10 µg NO2/ml): Pipette 5 ml of the stock solution
into a 500 ml volumetric flask and dilute to volume with distilled water.
 Sodium Nitrate Solution (1 µg NO2/ml): Pipette 25 ml of the solution into a
250 ml volumetric flask and dilute with absorbing reagent solution, prepare
fresh daily.
 Sulphanilamide: Melting point 165 to 167°C
 Sulphanilamide Solution: Dissolve 20.0 gm of sulphanilemide in 700 ml of
distilled water. Add with mixing 50 ml of 85 percent phosphoric acid and
dilute to 1000 ml.
[Note: This solution is stable for one month, if refrigerated.]

PROCEDURES:

Sampling:

 Assemble the sampling apparatus at the sampling site.

 Place 30 ml of absorbing solution in an impinger and sample for four hour at
the flow rate of 0.2 to 1 L/min.
 Connect the calibrated flow meter, set the flow rate as per method, measure
the flow rate before sampling.
Start sampling only after obtaining an initial flow rate of 1 liter per minute. Ensure
packing of ice around bubbler for low temperature to improve absorption efficiency.

 Record the exact sampling time by recording initial time (ti) and final time (tf)
of sampler.
 Measure and record the flow rate (fi) before the sampling and flow rate (ff)
after the sampling.
 Seal the collected samples after making it up to 30 ml using distilled water.
 Mark identification number on sample collection bottle and transport to the
laboratory for analysis.

Analysis:

 Pipette out 10 ml of the collected sample into 50 ml volumetric flask. Now.

 Add 1 ml of hydrogen peroxide solution, 10 ml sulphanilamide solution, and
1.4 ml of NEDA solution using pipette, with through mixing after the addition
of each reagent and make up to 50 ml with distilled water.
 Prepare a blank in the same manner using 10 ml of un-exposed absorbing
reagent.
 After a 10 min colour development interval, measure and record the
absorbance at 540 nm against the prepared reagent blank.
 Use distilled water; not the reagent blank, as the optical reference.
 Determine NO2 from the calibration curve.

Calibration:

Preparation of Standard:

 Prepare calibration curve using 1 µg/ml working standards in accordance

with the above analytical procedure.
 Pipette 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 15, 20 ml of working standard
solution (sodium Nitrate Solution,1 µg NO2/ml) in 50 ml volumetric flask.
 Pipette out 10 ml absorbing solution in one volumetric flask as a blank and
pipette out 10 ml absorbing solution to all other volumetric flask.
 Now add 1 ml of hydrogen peroxide solution, 10 ml sulphanilamide solution,
and 1.4 ml of NEDA solution using pipette, with through mixing after the
addition of each reagent and make up to 50 ml with distilled water.
 Take optical density at lamda 540 nm on spectrophotometer.
Standard Curve:

 Plot Plot a curve absorbance (Y axis) versus concentration (X axis). Draw a

line of best fit and determine the slope.
 The reciprocal of slope gives the calibration factor (CF).

CALCULATION:

Air Volume: Calculate the volume of air drawn for sample as follows:

fi + ff
Va= × (tf –ti) × 60 × 10-3
2

Where:

Va : Volume of air sample, (m3)

fi : Air flow rate before sampling, (LPM)
ff : Air flow rate after sampling, (LPM)

ti :Initial time, (Hr)

tf : Final time, (Hr)

10-3: Conversion of litre to m3

60 : Conversion of hours to min

NO2 Concentration in Analyzed Sample: Determine µg NO2/ml graphically from the

calibration curve or compute from the slope and intercept values.

NO2 Concentration in Air Sample: Calculate NO2 as µg per cubic meter of air as
follows:
(As – Ab) × CF × D × Vs
NO2 (µg/m3) = V × 0.82 × Vt
Where:

As : Absorbance of sample, (µg)

Ab : Absorbance of reagent blank, (µg)
CF : Calibration factor
D : Dilution factor (D:1for no dilution, D:2 for 1:1 dilution)
Va : Volume of air sample (m3)
Vs : Volume of sample (ml)
Vt : Volume of aliquot taken for analysis (ml)
0.82: Sampling efficiency

Conversion of µg/m3into ppm

NO2 (ppm) = (NO2µg/m3) ×5.32 × 10-4

EXP 2

Aim: Stack Monitoring: Concentration of particulate matter present in air (PM 10 & PM 2.5), SO2 and NOx
and other parameters.

Reference: IS 11255 (Part 1): 1985

Principle: The determinations of the particulate concentration consist essentially of

Sampling iso-kinetically a measured amount of gas from the flue and separating the
particles from the gas and hence determining the particulate concentration.

APPARATUS: -

 Stack Monitoring Kit

 Sampling Probe
 Suction Pump
 Nozzle
 Pitot Tube
 Thimble
 Thimble Holder

Stack Monitoring:

Flue gases in chimney are the mixture of hot gases and particulates. Flue gas mixture
from such stacks is sucked using Yash Stack Sampler at Iso Kinetic flow rates this
results extraction of representative mixture of flue gases which passes immediately after
their suction through a thimble (a kind of filter where several gram of particulates can
be accumulated without being lost while sampling) particulate free hot gases are divided
into two parts before bubbling through impinger where water and absorbing solution
are kept.

Distribution of hot gases is controlled by the needle valve of two capacities rotameter.
Major part is cooled just by passing them through a large capacity impinger where
chilled water is placed while gases at the rate of 2 lpm are taken directly into 120 mL
impingers where absorbing solutions are kept. All impingers are kept in ice tray where
ice is absorbing solutions are kept. All impingers are kept in ice tray where ice is packed
to ensure effective cooling and better absorption. Moisture of gases is removed before
passing cool gases through rotameter with the help of silica gel impinger bottle.

A thermocouple and digital pyrometer have been provided to measure the stack gas
temperature. A standard S-type pitot tube is used to measure the velocity of flue gases
in the stack. Differential pressure generated across the Pitot tube ends is measured on a
digital manometer filled in the panel of stack monitor.
The stack gas velocity (in m/s) can be calculated at prevailing conditions using Barnauli
equation given below.

V = K √ 2G ΔP Dm/Ds

Where,

V : Velocity of flue gas in chimney m/s

K : Pitot calibration constant

G : Gravitational constant 9.81 m/s2

ΔP : Differential pressure in m of Wc

Dm : Density of manometer fluid in kg/m3 (for water 1000 kg/m3, red oil800 kg/m3)

Ds: Stack gas density in kg/m3 at stack temperature and at stack absolute
pressure.

The stack gas density is a function of the molecular weight of gases comprising the flue
gas, the static pressure inside the dust/chimney and the temperature of flue gas.
Therefore find out static pressure flue gas composition of stack gases including
moisture to find out density of stack gases. However, in most of the common situations
the molecular weight of stack gas is practically the same as that of air while the static
pressure is close to atmospheric pressure. Hence stack gas density can be approximated
by following relation where significant errors are not expected. This is true if moisture
content is not more then 2-3%.

Ds = Da * Ta / Ts

Where Da is density of atmospheric air at ambient temperature Ta (273+25°C) and

Ts is the temperature of stack gas. Both Ta and Ts are in degrees Kelvin.

Da = 1.25 kg/m3, at (273+25) °K

Aerodynamic drag along the stack wall, damper vanes, right angle bends, side entry
ducts etc., cause variation in the flow rate across the cross- section of the
duct/chimney. Hence air velocity measurements must be carried out at number of
traverse point. Traverse points can be decided based on diameter of stack.

Travers Point Multiples to Determine Minimum Numbers of Traverse Point

Required When Port Hole Is Made At the Stack as Per Requirement:
Inside Diameter of Stack or Duct (m) Number of Points
I.D. ≤ 0.3 4
0.3 ≤ I.D. ≤ 0.6 8
0.6 ≤ I.D. ≤ 1.2 12
1.2 ≤ I.D. ≤ 2.4 20
2.4 ≤ I.D. ≤ 5 32

The velocity measure at each point is used to calculate ISOKINETIC sampling rate
needed for sampling at monitored traverse point for a nozzle at known temperature. A
set of three nozzles with different diameter is provided. The rate of sampling which
would achieve isokinetic condition for a nozzle of cross area An is given by relation
below:

Qs = V * An * 60* 1000

Where,
Qs : Rate of sampling from stack in LPM
Cross-Sectional
V : Stack gas velocity in m/sec Dia. of Nozzle
area in m2
An : Area of Nozzle in m2
1/8 0.0000079132
60 : Conversion factor second to minute
103 : Conversion factor m3 to liter 1/4 0.0000316531
3/8 0.00007122
However stack gases cool down as they pass through the sampling train and the rate of
flow indicated by the flow meter must correspondingly be corrected as per gas laws. It is
assumed a seen that if ice is used in cold box and stack temperature is 100-250° C.
Temperature at rotameter inlet is seen 25˚ C ± 2° C.
Therefore,

Q’s = Qs (25+ 273)/ Ts

Q’s = Iso kinetic sampling rate to be set in flow meter in lpm where it is assumed that
the flue gases will cool down up to 25° C when reaching to rotameter.
Since the flow meter of stack gases varies across the cross- section of the duct/chimney,
the particulate concentration too is likely to vary and must be sampled at different
traverse points with corresponding change in sampling rate to maintain isokinetic
conditions.
 Vstd = Vm (Pbar - Pm) (273+25)/ Pstd (273+Tm)

Where,
Vstd : Volume of dry gas through the sampling train (25°C, 760 mm Hg)
Vm : Volume calculated by sampling rate and time (Qs x t)
Pstd : Standard pressure (760 mm of Hg)
Pm : Static pressure in sampling train (mm of Hg)
Pbar : Barometric pressure at the metering point (mm of Hg)
Tm : Temperature of gas at dry gas meter condition (°C)

Dust Concentration

SPM: (Wf – Wi) / Vstd

Where
Wi : Initial weight of thimble (gm)
Wf : Initial weight of thimble (gm)
Vstd : Volume of dry gas through the sampling train (25°C, 760 mm Hg), Nm3

SULPHUR DIOXIDE (SO2)

Aim: Measurement of sulphur dioxide emission from stationary sources.

Reference: IS 11255 (Part 2): 1985

Method: Thorin Method

Principle: A gas sample is extracted from the sampling point in the stack. The acid
mist, including sulphur trioxide, is separated from the sulphur dioxide and the sulphur
dioxide fraction is measured by the barium thorin titration method.

APPARATUS: -

Apparatus for Sampling

 Probe: Chemical resistant glass, 5 to 6 mm ID, with a heating system to prevent

condensation and filtering medium to remove particulate matter including Sulphuric
acid mist.
 Dust Trap: For low dust concentration (up to 1 g/m3N ) are a standard large
impinger with glass wool packed in top to prevent acid mist carry over. For high dust
concentrations, use an appropriate thimble.
 Impingers: Three standard large impingers
 Drying Tube: Packed with 1-3 mm size indicating type silica gel, or equivalent, to
 dry the sample.
 Valve: Needle valve or equivalent, to adjust flow rate accurately in the range of 2-5
l/min.
 Pump: Leak-free, vacuum type.
 Rotameter: Rotameter or other suitable device, to measure flow rate in the range of
0-10 l/min.
 Dry Gas Meter: Sufficiently accurate to measure the sample volume within 1
percent.

Apparatus for Sample Recovery

 Glass Wash Bottles: Two.

 Polyethylene Storage Bottles: To store impinger samples.

Apparatus for Sample Analysis

 Pipettes: 5 ml and 10 ml sizes (0.1 ml division) and 25 ml (0.2 ml division)

 Volumetric Flasks: 50 ml, 100 ml & 1000 ml.
 Burettes: 5 ml and 50 ml.
 Long-necked Flask: 125 ml.

REAGENTS:

Reagents for Sampling:

 Water-Deionized or Distilled: Deionized water is preferable for sharp end points.

 Iso-propanol, 80 Percent: Mix 80 ml of iso – propanol with 20 ml of distilled H2O.
 Hydrogen Peroxide (3%): Dilute 100 ml of 30 percent hydrogen peroxide to 1000 ml
with distilled water. [Prepare fresh daily.]

Reagent for Sample Recovery

 Water-Deionized or Distilled: Deionized water is preferable for sharp end points.

 Iso-propanol, 80 Percent: Mix 80 ml of iso – propanol with 20 ml of dist. H2O

Reagent for Sample Analysis

 Water-Deionized or Distilled: Deionized water is preferable for sharp end points.

 Iso-propanol,
 Thorin Indicator: Dissolve 0.20 g in 100 ml distilled water.
 Barium Perchlorate (0.01 N): Dissolve 1.95 g of barium perchlorate Ba (ClO 4)2.
3H2O in 200 ml distilled water and, dilute to 1 litre with iso-propanol. Standardize
with sulphuric acid. Barium chloride may be used.
 Sulphuric Acid, Standard (0.01 N): Standardize to ± 0.0002 N against 0.01 N NaOH
which has previously been standardized against potassium acid phthalate (primary
standard grade).
PROCEDURES:

Preparation of Collection Train:

 Pour 15 ml of 80 percent iso-propanol into the impinger and 15 ml of 3 percent

hydrogen peroxide into each of the first two impingers.
 Leave the final impinger dry.
 Check the sampling train for leakage at the sampling site by plugging the probe inlet
and pulling a vacuum corresponding to 250 mm mercury column.
 A leakage rate not in excess of 1 percent of the sampling rate is acceptable.
 Carefully release the probe inlet plug and impingers and add more ice during the run
to keep the temperature of the gases leaving the last impinger at 20°C or less.


Sample Collection:

 Adjust the sample flow rate in the range 2 to 5 litres/minutes.

 To begin sampling, position the tip of the probe at the first sampling point and start
the pump.
 At the conclusion of each run, turn off the pump and record the final readings.
Remove the probe from the stack and disconnect it from the train.

Sample Recovery:

 Disconnect the impingers after purging. Discard the contents of the mist impinger
(with the glass wool).
 Pour the contents of the other impingers into a polyethylene shipment bottle.
 Rinse the three midget impingers and the contacting tube, with distilled water and
add these washings to the same storage container.

Sample Analysis:

 Transfer the contents of the storage container to a 50 ml volumetric flask.

 Dilute to the mark with deionized, distilled water.
 Pipette a 10 ml aliquot of this solution into a 125 ml erlenmeyer flask.
 Add 40 ml of iso-propanol and two to four drops of thorin indicator.
 Titrate to a pink end point using 0.01 N barium perchlorate.
 Run a blank with each series of samples.
 CALCULATION:

Dry Gas Volume:

Correct the sample measured by the dry gas meter to normal conditions (298 K and
101 kPa) by using the following equation.

V (TN) × (P)
VN =
(T) × (PN)
Where:

VN : Volume of gas sample through the dry gas meter (normal condition), (m3)
V : Volume of gas sample through the dry gas meter (meter condition), (m3)
TN : Absolute temperature at normal conditions (298 K)
T : Average dry gas meter temperature (K)
P : Absolute meter pressure (kPa)
PN : Absolute pressure at normal conditions (101 kPa)

Flow rate × Time

V=
1000

Sulphur Dioxide Concentration:

Calculate the concentration of sulphur dioxide using the following equation.

0.032× (V-Vb) × N × Vso/ Va

SO2 (gm/Nm3) =
VN
Where:
V : Volume of barium perchlorate titrate used for sample, (ml)
Vb : Volume of barium perchlorate titrate used for blank, (ml)
N : Normality of barium perchlorate titrate
VSO : Total solution volume of sulpher dioxide (50 ml)
Va : Volume of sample aliquot titrted(10 ml)
VN : Volume of gas sample through the dry gas meter (Normal Condition) (m 3)

NITROGEN DIOXIDE (NO2)

Aim: Measurement of Nitrogen dioxide emission from stationary sources.

Reference: IS11255 (Part 7): 2005

Principle: A grab sample is collected in a dilute sulphuric acid-hydrogen peroxide

absorbing solution, and the nitrogen oxides, except nitrous oxide, are measured
colorimetrically using the phenoldisulphonic acid(PDS) procedure.

APPARATUS: -

Apparatus for Sampling

 Probe: Borosilicate glass tubing sufficiently heated to prevent water condensation

and equipped an in-stack or out-stack filter to remove particulate matter (a plug of
glass wool is satisfactory for this purpose).
 Collection Flask: Two liter borosilicate round bottom flask, with short neck and
24/40 standard taper opening, protected against implosion or breakage.
 Flask Valve: T-bore stopcock connected to a 24/40 standard taper joint.
 Temperature Gauge: Dial type thermometer or other temperature gauge, capable to
measuring 1˚C on intervals from -5 to 50˚C.
 Vacuum Line: Tubing capable of withstanding a vacuum of 75 mm Hg absolute
pressure, with ‘T’ connection and T-bore stopcoke.
 Vacuum Gauge: U-tube manometer, 1 m high, with 1 mm divisions, or other gauge
capable of measuring pressure to within ± 2.5 mm Hg.
 Pump: Capable of evacuating the collection flask to a pressure equal to or less than
75 mm Hg absolute.
 Sueeze Bulb: one way
 Stopcoke
 Barometer: Mercury, aneroid, or other barometer capable of measuring atmospheric
pressure to within 2.5 mm Hg.
 Porcelain Evaporating Dishes: 175 to 250 mt capacity with lip for pouring, one for
each sample and each standard.
 Steam Bath: Low temperature ovens or thermostatically controlled-hot plates kept
below 70°C.
 Policeman Porcelain: One for each sample and each standard
 Graduated Cylinder: 100 ml with 1 ml divisions.
 Spectrophotometer: To measure absorbance at410nm.
 pH Meter& pH Paper
 Analytical Balance

REAGENTS:

 Absorbing Solution: To prepare the absorbing solution, cautiously add 2.8 ml

concentrated H2SO4 to 1000 ml of distilled water. Mix well and add 6 ml of 3 percent
hydrogen peroxide solution, freshly prepared from 30 percent hydrogen peroxide
solution. [Note: The absorbing solution should be used within 1 week of its
preparation. Do not expose to extreme heat or direct sunlight.]
 Sodium Hydroxide (1 N): Dissolve 40 gm NaOH in distilled water and dilute to 1000
ml.
 Fuming Sulphuric Acid: 15 to 18 percent by weight free sulphur trioxide. [Note:
Highly corrosive, handle with care]
 Phenol: White solid.
 Sulphuric Acid: Concentrated 95 percent minimum assay.
 Potassium Nitrate: Dried at 105°C to 110°Cfor a minimum period of 2 h just prior
to preparation of standard solution.
 StandardKNO3Solution: Dissolve 2.198 gm dried potassium nitrate (KNO3) in
distilled water and dilute to 1000 ml with distilled water in a 1000 ml volumetric
flask.
 Working Standard KNO3Solution: Dilute 10 ml of the standard solution to 100 ml
with distilled water. One millilitre of the working standard solution is equivalent to
100 μg nitrogen dioxide (NO2).
 Phenoldisulphonic Acid Solution: Dissolve 25 gm of pure white phenol in 150 ml
concentrated sulphuric acid on a steam bath. Cool, add 75 ml fuming sulphuric
acid, and heat at 100°C for 2 h. Store in a dark, stoppered bottle.

PROCEDURES:

Sampling:
 Pipette 25 ml of absorbing solution into a sample flask, retaining a sufficient
quantity for use in preparing the calibration standards.
 Insert the flask valve-stopper into the flask with the valve in the 'purge' position.
 Assemble the sampling train and place the probe at the sampling point. Make sure
that all fittings are tight and leak-free and that all ground glass joints have been
properly greased with a high vacuum, high temperature chlorofluorocarbon based
stopcock grease.
 Turn the flask valve and the pump valve to their 'evacuate' position. Evacuate the
flask to 75 mm Hg absolute pressure, or less. Evacuation to a pressure approaching
the vapour pressure of water at the existing temperature is desirable.
 Turn the pump valve to its ‘vent’ position and tum off the pump.
 Check for leakage by observing the manometer for any pressure fluctuation. Any
variation greater than 10 mm Hg over a period of 1 minute is not acceptable, and the
flask is not to be used until the leakage problem is corrected.
 Pressure in the flask is not to exceed 75 mm Hg absolute at the time sampling is
 commenced.

 Record the volume of the flask and valve (Vi), the flask temperature (Ti) and the
barometric pressure.
 Tum the flask valve counter clockwise to its 'purge' position and do the same with
the pump valve.
 Purge the probe and the vacuum tube using the squeeze bulb. If condensation
occurs in the probe and the flask area, heat the probe and purge until the
condensation disappears.
 Next, tum the pump valve to its 'vent' position. Turn the flask valve clockwise to its
‘evacuate’ position and record the difference in the mercury levels in the manometer.
 The absolute internal pressure in the flask (Pi) is equal to the barometric pressure
less the manometric reading.
 Immediately tum the flask valve to the 'sample' position and permit the gas to enter
the flask until pressures in the flask and sample line (that is duct, stack) are equal.
 This will usually require about 15 s; a longer period indicates a 'plug' in the probe,
which must be corrected before sampling is continued.
 After collecting the sample, turn the flask valve to its 'purge' position and disconnect
the flask from the sampling train. Shake the flask for at least 5 min.
 If the gas being sampled contains insufficient oxygen for the conversion of NO to NO 2
then oxygen shall be introduced into the flask by one of the following three methods:
1) Before evacuating the sampling flask, flush with pure cylinder oxygen, then
evacuate flask to 75 mm Hg absolute-pressure less; 2) Inject oxygen into -the flask
after sampling; or 3) Terminate sampling with minimum 50 mm Hg vacuum
remaining in flask, record the final pressure and vent the flask to atmosphere until
the flask pressure is equal to the atmospheric pressure.

Sample Recovery:
 Let the flask-set for a minimum of 16 h and then shake the contents for 2 min.
 Connect the flask to mercury filled U-tube manometer.
 Open the valve-from the flask to the manometer and record the flask temperature
(Tf), the barometric pressure, and the difference between the mercury levels in the
manometer.
 The absolute internal pressure in the flask (Pr) is the barometric pressure less the
manometer reading.
 Transfer the contents of the flask to a leak-free polyethylene bottle.
 Rinse the flask twice with 5 ml portions of distilled water and add the rinse water to
the bottle.
 Adjust the pH to between 9 and 12 by adding sodium hydroxide (l N), dropwise
(about 25 to 35 drops).
 Check the pH by dipping a stirring rod into solution and then touching to the pH test
paper.
 Remove as little material as possible during this step. Mar-k the height of the liquid
level so that the container can be checked for leakage after transport. Label the
container to clearly identify its contents.
Sample Analysis:
 Note the level of the liquid in container and confirm whether or not any sample was
lost during shipment; note this on analytical data sheet.
 Immediately prior to ana1ysis, transfer the contents of the shipping container to a 50
ml volumetric flask and rinse the container twice with 5 ml portions of distilled
water.
 Add the rinse water to the flask and dilute to the mark with distilled water; mix
thoroughly.
 Pipette a 25 ml aliquot into the porcelain evaporating dish. Return any unused
portion of the sample to the polyethylene storage bottle.
 Evaporate the 25 ml aliquot to dryness on a steam bath and allow to cool.
 Add 2 ml phenoldisulphonic acid solution to the dried residue and triturate
thoroughly with a polyethylene policeman. Make sure the solution contacts all
residue.
 Add 1 ml distilled water and four drops of concentrated sulphuric acid.
 Heat the solution on a steam bath for 3 min with occasional stirring. Allow the
solution to cool, add 20 ml distilled water, mix well by stirring and add concentrated,
ammonium hydroxide, drop wise, with constant stirring, until the pH is 10 (as
determined by pH paper). If the sample contains solids, these shall be removed by
filtration as follows:
 Filter through Whatman No. 41 or equivalent into a 100 ml volumetric flask, rinse
the evaporating dish with three 5 ml portions of distilled water; filter these three
rinses.
 Wash the filter with at least three 15 ml portions of deionized, distilled water. Add
the filter washings to the contents of the volumetric flask and dilute to the mark with
distilled water. If the solids are absent, transfer the solution directly to the 100 ml
volumetric flask and dilute to the mark with distilled water.
 Mix the contents of the flask thoroughly and measure the absorbance at the 410 nm
wavelength used for the standards using the blank solution as a zero reference.
Dilute the sample and the blank with equal volumes of distilled water.

Determination of Calibration Curve:

 Add 0 ml, 2 ml, 4 ml, 6 ml and 8 ml of KNO3 working standard solution (1 ml = 100g
of NO2) to a series of fine 50 ml volumetric flasks.
 To each flask, add 25 ml absorbing solution, 10 ml distilled water and sodium
hydroxide (1 N) drop wise until the pH is between 9 and12 (about 25 to 35 drops
each). Dilute to mark with distilled water.
 Mix thoroughly and Pipette 25 ml aliquot of each solution into a separate porcelain
evaporating dish. Beginning from evaporation step follow the analysis procedure as
given in above until the solution has been transferred to the 100-ml volumetric flask
and dilute to mark.
 Measure the absorbance of each solution at 410 nm wavelength on a
spectrophotometer.
  Plot the absorbances of the solutions ordinates against the concentration. A linear
relationship is obtained.

 Calculate the calibration factor (Kc) by taking the reciprocal of the slope of the line.

CALCULATION:

Sample Volume Corrected to Standard Conditions: Carry out the calculations as

given below:

Vsc = (Tstd/Pstd) (Vt-Va) (Pf/Tf-Pi/Pi/Ti) = K1 (Vt-25) (Pf/Tf-Pi/Ti)

Where:
Pf :Final absolute pressure of flask, (mmHg)

Pi :Ininal absolute pressure of flask, (mmHg)

Pstd : Standard absolute pressure of flask, (760 mmHg)

Tf : Final absolute temperature of flask (K)

Ti :Iniinal absolute temperature of flask (K)

Tstd : Standard absolute temperature of flask (298.15 K)

Vsc :Sample volume at standard conditions (ml)

K1 : 0.392 K/mm Hg

Vt : Volume of flask and valve (ml)

Va : Volume of absorbing solution (25 ml)

NO2 Concentration Corrected to Standard Conditions:

Carry out the calculations as given below:

= (As-Ab)* Kc* 1000*2*F
Vsc

Where:
As : Absorbance of
the sample Ab :
Absorbance of the
blank
C : Concentration of NOx as NO2, corrected to standard conditions, mg/Nm3
F : Dilution factor (that 25/5, 25/10, etc) required only, if sample
dilution was needed to reduce the absorbance into the range of
calibration
Kc : Spectrophotometer calibration factor
2 : 50/25 the aliquot factor [Note: If other than 25 ml aliquot is used for
analysis, the factor 2 must be replaced by corresponding factor]

EXP 3
BIOCHEMICAL OXYGEN DEMAND TEST

Aim of the Experiment:

To determine the Biochemical Oxygen Demand (BOD) of a given wastewater sample.

Apparatus:

1. BOD bottle (300 ml Capacity)

2. Incubator to control the temperature
3. Volumetric flasks
4. 50 ml Burette
5. 500 ml conical flask

Reagents:
1. Manganese Sulphate solution.
2. Alkali Iodine solution (Azide).
3. Concentrated Sulphuric acid.
4. Standard Sodium Thiosulphate solution of 0.025N
5. Starch solution.

Theory:

Microorganisms such as bacteria are responsible for decomposing organic matter.

When organic matter such as dead plants, leaves, grass clippings, manure, sewage, food waste
is present in a wastewater, the aerobic bacteria will start the oxidation of these wastes. When
this happens, much of the available Dissolved Oxygen (DO) is consumed by aerobic bacteria,
robbing other aquatic organisms of the oxygen they need to live. The biochemical oxygen
demand is measure of oxygen utilized by aerobic micro-organisms during biological oxidation
of organic matter. Generally, when BOD levels are high, there will be low DO levels.

Organic matter + O2 CO2 + new bacterias + H2O + Heat Drinking water must have a BOD of
less than 01 mg/l and the water is considered fairly up to 03 mg/l of BOD, but when the BOD
value ≥ 05 mg/l the water is doubtful in purity.

Ordinary domestic sewage may have a BOD of 200 mg/l. As per CPCB standards the treated
or untreated sewage to be discharged into surface water body must a have of BOD of less than
30 mg/l

Procedure: Part A:

Dilution

1. Place the desired volume of distilled water in a 05 liter conical flask. Aeration is done by
bubbling compressed air through distilled water.
2. Add 01 ml of manganous sulfate (MgSO4) solution, 01 ml of calcium chloride (CaCl2) and
01 ml of ferric chloride (FeCl3) solution for every liter of distilled water.
3. In the case of the wastewater samples, which are not expected to have sufficient bacterial
population, add seed to the distilled water. Generally 2 ml of settled sewage is sufficient for
1000 ml of distilled water as seed.
4. Highly acidic or alkaline samples are to be neutralized to pH of around 7.0.
5. Add 2 or 3 ml of sodium thiosulfate (Na2S2O3) to destroy residual chlorine if any.
6. Take sample as under: Strong wastes: 0.1, 0.5, or 1% Settled domestic sewage: 1.0, 2.5, or
5% Treated effluents: 5.0, 12.5 or 25% River water: 25 to 100%
7. Dilute the sample with distilled water and mix the contents well.

Part B: Titration
1. Take samples in 02 BOD bottles of 300 ml capacity.
2. Fill another 02 BOD bottles with distilled water (blank).
3. Immediately find initial DO of 01 bottle with distilled water in it and 01 bottle with diluted
wastewater sample in it by modified Azide method or Winkler’s method (same procedure as
used in DO determination).
Incubate the remaining 02 bottles by keeping them in an incubator for 5 days (120 hours) at
200C or for 3 days (72 hours) at 27oC and find out the final DO of the distilled water and
water/wastewater samples by modified Azide method or Winkler’s method.

Results:

Biological Oxygen Demand of the given sample __________ mg/l

Calculations:
1. Initial DO present in diluted wastewater (W0) = _____________ mg/l
2. Final DO present in diluted wastewater (W3) = _____________ mg/l
3. Initial DO present in distilled water (D0) = _____________ ___ mg/l
4. Final DO present in distilled water (D3) = ______________ ___ mg/l
 (Wo – W3) - (Do – D3) x Volume of BOD bottle 300 ml
=______________________________________________
ml of sample taken in BOD bottle

BOD of the sample = (Initial DO – Final DO) x Dilution ratio in mg/l

EXP 4 CHEMICAL OXYGEN DEMAND TEST

Aim of the Experiment:

To determine the Chemical Oxygen Demand (COD) of a given wastewater sample.

Methodology:
Titration and Open reflux method.

Apparatus:
1. COD digester
2. 250 ml conical flask
3. 50 ml burette
4. Pipettes, etc.

Reagents used:
1. Potassium dichromate solution
2. Ferrous ammonium sulphate (FAS) solution of 0.25 N.
3. Ferrion red indicator
4. Mercuric sulphate
5. Concentrated Sulphuric acid

Theory: Chemical oxygen demand (COD) is the oxygen required for chemical oxidation of
organics & inorganic impurities by strong oxidizing agents like potassium dichromate
(K2Cr2O7) under acidic conditions. The basis for the COD test is that nearly all organic
compounds can be fully oxidized to carbon dioxide with a strong oxidizing agent under acidic
conditions. The major advantage of COD test is the short time required for determination of
total O2 required for oxidation. COD test requires 03 hours instead of 3 or 5 days as needed
for measurement of BOD. The COD test is much more useful than BOD test for estimating
strength of certain industrial effluents of both organic type (pesticide industries) and inorganic
type (metallurgical industries) which contain toxic chemicals.
Procedure:

1. Take 20 ml of sample in a clean conical flask.

2. Add 10 ml K2Cr2O7 to conical flask and add 30 ml of concentrated H2SO4 slowly
containing Ag2SO4 and mix thoroughly. Add 0.4 gram or pinch of mercuric sulfate (HgSO4).
Mix the contents thoroughly.
3. Transfer the sample into the glass digester of COD apparatus and connect the air vessel.
4. Reflux the sample for 2½ hours at a temperature of ±1000C. Allow it for some time for
cooling to room temperature.
5. Dilute the sample with distilled water to make it up to 100 ml.
6. Titrate excess of K2Cr2O7 with 0.25 N FAS as a titrant with 2 to 3 drops of Ferroin
indicator until color changes from greenish blue to wine red indicating the end point of
titration.
7. Note down the volume of titrant used for given sample as V1 ml and for distilled water
(blank) as V2 ml.

RESULTS .

Tabulation:

SAMPLE INDICATOR BURRETTE READINGS VOL. OF

SL NO USED USED FAS USED
Waste water Ferrion red FR IR FR-IR
sample indicator

Calculations:

Chemical Oxygen Demand of the given sample:

COD = (V1 – V2) x N x 8x 1000 mg / ml of sample taken ________ mg/l

EXP: 5 Type- II settling of particle sedimentation

PENDING

EXP: 6 Break point chlorination test

Aim of the Experiment:

To determine the Break point chlorination test of a given sample.

Apparatus:
Erlemeyer flask (250 mL)
Bottle
Beaker (250 mL)
Measuring cylinder
Dropper
Stirrer

Reagents:
Starch Indicator
Standard 0.025 N Sodium thiosulfate
Potassium Iodine crystal
Concentrated Acetic Acid
Chlorine water

Procedure:

1. Place 200-mL portion of the water to be chlorinated in each of six 250-mL flasks.
2. Add required quantity (as instructed by your teacher) of "chlorine water" (stock solution of
bleaching powder in water) in each of the flasks. The chlorine content of the "chlorine
water" (determined earlier in the laboratory) would be provided to you by your teacher.
Calculate the chlorine dose for each of the six flasks.
3. Shake each flask gently and allow to stand for 30 minutes.
4. Determine residual chlorine of water from each flask by the starch-iodine method as
described below:

Starch-Iodine Method:

The starch-iodine method is based on the oxidizing power of free and combined chlorine
residuals to convert iodide ion into free iodine at pH 8 or less, as shown below.
Cl2 + 2I- = I2 + 2 Cl-
In the starch-iodine method, the quantity of chlorine residuals is determined by measuring the
quantity of iodine by titration with a reducing agent sodium thiosulfate (Na2S2O3). The end
point of titration is indicated by the disappearance of blue color, produced by the reaction
between iodine and starch (which is added as indicator during the titration),
I2 + 2 Na2S2O3 = Na2S4O6 + 2 Nal
or, I2 + 2S2O32- = S O 2- + 2I-
4 6color
I2 + starch = blue
(Qualitative test for the presence of iodine/chlorine)

The titration is carried out at pH 3 to 4, because the reaction with thiosulfate is not
stoichiometric at neutral pH due to partial oxidation of the thiosulfate to sulphate.

Procedure for determination of residual chlorine concentration:

1. Place 200 mL of the sample in an Erlenmeyer flask.

2. Add 'about 1g of potassium iodide (estimated on a spatula) and 2 mL of concentrated
Acetic acid to the water.
3. Add 0.025 N sodium thiosulfate drop by drop from a burette until the yellow color almost
disappears.
4. Add 1 mL of starch solution to the water.
5. Continue addition of standard sodium thiosulfate (Na2S2O3) solution until the blue color
 just disappears.
6. Record the quantity (in mL) of sodium thiosulfate (Na2S2O3) solution used.

Calculation:

Residual chlorine (mg/L) = mL of 0.025N sodium thiosulfate used x M.F.

M.F. =

Table

Observation Chlorine Residual

No. pH Dose Chlorine
(initial) (mg/L) (mg/L)

Introduction:

Chlorination of public water supplies and polluted waters serves primarily to destroy or de-
activate disease-producing microorganisms. Disinfection with chlorine is widely practiced.
Chlorination may produce some adverse effects including taste and odor problem. in recent
years, chlorination has been found to produce trihalomethanes (THMs) and other organics of
health concern (THMs are suspected human carcinogens). Thus, use of alternative
disinfectants, such as chlorine dioxide and ozone that do not cause this particular problem, is
increasing.
Theory:
Disinfectant capabilities of chlorine depend on its chemical form in water, which in turn is
dependent on pH, temperature, organic content of water, and other water quality factors.
Chlorine is used in the form of free chlorine [e.g., chlorine gas] or as hypochlorites [e.g.,
NaOCl and Ca(OC1)2]. Chlorine applied to water either as free chlorine or hypochlorite
initially undergoes hydrolysis to form free chlorine consisting of aqueous molecular chlorine,
hypochlorous acid and hypochlorite ion.

Chlorine gas rapidly hydrolyzes to hypochlorous acid according to:

Cl2 + H2O = HOCl + H+ +Cl– 14.1
Aqueous solutions of sodium or calcium hypochlorite hydrolyze too:
Ca(OCl)2 + 2H2O = Ca2+ + 2HOCl + 2OH– 14.2
NaOCl + H2O = Na+ + HOCl + OH– 14.3
Hypochlorous acid is a weak acid and will disassociate according to:
HOCl⇔H+ +OCl– 14.4

The two chemical species formed by chlorine in water, hypochlorous acid (HOCl) a
n
d
hypochlorite ion (OCl–), are commonly referred to as “free” or “available” chlorine.

Figure 14.1: Distribution of Chlorine species at 250C

Figure 13.1 shows that Cl2 can be significantly at low pH values (below pH 2); while HOCl is
dominant between pH 3 and 6. Between pH 6 and 9, the relative fraction of HOCl decrease,
while the corresponding fraction of OCl- increases. In waters with pH between 6.5~8.5, the
reaction is incomplete and both species (HOCl and OCl-) will be present. Hypochlorous acid
is the more germicidal of the two, especially at short contact time. The dissociation of HOCl is
also temperature dependent. The effect of temperature is such that at a given pH, the fraction
of HOCl will be lower at higher temperatures.

Reactions of Chorine with Impurities in Water:

Reactions with Ammonia:
Free chlorine reacts readily with ammonia and certain nitrogenous compounds to form what
are collectively known as "combined chlorine". The inorganic chloramines consist of three
species: monochloramine (NH2CI), dichloramine (NHCl2) and trichloramine or nitrogen
trichloride (NCI3). The presence and concentrations of these combined forms depend on a
number of factors including the ratio of chlorine to ammonia-nitrogen, chlorine dose,
temperature, pH and alkalinity.
NH3 + HOCI = NH2C1 + H2O; pH 4.5 to 8 14.5
NH2CI + HOCI = NHCl2 + H2O; pH 4.5 to 8 14.6
NHCl2 HOCI = NCI3 + H2O; pH < 4.5 14.7
In addition to chlorinating ammonia, chlorine also reacts to oxidize ammonia to chlorine-free
products (e.g., nitrogen gas and nitrate) as shown below.
3 Cl2 + 2 NH3 = N2 (g) + 6H+ + 6 CI- 14.8
4C12 + NH3 + 3H2O = 8C1- + NO3- + 9H+ 14.9
The mono- and dichloramines have significant disinfecting power and are therefore of interest
in the measurement of chlorine residuals. Combined chlorine in water supplies may be formed
in the treatment of raw waters containing ammonia; chlorinated wastewater effluents, as well
as certain chlorinated industrial effluents normally contain only combined chlorine.

Reactions with Other Impurities:

Chlorine combines with various reducing agents and organic compounds thus increasing the
chlorine demand which must be satisfied before chlorine is available to accomplish
disinfection.
Fe2+, Mn2+, NO2-, and H2S are examples of inorganic reducing agents present in water
supplies that will react with chlorine. Chlorine can react with phenols to produce mono-, di-,
or trichlorophenols, which can impart tastes and odors to waters, Chlorine also reacts with
humic substances present in water to form trihalomethanes (THMs, e.g., chloroform,
brornoform, etc.) which are suspected human carcinogens (Note: According to USEPA,
maximum allowable level of THMs in drinking water is 100 µg/L).

Break Point Chlorination

If chlorine is added to water containing reducing agents and ammonia (either naturally present
or added to water to produce combined chlorine), a hump-shaped breakpoint curve is produced
as shown in following figure. The different segment of the curve is described as follows:

a. If the water is free of ammonia and other compounds that may react with chlorine, the
application of chlorine will yield free available chlorine residual of same concentration.
This is denoted by the ‘no demand line’ or the "zero demand line" (see Fig.).
b. Chlorine first reacts with reducing agents such as H2S, Fe-2+, Mn2+ and develops no
measurable residual as shown by the portion of the curve from Origin up to point A.
Figure 14.2: Generalized curve obtained during breakpoint chlorination of water sample
containing ammonia

c. Addition of chlorine beyond point A results in forming mainly mono- and di- chloramines.
With mole ratios of chlorine to ammonia up to 1:1 [i.e., C12:NH3-N = 1:1], both mono
and di-chloramines are formed. Chloramines thus formed are effective disinfectants and
are shown as combined available chlorine residual in figure (From A to B).
d. Further increase in the mole ratio of chlorine to ammonia result in formation of some
trichloramine and oxidation of part of ammonia to N2 and NO3-. These reactions are
essentially complete when 1.5 mole of chlorine has been added for each mole of ammonia
nitrogen originally present in water [i.e., C12:NH3-N = 1.5:1]. This is represented by the
portion of the curve from B to C.
e. Addition of chlorine beyond point C would produce free chlorine residuals and is referred
to as "breakpoint chlorination". In other words, chlorination of water to the extent that all
ammonia is converted to N2 or higher oxidation state is referred to as "breakpoint
chlorination'.
f. Addition of chlorine beyond point C would produce free chlorine residuals and is
referredto as "breakpoint chlorination". In other words, chlorination of water to the extent
that all ammonia is converted to N2 or higher oxidation state is referred to as "breakpoint
chlorination".
Environmental Significance:
Breakpoint chlorination is required to obtain a free chlorine residual for better disinfection if
ammonia is present in a water supply. While free chlorine residuals have good disinfecting
powers, they are usually dissipated quickly in the distribution system. For this reason, final
treatment with ammonia if often practiced to convert free chlorine residuals to longer-lasting
combined chlorine residuals. The difference between the amount of chlorine added to the
water and the amount of residual chlorine (i.e., free and combined available chlorine
remaining) at the end of a specified contact period is termed as "chlorine demand'.

Determination of Total and Fecal

Coliformin water

Introduction:
A variety of different microorganisms are found in untreated water. Most of
these organisms do not pose a health hazard to humans. Certain organisms,
referred to as pathogens, cause disease to humans which include species of
bacteria, viruses and protozoa. These organisms are not native to aquatic systems
and usually require an animal host for growth and reproduction. Pathogens are
likely to gain entrance sporadically, and they do not survive for very long period
of time; consequently they could be missed in a sample submitted to the
laboratory. Although it is possible to detect the presence of various pathogens in
water, the isolation and identification of many of these is often extremely
complicated, time-consuming and expensive proposition. Hence in most cases
(except when presence of any particular microorganism is suspected) the
microbiological quality of water is checked using some indicator organisms.

An indicator organism is one whose presence presumes that contamination has

occurred and suggests the nature and extent of the contaminants. An indicator
organism should be a microorganism whose presence is evidence of fecal
contamination of warm blooded animals. Indicators may be accompanied by
pathogens, but typically do not cause disease themselves. The ideal indicator
organism should have the following characteristics:

 Always be present when pathogens are present

 Always be absent where pathogens arc absent
 Numbers should correlate the degree of pollution
 Be present in greater number than pathogens
 There should be no after-growth or re-growth in water
 There should be greater or equal survival time than pathogens
 Be easily and quickly detected by simple laboratory tests
 Should have constant biochemical and identifying characteristics
 Harmless to humans
No organisms or group of organisms meet all of these criteria; but the coliform
bacteria fulfill most of them, and this group is most common indicator used in
microbial examination of water. Total coliforms are grouped into two categories
(1) Fecal coliform (thermo-tolerant coliform-) and (2) Non- Fecal coliform

Total coliforms are defined as gram negative bacteria which ferment lactose at
35° or 37° C with the production of acid, gas and aldehyde within 24 or 48 hours.
Fecal coliform are a subgroup of total coliforms, which live in the warm blooded
animals and have the same properties as thetotalcoliform but tolerate and grow at
higher selective temperature range of 44° to 44.5°C. In addition, they form indole
from tryptophan. And these combined properties, when positive, are regarded as
presumptive Escherichia coli (presumptive E. coli). Some coliform species are
frequently associated with plant debris or may be common inhabitants in soil or
surface waters which arc called non-fecal coliforms.

Total coliform (TC) = Fecal coliform (FC) + Non-fecal coliform

Thus, the total coliform group should not be regarded as an indicator of
organisms exclusively of fecal origin. The use of total coliforms as an indicator
may therefore be of little value in assessing the fecal contamination of surface
water, unprotected shallow wells etc. where contamination by coliforms of non-
fecal origin can occur. The measurement of total coliforms is of particular
relevance for treated and / or chlorinated water supplies; in this case the absence
of total coliforms would normally indicate that the water has been sufficiently
treated / disinfected to destroy various pathogens. Measurement of focal
coliforms is a better indicator of general contamination by material of fecal
origin. The predominant species of fecal coliform group is Escherichia coil(E.
coil), which is exclusively of fecal origin, but strains of Klebsella pneumonia and
Enterobacterspecies may also be present in contaminated water.
Using coliform as indicators of the presence and absence of pathogens
sometimes may cause the following drawbacks:
 False positive result can be obtained from the bacterial genus aeromonas,
which can biochemically mimic the coliform group
 False negative result can be obtained when conforms are present along
with high population of other bacteria. The latter bacteria can act to
suppress coliform activity.
 A number of pathogens have been shown to survive longer in natural
waters and / or through various treatment processes than coliform.
But the use of coliforms was established first and there does not appear to be any
distinct advantages to warrant shifting to other indicator organisms. Since
bacteria are used as indicator organisms, the microbiological examination of
water is commonly called bacteriological examination.

Apparatus:
1. Petri Dish
2. Incubator
3. Measuring Cylinder, beaker, dropper etc.
Reagents:
1. Appropriate culture medium (broth)
2. Distilled water

Methods of Bacteriological Examination of Water:

Basically there two methods of bacteriological analysis of water: (a) Multiple
Tube or Most Probable Number (MPN) method, and (b) Membrane Filter
(MF) method.
(a) Multiple Tube/ Most Probable Number (MPN) method:
MPN is a procedure to estimate the population density of viable microorganisms
in a test sample. It’s based upon the application of the theory of probability to the
numbers of observed positive growth responses to a standard dilution series of
sample inoculums placed into a set number of culture media tubes. Positive
growth response after incubation may be indicated by such observations as gas
production in fermentation tubes or visible turbidity in broth tubes, depending
upon the type of media employed.
(b) Membrane Filter Method:
In contrast to the multiple-tube (MT) method, the membrane filter (IVIF) method
gives a direct count of total coliforms and fecal coliforms present in a given
sample of water. The method is based on the filtration of a known volume of
water through a membrane filter consisting of a cellulose compound with a
uniform pore diameter of 0.45 µm; the bacteria are retained on the surface of the
membrane filter. When the membrane containing the bacteria is incubated in a
sterile container at an appropriate temperature with a selective differential culture
medium, characteristic colonies of coliforms and fecal coliforms develop, which
can be counted directly. This technique is popular with environmental engineers.
This method is not suitable for turbid waters, but otherwise it has several
advantages. Its particular advantages and limitations are as follows:
Advantages:
 Results are obtained more quickly as the number of coliforms can be
assessed in less than 24 hours, whereas the multiple tube technique
requires 48 hours both for a negative or a presumptive positive test;
 Saving in work, certain supplies and glassware;
 Method gives direct results;
 Easy to use in laboratories, or even in the field if portable equipment is used.

Disadvantages:
 High turbidity caused by clay, algae, etc. prevents the filtration of a
sufficient volume of water for analysis and it may also produce a deposit
on the membrane which could interfere with bacterial growth;
 Presence of a relatively high non-coliform count may interfere with the
determination of coliforms:
 Waters containing particular toxic substances which may be absorbed by
the membranes, can affect the growth of the coliforms.

Test Procedure (For MF method):

This section describes the general procedures, it should be noted that different
types of filtration units and equipment are available in the market for performing
the tests.
Determination of Total Coliforms (TC):
1. Connect the Erlenmeyer (side-arm) flask to the vacuum source (turned off)
and place the porous support in position. if an electric pump is used, it is
advisable to put a second flask between the Erlenmeyer and the vacuum
source; this second flask acts as a water trap and thus protects the electric
pump.
2. Open a Petri-dish and place a pad in it.
3. 'With a sterile pipette add 2 mL of selective broth (culture) medium to saturate the pad.
4. Assemble the filtration unit by placing sterile membrane filter on the porous
support, using forceps sterilized earlier by flaming.
5. Place the upper container in position and secure it with the special clamps.
The type of clamping to be used will depend on the type of equipment.
6. Pour tide volume of sample chosen as optimal, in accordance with the type
of water, into the upper container. If the test sample is less than 10 mL, at
least 20 ml of sterile dilution water should be added to the top container
before filtration applying the vacuum.
7. After the sample has passed through the filter, disconnect the vacuum and
rinse the container with 20-30 mL of sterile dilution water. Repeat the
rinsing after all the water from the first rinse has passed through the filter.
8. Take the filtration unit apart and using the forceps, place the membrane filter
in the Petri- dish on the pad with the grid side up. Make sure that no air
bubbles are trapped between the pad and the filter.
9. Invert the Petri-dish for incubation.
10. Incubate at 35°C or 37°C for 18-24 hours with 100% humidity (to ensure
this, place a piece of wet cotton wool in the incubator). If ointment
containers or plastic dishes with tight-fitting lids are used, humidification is
not necessary.

Bacterial Colony observation:

Colonies of coliform bacteria are a medium red or dark red color, with a greenish
gold or metallic surface sheen. This sheen may cover the entire colony or appear
only in the centre of the colony. Colonies of other types should not be counted.
The colonies can be counted with the aid of a lens. The number of total coliforms
per 100 mL is then given by:

Determination of Fecal Coliforms (FC):

The procedure for fecal coliforms is similar to that used for determining total
coliforms. Filter the sample as described, and place the membrane filler on the
pad saturated with appropriate culture medium.
1. Place the dishes in an incubator at 44±0.5 °C for 24 hours at 100%
humidity. Alternatively, tight-fitting or sealed Petri-dishes may be
placed in water-proof plastic bags for incubation.
2. Submerge the bags in a water-bath maintained at 44±0.5°C for 24 hours. The
plastic bags must be below the surface of the water throughout the
incubation period. They can be held down by means of a suitable weight,
e.g., a metal rack.

Bacterial Colony observation:

Colonies of fecal coliform bacteria are blue in color. This color may cover the
entire colony, or appear only in the center of the colony. Colonies of other types
should not be counted. The colonies can be counted with the aid of a lens. The
number of fecal coliforms per 100 ml is then given by:

Calculation:

Total coliform (CFU/ 100 mL) =

Fecal coliform (CFU/ 100 mL) =

Table
 Observation Total Coliform Unit Fecal Coliform Unit
No. per 100 ml per 100 ml

EXP 7 Media preparation, Inoculation and Plate count test

Apparatus:
1. Autoclave of sufficient size and number. Used for sterilization of media and
for discarded plates / used media, etc (with calibrated thermometer and
pressure gauge).
2. Anaerobic jars or incubators with equipment and material for obtaining
anaerobic conditions.
3. Balance sensitive to 0.1 g with 200 g load.
4. Blenders with steel jar and lid / Stomacher.
5. Bunsen burners.
6. Colony Counter (Quebec or equivalent).
7. Dilution and media storage bottles. 120, 300, 600 and 1500 ml in capacity.
8. Durham’s tubes
9. Glass test tubes 16 x150mm
Rimless
10.10.Plastic caps for test tubes
11.Serological test tubes
12. Hot air ovens used for sterilization of glass and metal ware. They should
have a thermostat range between 150-185oC.
13. Hockey sticks: Glass bent rods (or suitable plastic make) with fine polished
edges, 3-4 mm diameter, 15-20 cms long with angled spreading surface 45-
55 mm long or disposable plastic material.
14. Howard Mold Counting
 Chamber 15.Haemocytometer.
Medium:
1. Plate count agar;
2. Peptone water 0.1%,
3. Overlay Medium (Agar Medium)
Principle:
Plate count agar (PCA) is a general purpose growth medium commonly used to assess "total" or
“viable” bacterial growth of a water sample. The number of microorganisms per milliliter of
sample is calculated from the number of colonies obtained on PCA plate from selected dilution.
Poured plates are prepared using a specified culture medium and a specified quantity of the sample.
The plates are aerobically incubated at two different temperatures i.e. 37°C for 24 hr and 20 – 22°C
for 72 hr

Procedure:

1. Disinfect the surface of the bottle/pouch/cups containing sample with 70% ethanol.
Thoroughly mix the sample by vigorous shaking to achieve uniform distribution.

2. Aseptically inoculate 1ml of the water sample using sterile pipette into sterile petri plates in
duplicate in two sets. The petri plates should be labeled with the sample number, date and
any other desired information.

3. Pour into each plate 15–18 mL of the molten sterilized PCA media (cooled to 44°C–47°C).
Avoid pouring of molten medium directly onto the inoculum. Mix the media and inoculum
by swirling gently clockwise and anti-clockwise so as to obtain homogenous distribution of
inoculum in the medium.

4. Allow to cool and solidify. In case, where in sample microorganism having spreading
colonies is expected, add 4ml of overlay medium onto the surface of solidified plates.

5. After complete solidification, invert the prepared plates and incubate one set at 370C for 24
hr and other set at 20 – 22°C for 72 hr.

6. After specified incubation period count all colonies including pinpoint colonies. Spreading
colonies shall be considered as single colony. If less than one quarter of dish is overgrown by
spreading, count the colonies on the unaffected part of the dish and calculate corresponding
number in the entire dish. If more than one quarter is overgrown by spreading colonies
discard the plate.

7. Calculation & Expression of results:

Case 1: Plates having microbial count between 10 and 300cfu

[Colonies Plate1 + Colonies Plate 2]

N=-------------------------------------------
2
 Case 2: Plates having microbial count less than 10cfu but at least 4. Calculate the results as
given in Case 1.

Case 3: If microbial load is from 3 to 1 then reporting of results shall be: “Microorganisms
are present, but, less than 4 per mL”

Case 4: When the test sample/plates contains no colonies then reporting of results shall be:
“Less than 1 cfu/ml”.

EXP 8
MEMBRANE-FILTER TECHNIQUE

APPARATUS AND MATERIALS

1. Dilution bottles or tubes:

Use bottles or tubes of resistant glass, preferably borosili- cate glass, closed with
glass stoppers or screw caps equipped with liners that do not produce toxic or
bacteriostatic compounds on sterilization.

Do not use cotton plugs as closures. Mark graduation levels indelibly on side of
dilution bottle or tube. Plastic bottles of nontoxic material and acceptable size may
be substituted for glass, provided that they can be sterilized properly.

2. Pipets and graduated cylinders:

i. Use pipets of any convenient size, provided that they deliver

the required volume accurately and quickly. The error of calibration for a
given manufacturer's lot must not exceed 2.5%. Use pipets having
graduations distinctly marked and with unbroken tips. Bacteriological-
transfer pipets or pipets conforming to the APHA standards given in the
latest edition of Standard Methods for the Examination of Dairy Products
may be used. Optimally, protect the mouth end of all pipets by a cotton plug
to eliminate hazards to the worker or possible sample contamination by
saliva.

ii. Use graduated cylinders meeting ASTM Standards (D-86

and D216) and with accuracy limits established by the National Bureau of
Standards where appropriate.

3. Containers for culture medium:

 i. Use clean borosilicate glass flasks presterilized to reduce
bacterial contamination. Any size or shape of flask may be used, but
Erlenmeyer flasks with metal caps, metal foil covers, or screw caps provide
for adequate mixing of the medium and are convenient for storage.

4. Culture dishes
5. :

i. Use Petri-type dishes, 60 by 15 mm, 50 x 12 mm, or other

appropriate size. The bottoms of the dishes should be flat and large enough
so that the absorbent pads for the culture nutrient will lie flat. Wrap clean
culture dishes before sterilization, singly or in convenient numbers, in metal
foil if sterilized by dry heat, or in suitable paper substitute when autoclaved.
If glass Petri dishes are used, use borosilicate or equivalent glass. Because
covers for such dishes are loose fitting, take precautions to prevent possible
loss of medium by evaporation, with resultant change in medium
concentration, and to maintain a humid environment for optimal colony
development.

ii. Disposable plastic dishes that are tight fitting and meet the
specifications noted above also may be used. Suitable sterile plastic dishes
are available commercially.

SUMMARY OF METHOD

1. A predetermined amount of sample is filtered through a membrane filter which retains the
bacteria found in the sample.

2. In the two-step enrichment procedure, the filters containing bacteria are placed on an
absorbent pad saturated with lauryl tryptose broth and incubated at 35°C + 0.5°C for 2 hr.
The filters are then transferred to an absorbent pad saturated with M-Endo media or to a
dish containing M-Endo agar and incubated for another 21 + 1 hr at 35°C + 0.5°C. Sheen
colonies are then counted under magnification and reported per 100 mL of original sample.

3. A more detailed treatment of this method is presented in Standard Methods for the
Examination of Water and Wastewater and in Microbiological Methods for Monitoring the
Environment (see References, Section 10.0).

INTERFERENCES

1. The presence of residual chlorine or other halogen can prevent the continuation of
bacterial action. To prevent this occurrence, sodium thiosulfate should be added.
 2. Water samples high in copper, zinc, or other heavy metals can be toxic to bacteria.
Chelating agents such as ethylenediaminetetraacetic acid (EDTA) should only be
added when heavy metals are suspected of being present.

3. Turbidity caused by the presence of algae or other interfering material may not permit
testing of a sample volume sufficient to yield significant results. Low coliform
estimates may be caused by the presence of high numbers of noncoliforms or of toxic
substances.

4. Samples containing large amounts of suspended solids will interfere with colony
growth and with the subsequent counting of colonies on the filter membrane. When
this is the case, use Method 9131.

5. Disposable plastic dishes that are tight fitting and meet the specifications noted above
also may be used. Suitable sterile plastic dishes are available commercially.
Filtration units:

1. The filter-holding assembly (constructed of glass, auto- clavable plastic,

porcelain, or any noncorrosive bacteriologically inert metal) consists of a
seamless funnel fastened by a locking device or held in place by magnetic force
or gravity. The design should be such that the membrane filter will be held
securely on the porous plate of the receptacle without mechanical damage and
all fluid will pass through the membrane during filtration.

2. Separately wrap the two parts of the assembly in heavy wrapping paper for
sterilization by autoclaving and storage until use. Alternatively, treat
unwrapped parts by ultraviolet radiation before using them. Field units may be
sanitized by igniting methyl alcohol or immersing in boiling water for 5 min.
Do not ignite plastic parts.

For filtration, mount receptacle of filter-holding assembly in a 1-liter

filtering flask with a side tube or other suitable device such that a pressure
differential can be exerted on the filter membrane. Connect flask to an electric
vacuum pump, a filter pump operating on water pressure, a hand aspirator, or
other means of securing pressure differential. Connect an additional flask
between filtering flask and vacuum source to trap carry- over water.

3. Filter membranes:

i. Use membrane filters with a rated pore diameter such

that there is complete retention of coliform bacteria (0.45 + 0.02
um). Use only those filter membranes that have been found,
through adequate quality control testing and certification by the
 manufacturer, to exhibit full retention of the organisms to be
cultivated, stability in use, freedom from chemical extractables
inimical to the growth and development of bacteria, a
satisfactory speed of filtration, no significant influence on
medium pH, and no increase in number of confluent colonies or
spreaders. Preferably, use membranes grid-marked in such a
manner that bacterial growth is neither inhibited nor stimulated
along the grid lines. Store membrane filters held in stock in an
environment without extremes of temperature and humidity.
Obtain no more than a year's supply at any one time.

ii. If presterilized membrane filters are to be used, use

those for which the manufacturer has certified that the
sterilization technique has neither induced toxicity nor altered
the chemical or physical properties of the membrane. If the
membranes are sterilized in the laboratory, remove the paper
separators -- but not the absorbent paper pads -- from the
packaged filters. Divide filters into groups of 10 to 12, or other
convenient units, and place in 10-cm Petri dishes or wrap in
heavy wrapping paper. Autoclave for 10 min at 121°C. At the
end of the sterilization period, let the steam escape rapidly to
minimize accumulation of water condensation on filters.
b. Absorbent pads:

i. Absorbent pads consist of disks of filter paper or

other material known to be of high quality and free of sulfites or
other substances that could inhibit bacterial growth. Use pads
approximately 48 mm in diameter and of sufficient thickness to
absorb 1.8 to 2.2 mL of medium. Presterilized absorbent pads or
pads subsequently sterilized in the laboratory should release less
than 1 mg total acidity (calculated as CaCO3) when titrated to
the phenolphthalein end point, pH 8.3, using 0.02 N NaOH.
Where there is evidence of absorbent pad toxicity, presoak pads
in Type II water at 121°C (in an autoclave) for 15 min, decant
the water, and repackage pads in a large Petri dish for
sterilization and subsequent use. Sterilize pads simultaneously
with membrane filters available in resealable Kraft envelopes or
separately in other suitable containers. Dry pads so they are free
of visible moisture before use. See sterilization procedure
described above for membrane filters.

ii. As a substrate substitution for nutrient-saturated

absorbent pads, 1.5% agar may be added to the total coliform M-
Endo broth medium.
 c. Forceps:

i. Forceps should be round-tipped, without corrugations

on the inner sides of the tips. Sterilize before use by dipping in
95% ethyl or absolute methyl alcohol and flaming.

d. Incubators

Use incubators to provide a temperature of 35 + 0.5°C and to maintain a high level of humidity
(approximately 90% relative humidity).

e. Microscope and light source:

Count membrane-filter colonies with a magnification of 10 to15 diameters and a light source adjusted
to give maximum sheen discernment. Optimally, use a binocular wide-field dissecting microscope.
However, a small fluorescent lamp with magnifier is acceptable. Use cool-white fluorescent lamps. Do not
use a microscope illuminator with optical system for light concentration from an incandescent light source
for coliform colony identification on Endo-type media.

REAGENTS

M-Endo medium:

Components of the medium are:

Tryptose or polypeptone 10.0 g

Thiopeptone or thiotone 5.0 g
Casitone or trypticase 5.0 g
Yeast extract 1.5 g
Lactose 12.5 g
Sodium chloride, NaCl 5.0 g
Dipotassium hydrogen
phosphate, K2HPO4 4.375 g
Potassium dihydrogen
phosphate, KH2PO4 1.375 g
 Sodium lauryl sulfate 0.050 g
Sodium desoxycholate 0.10 g
Sodium sulfite, Na2SO3 2.10 g
Basic fuchsin 1.05 g
Distilled (Type II) water 1 liter

Rehydrate in 1 liter Type II water containing 20 mL 95% ethanol. Heat to boiling in a water bath
to avoid degradation of carbohydrates, promptly remove from heat, and cool to below 45°C. Do
not sterilize by autoclaving. Final pH should be between 7.1 and 7.3.

Store finished medium in the dark at 2 to 10°C and discard any unused medium after 96 hr.
Medium is light sensitive.
NOTE: This medium may be solidified by adding 1.2% to 1.5% agar before boiling. Lauryl
tryptose broth.

SAMPLE COLLECTION, PRESERVATION, AND HANDLING

1. All samples must have been collected using a sampling plan.

2. Clean all glassware thoroughly with a suitable detergent and hot water, rinse with hot
water to remove all traces of residual washing compound, and finally, rinse with distilled
(Type II) water. If mechanical glassware washers are used, equip them with influent
plumbing of stainless steel or other nontoxic material. Do not use copper piping to
distribute Type II water. Use stainless steel or other nontoxic material for the rinse-water
system.
3. Sterilize glassware, except when in metal containers, for not less than 60 min at a
temperature of 170°C, unless it is known from recording thermometers that oven
temperatures are uniform, under which exceptional condition use 160°C. Heat glassware in
metal containers to 170°C for not less than 2 hr.
4. Sterilize sample bottles not made of plastic, as above, or in an autoclave at 121°C for 15
min.
5. For plastic bottles that distort on autoclaving, use low- temperature ethylene oxide gas
sterilization. If water containing residual chlorine and other halogens is to be collected, add
sufficient Na2S2O3 to clean sample bottle before sterilization to give a concentration of
about 100 mg/L in the sample. To a 120-mL bottle add 0.1 mL 10% solution of
Na2S2O3 (this will neutralize a sample containing about 15 mg/L residual
chlorine). Stopper bottle, cap, and sterilize by either dry or moist heat, as directed
previously.

6. Collect water samples high in copper or zinc and wastewater samples high in heavy metals
in sample bottles containing a chelating agent that will reduce metal toxicity. This is
particularly significant when such samples are in transit for 4 hr or more. Use 372 mg/L of
the tetrasodium salt of ethylenediaminetetraacetic acid (EDTA). Adjust EDTA solution to
pH 6.5 before use. Add EDTA separately to sample bottle before bottle sterilization (0.3
mL 15% solution in a 120-mL bottle) or combine it with the Na2S2O3 solution before
addition.

7. When the sample is collected, leave ample air space in the bottle (at least 2.5 cm) to
facilitate mixing by shaking, preparatory to examination. Be careful to take samples that
 will be representative of the water being tested and avoid sample contamination at time of
collection or in period before examination.

a. Keep sampling bottle closed until the moment it is to be filled. Remove

stopper and hood or cap as a unit, taking care to avoid soiling. During sampling, do not
handle stopper or cap and neck of bottle, and protect them from contamination. Hold
bottle near base, fill it without rinsing, replace stopper or cap immediately, and secure
hood around neck of bottle.

b. Start bacteriological examination of a water sample promptly after

collection to avoid unpredictable changes. If samples cannot be processed within
1 hr of collection, use an iced cooler for storage during transport to the laboratory.

c. Hold temperature of all stream pollution samples below 10°C during a

maximum transport time of 6 hr. refrigerate these samples upon receipt in the laboratory
and process within 2 hr. When local conditions necessitate delays in delivery of samples
longer than 6 hr, make field examinations using field laboratory facilities located at the
site of collection or use delayed-incubation procedures.
7.0 PROCEDURES

7.1 Selection of sample size:

7.1.1 Size of sample will be governed by expected bacterial density, which in

finished-water samples will be limited only by the degree of turbidity.

7.1.2 An ideal sample volume will yield growth of about 50 coliform colonies
and not more then 200 colonies of all types. Examine finished waters by filtering duplicate
portions of the same volume, such as 100 to
500 mL or more, or by filtering two diluted volumes. Examine other waters by filtering
three different volumes, depending on the expected bacterial density. When less than 20
mL of sample (diluted or undiluted) is filtered, add a small amount of sterile dilution water
to the funnel before filtration. This increase in water volume aids in uniform dispersion of
the bacterial suspension over the entire effective filtering surface.

7.2 Filtration of sample:

7.2.1 Using sterile forceps, place a sterile filter over porous plate of receptacle,
grid side up. Carefully place matched funnel unit over receptacle and lock it in place. Filter
sample under partial vacuum. With filter still in place, rinse funnel by filtering three 20- to
30-mL portions of sterile dilution water. Unlock and remove funnel, immediately remove
filter with sterile forceps, and place it on sterile pad or agar with a rolling motion to avoid
entrapment of air.

7.2.2 Use sterile filtration units at the beginning of each filtration series as a
minimum precaution to avoid accidental contamination. A filtration series is considered to
be interrupted when an interval of 30 min or longer elapses between sample filtrations.
After such interruption, treat any further sample filtration as a new filtration series and
sterilize all membrane-filter holders in use.

7.2.3 Decontaminate this equipment between successive filtrations by use of

flowing steam, boiling water, or, if available, an ultraviolet sterilizer. When using the UV
sterilization procedure, a 2-min exposure to UV radiation is sufficient and should kill
99.9% of all bacteria. Eye protection is recommended to protect against stray radiation
from a non- light-tight sterilization cabinet. This UV equipment is not commercially
available and is not required, although its use is recommended.

7.3 Two-step enrichment technique:

7.3.1 Place a sterile absorbent pad in the upper half of a sterile culture dish and
pipet enough enrichment medium (1.8 to 2.0 mL lauryl tryptose broth) to saturate pad.
Carefully remove any surplus liquid.
Aseptically place filter through which the sample has been passed on pad. Incubate filter,
without inverting dish, for 1.5 to 2 hr at 35 + 0.5°C in an atmosphere of at least 90%
relative humidity.

7.3.2 Remove enrichment culture from incubator, lift filter from enrichment
pad, and roll it onto the agar surface. Incorrect filter placement is at once obvious, because
patches of unstained membrane indicate entrapment of air. Where such patches occur,
carefully reseat filter on agar surface. If the liquid medium is used, prepare final culture by
removing enrichment culture from incubator and separating the dish halves. Place a fresh
sterile pad in bottom half of dish and saturate it with 1.8 to 2.0 mL of final M-Endo
medium. Transfer filter, with same precautions as above, to new pad. Discard used pad.
With either the agar or the liquid medium, invert dish and incubate for 20 to 22 hr at 35 +
0.5°C.

7.4 Counting:

7.4.1 The typical coliform colony has a pink to dark-red color with a metallic
surface sheen. The sheen area may vary in size from a small pinhead to complete coverage
of the colony surface. Count sheen colonies with the aid of a low-power (10 to 15
magnifications) binocular wide-field dissecting microscope or other optical device, with a
cool-white fluorescent light source directed above and as nearly perpendicular as possible
to the plane of the filter. The total count of colonies (coliform and noncoliform) on Endo-
type medium has no relation to the total number of bacteria present in the original sample
and, so far as is known, no significance can be inferred or correlation made with the quality
of the water sample.

7.5 Calculation of coliform density:

7.5.1 Report coliform density as (total) coliforms/100 mL. Compute the count,
using membrane filters with 20 to 80 coliform colonies and not more than 200 colonies of
all types per membrane, by the following equation:

(Total) coliform colonies/100 mL: coliform colonies counted* 100 / mL sample filtered

7.5.2 Water of drinking-water quality:

7.5.2.1 With water of good quality, the number of coliform colonies will be
less than 20 per membrane. In this event, count all coliform colonies and use the
formula given above to obtain coliform density.

7.5.2.2 If confluent growth occurs, that is, growth covering either the entire
filtration area of the membrane or a portion thereof, and colonies are not discrete,
report results as "confluent growth with or without coliforms." If the total number
of bacterial
 colonies, coliforms plus noncoliforms, exceeds 200 per membrane, or if the
colonies are too indistinct for accurate counting, report results as "too numerous to
count" (TNTC). In either case, request a new sample and select more appropriate
volumes to be filtered per membrane, remembering that the standard drinking-water
portion is 100 mL. Thus, instead of filtering 100 mL per membrane, 50-mL portions
may be filtered through each of two membranes, 25-mL portions may be filtered
through each of four membranes, etc. Total the coliform counts observed on the
membranes and report as number per 100 mL.

7.5.3 Water of other than drinking-water quality:

7.5.3.1 As with potable water samples, if no filter has a coliform count

falling in the ideal range, total the coliform counts on all filters and report as
number per 100 mL. For example, if duplicate 50-mL portions were examined and
the two membranes had five and three coliform colonies, respectively, report the
count as eight coliform colonies per 100 mL, i.e.,

(5 + 3) x 100
(50 + 50)
7.5.3.2 Similarly, if 50-, 25-, and 10-mL portions were examined and the
counts were 15, 6, and 1 coliform colonies, respectively, report the count as 25/100
mL, i.e.,
(15 + 6) x 100
(50 + 25 + 10)
7.5.3.3 On the other hand, if 10-, 1.0-, and 0.1-mL portions were examined
with counts of 40, 9, and 1 coliform colonies respectively, select only the 10-mL
portion for calculating the coliform density because this filter had a coliform count
falling in the ideal range. The result is 400/100 mL, i.e.,
(40 x 100)
10
In this last example, if the membrane with 40 coliform colonies also had a total
bacterial colony count greater than 200, report the coliform count as 400/100 mL.

7.5.4 Statistical reliability of membrane filter results: Although the statistical

reliability of the membrane filter technique is greater than that of the MPN procedure,
membrane counts really are not absolute numbers. Table 1 illustrates some 95%
confidence limits.
 TABLE 1. 95% CONFIDENCE LIMITS FOR MEMBRANE-FILTER
RESULTS USING 100-mL SAMPLE

Number of Coliform 95% Confidence Limits

Colonies Counted Lower Upper
1 0.05 3.0
2 0.35 4.7
3 0.81 6.3
4 1.4 7.7
5 2.0 9.2

8.0 QUALITY CONTROL

8.1 Extensive quality control procedures are provided in Part IV of U.S. EPA, 1978
(see Section 10.0, References). These procedures should be adhered to at all times.

8.2 Samples must be maintained as closely as possible to original condition by careful

handling and storage. Sample sites and sampling frequency should provide data representative of
characteristics and variability of the water quality at that site. Samples should be analyzed
immediately. If this is not practical, they should be refrigerated at a temperature of 1-4°C and
analyzed within 6 hr.

8.3 Quality control of culture media is critical to the validity of microbiological

analysis. Some important factors to consider are summarized below:

8.3.1 Order media to last for only 1 yr; always use oldest stock first. Maintain
an inventory of all media ordered, including a visual inspection record.

8.3.2 Hold unopened media for no longer than 2 yr. Opened media containers
should be discarded after 6 months.

8.3.3 When preparing media, keep containers open as briefly as possible.

Prepare media in deionized or distilled (Type II) water of proven quality. Check the pH of
the media after solution and sterilization; it should be within 0.2 units of the stated value.
Discard and remake if it is not.

8.3.4 Autoclave media for the minimal time specified by the manufacturer,
because the potential for damage increases with increased exposure to heat. Remove sterile
media from the autoclave as soon as pressure is zero. Effectiveness of the sterilization
should be checked weekly, using strips or ampuls of Bacillus stearothemophelus.
 8.3.5 Agar plates should be kept slightly open for 15 min after pouring or
removal from refrigeration to evaporate free moisture. Plates must be free of lumps,
uneven surfaces, pock marks, or bubbles, which can prevent good contact between the agar
and medium.

8.3.6 Quality control checks of prepared media should include the incubation
of 5% of each batch of medium for 2 days at 35°C to inspect for growth and
positive/negative checks with pure culture.

8.4 Analytical quality control procedures should include:

8.4.1 Duplicate analytical runs on at least 10% of all known positive samples
analyzed.

8.4.2 At least one positive control sample should be run each month for each
parameter tested.

8.4.3 At least one negative (sterile) control should be run with each series of
samples using buffered water and the medium batch used at the beginning of the test series
and following every tenth sample. When sterile controls indicate contamination, new
samples should be obtained and analyzed.

8.4.4 The Type II water used should be periodically checked for

contamination.

8.5 Quality control specifications for membrane filters:

8.5.1 Membrane filters can be purchased sterile or packaged for sterilization.

They can be sterilized by autoclaving, ethylene oxide, or irradiation. Membrane
manufacturers should certify that their membranes meet stated specifications on sterility,
retention, recovery, pore size, flow rate, pH, total acidity, phosphate, and other
extractables.

8.5.2 Membrane performance should be tested to ensure proper results. Each

lot ordered should be inspected for proper shape, grid lines, diffusability, and correct
colony development. Membranes containing sizable areas with no colony development are
questionable.
EXP 10: Noise Isopleths in Institution or Industry

1. Aim: Noise Isopleths in Institution or Industry

2. Equipment
Sound level meter and calibrator, sound source, measuring tape, markers

3. Measurements using sound power source

 3.1 Do the battery test and performance check of the SLM.
3.2 Set the sound power source in the centre of the class room and, using the wideband spectrum, adjust the

output to a convenient level.

3.3 Measure the noise level in dB(A) at 1m, 2m and 4m from the center of the source.

3.4 Determine the linear frequency spectrum at 2m from the center of the source.

3.5 Do not adjust the output setting! But do either part a) or part b)

a) relocate the sound source to a more reverberant space such as stair well or foyer OR

b) relocate the sound source to a less reverberant space such as in the open well away from reflecting surfaces.

3.6 Measure the noise level in dB(A) at 1m, 2m and 4m from the center of the source.

4. Report
Produce a report summarizing the results of the measurements and include:

4.1 A summary table showing the change in sound pressure level with distance in the two environments.

Comment on your findings and compare with general guidelines on reduction with distance from a source.

4.2 A calculation of the overall A weighted level and comparison with the value measured at that same

location.

4.3 A chart of the octave band frequency spectrum in terms of dB. Use the values for the A weighting to

calculate the A weighted frequency spectrum. Plot these A weighted values on the chart and comment on the shape of

the two curves.

Report on Basic Sound Level Measurement

Sound Source: ……………………………………………………………………………

Sound Level Meter:

Distance 1m 2m 4m

Sound Pressure
Level, dB(A) in
classroom

Sound Pressure
Level, dB(A) in
alternate space

EXP 11 TCLP – Leachate from Landfills

Aim: TCLP – Leachate from Landfills, Toxicity Characteristic Leaching Procedure Method Description and
Sample Preparation. The Toxicity Characteristic Leaching Procedure (TCLP).

EQUIPMENT/APPARATUS

The following are standard materials and equipment required for soil pH determination:
1. Agitation Apparatus - The agitation apparatus will be capable of rotating the extraction vessels in an end over
end fashion at 30 ± 2 rpm.
2. Extraction Vessels
a. Inorganic Analytes - Borosilicate glass bottles or polyethylene bottles shall be used for either organic
or inorganic analytes.
b. Organic Analytes - Borosilicate glass bottles shall be used for organic analytes.
3. Filtration Device, a 316 stainless steel or polytetrafluoroethylene (PTFE) lined pressure filtration device (filter
holder) will be used to filter all samples and extracts. This device will be capable of holding an internal
pressure of 100 psi.
4. Vacuum/Pressure Pump, a dedicated vacuum/pressure pump for each filtration device. The pressure pump
will be used to provide 60 ± 5 psi to the filtration device for pressure filtering of all samples and extracts.
5. Filters, 0.7 µm borosilicate glass. When evaluating the mobility of metals, each filter shall be acid washed
prior to use by rinsing with 1N nitric acid followed by three consecutive rinses with deionized distilled water
(a minimum of 1 L per rinse). Glass fiber filters are fragile and should be handled with care.
6. pH Meter, accurate to ± 0.05 units at 25oC
7. Laboratory balance, accurate to within ± 0.01 grams
8. Beakerlenmeyer flask, glass, 500 mL
9. Watch glasses, appropriate diameter to cover beaker or Erlenmeyer flask.
10. Magnetic Stirrer
 11. Extraction Fluid Containers, 20-Liter, glass construction for inorganic or organic analysis.
Polyethylene containers may only be used for inorganic analytes.

REAGENTS

1. Reagent grade chemicals shall be used in all tests. Unless otherwise indicated, all chemicals will conform to the
specifications of the Committee on Analytical Reagents of the American Chemical Society (ACS), where such
specifications are available. Other grades may be used, provided it is first ascertained that the reagent is of
sufficiently high purity to permit its use without lessening the accuracy of the determination.

2. Deionized (DI) water, prepared using an appropriate filtration purification system, capable of producing
American Society of Testing and Materials (ASTM) Type II water or equivalent
3. Hydrochloric acid (1N) HCl, made from ACS reagent grade
4. Nitric acid (1N) HNO3, made from ACS reagent grade
5. Sodium Hydroxide (1N), NaOH, made from ACS reagent grade
6. Glacial acetic acid, CH3CH2OOH, ACS reagent grade
7. Extraction Fluid, prepared in 1-L batches or it may be made in larger batches as required by the total number of
extractions required. For larger batches, the following procedures should be scaled by the total number of
required extractions.
a. Extraction Fluid #1 - Add 5.7 mL of glacial CH3CH2OOH to 500 mL of reagent water. Add 64.3 ml of
1N NaOH, and dilute to a volume of 1L. When correctly prepared, the pH of this fluid will be 4.93 ± 0.05.
b. Extraction Fluid #2 - Dilute 5.7 mL of glacial CH3CH2OOH with reagent water to a volume of 1L.
When correctly prepared, the pH of this fluid will be 2.88 ± 0.05.

1.0 PROCEDURES

1.1 Preliminary Evaluations

Preliminary evaluations shall be performed on a minimum 100 gram aliquot of waste. This
aliquot may not actually undergo extraction. These preliminary evaluations include: (1)
determination of the percent solids (Section 7.1.1.); (2) determination of whether the waste
contains insignificant solids and is, therefore, its own extract after filtration (Section 7.1.2); (3)
determination of whether the solid portion of the waste requires particle size reduction (Section 7.1.3); and (4)
determination of which of the two extraction fluids are to be used for the TCLP extraction of the waste (Section
7.1.4).

1. Preliminary determination of percent solids: Percent solids is defined as that fraction of a waste sample (as a
percentage of the total sample) from which no liquid may be forced out by an applied pressure, as described
below.

a. If the waste will obviously yield no liquid when subjected to pressure filtration (i.e.,is 100% solids proceed to
section 7.1.3.

b. If the sample is liquid or multi-phase, liquid/solid separation to make a preliminary

determination of percent solids is required. This involves the filtration device
described in Section 5.3 and outlined in Sections 7.1.1.3 through 7.1.1.9.

c. Pre-weigh the filter and the container that will receive the filtrate.

a. Assemble the filter holder and filter following the manufacturers’ directions. Place
the filter on the support screen and secure.

b. Weigh out a sub-sample of the waste (100 gram minimum) and record the weight.

c. Allow slurries to stand to permit the solid phase to settle. Wastes that settle slowly
may be centrifuged prior to filtration. Centrifugation is to be used only as an aid to
filtration. If used, the liquid should be decanted and filtered followed by filtration of
the solid portion of the waste through the same filtration system.

d. Transfer the waste sample to the filter holder (liquid and solid phases). Spread the
sample evenly over the surface of the filter. If filtration of the waste at 4 oC reduces
the amount of expressed liquid over what would be expressed at room temperature,
then allow the sample to warm to room temperature in the device before filtering.

Gradually apply gentle pressure of 1-10 psi, until air or pressurizing gas moves
through the filter. If this point is not reached under 10 psi, and if no additional liquid
has passed through the filter in any 2 minute interval, slowly increase the pressure in
10 psi increments to a maximum of 50 psi. After each incremental increase of 10
psi, if the pressurizing gas has not passed through the filter, and if no additional
liquid has passed through the filter in any 2 minute interval, proceed to the next 10
psi increment. When the pressurizing gas begins to move through the filter, or when
liquid flow has ceased at 50 psi (i.e., filtration does not result in any additional
filtrate within any 2 minute period), stop the filtration.

NOTE: Instantaneous application of high pressure can degrade the glass fiber filter
and may cause premature plugging.

e. The material in the filter holder is defined as the solid phase of the waste, and the
filtrate is defined as the liquid phase.

NOTE: Some wastes, such as oily wastes and some paint wastes, will obviously
contain some material that appears to be a liquid. Even after applying pressure
filtration, as outlined in Section 7.1.1.7, this material may not filter. If this is the
case, the material within the filtration device is defined as a solid. Do not replace the
original filter with a fresh filter under any circumstances. Use only one filter.

f. Determine the weight of the liquid phase by subtracting the weight of the filtrate
container (see Section 7.1.1.3) from the total weight of the filtrate filled container.
Determine the weight of the solid phase of the waste sample by subtracting the
weight of the liquid phase from the total waste sample, as determined in 7.1.1.5.

Record the weight of the liquid and solid phases. Calculate the percent solids as
follows:

Percentage of solids = Weight of solids x 100 / Total Weight of Waste

2. If the percent solids determined in Section 7.1.1.9 is equal to or greater than 0.5%, then
proceed to Section 7.1.3 to determine whether the solid material requires particle size
reduction. If the percent solids determined in Section 7.1.1.9 is less than 0.5% then
proceed to Section 7.2.9 to perform the TCLP.

3. Determination of whether the waste requires particle size reduction - Any material which
does not pass through a 9.5mm (0.375 inch) standard sieve requires size reduction. The
material should be prepared for extraction by crushing, cutting or grinding the waste to a
particle size as described above.

4. Determination of appropriate extraction fluid - If the solid content of the waste is greater
than or equal to 0.5%, determine the extraction fluid as follows:
 Weigh out a small sub-sample of the solid phase of the waste, reduce the solid to a
particle size of approximately 1mm or less, and transfer 5.0 grams of the solid phase
of the waste to a 500 mL beaker or Erlenmeyer flask.
 Add 96.5 mL of reagent water to the beaker, cover with a watch glass, and stir
vigorously for 5 minutes using a magnetic stirrer. Measure and record the pH. If the
pH is <5.0, use extraction fluid #1. Proceed to Section 7.2.
 If the pH is >5.0, add 3.5 mL 1N HCl, slurry briefly, cover with a watch glass, heat
to 50 oC, and hold at 50 oC for 10 minutes.
 Let the solution cool to room temperature and record the pH. If the pH is <5.0, use
extraction fluid #1. If the pH is >5.0, use extraction fluid #2. Proceed to Section 7.2.

5. If the aliquot of waste used for the preliminary evaluation (Sections 7.1.1 - 7.1.4) was
determined to be 100% solid at Section 7.1.1.1, then it can be used for the Section 7.2
extraction (assuming at least 100 grams remain).

Procedure When Volatiles are Not Involved

A minimum sample size of 100 grams (solid and liquid phases) is recommended. In some cases, a larger sample
may be appropriate, depending on the solids content of the waste sample (percent solids, see Section 7.1.1),
whether the initial liquid phase of the waste will be miscible with the aqueous extract of the solid, and whether
inorganics, semi-volatile organics, pesticides, and herbicides are all analytes of concern. Enough solids should be
generated for extraction such that the volume of TCLP extract will be sufficient to support all of the analyses
required. If the amount of extract generated by a single TCLP will not be sufficient to perform all of the analyses,
more than one extraction may be performed and the extracts from each combined and aliquoted for analysis.

1. If the waste is 100% solids, weigh out a sub-sample of the waste (100 gram minimum)
and proceed to Section 7.2.9.
2. If the waste is liquid or multi-phase, liquid/solid separation is required. This involves the
filtration device described in Section 5.3.2 and is outlined in Sections 7.2.3 to 7.2.8
3. Pre-weigh the container that will receive the filtrate.
4. Assemble the filter holder and filter following the manufacturer’s instructions. Place the
filter on the support screen and secure. Acid wash the filter if evaluating the mobility of
metals (see Section 5.4).
5. Weigh out a sub-sample of the waste (100 gram minimum) and record the weight.
If the waste contains <0.5% dry solids (Section 7.1.2), the liquid portion of the waste,
after filtration, is defined as the TCLP extract. Therefore, enough of the sample should
be filtered so that the amount of filtered liquid will support all of the analyses required of
the TCLP extract. For wastes containing >0.5% dry solids, use the percent solids information obtained in Section
7.1.1 to determine the amount of sample to be filtered to produce 100 grams of solids per required TCLP
extraction.
6. Allow slurries to stand to permit the solid phase to settle. Wastes that settle slowly may
be centrifuged prior to filtration. Use centrifugation only as an aid to filtration. If the
waste is centrifuged, the liquid should be decanted and filtered followed by filtration of
the solids portion of the waste through the same filtration system.
7. Transfer the waste sample to the filter holder (liquid and solid phases). Spread the
sample evenly over the surface of the filter. If filtration of the waste at 4 oC reduces the
amount of expressed liquid over what would be expressed at room temperature, then
allow the sample to warm to room temperature in the device before filtering.
Gradually apply gentle pressure of 1-10 psi, until air or pressurizing gas moves through
the filter. If this point is not reached under 10 psi, and if no additional liquid has passed
through the filter in any 2 minute interval, slowly increase the pressure in 10 psi
increments to a maximum of 50 psi. After each incremental increase of 10 psi, if the
pressurizing gas has not passed through the filter, and if no additional liquid has passed
through the filter in any 2 minute interval, proceed to the next 10 psi increment. When
the pressurizing gas begins to move through the filter, or when liquid flow has ceased at
50 psi (i.e., filtration does not result in any additional filtrate within any 2 minute period),
stop the filtration.

NOTE: Instantaneous application of high pressure can degrade the glass fiber filter and
may cause premature plugging.

8. The material in the filter holder is defined as the solid phase of the waste, and the filtrate
is defined as the liquid phase. Weigh the filtrate. The liquid phase may now be either
analyzed (See section 7.2.12) or stored at 4 oC until time of analysis.

NOTE: Some wastes, such as oily wastes and some paint wastes, will obviously contain
some material that appears to be a liquid. Even after applying pressure filtration, as
outlined in Section 7.1.1.7, this material may not filter. If this is the case, the material
within the filtration device is defined as a solid. Do not replace the original filter with a
fresh filter under any circumstances. Use only one filter.

9. If the waste contains <0.5% dry solids (See section 7.1.2), proceed to Section 7.2.13. If
the waste contains >0.5% dry solids (see Section 7.1.1 or 7.1.2), and if particle size
reduction of the solid was needed in Section 7.1.3, proceed to Section 7.2.10. If the
waste as received passes a 9.5 mm sieve, quantitatively transfer the solid material into the
extractor bottle along with the filter used to separate the initial liquid from the solid
phase, and proceed to 7.2.11.

10. Prepare the solid portion of the waste for extraction by crushing, cutting, or grinding the
waste to a particle size as described in Section 7.1.3. When the particle size has been
appropriately altered, transfer the solid material into an extractor bottle. Include the filter
used to separate the initial liquid from the solid phase.

11.Determine the amount of extraction fluid to add to the extractor vessel as follows:

Weight of Extraction Fluid = 20 * %Solids Weight of Waste Filtered / 100

Slowly add this amount of appropriate extraction fluid (see Section 7.1.4) to the extractor
vessel. Apply Teflon tape to the threads of the bottle, and close tightly. Secure in the
rotary agitation device, and rotate at 30 ± 2 rpm for 18 ± 2 hours. Ambient temperature
of the room shall be maintained at 23 ± 2 oC during the extraction period.
NOTE: As agitation continues, pressure may build up within the extractor bottle for
some types of wastes (e.g., limed or calcium carbonate containing waste may evolve
gases such as carbon dioxide). To relieve excess pressure, the extractor bottle may be
periodically opened (e.g., after 15 minutes, 30 minutes, and 1 hour) and vented into a
hood.

12. Following the 18 ± 2 hour extraction, separate the material in the extractor vessel into its
component liquid and solid phases by filtering through a new glass fiber filter, as outlined
in Section 7.2.7. For final filtration of the TCLP extract, the glass fiber filter may be
changed, if necessary, to facilitate filtration. Filter(s) shall be acid washed (see Section
4.4) if evaluating the mobility of metals.

13. Prepare the TCLP extract as follows:

 If the waste contained no initial liquid phase, the filtered liquid material obtained
from Section 7.2.12 is defined as the TCLP extract. Proceed to Section 7.2.14.
 If compatible (e.g., multiple phases will not result on combination), combine the
filtered liquid resulting from Section 7.2.12 with the initial liquid phase of the waste
obtained in Section 7.2.7. This combined liquid is defined as the TCLP extract.
Proceed to Section 7.2.14.
 If the initial liquid phase of the waste, as obtained from Section 7.2.7, is not or may
not be compatible with the filtered liquid resulting from Section 7.2.12, do not
combine these liquids. Submit these two liquids separately, and notify the laboratory
that they are both considered the TCLP extract for that sample.

 Following collection of the TCLP extract, the pH of the extract should be recorded.
Immediately aliquot and preserve the extract for analysis. Metals aliquots must be
acidified with nitric acid to pH <2. If precipitation is observed upon addition of
nitric acid to a small aliquot of the extract, then the remaining portion of the extract
for metals analyses shall not be acidified and the extract shall be analyzed as soon as
possible. All other aliquots must be stored under refrigeration (4 oC) until analyzed.

CALCULATIONS

This section is not applicable to this SOP.

 All data must be documented on TCLP Extraction Log/TCLP Extraction Fluid Preparation Log data sheets
(Appendices A and B, respectively) or in site or laboratory notebooks.
 All instrumentation must be operated in accordance with the manufacturer’s instructions. Equipment check-
out procedures and calibration activities must be performed prior to commencing this procedure.
 A minimum of one blank (using the same extraction fluid as used for the samples) must be analyzed for
every 20 extractions that have been conducted in the extraction vessel.
 Duplicate samples should be processed with the frequency of one in twenty samples. Duplicate. Samples
will be used to determine precision.
 Samples must undergo TCLP extraction within the following time periods:

SAMPLE MAXIMUM HOLDING TIMES (DAYS)

 From: Field From: TCLP From: Total
collection Extraction Preparative Elapse
To: TCLP To: Extraction d
Extraction Preparative To: Time
Extraction Determinative
Analysis

Semi 14 NA 40 61
volatiles

Mercury 28 NA 28 56

Metals 180 NA 180 360

(except
mercury)

RESULTS

For the pH meter, ± 0.1 pH unit represents the limit of accuracy under normal conditions, especially for
measurement of water and poorly buffered solutions (1)

Sample pH

EXP 12

Aim: Micrometeorology –Wind Direction, Wind speed, Humidity Temperature, Rainfall.

Wind Speed and Wind Direction Measurement

Wind measurements are of primary importance in the diffusion and transport of atmospheric
pollutants. These measurements include wind speed, wind direction, and turbulence or gustiness.
There are many wind-measuring systems commercially available. Some are ruggedly
constructed, designed for a wide range of applications, and require minimum attention. The more
delicate instruments, such as those used for measuring small-scale turbulence, can only be used
during periods of favorable weather.

Almost any anemometer or wind vane will provide some information on wind characteristics.
However, the quality of wind data depends directly upon how well the sensor is maintained and
how the measuring equipment functions as a system. Not only must the dynamic characteristics
of the sensor match program data requirements, but the sensor must also interface without
degradation of important performance characteristics with the total data acquisition system,
which may include the transducer, signal conditioner, telemetry, data processor, and readout
device.

Types of Sensors/Instrumentation

1. Anemometers - A number of wind speed sensors operating on a variety of physical

principles are available commercially. The rotational cup and propeller anemometers are
the most commonly employed wind speed sensors. Some sensors, called sonic
anemometers, use ultrasound to determine horizontal wind speed and direction. This type
of sensor’s measurement is based on transit time, the time it takes for the ultrasound to
travel from one transducer to another. Sensors with other designs are generally used in
specialized studies.

2. Wind Vanes - Wind-direction-measuring sensors are operated by wind exerting pressure on

a surface that rotates about a fulcrum. The standard wind vane measures only the horizontal
wind direction, but a bi-directional vane is free to move through 360 degrees horizontally
as well as ±50 degrees or more from the horizontal. The shape and design of the vane
surface may vary with the manufacturer. As with the wind speed, the ultrasound sensor will
also measure wind direction.

3. Combination Wind Sensors - Two types of sensors incorporate direction and speed
measuring capability in a single mechanical device. The propeller vane sensor
measures two-dimensional wind (horizontal) and the propeller bi-vane sensor measures
three- dimensional wind (horizontal and vertical).

4. Wind Component Anemometers - These can be used to determine the wind speed and
direction(s), using simple trigonometry. These instruments include the x, y, z prop (often
called u, v, w), the sonic, the vortex shedding, and the ion flow anemometers.

5. Transducers - Transducers convert the parameter being measured by a sensor into an

electrical signal. This chapter will discuss transducers used in rotation-type wind speed
sensors and vane type direction sensors only.

The rotary motion of wind speed cups and propellers is most often converted to a voltage or a
frequency. Both alternating current (AC) and direct current (DC) generators are used, but the
latter is used more often. Frequency-type devices, sometimes called light choppers, have
advantages in that they are almost frictionless, operate at lower wind speeds, and produce signals
that can be transmitted without loss over long distances. Typically, this type of transducer is
made to interrupt the light of a Light-Emitting Diode (LED) at a rate of 1 to 132 times for each
rotation of the sensor. Units that interrupt the light once per revolution of the anemometer shaft
are usually used to measure wind run (the distance or length of flow of the air past a point during
a given interval of time), thus producing longer time averages. A single sealed glass switch, used
in combination with a magnet on the shaft of the anemometer, is also a frequency-type
transducer for wind speed or wind run. Another frequency-type device now used in anemometers
is the Hall-effect generator. This works using electrical polarization of a conducting plate
moving through a magnetic field. Mechanical anemometers with wiper-type contacts, still in use
for climatological studies, are attractive alternatives because of their simplicity but are of limited
use in pollution studies.
Wind direction transducers are of several basic types: wiper or sealed contact switches, single or
double potentiometers, and DC or AC synchronous motors. Some of the more sophisticated
transducers operate on the principal of capacitance with outputs in frequency form. Wire-wound
and carbon-deposited potentiometers are used most frequently.

6. Signal Conditioners - In most electronic systems, the signal from the meteorological
parameter being measured is transmitted from the transducer to a signal conditioner or
readout device, with power applied at some or all of these steps. The signal conditioner
converts the transducer output into an electrical quantity suitable for the proper operation of
the readout equipment whether it is a chart recorder or a data acquisition system. The signal
conditioner may vary from a simple resistance network or impedance-matching device to an
amplifier, an analog-to-digital converter, or, as in the case of the photoelectric speed
transducer, a frequency-to-voltage converter. Signal conditioners are devices that convert
from 360 to 540 degrees of wind direction and provide "average" wind or wind with "time-
weighted" average. Scalers that compress, expand, or change the transducer signal from
electrical to engineering-equivalent units are also signal conditioners.

7. Readout Devices - Digital displays are popular for logging and displaying wind data. They
come in a wide range of designs and are usually elaborate systems that include
analog-to-digital conversion, integration over specified intervals, and memory or storage
capability. The output format of these systems should be compatible with the computer used
for data processing. The data generated provide the user with better accuracy compared to
data reduction from strip chart recorders.

Direct writing (D'Arsonval movement) galvanometric recorders with strip charts are used
frequently for wind measurements. They are used because of their reliability and their speed of
response. The chart drive mechanisms are available as hand-wound, spring-driven,
battery-powered, or AC powered. It is important to select a recorder in which the damping
characteristics of the galvanometer do not degrade the response of the sensor and are in
compliance with the frequency response required in the study. Most galvanometric recorders
used for wind measurements are continuous curve-tracing recorders. The chopper-bar type,
designed to make an imprint on pressure-sensitive paper each time the meter pointer is clamped
against a sharp-edged platen, produces a record that is non-continuous. Imprints are usually
made every 2 seconds, so in rapidly varying winds the record appears scattered. Some
manufacturers of meteorological instruments supply a recorder with a built-in signal conditioner
that reduces the scatter through the use of either unspecified or selective time constants on the
response of the galvanometer. This conditioner must be closely evaluated if calculation of the
standard deviation of the measurement is an objective in analyzing the record.

Potentiometric recorders, used frequently for wind measurement, may not have the rapid
response characteristics of galvanometric recorders. However, as a null-device, the input
impedance is high when the stylus is at equilibrium; therefore, errors due to electrical loading of
the sensor output and errors due to voltage drops in long signal leads are minimized.
Multi-point potentiometric recorders, which sequence through a series of inputs of scheduled
cycles, must not be used for recording wind. Instantaneous samples of wind direction or wind
speed once every few minutes are of little use for most air pollution investigations.
Curve-tracing potentiometric recorders are useful and are available with charts that require ink or
with inkless charts that operate with a heated stylus.

Wind Sensor Characteristics

1. Cup Anemometers - These anemometers have complex-shaped cups. The net torque (lift
greater than drag) causes a rate of rotation roughly proportional to wind speed. The cups
respond to any horizontal wind direction, which is an advantage, but they are also
responsive to the vertical component of the wind. In turbulent flow, the output (average
speed) may be closer to the total speed than to the presumed horizontal component
(MacCready, 1966). The design of the anemometer cup assembly and the material from
which it is constructed are important in determining the starting threshold, dynamic
response, linearity, and durability of the instrument. The ratio between cup diameter and
cup wheel diameter influences the calibration curve (Gill, 1973). Starting threshold is
defined as the lowest wind speed at which the rotating cups meet the accuracy
specifications (Lockhart, 1970). In order to define the smallest eddy size to which cups will
be responsive, a dynamic characteristic known as the “distance constant” must be known.
This is determined in a wind tunnel by measuring the time for the cups to reach 63 percent
of the tunnel speed after being released from a non-rotating condition. The distance
constant in meters may be expressed as a time constant in seconds at a given wind speed by
dividing by that wind speed in meters per second. USEPA does not specify a distance
constant for anemometers. USEPA does suggest a distance constant of <5 meters at 1.2
kg/m. The reason why the distance constant is included is to urge users to buy high quality
responsive sensors. Heavy sensors with long distance constants are more likely to produce
over-speed errors, which overstate the average wind speed. If they are used to measure
turbulence, they will underestimate sigma theta (standard deviation of the wind direction)
because of a failure to respond properly to eddy sizes smaller than twice the distance
constant.

2. Propeller Anemometers - Propeller anemometers, especially helicoid types, are sensors with
rotation rates linearly proportional to the wind speed over a wind speed range (Gill, 1973).
Propeller anemometers must be oriented into the wind. The error from the failure of the
vane to perfectly orient the propeller is small since propellers have a nearly cosine response,
i.e., the propeller turns at a rate almost directly proportional to the wind component parallel
to its axis. Like the cup anemometer, the propeller anemometer is a first-order, non-
oscillatory system whose dynamic characteristics can be described by the distance constant
referenced above. Fixed axis propeller anemometers are designed to measure two or three
components of wind simultaneously at a point in space. They represent a special type of
propeller anemometer for the direct measurement of turbulence (Fill, 1975). Three helicoid
anemometers in an orthogonal array measure the wind for the axes u-v-w. Each propeller
turns at a rate almost proportional to the wind component parallel to its axis. Cosine
response, although not yet perfected, is critical in this equipment. However, this device does

not have the static balance (a two axis wind vane), which is also used for the turbulence
measurements. Under conditions of rain, snow, and heavy dew, the bi-vane imbalance may
 produce unacceptable errors.

3. Wind Vanes - Wind vanes have a damped, oscillatory motion. This characteristic second
order response is the result of such factors as weight of materials, shape and size of vane,
and location and weight of counter-balance. One indication of the performance of the vane
is the starting threshold. As described by Finkelstein (1981), this is the lowest speed at
which a vane released from a position 10 degrees off the centerline in a wind tunnel moves
to within 5 degrees of center. There are several dynamic characteristics, identifiable as
constants, which can be used to define the performance of a wind vane (MacCready, 1965,
and Wieringa, 1967) in response to a step function. These include damping ratio, damped
wavelength, undamped wavelength, and delay distance. Undamped wavelength is used in
determining the dynamic response of a wind vane to sinusoidal wind direction fluctuations
(varies to a sine curve).

4. Damping Ratio - The damping ratio is a constant that is dimensionless and independent of
wind speed. It is calculated from the relative amount of overshoot on each of two successive
half cycles of a decaying oscillation.

5. For most operational programs, a damping ratio of 0.4 or greater is recognized as satisfactory.

6. Damped Wavelength - The damped wavelength is easily determined by multiplying the time
for one complete oscillation by the wind speed in the wind tunnel.

7. Delay Distance - Delay distance is another observed measure of the response of a vane to a
step change. Delay distance is based on the time required for a vane to reach 50 percent of
the distance from an initial displacement, to 10 degrees toward the centerline on the first
swing. This is multiplied by the tunnel speed to obtain the delay distance.

8. Sonic Anemometers - Sonic anemometer systems are based on the principle that wind
changes the transit time of a sound pulse across a fixed distance. Sonic systems can be
designed in two dimensions for horizontal wind speed and direction as a replacement for the
cup and vane or propeller units, or in three dimensions for both horizontal and vertical wind
measurements. For those applications where the contribution of small eddies is important,
sonic systems are an excellent choice.
 Wind Data Requirements

Any data requirement should be expressed in the context of all applications for which the data
may be used. Wind data are used in environmental monitoring for source location, transport and
dilution modeling, and as a diffusivity indicator. The principal specifications are dynamic range
(most important is the threshold) and dynamic performance (distance constant for speed and
damping ratio and delay distance for direction). It is also important to specify the averaging time
and method in order to judge the adequacy of the measuring and recording system. For most
atmospheric dispersion studies, a starting speed (the speed required to start motion of the sensor)
of 0.5 m/s or less is appropriate for both vanes and anemometers. Wind vanes should have a
damping ratio of 0.4 or greater and a delay distance of 5 m or less. Anemometers should have a
distance constant of 5 m or less. For climatological studies, less sensitive instruments may be
used.

Averaging for wind speed may be done by scalar methods (dilution) or vector methods
(transport) and should represent one hour. Wind direction should be averaged by obtaining the
resultant vector direction for the hour. Sigma theta should represent 3 to 10 minutes if stability
categories are to be selected, but should represent the hour when the preferable direct
calculations are made for diffusion (Strimaitis, 1981).

Procurement

For research projects it might be possible to purchase instruments with better specifications or of
more recent design. But in operational programs, only field-tested and time-proven instruments
with known performance records should be purchased.

Caution should be used in purchasing components as opposed to total systems. It is important to

match the dynamic characteristics of the wind sensors and the electrical characteristics of the
transducers with the readout device. For digital systems, special attention should be given to
sampling and averaging times as well as instantaneous, as opposed to integrated, data
acquisition.

Acceptance Testing

The supplier should provide all calibration certification data for the equipment, including curves
and specifications. A table or formula should be provided which relates the rate of rotation of the
anemometer shaft (or frequency for light-chopper sensors if the number of pulses per revolution
is also given) to wind speed (or output voltage given to a voltage to speed range relationship).
This formula or table will usually relate to a nominal propeller or cup. The specific propeller or
cup assembly should have a permanent identification code (serial number). In those cases when
wind tunnel calibration data are provided, this identification is required.

The acceptance test for the direction vane should include a measure of how well the sensor
represents the relative position of the vane to the sensor housing. Four points 90 degrees apart
can be easily bench-tested by drawing perpendicular lines crossing at the vane rotation axis and
holding the vane shaft parallel to the lines. The manufacturer's method of coping with the
discontinuity between 360 degrees and 1 degree should be checked to ensure that the method
meets specifications.

Calibration

Calibration for wind speed and wind direction may be performed using several methods.
However, specific dynamic response characteristics such as threshold speeds, damping ratios,
delay distances, and distance constants can only be checked in a wind tunnel. Instruments should
be returned to the manufacturer or a properly equipped wind tunnel facility for this type of
calibration check. Sonic anemometers should be returned to the manufacturer for calibrations. In
addition, prior to any calibration adjustments, the operational period must be verified.

1. Wind Speed - Calibration of wind speed sensors will normally be performed using a
certified synchronous motor or a certified selectable speed anemometer drive. Other
methods for calibration may include using a photo tachometer, number of turns per minute,
or by using a calibrated collocated sensor. These other methods introduce more error and
may not be as convenient as using the synchronous motor or selectable speed anemometer
drive. Therefore, it is recommended that a synchronous motor or selectable speed
anemometer drive be used. This synchronous motor/selectable speed anemometer drive
must be certified against the Quality Assurance Section's photo tachometer annually to
ensure the revolutions per minute (rpm) are accurate. The calibration can be performed
when the sensor is on top of the tower or the sensor may be removed from the tower and the
calibration performed at ground level. The calibration must include a zero check and at least
2 different speeds. One speed should represent low wind speed (e.g., within 10 mph) and
one at high wind speed (e.g., at least 20 mph). In order to use the synchronous motor or
selectable speed anemometer drive, the cups or propeller will need to be removed. The cups
or propeller must be inspected for any type of damage that could alter the wind speed. If any
damage is apparent, the data collected will be suspect and the sensor should not be used
until repairs are made. The conversion of rpm to mph will depend on the type of sensor.
The formula for this conversion should be available in the sensor's maintenance book. A
torque test is recommended to determine the wind speed needed for the sensor to respond.

2. Wind Direction - The calibration of wind vanes will consist of aiming the vane at a known
direction, performing a bench linearity check, and a torque test. A telephone pole or any
other distant object that will remain stationary could be used as the reference point once
the degrees from the vane to this reference point are determined. The degrees to and from
the object can be determined by several methods. An acceptable method of establishing
true north is by the location of the sun at true solar noon. Another acceptable procedure for
determining true north involves shooting the North Star with a first-order theodolite. Any
textbook or handbook on land surveying will describe the technique and contain all the
necessary tables. Another method for determining true north involves using a magnetic
compass. However, caution should be used so that the magnetic compass is not influenced
by the strong permanent magnet in the wind recorder, iron metal objects, or the
electromagnetic field. Do not forget to allow for magnetic declination from true north. In
 addition, alignment in the vertical is equally important. Studies have indicated that vertical
misalignment of 1 degree may yield data errors of 10 percent or greater in measurement of
turbulent parameters (Pond, 1968; Deacon, 1968; and Kraus, 1968). Vertical alignment
should be established with a good carpenter's level or torpedo level at two points that are 90
degrees apart in the horizontal. Factory calibrated sonic anemometers should be attached
securely and aligned 180 degrees from true north, using any of the methods described
above.

The bench linearity check involves moving the vane in known degree increments, such as
advancing the vane exactly 90 degrees for every point and waiting for the response to stabilize,
and recording the degree output. Enough increments should be used to include the full scale of
the sensor (this will allow all potentiometers to be evaluated). A torque test is recommended to
determine the wind speed needed for the vane to respond. There will be a bias to the direction
recorded due to the wind vane not pointing in the correct direction at the recorded time (see
Form 7) if the torque test fails.

Frequency

Calibrations must be performed at least annually for State/Local Air Monitoring Station
(SLAMS) sites, semi-annually for National Core multi-pollutant monitoring stations (NCORE)
and Photochemical Assessment Monitoring Stations (PAMS) sites, and quarterly for PSD sites.
In addition, calibrations must be performed if the sensor fails an audit or if the sensor is
damaged and repaired. Sonic anemometers should be sent to the manufacturer on an annual basis
or if the unit is damaged or fails an audit.

Limits

1. Wind Speed - Calibration limits for wind speed are ±0.50 mph up to 10.0 mph and ±5% if
the speed is >10.0 mph. If these limits are not met, the sensor will require maintenance and
another calibration until these limits are met. In addition, the sensor must respond within the
torque limit. The torque limit will depend on the type of sensor and should be found in the
sensor's maintenance manual. The data collected from an out-of-calibration sensor are
invalid back to the previous audit or calibration, or if it can be determined by analyzing the
data when the unit malfunctioned.

2. Wind Direction - Calibration limits for wind direction are ±5.0 degrees when the vane is
pointing to a standard reference point. In addition, a bench linearity check must be made
with no more than a 10.0 degree spread between the highest negative and highest positive
difference between the response and the known degrees. If either one or both of these
calibration checks fail, maintenance is required and both checks must be repeated. In
addition, a torque test is recommended. The data collected from an out-of-calibration sensor
are invalid back to the previous audit or calibration, or if it can be determined by analyzing
the data when the unit malfunctioned.
1. Wind Speed - An audit for wind speed consists of repeating the calibration procedure
without making any adjustments. The audit must provide enough information to determine
the validity of the data. Any of the methods mentioned in Section 3.6 may be used to
perform the audit, but a synchronous motor/selectable speed anemometer drive is preferred.

2. Wind Direction - An audit for wind direction consists of repeating the calibration procedure
without making any adjustments. The audit does not require every calibration check. The
linearity check is optional for the audit, but the vane must be challenged with at least 2
reference points (e.g., the primary reference point, and a 180 degree swing from the primary
reference point.

Frequency

Audits for wind speed and wind direction must be performed once a year. Ideally an audit should
be performed 6 months after a calibration. This will allow the sensors to be evaluated twice a
year. If a problem does occur, the amount of data which would be affected will be limited.

Result:

Site:

Sensors 10 meters above

ground in open, level terrain

Humidity
Humidity is a general term for the water vapor content of air. Other, more specific, terms for
humidity include: absolute humidity, relative humidity, specific humidity, mixing ratio, and dew
point (Huschke, 1959). This section discusses the measurements of relative humidity and dew
point. Relative humidity (RH) is a dimensionless ratio of the actual vapor pressure of air to the
saturation vapor pressure at a given dry-bulb temperature. Dew point is the temperature to which
air must be cooled, at constant pressure and constant water vapor content, to be saturated with
respect to liquid water. Frost point is the temperature below 0 degrees Celsius at which air is
saturated with respect to ice. There are many ways to measure the water vapor content of the
atmosphere. These can be classified in terms of six physical principles (Middleton and Spilhaus,
1953). Three principles are listed below with instruments used with each:

1. Reduction of temperature by evaporation - Psychrometer

2. Formation of dew or frost by artificial cooling - Cooled mirror surfaces

3. Diffusion of moisture through porous membranes - Diffusion hygrometers

Types of Sensors/Instrumentation
Psychrometry identifies a basic technique for deriving both relative humidity and dew point
temperature from a pair of thermometers - a dry-bulb thermometer that measures the ambient
temperature, and a wet-bulb thermometer. The reservoir of the wet-bulb thermometer is covered
with a muslin wick. When the wick is moistened and the thermometer ventilated, the
indicated temperature is related to the amount of evaporation cooling that can take place at the
existing ambient temperature, water vapor partial pressure, and the atmospheric pressure. A
psychrometer uses this principle to obtain relative humidity and dew point.

1. Psychrometers - The temperature sensors in a sling psychrometer are usually mercury or

alcohol filled thermometers. The same is true of portable motor-operated psychrometers, but
the psychrometric principle has been used with sensors made of thermocouples, wire-wound
resistance thermometers, thermistors, and bi-metal thermometers. Relative humidity and
dew point are easily determined by observing the wet-bulb depression, which is the
difference between the dry-bulb and the wet-bulb temperatures and then referring to a
psychrometric table or chart. One must be certain to use the chart values corresponding to
the correct atmospheric pressure range of the location where the observation is taken.
Computer programs are also available to calculate out the humidity values. These programs
may offer better accuracy than look-up tables.

More measurements of atmospheric water vapor have probably been made with the sling
psychrometer than by any other manual method. When properly used and read, the technique is
reasonably accurate. The most common errors encountered using this method are from radiation,
rapid changes in conditions during reading, and parallax. The Assmann psychrometer
continuously aspirates the thermometers and protects them from radiation. This will allow time
and accessibility for a careful reading to avoid parallax (keep the eye perpendicular to the
meniscus for best results). For good accuracy, particularly where a variety of observers are
taking measurements, an Assmann or equivalent type psychrometer is recommended. One should
use the psychrometric tables with dew point values for the altitude (pressure) of the location
where measurements are being made.

2. Electrical Hygrometers - The resistance and capacitance of thin hygroscopic films on

electrical hygrometers are affected by the presence of moisture. Measurement circuits
provide the instrument output with scaled voltages and readouts of atmospheric moisture
content. Corresponding temperature measurement is included within the instruments to
calculate the results expressed in variables other than actual moisture content. Electrical
hygrometers are considerably less expensive than some other automated relative humidity
measurement methods and are readily adaptable to portable, hand-held units suitable for
temporary measurements and transfer standard use.

3. Cooled-Mirror Hygrometer - In the early 1960's, the technique of detecting the dew point on
a cooled mirror surface evolved into a production-type unit. This unit was operated
automatically and had an optical dew-sensing system that incorporated thermoelectric
cooling (Francisco and Beaubien, 1963). Of four meteorological grade, thermoelectric,
cooled-mirror dew point instruments investigated (Mazzarella, 1977), three operate in the
range from -50 degrees to +50 degrees Celsius. Linear thermistors are used to measure the
mirror temperature in three of the units; a platinum wire sensor is used in the other. All are
designed with simultaneous linear output signals for Tdp (dew point temperature) and T
(ambient temperature). Two of the manufacturers make claims of NIST-traceability with
stated dew point accuracies ranging from ±0.2 degrees to ±0.4 degrees Celsius and ambient
temperature accuracies ranging from ±0.1 degrees to ±0.5 degrees Celsius. All incorporate
 some form of standardization that involves clearing the mirror by heating, either
automatically or manually. Although complex in design and operation, this type of cooled-
mirror hygrometer is considered to be a functional standard.

Sensor Characteristics
Although the psychrometer is considered the most practical and widely used instrument for
measuring humidity, two major problems are associated with wet- and dry-bulb psychrometry
involving the accuracy of the thermometers and the cumulative errors related to operating
technique (Quinn, 1968). An accuracy of ±1 percent at 23 degrees Celsius and 50 percent RH
requires thermometers with relative accuracy of ±0.1 degrees Celsius. The commonly used
thermometers with 0.5 degree Celsius divisions introduce an uncertainty of ±5 percent RH. This
assumes that the readings were taken at the maximum wet-bulb depression, a difficult task with a
sling psychrometer.

Electrical hygrometers use capacitive and resistive sensors which respond better to relative
humidity than to dew point. Specifications for both types of sensors are similar. Capacitive
sensors are most linear at low humidity levels and can tolerate condensation, although calibration
shifts can occur. Resistive sensors are most linear at high humidity levels and cannot tolerate
condensation, although some have automatic protection from saturation conditions. Dew point
impedance sensors use a slightly different element; they measure absolute rather than relative
humidity. The sensors are covered by membranes that are readily porous to moisture, although
the membranes thermally insulate the sensor, causing some lag time in measurement.

The optical chilled (cooled) mirror technique of measuring dew point is a fundamental
measurement. No calibration is required for the fundamental dew generating process. The
measurement is the temperature of the surface at which the dew forms, and as with any electrical
temperature measurement system, calibration is required. A balancing of the optical system to
correct for changing the dry mirror reflectance that might result from contamination follows the
process of periodically heating the mirror to a temperature above the dew (or frost) point. In the
better instruments, automatic balancing is programmable in terms of frequency and length of
time. It can also be accomplished manually.

Sensor Housings and Shields

Psychrometers of all types should be acclimated to the environmental conditions in which the
measurements are to be made. In most cases, psychrometers should be stored in a standard
instrument shelter so that the mass of the thermometers, and especially the mass of the housing,
adjusts to the temperature of the air. Psychrometers with a stored water supply, such as those on
a tower, must be shielded from solar radiation.

Electrical hygrometers should be aspirated and shielded from solar radiation.

Manufacturers of optical cooled-mirror dew point and temperature monitoring equipment
provide housings for the sensors. These housings include forced ventilation and shielding from
solar radiation.

Data Requirements
Electrical hygrometers used for meteorological monitoring applications have time constants
generally longer than air temperature systems. The usual data of interest are hourly average
values. Data should be reported in terms of the condition measured, dew point temperature,
relative humidity, or wet-bulb and dry-bulb temperature. Computer programs may be used to
convert among these if all the relevant variables are known. The station elevation may be used to
estimate a nominal pressure if a measurement is not available. The temperature needed to
convert a relative humidity measurement to dew point temperature is the temperature at the
relative humidity sensor surface. This may not be the same temperature as that measured at some
other location. On the other hand, the dew point temperature is a fundamental measure of the
amount of water vapor in the air and is independent of air temperature. Relative humidity
calculations can therefore be made given the dew point temperature and any temperature
measurement point in the same general air mass.

Psychrometers are convenient devices for making spot checks on the performance of other
devices, especially those that are permanently installed, providing the check is done under
reasonably steady overcast conditions. The psychrometric technique built into tower installations
presents servicing problems, especially at temperature extremes. High temperatures cause rapid
evaporation. Low enough temperatures cause freezing.

Both the dew cell and the cooled-mirror type instruments are useful for 10-meter or taller tower
pollution studies. For these installations, the sensors must be housed in the recommended shields
with little, if any, aspiration for the dew cell and the recommended rate of aspiration for the
cooled-mirror design.

Procurement
Sling psychrometers and aspirated psychrometers with thermometers shorter than 10 inches do
not have sufficient resolution to provide the accuracy required checking other instruments.
Equally important, the thermometers should have etched stems; i.e., the scale markings should be
etched on the glass. Reliable thermometers are factory calibrated at a minimum of two
temperatures, and usually at three. Thermometers calibrated with NIST-traceable standards are
required.

Optical cooled-mirror dew point systems are now available commercially from several
manufacturers, all whom currently incorporate either linear thermistors or platinum resistance
temperature devices.
 Calibration
Prior to any calibration adjustments, the operational period must be verified.

1. Dew Point - Calibration of dew point sensors consists of using a psychrometer with
sufficient resolution to eliminate errors. The psychrometer could be a sling, motorized type,
or an electronic sensor. If applicable, the thermometers need to be certified prior to use and
every year thereafter against an NIST-traceable thermometer. If a sensor is used, it must be
certified every year with the appropriate NIST standard (see Chapter 6, QA Manual).
Temperature readings must be as accurate as possible. Care must be taken to make sure a
stable reading is obtained for the wet-bulb temperature. In addition, the calibration needs to
be performed in a shaded area or a place where no sunlight hits the thermometers/sensor and
as close to the dew point sensor as possible. It is advisable to obtain several dew point
measurements in as small a time period as possible and to take the sensor reading as close to
the calibration reading as possible. This will help improve the precision and accuracy of the
values. The dew point values must be obtained from the appropriate table/chart or by using a
computer program.

2. Relative Humidity - The procedure for performing a calibration on relative humidity sensors
is the same as the dew point calibration (see Form 13).

Frequency
Calibrations must be performed at least annually for SLAMS sites, semi-annually for NCORE
and PAMS sites, and quarterly for PSD sites. In addition, calibrations must be performed if the
sensor fails an audit or if the sensor is damaged and repaired.

Calibration of micrometeorological humidity sensors should be performed at the initial startup

sampling. Should a micrometeorological site be put into operation on a permanent basis,
humidity sensor calibrations must be performed annually with preventive maintenance
performed in accordance with Section 5.7. However, micrometeorological sites often operate on
a short-term basis. Therefore, annual calibrations may not be applicable. For short-term
monitoring applications, an audit must be performed prior to the dissolution of the site to confirm
the validity of the data for any given operational period.

Limits
Calibration limits for dew point are ±1.5 degrees Celsius dew point for PSD sites. No calibration
criteria currently exists for dew point at SLAMS or NCORE sites. Calibration limits for relative
humidity are ±7%RH at PSD and NCORE sites and ±10%RH at SLAMS sites. If these limits are
not met, the sensor will require maintenance and another calibration until this limit is met. The
data collected from a sensor that is out-of-calibration are invalid back to the previous audit or
calibration, or if it can be determined by analyzing the data when the unit malfunctioned.
 Field Operation and Preventive Maintenance
Field calibration checks should be made at least monthly on dew cell type units. The use of gold
wire windings around the LiCl cylinder minimizes corrosion problems in polluted atmospheres.
Periodic removal and flushing of old lithium chloride, followed by recharging with a fresh
solution, improves data reliability. Once a mercury or alcohol liquid-in-glass thermometer is
calibrated, there is no need for recalibration, unless it is to be used for reference or as a transfer
standard. Errors in wet-bulb temperatures are most frequently the result of an improperly
installed or dirty muslin wick, the repeated use of tap water instead of distilled water, or human
error in reading. Wicking material used on psychrometers must be washed to remove traces of
sizing and fingerprints. Once cleaned, the material is tied at the top of the thermometer bulb and
a loop of thread placed around the bottom so the thermometer bulb is tightly covered. To prevent
solid materials from collecting on the cloth and prevent proper evaporation, the wick should be
wetted with distilled water. Slinging or motor aspiration should be done in the shade, away from
reflected or scattered radiation, at a ventilation rate of about 3 to 5 m/s. Many technique-related
errors are minimized by using an Assmann-type, motor-operated psychrometer providing the
instrument is allowed to assume near ambient conditions prior to use.

The cooled-mirror instruments require no calibration except for the minor temperature sensor.
Depending on environmental conditions, the mirror is easily cleaned with a cotton tipped swab
dipped in the recommended cleaning fluid, usually a liquid with an alcohol base. While the
accuracy of a psychrometer is inferior to that of the optical chilled mirror system, an occasional
check at the intake to the sensor shield is recommended under the provisions specified earlier.
All operational and preventive maintenance activities should be entered in a logbook.

Frequency
Dew point and relative humidity audits must be performed semi-annually at PSD and PAMS
sites, annually at SLAMS and NCORE sites. Ideally audits should be performed 6 months after a
calibration. This will allow the sensors to be evaluated twice a year. If a problem exists, the
amount of affected data will be limited.

Should a micrometeorological site be put into operation on a permanent basis, humidity sensor
audits must be performed annually with preventive maintenance performed in accordance with
Section 5.7. However, micrometeorological sites often operate on a short-term basis. Therefore,
annual audits may not be applicable. In that circumstance, an audit must be performed before
dissolution of the site to confirm the validity of the data for any given operational period.
 Results
Site:

Ground beneath sensor unwatered

short grass or natural earth :

Sensor above open, level ground

9 meters diameter :

Temperature
Temperature is not simply an isolated piece of meteorological data, but it is used in the
measurement and determination of a number of other atmospheric parameters such as vertical
temperature gradient (stability), relative humidity, and gaseous pollution concentrations. Several
types of sensors and recorders are used routinely in a variety of combinations to acquire
temperature data. The temperature sensors listed below are used most often for environmental
measurements.

Types of Sensors/Instrumentation
1. Linear Thermistors - Thermally sensitive resistors or thermistors are electronic
semiconductors that are made from metallic oxides. The resistance of a thermistor varies
inversely with its absolute temperature. Linear thermistors are a composite of two or more
thermistors and fixed resistors designed to produce a linear response over a wide
temperature range. In system configuration, the thermistor is connected to a bridge circuit or
some suitable signal conditioning circuit. When a low voltage is applied to the thermistor,
the output from a bridge circuit is a voltage that varies directly with the temperature of the
sensor. Linear thermistors are particularly well suited to remote sensing applications
because of their ratio of resistance change to temperature change. Coefficients on the order
of 125 ohm/degree Celsius are common (Lockhart and Gannon, 1978), reducing the impact
of lead resistance errors. A lead resistance of 12.5 ohms would be required to produce a 0.1
degree Celsius temperature measurement error. The thermistor or thermistor composite may
be packaged as a glass-covered bead or encased in a stainless steel sheath. The latter is
common and best for monitoring applications.

2. Resistance Temperature Detectors - A Resistance Temperature Detector (RTD) operates on

the principle that the electrical resistance of a pure metal increases as temperature increases.
RTD sensors are made using silver, copper, nickel, and platinum wire. Platinum wire is the
best material to use because of its superior linearity and stability characteristics, high
sensitivity, and resistance to corrosion. The RTD probe is encased in a protective stainless
steel housing composed of an insulating core wrapped with platinum wire. The resistance of
the wire is measured using a bridge circuit and a signal conditioner.

The RTD operates at a much lower resistance to temperature ratio than the linear thermistor.
RTDs used most often in meteorological work have a coefficient of resistance change on the
order of 0.4 ohms per degree Celsius and are more sensitive to lead resistance errors than are
thermistors. "Three wire" and "four wire" systems are configurations that automatically
 compensate for lead resistance.

3. Liquid-In-Glass-Thermometers - Most of these thermometers are 10.5 inch 10 gram glass

tubes with a uniform capillary bore and a liquid reservoir or bulb at the bottom.
Thermometers are usually mounted on a stainless steel back with the bulb protruding to
allow free air circulation. Volumetric expansion and contraction of the operating liquid
provides the measure of temperature. The liquid is usually mercury but may be a mixture of
ethyl alcohol and red dye. Thermometers filled with alcohol are called spirit thermometers.
Though inherently less accurate than a mercury thermometer, the spirit thermometer must
be used where extremely cold temperatures are anticipated because mercury will freeze at -
38 degrees Celsius.

Sensor Characteristics

Accuracy
A high-quality mercury thermometer, properly calibrated, will provide temperature data of
sufficient accuracy for most atmospheric measurement programs. Over the years, the capillary
bore will tend to contract slightly, causing the zero point to rise. Because the thermal response of
mercury is essentially linear, this error is correctable through calibration. The most serious errors
incurred in using a mercury thermometer are usually due to improper exposure and/or observer
error (e.g., parallax error in reading the temperature). Typical commercial platinum RTD sensors
follow the platinum characteristic that represents a resistance accuracy of ±0.26 degrees Celsius
at 0 degrees Celsius and ±0.38 degrees Celsius at 50 degrees Celsius. Linear thermistors are
available commercially with an accuracy of ±0.15 degrees Celsius over the range of temperatures
normally encountered in environmental measurements (Lockhart and Gannon, 1978). Ordinary
commercial thermocouples are only accurate to ±1 degree Celsius (Wang, 1975). Accuracy of a
measurement system is limited more by such factors as sensor exposure, improper coupling, and
signal interference than by accuracy of the sensor. For air quality monitoring applications, an
accuracy (including coupling error) of better than 0.5 degrees Celsius is easily achievable and
adequate (Strimaitis, 1981).

Linearity
There are no linearity problems with any of the electronic or liquid-in-glass sensors within the
extremes of tropospheric temperature. Some linearity problems occur in the deformation-type
sensors, particularly at extreme temperatures.

Response Time
A certain amount of caution must be exercised when assessing the value of quick response time
in taking stationary environmental temperature measurements. At the warmest time of the day, a
continuous temperature trace may show variations of up to 1 degree Celsius in a 30-second
period. Care must be taken in evaluating instantaneous temperature data from a thermistor or
RTD, both of which may have time constants as short as 5 seconds or less. For both of these
sensors, temperature values should be the average of a series of readings taken over a minimum
period of 1 minute to avoid "noise" errors. Usually the stainless steel sheath, which protects the
sensitive element for field monitoring use, adds to the time constant mechanically thereby
providing both ruggedness and a more useful time constant.

Precision
Most good quality electrical temperature systems used for monitoring in the atmosphere will
have an output with better resolution and precision than accuracy. The application of the data
may take advantage of this fact. For example, a temperature difference between two levels on a
tower may (and should) be calibrated around the precision of each sensor, yielding an accuracy
of a difference on the order of the precision of the sensor (0.1 degree Celsius for example). This
statement must be qualified to either ignore the coupling error or assume each sensor will suffer
the same coupling error and, therefore, not influence the difference measurement. The latter is a
reasonably good assumption when identical radiation shields are used.

The accuracy of the temperature differential measurement system, excluding coupling errors,
may be as good as the resolution or precision if the effort is made to either adjust the circuits or
correct the data to achieve it. The nominal values quoted above reflect interchangeability with
circuits adjusted to the nominal values given in the manufacturer's manual. Given sensor
stability, the accuracy will be as good as the calibration.

Durability
All of the electronic temperature sensors are fairly rugged and stable. Liquid-in-glass
thermometers obviously require careful handling. The deformation thermographs also require
delicate handling as any physical damage will alter the calibration.

Radiation Shields
The most serious temperature measurement problem encountered in environmental thermometry
is radiation error, which can amount to several degrees Celsius at midday. Early attempts to
combat radiation error took the form of instrument shelters, such as the all wood, louvered,
double roofed cotton region type. Much better results in reducing radiation error have been
achieved with aspirated radiation shields.

1. Naturally Ventilated Radiation Shields - The most common type of the naturally ventilated
radiation shield employs a vertically mounted sensor with a 360 degree ventilation exposure.
The housing is topped with single or multiple polished domes for solar radiation shielding
and has lower circular shields to block terrestrial and reflected radiation. The vane oriented
radiation shield directs the ambient wind to the temperature sensor. A swivel mounted, white,
horizontal, double walled tube is equipped with a large vane that keeps the housing opening
pointed into the wind. Unfortunately, these shields allow large errors when the wind is light.
Both of these naturally ventilated radiation shields will accept a variety of electronic
temperature sensors, including the thermistor, thermocouple, and RTD.
2. Mechanically Aspirated Radiation Shields - Mechanically aspirated radiation shields can be
used where power is readily available to drive the blower. The motor draws ambient air
across the sensor at an average flow of 5 m/s. The radiation shield may take a number of
forms, but common features include double-wall construction and white paint. Some use
thermos bottle type wall construction. Some are mounted horizontally with the air intake
facing north, while on others, the tube is mounted vertically with the air intake facing
downward. Some vertical units also employ domed polished radiation shields for both sky
and ground radiation shielding (McTaggart-Cowan and McKay, 1976). These mechanically
aspirated radiation shields will accommodate the full range of electronic temperature sensors
along with a variety of dew point and humidity sensors. It is desirable that blowers be wired
to signal whether or not they are operating, particularly in tower installations.

Temperature Data Requirements

In general, measuring temperature with accuracy greater than 0.5 degrees Celsius may not be
necessary. Extreme daytime horizontal temperature gradients have been documented (Hoffmann,
1965, and Department of the Army, 1975). Certain circumstances require more accuracy or
relative accuracy for differential temperature measurements. For example, a temperature gradient
measurement requires a relative accuracy of temperature or an absolute accuracy of temperature
difference of 0.1 degrees Celsius. This is true for the low level, local surface gradient
measurement between 2 meters and 10 meters above the ground or for the more conventional
measurement between 10 meters and 60 meters. Also, if humidity is measured by the wet-bulb
and dry-bulb difference, sensors should be matched so that small differences are meaningful.
The atmosphere is a turbulent, differentially heated fluid should remind any investigator that a
temperature recorded to the nearest tenth of a degree is representative of only a small volume of
air for a very short period of time. All routine monitoring applications of temperature data will
be expressed as a long-term average such as in 1-hour increments.

Calibration
Temperature sensors may be calibrated in the laboratory but must be calibrated in the field after
the sensor is installed. Performing both an in-house and then a field calibration may provide
better precision and accuracy from the sensor; however, only the field calibration is required.
The calibration must consist of submerging the sensor into a water bath or a calibration chamber.
In addition, a check should be made after the sensor is placed in the shelter. The sensor must be
compared against a thermometer that is traceable to a National Institute of Standards and
Technology (NIST) thermometer or some other temperature transfer standard. The NIST-
traceable thermometer should be graduated in 0.1 degree Celsius increments. In addition, prior to
any calibration adjustments, the operational period must be verified.

Procedure
The sensor and the thermometer will both be placed in a water bath or calibration chamber.
When using a water bath, uniform temperature throughout the liquid can be accomplished by
using a magnetic stirrer. Three or more temperatures must be evaluated spaced across the range
of the sensor. The first at room temperature (e.g., 25 degrees Celsius), the second at freezing
point (e.g., 0 degrees Celsius), and a third higher temperature (e.g., 45 degrees Celsius). When
using a water bath, the higher temperature would be produced using a heat pad. While
performing the water bath calibration, the following three conditions must be met:

 the sensor and the thermometer must not touch each other or the wall of the container
holding them.
 both the thermometer bulb and the sensor must be submerged completely into the liquid
(ideally at the same water level), if in a liquid bath.
 the stirrer must operate continuously during the calibration to maintain uniform temperature.

After the calibration is performed and the results show the sensor meets calibration limits, it can
be placed in the shelter. A comparison should then be made with air temperature to ensure that
the blower is working correctly. Be sure that the thermometer is in a shaded area and
approximately at the same height as the sensor. In addition, the thermometer bulb must not come
into contact with the person holding the thermometer and it should be kept as far away from their
body as possible. If the sensor does not compare to the thermometer or represent the actual air
temperature, the comparison and/or calibration should be performed again (see Form 10).

Frequency
Calibrations must be performed at the start of sampling, and at least annually for SLAMS sites,
semi-annually for NCORE and PAMS sites, and quarterly for PSD sites. In addition, calibrations
must be performed if the sensor fails an audit or if the sensor is damaged and repaired.

Limits
The calibration limit for outdoor temperature is ±1.0 degrees Celsius for SLAMS sites, ±0.5
degrees Celsius for NCORE, PAMS, and PSD sites. If this limit is not met, the sensor will
require maintenance and another calibration until the limit is met. The data collected under out-
of-calibration conditions are invalid back to the previous audit or calibration, or if it can be
determined by analyzing the data, when the unit malfunctioned.

Preventive Maintenance
Most preventative maintenance on thermometry systems concerns housings and shelters.
Instrument shelters must be cleaned as often as local weather conditions require. The air
passageways and screens in radiation shields, both aspirated and naturally ventilated, should be
cleaned out at least once every month. The blower should be checked during each visit. For
remote sites or where data collection is critical, the blower should be changed periodically as
recommended by the manufacturer. Lubricate the aspirator system as required. The spare parts
inventory should include a sensor and blower. The most common cause of component failure is
lightning. Obviously a system damaged by lightning will require repairs and calibration. All
preventive maintenance activities should be entered into a site logbook.
 Frequency
A temperature audit must be performed every year. Ideally an audit
should be performed 6 months after a calibration. This will allow the
sensors to be evaluated twice a year. If a problem does occur, the amount
of data which would be affected will be limited.

In the case of micrometeorological sites, which often operate on a short

term basis, annual audits may not be applicable. However, a final audit
must be performed before the dissolution of the site to confirm the validity
of the data for any given operational period.

Result:
Site:

Ground beneath sensor un watered

short grass or natural earth

Sensor above open, level ground

9 meters diameter

Rainfall

Precipitation Measurements

In any method of precipitation measurement, the aim should be to obtain

a sample that is representative of the rainfall in the area. At the outset, it
should be recognized that the extrapolation of precipitation amounts from
a single location to represent an entire region is an assumption that is
statistically questionable. A network of stations with a density suitable to
the investigation is preferable. Two basic types of precipitation collectors
are non-recording and recording.

Types of Sensors/Instrumentation

1. Non-recording Gages - In its simplest form, a precipitation gage

consists of a cylinder, such as a can with straight sides, closed at one end
and open at the other. The depth of the liquid in the gage may be
measured with a measuring stick calibrated in subdivisions of centimeters
or inches.

To obtain greater resolution, as in the case of the standard 8-inch gage

made to National Weather Service Specification No. 450.2301, the gage
is constructed with a ratio of 10:1 between the area of the outside
collector cylinder and the inside measuring tube. The funnel attached to
the collector directs the precipitation into the tube and minimizes
evaporation loss. Amounts in excess of two inches of rainfall overflow
into the outer can, and all measurements of liquid and melted
precipitation are made in the measuring tube with a measuring stick. The
automatic wet/dry precipitation collector, available in several designs,
represents a specialized,non-recording instrument designed for programs
involving the chemical and/or radioactive analysis of precipitation. The
collector is built with a sensor that detects the onset and cessation of
precipitation and automatically releases a lid to open and cover the
collector. In one design, the lid can remain open during either wet or dry
periods. Another model is made with two collectors; the lid is made to
cover one bucket during periods of rain and snow. In equipment of this
kind involving precipitation chemistry, the volume of water in proportion
to the constituents collected with the water is important, so evaporation
must be kept to a minimum.
2. Recording Gages - Recording gages are of two basic designs based
on their operating principles; the weighing-type gage and the tipping
bucket-type gage. The former, when made to National Weather Service
Specification No. 450.2201, is known as the Universal gage, indicating
usage for both liquid and frozen precipitation. There are options for the
remote transmission of signals from this type of gage. The standard
National Weather
Service Tipping Bucket Rain Gage is designed with a 12-inch collector
funnel that directs the precipitation to a small outlet directly over two
equal compartments, or buckets, which tilt in sequence with each 0.01
inches of rainfall. The motion of the buckets causes a mercury switch
closure. Normally operated on 6 V DC, the contact closure can be
monitored on a visual counter and/or one of several recorders. The
digital-type impulse can also be used with computer-compatible
equipment.

Instrument Characteristics

The most accurate precipitation gage is the indicating-type gage;

however, the recording-type gage measures the time of beginning and
ending of rainfall and the rate of fall. The Universal weighing gage
incorporates a chart drum that is made to rotate by either an 8-day spring-
wound clock or a battery-powered clock. Recent developments include a
unit with a quartz crystal mechanism with gear shafts for a wide range of
rotation periods from half a day to one month.

1. Weighing Gage - The weighing gage is sometimes identified by the

name of its designer (Fergusson) and comes with one of two recording
mechanisms. In the single traverse unit, the pen moves from the base of
the drum to the top, typically a water equivalent of 6 inches. In a dual
traverse unit, the pen moves up and down for a total of 12 inches of
precipitation. A variation of the weighing gage, a "high-capacity" design
with dual traverse, will collect as much as 760 mm or 30 inches. To
minimize the oscillations incurred by strong winds on the balance
mechanism, weighing gages are fitted with a damper immersed in silicone
fluid. By incorporating a potentiometer in the mechanism, the gage
becomes a remote-transmitting unit, capable of providing a resistance, or
as another refinement, a voltage proportional to the amount of
precipitation collected. Linearity of response is usually a factory
adjustment involving the use of calibrated weights to simulate
precipitation amounts. In spite of the manufacturer's specifications, it is
doubtful that the gage can resolve
0.01 inches, especially when the bucket is nearly empty.

2. Tipping Bucket Gage - In the tipping bucket gage, the balance of the
buckets and leveling of the bucket frame are critical. Low voltage at the
gage is imperative for reasons of safety. Power is typically 6 V DC. The
signal is provided by a switch closure every time the bucket assembly tips
(0.01 inches rainfall per bucket). An event recorder calculates rain rates
with pens energized sequentially to improve resolution. The tipping
bucket (a mechanical device) takes time to go from one position to the
next. When the rate of fall is high, there is spillage and the unmeasured
precipitation falls into the reservoir. Where there is a need for greater
accuracy, the collected water is measured manually, and excess amounts
are allocated proportionately in the record. The accuracy of the gage is
given as 1% for rainfall rates of 1 in/hr. or less; 4% for rates of 3 in/hr.;
and 6% for rates up to 6 in/hr.

3.Windshields and Heaters - Accuracy of measurement for all types of

gages is influenced more by exposure than by variation in design.
Windshields represent an essential accessory to improve the catch of
precipitation, especially snow in windy conditions. The improved Alter
design, made of 32 free-swinging but separated leaves supported 1/2 inch
above the level of the gage collecting orifice, is an effective way to
improve the catch. In a comparison of shielded and unshielded 8-inch
gages, it has been shown that at a wind speed of 5 mph, the efficiency of
the unshielded gage decreases by 25 percent, and at 10 mph, the
efficiency of the gage decreases by 40 percent (Weiss, 1961).
 In below freezing conditions, when the catch in a gage is snow or some
other form of solid precipitation, it is necessary to remove the
collector/funnel of non-recording gages and the funnel in recording gages.
Some instruments are available with built-in heater elements that are
thermostatically controlled. An effective heater for conditions that are not
too severe is an incandescent lamp installed in the housing of the gage.
Caution should be exercised; too great a heat will result in evaporative
loss.
Precipitation Data Requirements

In research studies, especially those related to acid rain, the instrument

used most frequently is the Automatic Precipitation Collector with one or
two collecting buckets and a cover to prevent evaporation. In operational
activities, the choice is between the weighing gage and the tipping bucket
gage. For climatological surveys, the choice might include one of the
above gages as well as a non-recording type gage. The use of a
windshield is recommended to minimize the errors that result from windy
conditions if the application requires maximum accuracy.

The precipitation measurement made in air quality monitoring stations is

frequently used for descriptive purposes or for episodic analysis. If the
effort required to achieve the level of accuracy specified by most
manufacturers of electrical recording gages is more than the application
of the data can justify, a tolerance of 10 percent may be adequate.

Procurement

A variety of gages are available commercially. In general, gages which

conform to the standards established by the National Weather Service
result in the fewest problems. For example, there are numerous 8-inch
gages available, but those following NWS specifications are made only of
brass and copper, are more durable, and are reported to rupture less
frequently under extended freezing conditions than those made of
galvanized steel.

The procurement of a weighing-type gage should include a tripod

mounting base as well as a set of calibration weights. For locations that
may not be readily accessible, or locations with heavy precipitation, the
bucket of the weighing gage should have an overflow tube. If the
resolution of time is not too important, recording rain gages of the drum
type can be obtained with monthly rather than weekly mechanisms.
Unless the tipping bucket gage is equipped with a heater, it is of no use
for frozen precipitation.

Calibration
Prior to any calibration adjustments, the operational period must be
verified.

1. Precipitation Gages - Bench calibrations for gages should follow the

recommendation of the manufacturer. The electrical output gage or the
drum recording gage measures weight, whether total weight in the case of
the "weighing gage" or increments of weight in the case of the tipping
bucket gage. Density of water is assumed so the weight can be expressed
i in units of volume or depth assuming the area of the collector opening.
Calibrations of the measurement apparatus can be based upon the
introduction of known volumes of water. For rate-sensitive systems such
as the tipping bucket, the rate of simulated precipitation should be kept
less than one inch per hour.

Calibrations require properly leveled weighing systems (gages) whether

on the bench or in the field. Once the precipitation gage is installed in the
field, a final calibration check should be made prior to any sampling. For
the weighing bucket, standard weights or a suitable substitute can be used
to perform the calibration check. To check the operation of the tipping
bucket, the best approach is to put a known quantity of water in a can
with a small hole so that the slow flow can be timed. It may be necessary
to adjust the set-screws, which act as limits to the travel of the tilting
buckets. The average of a minimum of ten tips should be used. Three
different amounts of water should be used for the calibration.

Frequency

Calibration of precipitation gages must be performed at least annually at

SLAMS sites, semi- annually at NCORE and PAMS sites, and quarterly
at PSD sites. In addition, calibrations must be performed if the sensor
fails an audit or the sensor is damaged and repaired.

Calibration of micrometeorological precipitation gages should be

performed at the initial start-up sampling. Should a micrometeorological
site be put into operation on a permanent or long term basis, precipitation
gage calibrations must be performed annually with preventive
maintenance performed in accordance with Section 7.6. However, these
sites often operate on a short term basis. Therefore, annual calibrations
may not be applicable. In that circumstance, an audit must be performed
prior to the dissolution of the site to confirm the validity of the data for
any given operational period. In addition, calibrations must be performed
if the unit fails an audit or is damaged and repaired.

Limits
 The calibration limit for precipitation gages is ±10.0%. If this limit is not
met, the sensor will require maintenance and another calibration until the
limit is met. Data collected under out-of-calibration conditions are
invalid.

Frequency

Precipitation sensors should be audited every year. Ideally an audit should be performed 6
months after the calibration check. This will allow the sensor to be challenged twice a year.

Should a micrometeorological site equipped with precipitation gages be put into operation on
a permanent or long term basis, an audit must be performed every year. Ideally an audit
should be performed 6 months after a calibration check. This will allow the sensors to be
checked twice a year. If a problem does occur, the amount of data that would be affected will
be limited.

Result:
Site:

Mouth of rain gage mounted :

horizontal
:
Rain gage minimum 30 cm
above ground