Collection and Preservation of
Samples and Field Analysis
Presented at : Training Programme on QAQC in Water
Quality Monitoring and Assessment, Oct. 21‐22, 2010
BY
Dr. G.K.Khadse, Sr. Scientist
National Environmental Engineering Research Institute, Nagpur
(CSIR, New Delhi)
Step‐5: Sampling
Sample: A part of the whole, that can be
examined to see what the rest is like
Objectives of Water Sampling
Baseline Water quality
Impact assessment
Assessment of Technology
Performance assessment of treatment plant
Assessment of products and processes
Design of Water Sampling Programme
Objectives, parameters, levels of
accuracy and use of the data
Sampling Location
Sampling frequency
Sample collection
On Sport Analysis Sample Preservation
Analytical methods and
Instrumental Techniques
Data handling and reporting
Planning for Sampling
When planning a sampling programme the
number of sampling stations or wells that can
be sampled in one day is required.
For this is necessary to know the required time
needed for sampling, and other actions
required, at the site.
Since purging is a time consuming
activity an estimate of the required
purging time is a must to arrive at a
fair estimate of the sampling time.
Check list for the field visit Developing Country Times
9At least one day before sampling, make GOOD NEWS !!
NEW METHOD DEVELOPED FOR
sure that all the arrangements are made DESIGNING INTERMITTENT SYSTEMS
Claims that supply conditions will improve !
as per the check list. Claims to share water more fairly !
Improved pressure conditions for all !
9Make sure, how to reach sampling site(s).
9Take help of location map
9Field operator may make his/her own personal checklist.
9Decide on the number of each item that would be required
depending on the number of samples to be collected.
9It is always safer to carry a few numbers in excess.
9If the laboratory conducting analyses is different from the
laboratory doing sampling...informed the concerned laboratory
to receive samples on time, particularly those which would
need immediate attention.
9In case there is any deviation in the collection point, record it.
Checklist for Field Visit
• Itinerary for the trip (route, stations to • Personnel and sample transport
be covered, time schedule) arrangement
• Area map • Sampling site location map
• Icebox filled with ice or icepacks • Weighted bottle sampler
• BOD bottles • Rope
• Special sample containers: • Sample containers
bacteriological, heavy metals, etc.
• Sample preservatives (e.g. acid • Thermometer
solutions)
• Tissue paper • Other field measurement kit, as required
• Sample identification forms • Labels for sample containers
• Field notebook • Pen / pencil / marker
• Soap and towel • Match box
• Spirit lamp • Torch
• Drinking water • Knife
• First-aid box • Gloves and eye protection
• Dump sampler to check well conditions • Submersible pump and accessories
General Guidelines for Sampling
• Rinse the sample container 3 times
with the sample before it is filled
• Leave a small air space in the bottle to
allow mixing of sample at the time of
analysis.
• Label the sample container properly.
• The sample code and the sampling
date should be clearly marked
• Complete the sample identification form for each sample.
• The sample identification form should be filled for each sampling
occasion at a monitoring station.
• If more than one bottle is filled at a site, this is to be registered
on the same form.
• Sample identification forms should be kept in a master file at the
laboratory where the sample is analysed.
Surface Water Sampling
• Samples will be collected from well
mixed section of the river (main stream)
30 cm below the water surface using a
weighted bottle or DO sampler.
• Samples from reservoir sites will be collected from the outgoing
canal or water intake structure, in case water is pumped. When
there is no discharge in the canal, sample will be collected from the
upstream side of the regulator structure, directly from the reservoir.
• DO is determined in a sample collected in a DO bottle using a DO
sampler. The DO in the sample must be fixed immediately after
collection, using chemical reagents.
• DO concentration can then be determined either in the field or later,
in a level I or level II laboratory.
Groundwater Sampling
Samples for groundwater quality monitoring would be
collected from one of the following 3 types of wells:
• Open dug wells in use for domestic / irrigation
• Tube wells fitted with a hand pump or a power-driven
pump: domestic / irrigation
• Piezometers (built for recording of water level & WQM).
• Open dug wells, which are not in use (or abandoned), will not be considered as
WQM station. However, such wells could be considered for water level monitoring.
• Use a weighted sample bottle to collect sample from an open well about 30 cm
below the surface of the water.
• Samples from the production tube wells will be collected after running the well for
about 5 minutes.
• Non-production piezometers should be purged using a submersible pump. The
purged water volume should equal 4 to 5 times the standing water volume, before
sample is collected.
• For bacteriological samples, when collected from tubewells/hand pump, the outlet of
the pump should be sterilised under flame by spirit lamp before collection of sample.
Sample Labeling
Label the sample container properly,
preferably by attaching an appropriately
inscribed tag or label.
Alternatively, the bottle can be labelled
directly with a water-proof marker.
Information on the sample container or the tag should include:
• Sample code number (identifying location)
• Date and time of sampling
• Source and type of sample
• Pre-treatment or preservation carried out on the sample
• Any special notes for the analyst
• Sampler’s name
Sample Preservation and Transport
Samples for BOD and bacteriological analyses should
be stored at a temp. < 4oC (ice/cold packs) and in the
dark as soon as possible after sampling. Once in the
laboratory, samples should be transferred as soon as
possible to a refrigerator.
COD analysis is to be done on the day of collection or
they should be preserved below pH 2 by addition of
conc. H2SO4. This procedure should also be followed
for samples for ammoniacal nitrogen, total oxidised
nitrogen and phenol analysis.
For metals: Samples should be acidified to below pH 2 with conc. HNO3.
Such samples can then be kept up to 6 months before they need to be
analysed; mercury determinations should be carried out within 5 weeks.
Samples should be transported to concerned laboratory as soon as
possible, preferably within 48 hours. Analysis of bacteriological samples
should be started and analysed within 24 hours of collection.
Importance of the sampling procedures
Often the quality control aspects are only related to the
analytical part, whereas the control procedures for the
sampling are neglected.
There appears to be a need for a detailed description
of the sampling and preservation procedures.
The objective of sampling is to collect a portion of material from an environmental
compartment (either water, sediment or biota) small enough in volume to be conveniently
transported & handled in the laboratory, while still accurately retaining its representativity.
This implies that the relative proportions or concentrations of the components of interest
should be the same in the samples and original environment. This requires that the
sample will be handled and treated in such a way that no significant changes in
composition occur that may hamper proper analysis.
In other words, no addition (e.g. contamination), loss (e.g. adsorption to the wall of the
sample bottle) or deterioration (e.g. physico-chemical or biological degradation or
transformation) can be allowed.
TYPES OF SAMPLES
1) Grab (spot or catch) sample
A source is known to be fairly constant
- In composition
- Over a considerable period
- For substantial volume
- Over spatial dimensions
Condition of source
- Fairly steady = Represent the composition
- Varies with time = Suitable intervals
- Varies in space = Appropriate locations
2) Composite samples
In most cases, the samples refer to a mixture of spot samples
collected at the same sampling site at different times.
Useful technique in case of temporal variation in water quality.
The series of spot samples are not mixed but analyzed individually,
also information on the daily variability can be obtained, and
afterwards the average can be computed.
Sometimes the indication 'time-composite‘ (24-hour period) is used
to distinguish from 'location composite‘ sampling.
To evaluate the nature of special discharges (e.g. variable in
volume or irregular in time), sampling at time intervals representing
the period during which such discharges occur.
This type of sampling is also required to measure the flux of
pollution load discharged through a point source.
3) Integrated samples
Sometimes samples are collected at the same location but,
due to horizontal or vertical variation in the composition of
the river (or in water flow) or lake, they come from different
points in the cross-section that are regarded with a different
relative importance.
To evaluate the average composition, total load or mass
balance, integrated samples are collected, often in
proportion to the river flow of the areas of sample collection.
Collection of integrated samples may require equipment to
collect sample from various depths
4) In‐situ measurements
Some determinations are more
likely to be affected by sampling
and sample storage.
In several cases the expected
changes are so large, that it is
impossible to store the sampled
material for a correct analysis at
a later moment.
If possible, these parameters should be analyzed on the
sampling site or, even better, in-situ.
Important parameters: pH, DO, temperature, conductivity and
sometimes turbidity.
Require special measuring devices.
Numbers and diversity of organisms is also to be considered as
in situ analysis.
Contamination control
Contamination may occur from: sampling equipment, sample
bottle, preservatives, ambient atmosphere, personnel taking the
sample etc. Utmost care should be maintained during sampling
- in order to prevent contamination.
Often sampling bottles need to be cleaned in a special way,
depending on the parameter. To avoid cross-contamination,
the same bottles should be used only for identical selected
parameters.
• Separate sets of bottles should be used for natural waters and for effluents.
• To prevent contamination by the hands, plastic (PE) gloves are needed.
• Atmospheric dust and (exhaust) fumes are readily available to contaminate the
sample: minimum contact of the sample with the atmosphere is essential, here.
• The person taking a sample (and the analyst) should take care not to touch the
inside of bottle and cap.
• The sampling bottles should be kept clean from dust and dirt.
• Pipettes or pipette tips should (in the field and in the laboratory) only be used once.
Cleaning procedures
The cleaning of samplers, sampling bottles and other labware, different
cleaning procedures can be applied depending on the parameter
For heavy metals rinsing with 1:1 diluted HNO3(supra pure quality) for 1
week is needed, followed by 3 times washing with double distilled water.
Bottles for trace organic (chlorinated) compounds, like pesticides, should be
cleaned with the solvent used for extraction (also of high purity quality).
Samples for the general physical-chemical characterization allow less
vigorous methods. Thorough cleaning with water to remove particulates and
two times rinsing with distilled water will usually be sufficient.
Organisms that are to be preserved (alcohol, formalin)
should be stored in glass bottles.
All bottles should arrive at the sampling site in a fully
cleaned state, protected from accidental contamination.
The last cleaning step is (in most cases) rinsing 2-3
times with the water to be sampled.
Sample Containers
The sample containers needed for a sampling campaign are prepared
by the laboratory and given to the person collecting samples.
Table 4: Container Types and Volumes Needed for
Sampling
Analysis Container Volume Preservation
0 On-site analysis PE bowl ±200 --
1 General Glass, PE 1000 --
(SS TDS major ions)
2 COD, NH3, NO2-+NO3- Glass, PE 500 H2SO4, pH
3 o-PO4 Glass 100 --
4 BOD Glass, PE 1000 4oC, Dark
5 Coliforms Glass, PE 300 4oC, Dark
6 Heavy metals (Cd, Zn) Glass, PE 500 HNO3, pH
7 Mercury Glass 1000 HNO3, pH
8 Pesticides Glass, PE 1000 4oC, Dark
Reagent Solutions Instruments (required at field)
• Solutions & reagents for estimation of DO
• Buffer solutions (pH = 4, 7, & 9) to
standardise the pH meter
• Standard KCl solution (0.01M) to
standardise the EC meter.
• For preservation of certain samples,
conc. HNO3, conc. H2SO4, etc.
• Distilled water for rinsing equipment.
Instruments
• A (mercury) Thermometer
• pH meter
• EC meter.
Field Analysis
Measurements of colour, odour,
temperature, EC, pH and DO
are considered to be 'Field
Determinations' and should be
made as soon as possible after
collecting a sample.
Measurement of these parameters can be made in the field if
field meters are available. This is the best option, as the
analyses will be made immediately.
If samples are brought to the laboratory, the travel time should
be very short, so that parameter values do not change between
the time the sample is collected at the time of analysis.
Colour
Determining the colour in the field is relatively easy.
Pour an aliquot of approx.10ml of sample into a glass
test tube and judge the colour observed. Consider one
of the following options:
(1) Light brown
(2) Brown
(3) Dark brown
(4) Light green
(5) Green
(6) Dark green
(7) Clear
(8) Other specify
Odour
Determining the odour should always be done in the field, as soon
as possible after collecting a sample. After collection, fill a cleaned
odourless bottle half-full of sample, insert stopper, shake vigorously
for 2-3 seconds and then quickly smell the odour.
Alternatively, pour an aliquot of approx. 5ml of sample into a glass
test tube and judge the odour. Consider one of the following
options:
(1) Odour free
(2) Rotten eggs
(3) Burnt sugar
(4) Soapy
(5) Fishy
(6) Septic
(7) Aromatic
(8) Chlorinous
(9) Alcoholic
(10) U l t
Temperature
Water temperature should be measured in degrees Celsius, using
a mercury thermometer or a thermistor. Normally, if temp. is
measured electronically using a thermistor this device is built into
an instrument which is capable of making other water quality
measurements (e.g. pH and EC).
Whenever possible, the temperature should be measured by
directly dipping the thermometer in the natural body of water
being studied.
In case it is not possible, collect about 500 ml sample in a plastic
or glass container and measure temperature by immersing the
thermometer in the sample. Read the temperature after
equilibration.
Report the Temperature on the sample identification form in
degrees Celsius with 1 digit after the decimal point e.g. 13.2oC.
pH
Before measuring pH, it is necessary to calibrate the meter. This should be done at
least once per day, before the first pH measurement is attempted.
The procedure of this is as follows:
After removing their protective caps, the electrodes are rinsed in distilled water
and carefully blotted dry with soft absorbent paper. NOTE: needs careful handling
The electrodes are then placed in a fresh buffer solution and after stabilisation,
pH reading of the meter is adjusted to the pH of buffer solution (normally pH=7).
The electrodes are then rinsed again with distilled water and blotted dry.
If a pH measurement is not to be taken immediately, the electrodes should be
replaced in their protective caps. Normally, the glass electrode cap is filled with
distilled water before replacement to prevent the electrode drying out.
Report the pH on the sample identification form in pH units showing one digit
after the decimal point, e.g. 7.6.
Once calibrated, the pH meter can be used to measure the pH directly by placing the
electrodes in water sample immediately after it is obtained. Care should be taken to
ensure that the electrodes are rinsed with distilled water before and after each
determination and that distilled water is placed in to the glass electrode cap.
Electrical Conductivity (EC)
Before measuring conductivity it is necessary to calibrate the meter
at least once per day (before 1st measurement).
Calibration is achieved by determining the conductivity of a known,
fresh solution of KCl and adjusting the meter accordingly. In order to
ensure the conductivity reading is accurate, it is necessary to adjust
the conductivity reading to compensate for temperature changes.
Once calibrated, the conductivity of the water can be measured by
immersing electrode in a sample of water as soon as it is taken. It is
important to remember that conductivity meters often take some
minutes to stabilise. The reading must, be taken after stabilisation.
Report the EC at 25°C preferably in µmhos/cm with no figure after
the decimal point, e.g. 1135 µmhos/cm.
Field Data Protocol
a. Sampling team members
b. Date & time (24 hr method) of collection (time span
in case of composite sampling)
c. Nature of the sample: spot/composite/integrated
d. Results of performed in-situ analyses (water/air temp., DO, pH, EC, turbidity,
macrofauna composition, macrofauna diversity, and 24 hr oxygen P/R ratio)
e. Exact sampling location and depth of collection
f. Definition of sampling intervals and volumes in case of composite sampling
g.Maximum depth of the river, lake and current velocity in case of river
h.Weather conditions w.r.t. clouds, precipitation, wind (direction & force)
i. Consistency of sediment (sandy, silty etc.)
j. Comments on smell, colour, discharges etc.
k. Parameter(s) that will be analyzed
l. Sample bottle (number, type, material, volume, preservative)
m. The method of preservation/storage
Analytical result sheets
When offering the samples to the
analytical laboratory, each and every
series of replicate sample containers
has to be accompanied by a prefilled
"result sheet".
This sheet is marked with sample specifications identical to
the specs marked on the bottle. The individual parameters to
be measured in the sample are tabulated, together with the
units they should be reported in. The sheet leaves space for
the analytical lab to fill in the results of replicate analysis.
Special Care in Water Sampling
In specific situations
Inform concern agencies in advance and
keep record, if some dispute is anticipated
Do not inform the operating agency, if
sampling is for monitoring and/or quality
control
Get relevant details of technology and/or
installation, if sampling is for performance
assessment
Get information on sensitive issues before
preceding for filed sampling
Ensure infrastructural facilities and safety
in sampling programme
