Chapter 6 Laboratory Tests

6.1 Introduction

In order to get the qualitative results required for this project laboratory tests on the
water samples were done.

The tests were carried out at The National university of Science and Technology;
laboratory for the department of Civil and Water engineering. The test for nitrate was
carried out at the Criterion laboratories due to lack of reagent at the university lab.

Basically two different types of tests were carried out namely the Paqualab test on
coliforms which is a biological parameter, and the Palintest which gives the
concentrations of the physical parameter being tested.

6.2 The Palintest

The photometer is an instrument used for measuring light intensity. Light from an
incandescent lamp is passed through a test tube containing the sample solution, and
then through a coloured filter on to a photocell. Five built-in filters offer a choice of
wavelengths covering the visible spectrum. Light detected by the photocell is
recorded as a digital response. The display shows percentage transmittance (%T)
which is proportional to the incidence light that reaches the photocell.

In Palintest colour match tests, the photometer is used to measure the colour produced
when reagent tablets are added to the sample solution. The colour intensity is
proportional to the parameter under test.In order to obtain the test result; the
transmittance reading obtained is compared against the calibration chart of the
parameter test carried out.
6.2.1 Procedure

1. Place distilled water in a test bottle up to the 10ml mark and close the bottle
tightly. This will be the blank tube.
2. Place the sample in another test bottle up to the 10ml mark. This will be the
sample tube.
3. Crush and mix to dissolve the relevant tablet(s) in the sample tube for the
parameter being tested and stand a time specified by the test.
4. Select a wavelength as stated in the instruction of the test being carried out.
The wavelengths available on the photometer are 410, 490, 520, 570, and
640nm.
5. Place the blank tube in the test chamber of the instrument, (the chamber that
lights at the bottom when the on button is pressed.) and the sample tube in
the tube holder of the instrument.
6. Place the on button until the display reads 100 (100%T)
7. Quickly exchange the positions of the blank and the sample tubes such that
the blank will be in the tube holder and the sample tube in the test chamber.
8. Note the reading displayed when steady. (NB-machine turns off
automatically after 6-8 seconds.)
9. Compare the reading %T against appropriate calibration chart to get the
tested parameter’s concentration in mg/l.

6.3 Paqualab test for coliform count

6.3.1 Apparatus

Petri-dishes
Sterile pads
Forceps
Pipette
Incubator
Macconkey Broth
6.3.2 Procedure

1. Place a pad in the Petri-dish using forceps and spread the sample to be
tested on the pad using a pipette.
2. Using a pipette add 2ml of broth on to the pad. The broth is prepared as
stated in clause 6.3.3
3. Replace the lid of the Petri-dish and place the dish in the incubator.
4. Repeat steps 1-3 for all the samples to be tested.
5. Switch on the incubator an hour after the last sample has been taken.
Incubate for about 16hrs.
6. After the incubation period, remove the petri-dishes from the incubator and
count the colonies with a yellow colour and diameter 1mm.this reading will
be cfu/2ml because 2ml of sample was used therefore to get the value of
cfu/ml the number of counted colonies was divided by two.

6.3.3 Preparation of Broth

The broth used was Macconkey Broth. It is a powder purple in colour and indicates a
coliform presence by a colour change to yellow.

To prepare a single strength broth ;add 40g of powder to 1 litre of distilled water and
mix to dissolve.

6.3.4 Possible points of error

1. The water was not distilled

2. The apparatus was not sterilised since a steam steriliser is not available in the
lab.

3. The incubation period lasted for more than 16hrs (it was about 20hrs)