3 WHE+ Lab Manual

WATER HARMONY
ERASMUS+
Selected laboratory courses for graduate courses
on water supply and wastewater treatment
Lab Course
Version 2
June 2018
Lab course guide www.waterh.eu Page 1/128

PREFACE
The main goal of the Water Harmony Erasmus+ project is to develop improved learning
and teaching tools, methodologies and pedagogical approaches using best practices. As a
result, skills will be developed on multiple use of resources and long-term planning for
multiple benefits in such a way that project partners will have harmonized teaching and
pedagogical approaches in water related graduate education.
Development of teaching materials like lectures, text books and e-learning is a part of
the project. The development of lab courses is also an important part of this process,
especially to utilise some of the instruments purchased through the project. This booklet
presents selected lab courses drafted jointly by the project partners.
On behalf of the project I would like to thank all the partners for their excellent
contributions for this booklet.
Harsha Ratnaweera
Project Coordinator
Water Harmony Erasmus+

Content
1 Good Lab Practices .......................................................................................................................................... 1

1.1 Quantitative analysis. Spectrophotometric determination of Fe ions in ground water............................. 1
Lab purposes.................................................................................................................................................... 1
Theory .............................................................................................................................................................. 1
Equipment and reagents .................................................................................................................................. 4
Tasks performance order ................................................................................................................................. 5
Report content .................................................................................................................................................. 8
Discussion issues: ............................................................................................................................................ 9
References ..................................................................................................................................................... 10
1.2 Determination of alkalinity, acidity, hardness, pH, conductivity, turbidity and SUVA in samples of
natural water. ...................................................................................................................................................... 11
Lab purpose ................................................................................................................................................... 11
Theory ............................................................................................................................................................ 11
Equipment and reagents ................................................................................................................................ 11
Tasks performance order ............................................................................................................................... 11
Experiment 1. Determination of alkalinity of the water samples..................................................................... 11
Experiment 2. The determination of general acidity of water ......................................................................... 12
Experiment 3. The determination of hardness of water. ................................................................................ 13
Experiment 4. The determination of pH of water. ........................................................................................... 13
Experiment 5. The determination of conductivity of water. ............................................................................ 15
Experiment 6. The determination of turbidity of water.................................................................................... 16
Experiment 7. The determination of SUVA .................................................................................................... 16
Report content ................................................................................................................................................ 17
Discussion issues: .......................................................................................................................................... 19
References ..................................................................................................................................................... 20
2 Analytical procedures ..................................................................................................................................... 21
2.1 Determination of SS, BOD, COD of natural water. ................................................................................ 21
Lab purpose ................................................................................................................................................... 21
Theory ............................................................................................................................................................ 21
Experiment 1. The determination of dry residue in samples of natural water. ............................................... 22
Experiment 2. The determination of losses at the calcination of dry residue. ................................................ 22
Experiment 3. The determination of permanganate oxidation in natural water (Kubel’s method). ................ 22
Experiment 4. Chemical Oxygen Demand (COD). ......................................................................................... 23
Report content ................................................................................................................................................ 30
Discussion issues: .......................................................................................................................................... 30
References ..................................................................................................................................................... 30

2.2 Biochemical Oxygen Demand ............................................................................................................... 31
Theory ............................................................................................................................................................ 31
Dilution method of BOD5 determination ......................................................................................................... 31
Manometric method of BOD5 determination .................................................................................................. 32
3 Unit processes................................................................................................................................................ 37
3.1 Coagulation ........................................................................................................................................... 37
Lab purpose ................................................................................................................................................... 37
Theory ............................................................................................................................................................ 37
Report content ................................................................................................................................................ 40
Discussion issues ........................................................................................................................................... 41
References ..................................................................................................................................................... 41
3.2 Filtration ................................................................................................................................................. 42
Lab purpose ................................................................................................................................................... 42
Theory ............................................................................................................................................................ 42
Report content ................................................................................................................................................ 43
3.3 Rapid Sand Filtration and Ultrafiltration ................................................................................................. 45
Experiment 1: Direct filtration of a model suspension in a rapid sand filter.................................................... 45
Theory ............................................................................................................................................................ 45
Tasks .............................................................................................................................................................. 45
Experimental approach .................................................................................................................................. 46
Experiment 2: Cross-flow ultrafiltration of a model suspension. .................................................................... 48
Theory ............................................................................................................................................................ 48
Task................................................................................................................................................................ 48
Experimental approach .................................................................................................................................. 48
3.4 Adsorption on Activated Carbon ............................................................................................................ 51
Theory ............................................................................................................................................................ 51
Experiment 1: Determination of a single-solute isotherm............................................................................... 51
Experiment 2: Determination of the internal mass transfer coefficient. .......................................................... 53
Experiment 3: Adsorption of breakthrough curves of a single-solute in a fixed-bed granular activated
carbon column ................................................................................................................................................ 55
3.5 Membrane methods for cleaning water solutions: ultrafiltration, nanofiltration, reverse osmosis ......... 56
Lab purpose ................................................................................................................................................... 56
Theory ............................................................................................................................................................ 56
Report content ................................................................................................................................................ 61
References ..................................................................................................................................................... 62
4 Operation........................................................................................................................................................ 63
4.1 Microscopy of activated sludge ............................................................................................................. 63
Microscopy ..................................................................................................................................................... 63
Practice of microscoping ................................................................................................................................ 66
The preparations ............................................................................................................................................ 66
The investigation ............................................................................................................................................ 67
Assessment of the flocs ................................................................................................................................. 69
Bacteria .......................................................................................................................................................... 70
Vegetable Organism....................................................................................................................................... 74
Animal unicellular organism ........................................................................................................................... 75
Flagellates ...................................................................................................................................................... 75
Multicellular organism..................................................................................................................................... 80
Additional microscopic examinations ............................................................................................................. 80
References ..................................................................................................................................................... 83
4.2 Optimizing recovery of RO under different salinity ................................................................................ 84
Lab purpose ................................................................................................................................................... 84
Theory ............................................................................................................................................................ 84
Report content ................................................................................................................................................ 93
Questions ....................................................................................................................................................... 93
4.3 Impact of the main parameters on the effectiveness of coagulation and the determination of the
optimal dose of coagulant ................................................................................................................................... 95
Lab purpose ................................................................................................................................................... 95
Theory ............................................................................................................................................................ 95
Determination of the efficiency of removal of phosphates by coagulation method, selection of the optimal
dose of coagulant ........................................................................................................................................... 98
Experiment 1: Coagulation at pH 9 (NaOH substrate reagent). ..................................................................... 98
Experiment 2: Coagulation with the addition of CaO as a subletting reagent. ............................................... 98
Report content ................................................................................................................................................ 99
References ..................................................................................................................................................... 99
4.4 The dynamic exchange capacity of cation exchanger ......................................................................... 100
Lab purpose ................................................................................................................................................. 100
Theory .......................................................................................................................................................... 100
Equipment and reagents .............................................................................................................................. 101
Tasks performance order ............................................................................................................................. 101

Experiment 1. Ion exchange filtration of calcium chloride solution............................................................... 101
Experiment 2. The regeneration and washing up of the cation exchanger .................................................. 102
Report content .............................................................................................................................................. 102
Discussion issues ......................................................................................................................................... 104
References ................................................................................................................................................... 104
4.5 Evaluation of the carbonic-acid equilibrium ......................................................................................... 105
Lab purpose ................................................................................................................................................. 105
Theory .......................................................................................................................................................... 105
Equipment and reagents .............................................................................................................................. 113
Tasks performance order ............................................................................................................................. 113
Report content .............................................................................................................................................. 120
Discussion issues: ........................................................................................................................................ 120
References ................................................................................................................................................... 122

1 Good Lab Practices
1.1 Quantitative analysis. Spectrophotometric determination of Fe ions in

ground water.
Lab purposes
1. Introduction to spectrophotometric analysis.
2. Preparation of the calibration graph.
3. Learning how to use micropipettes and macropipettes.
4. Learning how to take ground water samples and their preparation.
5. Familiarization with the Fe determination in natural water samples.
Theory
Introduction
Iron (Fe) is the first element in Group VIII of the periodic table. It has an atomic number of 26,
an atomic weight of 55.85 and common valences of 2 and 3.
The average abundance of Fe in the earth’s crust is 6.22 %. In the soil, Fe ranges from 0.5 to
4.3 %. In streams, it averages about 0.7 mg/L, and in ground water, it is 0.1 to 10 mg/L.
Iron is present in the following minerals: hematite (Fe2O3), magnetite (Fe3O4), taconite (a low-
grade siliceous iron formation) and pyrite (FeS2).
The solubility of iron compounds in water is controlled by the carbonate concentration.
Because groundwater is often anoxic, any soluble iron in groundwater exists usually in the
ferrous (Fe2+) state, presumably as Fe(HCO3)2. On exposure to air or addition of oxidants,
ferrous iron oxidize to the ferric state (Fe3+) and may hydrolyze to form a red insoluble hydrated
ferric oxide. Oxidation and hydrolysis of Fe2+ occur by the following schema:
4Fe+2 + O2 + 2H2O = 4Fe+3 + 4OH-
4Fe+3 + 3H2O = Fe(OH)3 + 3H+

Or:
4 Fe(HCO3)2 + O2 + 10H2O = 4Fe(OH)3 + 8 H2CO3
Or
4 Fe(HCO3)2 + O2 + 2H2O = 4Fe(OH)3 + 8CO2
In the absence of complex forming ions, ferric ion is not significantly soluble unless pH is very
low.

Elevated iron concentration in water can cause stains in plumbing, laundry and cooking
utensils.
Allowable level of iron concentration in water for irrigation is 5 mg/L and in water for drinking -
0.2 mg/L in the USA, and 0.1 mg/L in EU.
High concentration of iron in drinking water (more than 0.3 mg/L) is inadvisable since it causes
turbidity of the water and results in an unpleasant taste.
Increased iron concentration causes stains on the washed underwear.
Some industries, e.g. paper, need water free of iron.
High iron concentration has a bad influence on the growth of some kinds of fish, mainly
salmonids.
Principle of determination
Quantitative analysis of Fe in water samples can be carried out by atomic absorption

spectrophotometric method, the inductively coupled plasma method and by the method of
absorption spectrophotometry.
Absorption spectrophotometry is based on a selective absorption of light radiation with
specified wavelength by the solution of analyzed substance. The photo-colorimetric analysis
utilizes the characteristic color of the ions. If the ions are uncolored, it is required to convert
them to a colored chemical compound.
For spectrophotometric determination of iron in samples of natural, tap, or sewage water, the
standard Phenanthroline Method is used. The principle of this method consists in following:
The Iron brought into solution is reduced to the ferrous (Fe+2) state by boiling with acid and
hydroxylamine. The equation for this reaction is:
2NH2OH + 4Fe3+ = N2O + 4Fe2+ + H2O + 4H+.
After that Fe2+ ions treat with 1,10 phenanthroline at pH 3,2 to 3,3. The molecules of
phenanthroline chelate atoms of ferrous Iron to form an orange-red complex as shown below:
In this reaction, n is equal to 3.

The excess of 1,10 phenanthroline in a water sample with pH range 2,9 – 3,5 insures rapid
colour development. (Colour standards are stable for at least 6 months).
Interference: The interfering substances are strong oxidizing agents, cyanide, nitrite,
phosphates, chromium, zinc, cobalt, copper, nickel, bismuth, cadmium, mercury, molybdate,
silver.
The initial boiling of selected samples with acid and hydroxylamine (pH = 3,2 – 3,3) and
following adding of excessive amounts of 1,10 phenanthroline eliminates the action of
interfering substances. This is the reason why the measurements should be carried out in an
excess of 1,10 phenanthroline and at recommended pH value.
Spectrophotometer
A spectrophotometer (photo-colorimeter) is an instrument designed to accurately measure the

absorbance of monochromatic light by a substance placed in the beam (cell) as shown below:
Light source → Monochromatic unit → Cell → Detector → Meter

Incident Beam (I0) → Absorbed Beam (It)
In the spectrophotometric study, the concentration of the iron (Fe2+) ions can be determined
using a method with a calibration graph. In this method, a dependency between concentration
and absorbance must be determined. For this purpose, colored standard solutions of known
concentration of iron Fe2+ are prepared and their optical densities measured at the chosen
wavelength. Usually, it is a linear dependence, described by a basic principle of absorption
spectrophotometry - the Lambert-Beer law, described by the equation:
where: A – absorbance,
I0 – intensity of light incident on a sample,
It – intensity of transmitted light,
a – molar absorption coefficient (constant value, dependent
on the light wavelength, properties of solved substance and
temperature),
b – thickness of the absorbing layer,
c – concentration of the absorbing substance.
As can be seen on the equation, absorbance is proportional to the concentration of absorbing
molecules and thickness of the layer.
Notice that as the concentration gets larger and larger, less and less light will pass through the
solution, until the concentration gets to a point where no light will pass through and the "law"
fails. It is always necessary to check whether the Lambert- Beer's law is obeyed and the plot
of absorbance vs. concentration yields a straight line over the range of concentrations studied
before an analysis of an unknown sample is made. Since neither cell path length b nor the
extinction coefficient a are changing over the course of the experiment, we can replace them
with a constant k, which will be determined from the slope of linear regression data for the
calibration curve.
In practice, the absorbance measurements carried out in the same cuvette or two identical
cuvettes, with the same thickness b.
Determination of unknown concentration in a sample (Cx) is done by reading value from the
calibration graph A=f(c). A series of standard solutions with known concentrations are used for
this purpose. The concentration of determined substance should be in a range of the standard
solutions. The analyzed sample and the standard solution should be prepared via the same
method.
Reading of concentration
A Ax of analysed substance
b (Cx) from the calibration
s graph A = f(c).
o
r Ax – absorbance of the
b sample.
a
Cx
Concentration
Figure 1: Calibration graph
Equipment and reagents
 Spectrophotometer UV-Vis or photo-colorimeter for use at 510 nm, providing a

light path of 1 cm or longer.
 Micropipettes and macropipettes,
 Nessler tubes, matched 100 mL, tall form,
 Acid-washed glassware (Washed with conc. HCl and rinse with reagent water before
use to remove deposits of iron oxide),
 Separatory funnels: 125 mL,
 HCl solution (0.1 M),
 Hydroxylamine solution (Dissolve 10g NH2OH*HCl in 100 mL water),
 Ammonium acetate buffer solution (Dissolve 250g NH4C2H3O2 in 150 mL water. Add
700 mL conc. acetic acid),
 Sodium acetate solution (Dissolve 200g NaC2H3O2*3H2O in 800 mL water),
 Phenanthroline solution (Dissolve 100g 1.10 phenanthroline monohydrate
C12H8O2*H2O in 100mL water by stirring and heating to 80oC. Do not boil. Heating is
unnecessary if two drops of conc. HCl added to the water).
 Stock iron solution (0,200 mg iron per 1mL), Weight 200.0 mg of “iron wire for
standardizing” and place in a 1000 mL volumetric flask. Dissolve the handling in 20 mL
6N sulphuric acid (H2SO4) and dilute to mark with water. (1 mL of stock iron solution
consists of 200 µg Fe).
 Standard iron solutions (10 µg/mL,). Pipet 50 mL stock solution into 1000 mL volumetric
flask and dilute to the marc with water.
Safety
According to safety regulations, it is obligatory to wear a lab coat and closed shoes in
the lab, safety glasses and gloves.
The wearing of safety glasses/goggles is mandatory at all time. Those students wearing
prescription glasses must wear goggles over their glasses. Students without prescription
lenses must wear the safety glasses provided. Contact lenses should not be worn in the lab.
Tasks performance order
1. Preparation of reference solutions
Prepare a series of color standards (range of Fe+2 concentration 0.01 -1.0 ppm) by
accurately pipetting calculated volumes of standard iron solutions to measure 1 to 100 µg
portions into 125-mL Erlenmeyer flasks and diluting to 50 mL by adding measured volumes of
water. Add 2 mL conc. HCl and 1 mL NH2OH*HCl solution. Transfer this mixture to 100 mL
volumetric flask. Add 10mL of NH4C2H3O2 buffer solution and 4 mL of phenanthroline solution
and dilute to mark with water. Mix thoroughly and allow 10 minutes stay for color development.

Table 1: Doses of reagents to prepare 10 standard solutions (including blank solution)
ranging from 1 to 100 µg Fe in final 100 mL volume (concentration of standard solutions - 0.01
– 1 mg/L of dissolved Fe)
Concentration Volume of Volume of Volume of Volume of Volume of the

of Fe3+ standard iron conc. HCl the the phenanthroline
No
solution with solution NH2OH*HCl NH4C2H3O2 solution
.
concentration solution buffer
(10 µg Fe per solution
(ppm, 1 mL)
(mL) (mL) (mL) (mL)
mg/L)
(mL)
1 0 (blank 0 2,0 1,0 10 4,0
solution)
2 0.01 0.1 2,0 1,0 10 4,0
3 0.02 0.2 2.0 1.0 10 4.0
4 0.05 0.5 2,0 1,0 10 4.0
5 0.10 1.0 2,0 1,0 10 4,0
6 0.20 2.0 2,0 1,0 10 4,0
7 0.40 4.0 2,0 1,0 10 4,0
8 0.60 6.0 2,0 1,0 10 4,0
9 0.80 8.0 2,0 1,0 10 4,0
10 1.00 10.0 2,0 1,0 10 4,0
2. The samples preparation
The samples must be collected into clean glass or polyethylene bottles and slightly acidify (2
mL conc. HCl per 100mL of water sample) directly after taking.
3. Determination of total iron:
Mix the water sample thoroughly and transfer 50.0 mL of this sample into 125 mL Erlenmeyer
flask. (If this sample volume contains more than 200 µg iron, use a smaller accurately
measured portion and dilute to 50 mL). Add 2 mL conc. HCl and 1 mL NH2OH*HCl solution.
Add a few glass beads and heat to boiling. Continue boiling until volume decreased to 15-20
mL. After cooling the product to room temperature, transfer it to 100 mL volumetric flask. Add
10 mL of NH4C2H3O2 buffer solution and 4 mL of phenanthroline solution. After that add distilled
water up to the mark of volumetric flask. Mix thoroughly and allow of 10 minutes stay for color
development. Determine the light absorbance of colored sample at λ=510 nm.

4. Determination of dissolved iron
Immediately after collection, filter the sample of water through 0,45 µm membrane filter into
vacuum flask, containing 1 mL of conc. HCl per 100mL of sample. Analyze filtrate for total
dissolved iron like described above. Namely, add to the filtrate1 mL NH2OH*HCl solution. After
that add a few glass beads and heat to boiling. Continue boiling until volume decreased to 15-
20 mL. After cooling the product to room temperature, transfer it to 100 mL volumetric flask.
Add 10 mL of NH4C2H3O2 buffer solution and 4 mL of phenanthroline solution. After that add
distilled water up to the mark of volumetric flask. Mix thoroughly and allow of 10 minutes stay
for color development. Determine the light absorbance of colored sample at λ=510 nm.
5. Determination of ferrous iron

Determine ferrous iron at sampling site because of the possibility of change in the ferrous-
ferric ratio (Fe+2 ↔ Fe+3) with time.
To determine ferrous iron only acidify the sample with 2 mL conc.HCl / 100mL sample at the
time of collection. Fill bottle directly from sampling source and stopper. Immediately withdraw
a 50 mL portion of acidified sample and add 20 mL of phenanthroline solution and 10 mL of
NH4C2H3O2 solution with vigorous stirring. Dilute to 100 mL and measure the color intensity
within 5 to 10 minutes.
Do not expose to sunlight. Color development is rapid in presence of an excess of
phenanthroline. The given phenanthroline volume is suitable for less than 50µg of total iron. If
larger amounts are present, use a correspondingly larger volume of phenanthroline or a more
concentrated reagent.
Calculate ferric (Fe+3) iron by subtracting ferrous iron concentration from the concentration of
total iron.
The Use of Spectrometer
1. Turn on the instrument and allow it to warm up for about twenty minutes.
2. Rotate the wavelength selector to 510 nm. Approach the wavelength from the low end
of the scale and stop at 510 nm. Do not search back and forth for the exact setting.
3. With the door of the sample holder closed, adjust the "Zero Adjust" knob to bring the
needle to zero on the percent transmission scale.
4. Fill a cuvette about half or two-thirds full with the blank solution, wipe it clean, and
insert into the sample holder. Close the sample door and adjust the "Light Control"
knob until the meter reads 100% transmittance.
5. Replace the sample blank with your first diluted standard iron solutions and record its
transmittance. Repeat with the rest of your series of diluted standard iron solutions.
Recheck the zero and 100% transmittance setting, using procedures 3 and 4 above,
to be certain no "drift" has occurred. If it has, redo the calibration solutions.
6. Record the spectrometer number on your report sheet, as this same spectrometer will
be used for your unknown samples.
Using the spectrophotometer (photo-colorimeter), carefully measure the percent transmittance
of 10 standard solutions and water probes. Record your results in the following table.

Table 2. Solutions transmittance percentage and absorbance.
Solution % Transmittance Absorbance (A=-logT)

0.00 mg/L Fe (blank) 100% 0
0.01 mg/L Fe
0.02 mg/L Fe
0.05 mg/L Fe
0.10 mg/L Fe
0.20 mg/L Fe
0.40 mg/L Fe
0.60 mg/L Fe
0.80 mg/L Fe
1.00 mg/L Fe
total iron
dissolved iron
ferrous iron
Report content
1. Describe briefly the abundance of iron distribution in Earth crust. In what state it is
present in natural water and why is required to purify water from the excess of iron?
2. Describe the principle of colorimetric method used for quantitative determination of iron
in water probes.
3. Describe the procedures of preparation of standard solutions and water probes for
execution of colorimetric measurements. Reproduce the Table 2 in report.

Plot the calibration graph of the function: the concentration of Fe in standard solutions vs.
absorbance ( A ) like shown below:
Calibration graph
1,2
1
ABSORBANCE
0,8
0,6
0,4
0,2
0
0 0,2 0,4 0,6 0,8 1 1,2
CONCENTRATION , MG/L

Draw the best fitting straight line through the points – this called the Beer-Lambert Law plot.
By using plotted line and measured data of light beam absorbance in studied samples of water,
calculate the concentration of Fe in each of them. Explain the cause of the differences in results
obtained.
Based on the literature, explain why the ground water samples require acidifying.
Discussion issues:
1. What is the purpose of using the hydroxylamine hydrochloride in procedures of preparation

the standards and water samples?
2. The reaction: Fe+2 + 2H2O → Fe(OH)2 + H+ occurs readily. Why would this be a problem in
the determination of the amount of Fe+2 ? What procedure was used in analytical experiments
to prevent this reaction from interfering with the determination of the percent Fe+2 in the
unknown sample?
3. Why the "blank" sample is necessary at performing of colorimetric measurements?
4. How to determine Fe+3 concentration in water samples using phenanthroline method?

References
1. Standard Methods for the Examination of Water and Wastewater. ISBN: 9780875532875.
Author: E.W. Rice, R.B. Baird, A.D. Eaton, editors. Publisher: American Public Health
Association, American Water Works Association, Water Environment Federation. Publication
date: 2017. AWWA catalog no: 10086. Media Type: HARDBACK

1.2 Determination of alkalinity, acidity, hardness, pH, conductivity, turbidity
and SUVA in samples of natural water.
Lab purpose
The aim of this work is to learn the standard methods of evaluation of the following water quality
indicators: pH, alkalinity, acidity, conductivity, hardness, turbidity and SUVA.
Theory
The alkalinity of natural water is caused by weak acid anions and OH- ions. Total alkalinity is
characterized by concentration of ions OHˉ, HCO3ˉ, CO32ˉ and anions of other weak acids.
The acidity of natural water with pH > 4,5 is caused by free carbon (IV) oxide, humiс and other
weak organic acids; with pH < 4,5 by strong acids and salts of strong acids and weak bases.
It has distinguished the total acidity, free acidity, free carbon (IV) oxide, aggressive carbon (IV)
oxide and also acidity independent from other weak non-volatile acids.

Equipment: pipettes (100 cm3) – 5 ones, bulbs for the titration (250 cm3) – 5 ones; pH-meter;
Burettes.
Reactants: hydrochloric acid solutions – 0,05 M and 0,1 M; sodium hydroxide solution 0,1 M;
indicators – methyl orange, phenolphthalein; EDTA standard solution, water samples.
Experiment 1. Determination of alkalinity of the water samples.
Alkalinity reflects the amount of substances in water that can be neutralized by strong acids.
These substances are hydrocarbonates, carbonates, and hydroxides. Depending upon
indicator used, one can evaluate the total alkalinity, carbonate alkalinity and alkalinity
stipulated by OHˉ ions.
When phenolphthalein is used as indicator, the titration of water sample with hydrochloric acid
standard solution until vanishing of red color of indicator, required the consumption of
hydrochloric acid standard solution V1. When methyl orange is used as indicator the titration
of water sample with hydrochloric acid standard solution until its colour just changes from
yellow to orange, required the consumption of hydrochloric acid standard solution V2. These
data allow to calculate the alkalinity of water sample due to presence of OHˉ ions (expenditure
of hydrochloric acid standard solution V1), and calculate the total alkalinity (TA) of water
samples with V1+V2 by equations:

PA (CaCO3, mg/L)=
TA (CaCO3, mg/L)=
Where, C is the hydrochloric acid standard solution concentration (mol/L); V is the water
sample volume (mL); 50 - (1/2 CaCO3) molar mass (g/mol).
To determine the composition of alkalinity:
Alkalinity consists of carbonate and hydroxide if V1> V2; alkalinity consists of carbonate and
bicarbonate if V1 <V2; alkalinity from carbon Acid salt composition; V1 = 0, alkalinity only
bicarbonate; V2 = 0, alkalinity only hydroxide.
Step 1. Determination of phenolphthalein alkalinity of water.
Step 2. Determination of total alkalinity of water.
Experiment 2. The determination of general acidity of water.
Acidity refers to the total amount of material in water that can neutralize with strong bases,
mainly indicating the presence of strong acids, weak acids, strong acids and weak bases, and
other substances in water. There are two ways to express acidity: phenolphthalein acidity (total
acidity) and methyl orange acidity.
Take a suitable amount of water (VmL) in 250 mL Erlenmeyer flask, diluted with carbon
dioxide-free water to 100 mL, add two drops of methyl orange indicator, titration with sodium
hydroxide standard solution until the solution color changed from orange to yellow orange,
count Sodium hydroxide standard solution dosage (V1). Calculate the Methyl orange acidity
(MOA) in water sample with V1.
Another sample of water (VmL) placed in 250mL Erlenmeyer flask, diluted to 100 mL with
carbon dioxide-free water, add 4 drops of phenolphthalein indicator, titration with sodium
hydroxide standard solution until the solution just changed to light red. Note the amount of
sodium hydroxide standard solution (V2). calculate the total acidity (TA) of water samples with
V2.
MOA (CaCO3, mg/L)=
TA (CaCO3, mg/L)=
Where, C is NaOH standard solution concentration (mol/L); V is the water sample volume (mL);
50 - (1/2 CaCO3) molar mass (g/mol).
Step 1. Determination of methyl orange acidity of water.
Step 2. Determination of total acidity of water.

Experiment 3. The determination of hardness of water.
Total hardness of water refers to the total concentration of calcium and magnesium ions in
water, including non- and carbonate hardness.
Step 1. Prepare three water samples.
Step 2. Add 5 ml ammonia - ammonium chloride buffer solution (pH = 10) and 2 or 3 drops of
chrome black T indicator to the samples.
Step 3. Titrating the samples with EDTA standard solution until the color turns from wine red
into pure blue.
Total permanent water hardness is calculated with the following formula:

TOTAL PERMANENT HARDNESS = CALCIUM HARDNESS + MAGNESIUM HARDNESS
The calcium and magnesium hardness are the concentration of calcium and magnesium ions
expressed as equivalent of calcium carbonate. The molar mass of CaCO3, Ca2+ and Mg2+ are
respectively 100,1 g/mol, 40,1 g/mol and 24,3 g/mol.
The ratio of the molar masses is:
So total permanent water hardness expressed as equivalent of CaCO3 can be calculated

with the following formula:
[CaCO3] = 2,5 [Ca2+] + 4,1 [Mg2+].
Experiment 4. The determination of pH of water.
In chemistry, pH (/piːˈeɪtʃ/) is a logarithmic scale used to specify the acidity or basicity of

an aqueous solution. It is approximately the negative of the base 10 logarithm of the molar
concentration, measured in units of moles per liter, of hydrogen ions. More precisely it is the
negative of the base 10 logarithm of the activity of the hydrogen ion. Solutions with a pH less
than 7 are acidic and solutions with a pH greater than 7 are basic. Pure water is neutral, at pH
7 (25 °C), being neither an acid nor a base. Contrary to popular belief, the pH value can be
less than 0 or greater than 14 for very strong acids and bases respectively.
Measurements of pH are important in agronomy, medicine, chemistry, water treatment, and
many other applications.
The pH scale is traceable to a set of standard solutions whose pH is established by
international agreement. Primary pH standard values are determined using a concentration
cell with transference, by measuring the potential difference between a hydrogen
electrode and a standard electrode such as the silver chloride electrode. The pH of aqueous
solutions can be measured with a glass electrode and a pH meter, or an indicator.

Equipment
 pHS-3E digital pH meter
 pH glass composite electrode
 Temperature compensation electrode
 Constant temperature water bath
Figure 1: Digital pH meter

Potentiometric determination of the solution pH: the glass electrode is the indicator electrode
(-), saturated calomel electrode as the reference electrode (+) to form the galvanic cell
at 25 °C, when the pH of the solution changed by 1 unit, the difference of electric potentials
between indicator and reference electrodes changed by 59.0 mV. In the measurement,
choosing the pH close to the water samples as the standard buffer solution to calibrate the pH
meter, and keep the solution temperature constant, in order to reduce the error caused by
liquid junction potential, asymmetry potential and temperature changes. Before the
determination of pH of water samples, two different pH buffer solution were used to calibrate.
After calibration with one pH buffer solution, the error should be within ± 0.1 pH when
determining the pH of another buffer solution that differs by about 3 pH units.
The calibrated pH meter allows direct determination of the pH of the water sample or solution.
Step 1. After rinsing the electrode and the plastic cup with distilled water, rinse them with a
standard buffer solution for 1 to 2 times and blot dry with filter paper.
Step 2. Perform the Calibration of the instrument with the standard buffer solution and
determine the pH of the standard buffer solution.
Step 3. Determination of HCl solution pH.
Step 4. Determination of NaOH Solution pH.
Step 5. Determination of KH2PO4 Solution pH.

Experiment 5. The determination of conductivity of water.
Conductivity (or specific conductance) of an electrolyte solution is a measure of its ability

to conduct electricity. The SI unit of conductivity is siemens per meter (S/m).
Conductivity measurements are used in routinely in many industrial
and environmental applications as a fast, inexpensive and reliable way of measuring the ionic
content in a solution. For example, the measurement of product conductivity is a typical way
to monitor and continuously trend the performance of water purification systems.
In many cases, conductivity is linked directly to the total dissolved solids (T.D.S.). High quality
deionized water has a conductivity of about 5.5 μS/m at 25 °C, typical drinking water in the
range of 5–50 mS/m, while sea water about 5 S/m[2] (or 50,000 μS/cm) (i.e., sea water's
conductivity is one million times higher than that of deionized water).
Equipment
 SLDS-Ⅰ digital conductivity meter
 DJS-1C type platinum black electrode
 Constant temperature water bath
Figure 2: Digital Conductivity meter

1. Conductivity G: For electrolyte solution, commonly using conductivity indicates the size of
its conductive capacity. Conductance G is the inverse of resistor R, namely, G = 1/R.
Conductivity unit is Siemens (S). 1S = 1Ω-1.
It represents conductivity of the cube conductor with an electrode area of 1 m2 and an electrode
pitch of 1 m in S ꞏ m-1. For electrolyte solution, let l/A = Kcell, Kcell is called the cell constant. So
κ = Gl/A = G Kcell
2. Molar Conductivity: Λm = κ/C (2).
The molar conductivity Λm of a dilute electrolyte with strong electrolyte has the following
relationship: Λm = Λ∞m- AC (3).

Where C is the concentration of the solution in units of mol ꞏ m-3, Λ ∞
m is the infinite dilution
molar conductivity.
Draw a straight line Λm to C, the intercept is Λ∞m.
Weak electrolyte in the infinite dilution of the solution can be fully ionized. The molar
conductivity of the solution at this time is Λ∞m = V + Λm, + + V- Λm, - (4)
Where Λm, + and Λm, - respectively, positive and negative ions of infinite dilution molar
conductivity.
According to the ionization theory, it can be assumed that the ionization degree α of the weak
electrolyte is equal: α = Λm/Λ∞m (5).
3. Weak Electrolyte Ionization Equilibrium Constant: Weak Electrolyte AB Ionization
Equilibrium Constant: Kθ = (Cα2)/Cθ (1-α). Therefore, the value of Kθ can be obtained by
experimentally measuring α. Substituting (4) into (6) yields
Kθ = C(Λ∞m)2/Λ∞mCθ (Λ∞m-Λm) or CΛm = (Λ∞m)2 KθCθ1/Λm - Λ∞mKθCθ, (7)
plot CΛm to 1/Λm, the slope of the straight line is (Λ∞m) 2 KθCθ, if you know Λ∞m value, Kθ can be
calculated.
Step 1. Turn on the conductivity meter switch, warm-up 5min.
Step 2. Determination of KCl solution conductivity.
Step 3. Determination of HAc Solution Conductivity.
Experiment 6. The determination of turbidity of water.
Turbidity is a water quality indicator that refers to the obstruction degree that occurs when the
light pass through the suspended solids in water
The suspended solids include soil, silt, micro granular organic matter, inorganic matter,
plankton, etc.
Experiment: Directly measure turbidity of water sample using the turbidimeter
Experiment 7. The determination of SUVA.
Equipment
 UV-spectrophotometer
 0.45 μm microfiltration membrane
Step 1. The determination of UV254 using UV-spectrophotometer.
Step 2. Filtering the water sample prior to determination of dissolved organic carbon.
Step 3. Calculating SUVA according to equation SUVA=UV254/DOC*100.

Report content
Calculation of total alkalinity and phenolphthalein alkalinity
First Second Third Average
V1 (mL)
PA(CaCO3, mg/L)
V2 (mL)
TA(CaCO3, mg/L)
Calculation of acidity of water

V1 (mL)
MOA CaCO3, mg/L)
V2 (mL)
TA (CaCO3, mg/L)
Calculation of hardness
Volume (ml) 1 2 3
Sample
Titration
Average total hardness=
Determination of HCI solution pH:

pH
C/(molꞏL-1)
1.00
0.10
0.01
0.001
0.0001

Determination of NaOH solution pH:
pH
C/(molꞏL-1)
1.00
0.10
0.01
0.001
0.0001
Determination of solution pH:
pH
C/(molꞏL-1)
1.00
0.10
0.01
0.001
0.0001
Determination of KCI solution conductivity:
κ (Sꞏm-1)
C/(molꞏm-3)
10.00
5.00
2.50
1.25
0.625
Determination of HAc solution conductivity:
κ (Sꞏm-1)
C/(molꞏm-3)
0.093
0.0465
0.02325
0.011625

Turbidity:
sample 1 2 3 4 5
NTU
SUVA:
sample 1 2 3 4 5
UV254(cm-1)
DOC (mg/L)
SUVA
Discussion issues:
1. What substances cause the alkalinity of water?
2. What kinds of acidity can be distinguished in natural water?
3. What substances cause availability of free and aggressive carbon acid in water?
4. What is the active water reaction?
5. What water indicators characterize dry residue and losses of residue at the calcination?
6. How to determine the content of organic substances in samples of natural water?
7. What total content of salts allows to recommend water for drinking?
8. What indicator of water quality is determined by the Kubel’s method?
9. The availability of which impurities in water characterizes the value of permanganate

oxidation?

References
1. Powell, Sheppard T., “Sodium Aluminate As An Adjunct To Alum For The Coagulation
Of Public Water Supplies.” American Journal of Public Health (NY). 1927 August; 17(8):
804–809.
2. Ravina, Louis, Everything You Want to Know About Coagulation and Flocculation,
Zeta-Meter, Inc., 4th Ed., 1993.
3. Reaves, Wm J, et.al., Can A Utility Afford To Operate Without On-Line Toc Analyzers
Considering The Requirements Of The D/DBP Rule? On-Line Vs. Grab Samples: What
Are The Advantages And Disadvantages? City Of Houston Public Works And
Engineering , Water Quality Control Group, Houston, Texas, 2001
4. Sawyer, Clair and McCarty, Perry, Chemistry of Environmental Engineering, 3rd ed.,
McGraw-Hill, 1978.
5. Schlicht, Arthur C., The Mc Clariflow - The Ultimate Upflow Solids Contact Clarifier,
Walker Process Equipment, November 1999.
6. Schreppel, Connie, “Plant Performance Picture Emerges with Instrumentation.”
Opflow, American Waterworks Association, January 2010.
7. Sobsey, Mark D., Managing water in the home: accelerated health gains from improved
water supply, World Health Organization, WHO/SDE/WSH/02.07.

2 Analytical procedures
2.1 Determination of SS, BOD, COD of natural water.
Lab purpose
The aim of work is to learn standard methods of the evaluation of salt content and organic
substances content in natural origins of water supply.
Theory
The quantity of salts, dissolved in natural waters, can be determined by dry residue and losses
at calcination. The dry residue is formed from the evaporation of some water volume, filtrated
previously through the paper filter. Filtrate consists of mineral salts and non-volatile organic
substances. The organic part of dry residue of water is determined by residue losses at
calcination process. The data about organic substances content in investigated water can be
obtained higer than the real magnitude if dissolved salts contain nitrogen, carbon etc.
The water is considered fresh, if it usually contents of 1000 mg/L of dissolved salts.
Chemical Oxygen Demand (COD) test determines the oxygen requirement equivalent of
organic matter that is susceptible to oxidation by means of a strong chemical oxidant. It is an
important, rapidly measured (COD) as indicator of measuring organic strength for streams and
polluted water bodies. The test can be related empirically to BOD, organic carbon or organic
matter in samples from a specific source taking into account its limitations. The test is useful
in studying performance evaluation of wastewater treatment plants and monitoring relatively
polluted water bodies. COD determination has advantage over BOD determination. COD
results can be obtained in 3 - 4 hrs as compared to 3 – 5 days required for BOD test. Further,
the test is relatively easy, precise, and is unaffected by interferences as in the BOD test. The
intrinsic limitation of the test lies in its inability to differentiate between the biologically
oxidisable and biologically inert material and to find out the system rate constant of aerobic
biological stabilization.
Equipment: water bath; porcelain cups; conic bulbs; pipettes; paper filters; funnels; burettes;
analytic balances; dry wardrobe; muffle furnace; desiccator.
Reactants: sulfuric acid solution (1:2) (one volume of 96 % mas. and two volumes of
distilled water); 0,01 M solution of potassium permanganate; 0,01 M solution of oxalate acid.

Experiment 1. The determination of dry residue in samples of natural water.
This determination is usually conducted in a day of picking the sample. Water is filtrated
through the paper filter. Porcelain cup is dried to the constant mass, cooled and weighed on
analytical balances. Filtrated water is evaporated on water bath, filled by distilled water (100 –
150 cm3). The cup with dry residue is dried at 100 – 150 0C to the constant mass, cooled and
weighed.
The content of dry residue is calculated by the formula:
where X – the content of dry residue, mg/L; m – the mass of cup, mg; m1 – the mass of cup
with dry residue, mg; V – volume of water sample, cm3.
Experiment 2. The determination of losses at the calcination of dry residue.
For the determination of this losses, the dry residue, obtained in previous experiment, is
calcined in muffle furnace at the temperature 600 0C for 1 hour, cooled in desiccator and the
cup weighed on analytic balances. Losses are calculated by the formula:
where X1 – the content of calcined sample, characterizing mineral impurities in water, mg/L;
m2 – the mass of cup with residue after calcination, mg.
The content of organic part of impurities is found by the formula:
where X2 – the content of organic impurities in water sample, mg/L.
Experiment 3. The determination of permanganate oxidation in natural water (Kubel’s

method).
The water sample (100 cm3), picked by pipette or volumetric flask, is poured to conic bulb with
glass pellets on the bottom, diluted sulfuric acid (5 cm3) and 10 cm3 of 0,01 M solution of
potassium permanganate is added. The bulb must be heat-resistant. The bulbs content is
achieved to boiling. It is boiled exactly 10 min. The 0,01 M solution of oxalate acid (10 cm3) is
added to the bulb with solution. Heated solution is titrated by 0,01 M solution of potassium
permanganate to pale pink color. The temperature of the sample shouldn’t be less than 80 0C.
Then 10 cm3 of 0,01 M oxalate acid solution is poured to the same bulb to another head liquid

and titrated by 0,01 M solution of potassium permanganate. The color shouldn’t disappear for
one minute.
The permanganate oxidation is calculated by the formula:
where X3 – the permanganate oxidation of water, mg of O2 on 1 L of water; V1 – total content

of 0,01 M potassium permanganate solution, taken for the titration, cm3; V2 – volume of
permanganate solution, taken for the titration of oxalate acid (10 cm3, the second titration),
cm3; V – volume of water sample, cm3; 0,08 – oxygen quantity, responding by 1 cm3 of 0,01 M
potassium permanganate solution, mg. (If the calculation of oxidation is in milligrams of the
permanganate per 1 L of water, it’s necessary to use 0,32 instead of 0,08 – the equivalent of
potassium permanganate, responding by 0,01 M solution).
Attention! The determination should be repeated, if water sample will become fulvous or
transparent during boiling.
Experiment 4. Chemical Oxygen Demand (COD).
Chemical oxygen demand (COD) is a measure of the capacity of water to consume oxygen
during the decomposition of organic matter and the oxidation of inorganic chemicals such as
ammonia and nitrite. COD measurements are commonly made on samples of waste waters or
of natural waters contaminated by domestic or industrial wastes. Chemical oxygen demand is
measured as a standardized laboratory assay in which a closed water sample is incubated
with a strong chemical oxidant under specific conditions of temperature and for a particular
period of time. A commonly used oxidant in COD assays is potassium dichromate
which is used in combination with boiling sulfuric acid ( ). Because this chemical oxidant
is not specific to oxygen-consuming chemicals that are organic or inorganic, both of these
sources of oxygen demand are measured in a COD assay.
Chemical oxygen demand is related to biochemical oxygen demand (BOD), another standard
test for assaying the oxygen-demanding strength of waste waters. However, biochemical
oxygen demand only measures the amount of oxygen consumed by microbial oxidation and is
most relevant to waters rich in organic matter. It is important to understand that COD and BOD
do not necessarily measure the same types of oxygen consumption. For example, COD does
not measure the oxygen-consuming potential associated with certain dissolved organic
compounds such as acetate. However, acetate can be metabolized by microorganisms and
would therefore be detected in an assay of BOD. In contrast, the oxygen-consuming potential
of cellulose is not measured during a short-term BOD assay, but it is measured during a COD
test.
Open Reflux method
I. Principle
The open reflux method is suitable for a wide range of wastes with a large sample size. The
dichromate reflux method is preferred over procedures using other oxidants (e.g. potassium

permanganate) because of its superior oxidizing ability, applicability to a wide variety of
samples and ease of manipulation. Oxidation of most organic compounds is up to 95-100% of
the theoretical value.
The organic matter gets oxidized completely by potassium dichromate with silver
sulphate as catalyst in the presence of concentrated to produce and . The
excess remained after reaction is titrated with ferrous ammonium sulphate
. The dichromate consumed gives the oxygen ( ) required for oxidation of
organic matter. The chemical reactions involved in the method are as under:
a)
b)
c)
II. Apparatus and equipment

a. 250 or 500mL Erlenmeyer flask with standard (24/40) tapered glass joints.
b. Friedrich’s reflux condenser (12 inch) with standard (24/40) tapered glass joints.
c. Electric hot plate or six-unit heating shelf.
d. Volumetric pipettes (10, 25, and 50mL capacity).
e. Burette, 50mL with 0.1mL accuracy.
f. Burette stand and clamp.
g. Analytical balance, accuracy 0.001 g.
h. Spatula.
i. Volumetric flasks (1000 mL capacity).
j. Boiling beads, glass.
k. Magnetic stirrer and stirring bars.
III. Reagents and standards

a. Standard potassium dichromate solution, 0.25N (0.04167 M): Dissolve 12.259 g
dried at 103°C for 24h in distilled water and dilute to 1000 mL. Add about 120mg sulphamic
acid to take care of 6 mg/L NO2 – N.
b. Sulphuric acid reagent: Add 10g of Ag2SO4 to 1000mL concentrated and let stand for
one to two days for complete dissolution.
c. Standard ferrous ammonium sulphate approx. 0.25N (0.25M): Dissolve 98g
in about 400mL distilled water. Add 20 mL concentrated and
dilute to 1000 mL.
d. Ferroin indicator: Dissolve 1.485 g 1,10-phenanthroline monohydrate and 695mg
in distilled water and dilute to 100 mL.
e. Mercuric Sulphates: , crystals, analytical grade
f. Potassium hydrogen phthalate (KHP) Standard: Dissolve 425 mg lightly crushed dried
potassium hydrogen phthalate in distilled water and dilute to 1000 mL.
This solution has a theoretical COD of 500 μg O2/mL. This solution is stable when refrigerated,
up to 3 months in the absence of visible biology.
IV. Sample collection, preservation and storage

Preferably collect samples in glass bottles. Homogenize samples containing settleable solids.
If there is delay in collection and analysis, preserve sample by acidification to pH ≤ 2 using
concentrated . Samples can be preserved for maximum 7 days.
V. Calibration
Since the procedure involves chemical oxidation of organic matter by potassium dichromate
as oxidizing agent, which is a primary standard, calibration is not required. For standardization
of ferrous ammonium sulphate, dilute 10mL standard to about 100 mL. Add 10mL of
concentration and allow it to cool. Titrate with ferrous ammonium sulphate ( ) is to
be standardized using 2 - 3 drops of ferroin indicator. (mL ) (0.25)
Normality of =
mL required
The deterioration of can be decreased if it is stored in a dark bottle.
VI. Procedure
Sample preparation: All samples high in solids should be blended for 2 minutes at high speed
and stirred when an aliquot is taken for analysis. Select the appropriate volume of sample
based on expected COD range, e.g. for COD range of 50-500 mg/L take 25 - 50 mL of sample.
Sample volume less than 25 mL should not be pipetted directly, but serially diluted and then a
portion of the diluted sample taken. Dilution factor should be incorporated in calculations.
a. 500 mL of sample diluted to 1000 mL = 0.5 mL sample/mL of diluent, 50 mL = 25 mL of
sample.
b. 100 mL of sample diluted to 1,000 mL = 0.1 mL sample/mL diluent, 50mL of diluent = 5 mL
of sample.
Reflux of samples:
a. Place 0.4 g in a 250 mL reflux sample.
b. Add 20mL sample or an aliquot of sample diluted to 20 mL with distilled water. Mix well.
c. Add clean pumic stones or glass beads.
d. Add 10 mL 0.25N (0.04167M) solution and mix.
e. Add slowly 30 mL concentrated containing mixing thoroughly. This slow
addition along with swirling prevents fatty acids to escape due to generation of high
temperature. Alternatively attach flask to condenser with water flowing and then add
slowly through condenser to avoid escape of volatile organic substance due to generation of
heat.

f. Mix well. If the colour turns green, either take fresh sample with lesser aliquot or add more
potassium dichromate and acid.
g. Connect the flask to condenser. Mix the contents before heating. Improper mixing will result
in bumping and blow out of flask content.
h. Reflux for a minimum of 2 hours. Cool and then wash down condenser with distilled water.
i. Disconnect reflux condenser and dilute the mixture to about twice its volume with distilled
water. Cool to room temperature and titrate excess with0.1M using 2-3 drops of
ferroin indicator. The sharp colour change from blue green to reddish brown indicates end-
point or completion of the titration. After a small time, gap, the blue-green colour may reappear.
Use the same quantity of ferroin indicator for all titrations.
j. Reflux blank in the same manner using distilled water instead of sample.
Alternate procedure for low COD samples less than 50 mg/L: Follow similar procedure with
two exceptions (a) use standard 0.025N (0.004167M) and (b) titrate with standardize
0.025M . The sample volume should be 5mL. Exercise extreme care with this procedure
because even a trace of organic matter on the glassware or from the atmosphere may cause
gross errors. Compute amount of to be added based on chloride concentrations. Carry
blank reagent through the same procedure.
VII. Calculations
COD as mg/L = (a –b) x N x 8000 / mL sample
Where, a = mL used for blank
b = mL used for sample
N = normality of
8000 = Milieq. wt. of O2 x 1000
VIII. Precision and Bias

Precision and bias: A set of synthetic samples containing potassium hydrogen phthalate with
a COD of 200 mg/L was analyzed in many labs with standard deviation of 13 mg/L in absence
of chloride.
Sources of Error:
a. The largest error is caused by using a non-homogeneous sample. Every effort should be
made to blend and mix the sample so that solids are never excluded from any aliquot.
b. Use volumetric flasks and volumetric pipettes with a large bore.
c. The oxidising agent must be precisely measured. Use a volumetric pipette and use
the same one each time if possible.
d. When titrating, be certain that the burette is clean and free of air bubbles.
e. Always read the bottom of the meniscus and position the meniscus of eye level.

Carry distilled water blank through a same procedure to nullify impurities if any. A standard
solution of glucose or potassium acid phthalates should be checked for precision and accuracy
every fortnight. Duplicate analysis is preferred.
Method Sensitivity:
1 Standard procedure is precise and accurate for COD of 50 mg/L or more
2 For low COD more sample volume and the dilute reagents are used
3 Interference by chloride needs to be handles very carefully to get accurate results
IX. Interferences
Fatty acids, straight chain aliphatic compounds, aromatic hydrocarbons, chlorides, nitrite and
iron interfere in the estimation. The interference caused by chloride can be eliminated by the
addition of mercuric sulphate to the sample prior to the addition of other reagents. About 1.0g
of mercuric sulphate is adequate to complex 100mg chloride ions in the sample in the form of
poorly ionized soluble mercuric chloride complex. Addition of to concentrated
as a catalyst stimulates the oxidation of straight chain aliphatic and aromatic compounds.
Nitrite nitrogen exerts a COD of 1.14 mg/mg .
Sulphuric acid at the rate of 10mg/mg may be added to solution to avoid
interference caused by . Aromatic hydrocarbons and pyridine are not oxidised under
any circumstances. Volatile organic compounds will react in proportion to their contact with the
oxidant. For complete oxidation of organic matter, it is necessary to take volumes of Sulphuric
acid and sample plus potassium dichromate in 3:2:1 ratio. However, to maintain the ratio, the
volumes and strength of oxidant/sample may suitable be varied.
X. Warnings
In carrying out the procedures, use proper safety measures, including protective clothing, eye
protection and a fume hood. Reagents containing heavy metals ( and ) should
be disposed of as toxic wastes. Use of such chemicals should be minimized whenever feasible.
XI. Pollution prevention and waste minimization

Since hazardous chemicals like silver and mercury salts, Sulphuric acid, dichromate are used
in the test, the quantity of such chemicals can be minimized by selecting minimum suitable
sample size. The liquid waste generated should be treated as hazardous waste. Adequate
dilution of such waste before final disposal is essential.
Closed reflux (titrimetric and colorimetric) method using COD digester
I. Principle
The closed reflux (titrimetric and colorimetric) method using metallic salt reagents are more
economical but require homogenization of samples to obtain reproducible results. This method
is conducted with ampules and culture tubes with pre-measured reagents which are available
commercially. Moreover, for performing the tests, instructions furnished by the manufacturer
are to be followed. Measurement of sample volume and reagent volume are critical. This
method is economical in the use of metallic salt reagents and generates a smaller quantity of
hazardous wastes.
The principle of the oxidation reaction is similar to open reflux method. Volatile organic
compounds are more completely oxidized in a closed system because of longer contact time
with oxidants. Digestion vessels with premixed reagents are also available from commercial
suppliers.
II. Apparatus and equipment

a. Digestion vessels with premixed reagents and other accessories commercially available.
b. Borosilicate culture tubes 16 x 100 mm, 20 x 150 mm or 25 x 150 mm with TF and lined
screw caps.
c. Borosilicate ampule 10 mL cap – 19 to 20 mm diameter.
d. Block heater to operate at 150 ± 2°C with holes to accommodate digestion vessels. Care
for culture tube caps required.
e. Micro-burette.
f. Ampule sealer.
III. Reagents and standards

Standard potassium dichromate digestion solution 0.01667M: Dissolve 4.903 g ,
primary standard grade, previously dried at 150 ℃ for 2 hours in 500 mL distilled water, add
167 mL conc. at the rate of 5.5g /kg . Let stand 1 to 2 d to dissolve and
mix.
Ferroin indicator solution: Dissolve 1.485 g 1,10-phenanthroline monohydrate and 695 mg
in distilled water and dilute to 100mL. This indicator solution may be purchased
already pre
Standard ferrous ammonium sulphates ( ) titrant, approximately 0.1M: Dissolve 39.2g
in distilled water. Add 20mL concentrated , cool and dilute
to100mL. Standardise this solution daily against standard .
IV. Calibration
Dilute 5mL standard digestion mixture to about 100mL. Titrate with FAS using 0.1 to
0.15mL. (2 to 3 drops) ferroin indicator.
Normality of solution
Volume of 0.01667 M solution treated, mL = x 0.1
Volume used in titration, mL

V. Procedure
a. Treatment of sample with COD of > 50mg/L.
b. Blend sample if the suspended matter is present.
c. Wash culture tubes and caps with 20% before first use.
d. Refer the following to select analytical parameters for proper sample and reagent volume.
e. Place sample in culture tube or ampule.
f. Add digestion mixture.
g. Carefully run sulphuric acid reagent down inside of vessel.
h. Tightly cap the tubes or seal ampules. Invert several times for proper mixing.
i. Place tubes or ampules in preheated reaction block digestor.
j. Reflux for 2h at 150°C behind a protective shield.
k. Cool to room temperature.
l. Remove caps and put TFE covered magnetic stirrer.
m. Titrate while stirring with using 1 or 2 drops of ferrous indicator.
n. The end points are from blue-green to reddish brown.
o. Reflux and titrate blank in a similar way with distilled water.
VI. Calculation
COD as mg O2/L = (A – B) x M x 8000 / ml sample
Where:
A = mL used for blank
B = mL used for sample
M = molarity of , and
8000 = milli equivalent weight of oxygen x 1000 mL/L
VII. Precision and Bias

A set of synthetic containing potassium hydrogen phthalate and was tested by 74
laboratories. At a COD of 200 mg /L in the absence of chloride, the standard deviation was
± 13 mg/L (coefficient of variation, 6.5%). At COD of 160 mg /L and 100 mg /L, the standard
deviation was ± 14 mg/L (coefficient of variation, 10.8%).
For quality control refer Open Reflux Method.
a. Preferable analyze duplicate samples.
b. Proper homogenization is essential for reproducible results.
c. Make volumetric measurement as accurate as possible.
d. Use class-A volumetric flask.

Table 1: Sample and reagent quantities for various digestion vessels
Culture Tubes: Quantity of samples, g

16 x100 mm 5.00 1.50 3.5 7.5
20 x 150 mm 2.50 3.00 7.0 15.0
25 x 125 mm 10.00 6.00 14.0 30.0
Standard10-mL 2.50 1.50 3.5 7.5
Ampules
Report content
COD and BOD of different samples of water

COD, mg O2 / L
Sample BOD, mg O2 / L
Permanganate Dichromate
1
2
3
4
Discussion issues:
1. What water indicators characterize dry residue and losses of residue at the calcination?
2. How to determine the content of organic substances in samples of natural water?
3. What total content of salts allows to recommend water for drinking?
4. What indicator of water quality is determined by the Kubel’s method?
5. The availability of which impurities in water characterizes the value of permanganate

oxidation?
References
1. Standard Methods for the Examination of Water and Wastewater; APHA, AWWA AND
WEF, 21st Edition, 2005.
2. Clair N. Sawyer, Perry Z. McCarty. Chemistry for Environmental Engineering, 4th
Edition, Gener F. Parkin.
3. American Society for Testing and Materials, (1195). Standard Methods for Chemical
Oxygen Demand (Dichromate Oxygen Demand of Water D1252-95, ASTM Annual
Book of Standard, American Testing and Materials, Philadelphia, PA.

2.2 Biochemical Oxygen Demand
Theory
As defined by the DIN EN 1899-1 the Biochemical Oxygen Demand (BOD) is the mass per
unit volume of oxygen for the oxidation of biochemically oxidizable substances contained in 1
L of sample water in n days, in general, n = 5; BOD5, possible n = 7) summarily consumed in
the metabolic activity of a corresponding microbiocenosis at 20 ° C.
The ratio of the theoretical biological oxygen demand, to chemical oxygen demand (COD),
characterizes the degradability of the ingredients of the water to be examined.
The BOD5 / COD ratio is changing during wastewater treatment as follows:
Inflow wastewater treatment plant BOD5 : COD: about 1: 2
Effluent secondary clarifier BOD5 : COD: about 1: 4
The dilution method is the officially approved method of measurement. Here shares of
wastewater sample are diluted with a specially prepared dilution water (see below). The
mixtures are incubated for 5 days at 20 ° C in dark environment.
BOD5 is calculated from the decrease in oxygen concentration in the dilutions as well as in
dilution water (O2 consumption).
The determination of BOD by a manometric method is easier by measuring the oxygen
consumption via pressure measurement (see below).
The determination of the BOD5 should only achieve on the oxygen demand for the oxidation
of carbon compounds. Nitrification causes an additional, poorly reproducible oxygen
consumption and is undesirable in the BOD determination. The addition of an inhibiter delays
the start of nitrification; usually Allylthiourea (ATU) is used for this purpose.
Dilution method of BOD5 determination
Experimental Procedure
Dilution water
The dilution water is composed as follows:
Adding 1 ml of the below mentioned salt solutions to
approx. 0.5 l of deionized water. To this solution, 5 ml inoculating water (domestic wastewater
- either sedimented or filtrated effluent of primary clarifier) are added.
At least 1 h aeration at 20 °C: The O2 concentration should be at least 8 mg / l. The water may
not be supersaturated with oxygen, therefore leave the solution in an open tank for 1 h. This
solution is to be used within 24 h after preparation.
Salt solutions
Phosphate buffer solution, pH 7.2
Magnesium sulfate heptahydrate solution 22.5 g/L
A solution of calcium chloride 27.5 g/L
Iron-(III)-chloride hexahydrate solution 0.25 g/L

 The sample is to be homogenized with an Ultra-Turrax for 30 seconds.
 A defined volume (see below) of homogenized wastewater sample is to be filled into a
graduated cylinder.
 Add 2 ml ATH solution (cATH = 1 g/L)
 Filling up to 1l with dilution water and mix.
 3 sample is to be prepared for each dilution series:
Effluent primary clarifier: (expected BOD5: 100-300 mg/L):
Sample volume: 20, 30, 50 ml
Effluent secondary clarifier: (expected BOD5: 5-10 mg / L)
Sample volume: 200, 300, 500 ml
Distribute the content of the glass cylinder on 3 Reagent bottles and seal immediately with a
plug - small volumes of liquid should leak out to prevent trapped air bubbles.
In addition, a blank sample is to be applied: Fill measuring cylinder with 1 L of dilution water
and add 2 ml ATU-solution, also spread over 3 reagent bottles and closed.
In one bottle of each dilution series, the initial oxygen content is to be measured. The remaining
2 bottles of each dilution series and the blank value at 20°C are thermostated in the incubator
for 5 or 7 days. After this time the final oxygen is to be measured and the BOD-value is to be
calculated.
Test results
Determining the BOD value according to the formula:
d
n p V e V
With BODn = Biological oxygen consumption after n days [mg/L O2]

Zp = O2-consumption of the diluted wastewater sample [mg / L]
ZV = O2-consumption of dilution water [mg / L]
d = sum of sample and dilution filled in the mixing tank [ml]
e = water sample charged into the mixing tank [ml]
The BOD value is to be indicated as an average of the individual dilutions. The dilution water
shall have an oxygen consumption of 0.5 to 1.5 mg/L.
Manometric method of BOD5 determination
Theory
The manometric BOD determination with the OxiTop system supplied by WTW, based on a
pressure measurement (differential measurement). The data acquisition via electronic
piezoresistive pressure sensors, that are integrated into the OxiTop measuring head. This
measuring head is so mounted on the measuring flask, that the sample contained therein is
completely sealed from the atmosphere.
On top of the sample is an airspace, which oxygen content is used for the oxygen supply of
the microorganisms in the sample. The released CO2 is absorbed in the airspace by potassium
or sodium hydroxide. By this means, a pressure loss happens which is proportional to the
oxygen consumption of the microorganisms. This pressure loss is read as a BOD-value at the
measuring head.
Accordingly, to the expected BOD, the water sample volumes (homogenized) used is listed in
the table below:
Table 1: Expected BOD(mg/L) range for corresponding volume (ml)
measuring range Filling volume (ml) conversion factor
BOD5 (mg / l) (Multiplied by the

indication value)
0 - 35 432 1
0 - 70 365 2
0-175 250 5
0-350 164 10
0-700 97 20
0-1750 43.5 50
0-3500 22.7 100
Experimental Procedure
The corresponding sample volume is filled into a brown glass bottle (500 ml). ATU from the
stock solution (c = 1 g/L) is added to the sample with the target to establish an
ATU-concentration of about 2 mg / l. In the bottle, a magnetic stirrer is given.
A rubber receptacle is filled with 2 NaOH pellets and placed on the bottle neck. Thereafter, the
OxiTop measuring head is screwed onto the bottle firmly.
For further information see instructions, "Operation OxiTop." (In chapter "Forms")
It is recommended to establish 2 bottles per sample.
Fig 1: Brown glass bottles with sample Fig 2: Thermostated oven (20 ° C) (BOD cabinet)

Experimental sheets :
Table 2: Datasheet for evaluating the BOD 5 dilution method

Table 3: Chart sheets for entering the BOD values of the Manometric Method.

Table 4: Quick Guide OxiTop measuring heads.

3 Unit processes
3.1 Coagulation
Lab purpose
The objectives of this lab work are to estimate the optimum dosage of coagulant (aluminum
sulfate or ferric sulfate) for the removal of suspended matter and to investigate the formation
of the flocs as a result of aluminum sulfate addition.
Theory
Coagulation and flocculation are important unit processes in water and wastewater treatment
plants. The purpose of coagulation/flocculation is to remove suspended matter, turbidity, color
and colloidal substances, which cannot be separated from the liquid by sedimentation alone
except by the use of reasonably long detention periods. In the coagulation/flocculation process,
the chemical reagents (called coagulants) are used to destabilize the suspended and colloidal
substances. This form the flocs and increase the floc size. Then, the flocs formed are
subsequently removed by sedimentation and/or filtration.
Coagulation involves the addition of chemicals to destabilize the suspended particles, colloidal
materials and macromolecules. Coagulants with positive charges, opposite to those of the
suspended solids are added to the water to neutralize the negative charges on dispersed non-
settable solids, such as clay and organic substances. Thereby, the zeta potentials are reduced
to achieve destabilization of particles.
Figure 1: Scheme of emergence micelle.

The destabilization of colloidal particles can be influenced by the double layer compression,
adsorption and charge neutralization, entrapment in precipitates (sweep flocculation) and
interparticle bridging. The destabilized small-suspended particles are capable of approaching,
aggregating and form a cluster. The slightly larger particles formed through this process are
called microflocs and are still too small to be visible to the naked eye. A high-energy, rapid-mix
to properly disperse the coagulant and promote particle collisions is needed to achieve good
coagulation and formation of the microflocs. Over-mixing does not affect coagulation but
insufficient mixing will leave this step incomplete. Proper contact time in the rapid-mix chamber
is typically 1 to 3 minutes.
Following coagulation, flocculation increases the particle size from submicroscopic microfloc
into larger flocs under gentle mixing conditions. Through mild agitation of 20 min to 60 min,
collisions of the microflocs and interaction with inorganic polymers formed by the coagulants
are cause them to bond together to achieve larger, visible flocs (called macroflocs). The
agitation should be gentle, in order to promote floc growth and not to break flocs already
formed. After the floc has formed and grown to its most effective size, the water passes through
a separation process (sedimentation or filtration) for solids removal.
The commonly used metal coagulants fall into two general categories: those based on
aluminum and those based on iron. The aluminum coagulants include aluminum sulfate,
aluminum chloride and sodium aluminate. The iron coagulants include ferric sulfate, ferrous
sulfate, ferric chloride and ferric chloride sulfate. Other chemicals used as coagulants include
hydrated lime and magnesium carbonate.
The effectiveness of aluminum and iron coagulants arises principally from their ability to form
multi-charged polynuclear complexes with enhanced adsorption characteristics. The nature of
the complexes formed may be controlled by the pH of the system.
When metal coagulants are added to water the metal ions (Al and Fe) hydrolyze rapidly but in
a somewhat uncontrolled manner, forming a series of metal hydrolysis species.
The efficiency of rapid mixing, the pH and the coagulant dosage determine which hydrolysis
species is effective for treatment.
There has been the considerable development of pre-hydrolyzed inorganic coagulants, based
on both aluminum and iron to produce the correct hydrolysis species regardless of the process
conditions during treatment. These include aluminum chlorohydrate, polyaluminum chloride,
polyaluminum sulfate chloride, polyaluminum silicate chloride and forms of polyaluminum
chloride with organic polymers. Iron forms include polyferric sulfate and ferric salts with
polymers. There are also polymerized aluminum-iron blends.
The principal advantages of pre-polymerized inorganic coagulants are that they are able to
function efficiently over wide ranges of pH and raw water temperatures. They are less sensitive
to low water temperatures; lower dosages are required to achieve water treatment goals; less
chemical residuals are produced; and lower chloride or sulfate residuals are produced,
resulting in lower final water TDS. They also produce lower metal residuals.
Pre-polymerized inorganic coagulants are prepared with varying basicity ratios, base
concentrations, base addition rates, initial metal concentrations, aging time and aging
temperature. Because of the highly specific nature of these products, the best formulation for

a particular water is case specific and needs to be determined by jar testing. For example, in
some applications alum may outperform some of the polyaluminum chloride formulations1.
PoIymers are a large range of natural or synthetic, water soluble, macromolecular compounds
that have the ability to destabilize or enhance flocculation of the constituents of a body of water.
Natural polymers have long been used as flocculants. For example, Sanskrit literature from
around 2000 BC mentions the use of crushed nuts from the Nirmali tree (Strychnos potatorum)
for clarifying water – a practice still alive today in parts of Tamil Nadu, where the plant is known
as Therran and cultivated also for its medicinal properties. In general, the advantages of natural
polymers are that they are virtually free of toxins, biodegradable in the environment and the
raw products are often locally available. However, the use of synthetic polymers is more
widespread. They are, in general, more effective as flocculants because of the level of control
made possible during manufacture.
Important mechanisms relating to polymers during treatment include electrostatic and bridging
effects. Figure shows schematic stages in the bridging mechanism. Polymers are available in
various forms including solutions, powders or beads, oil or water-based emulsions and the
Mannich types. The polymer charge density influences the configuration in solution: for a given
molecular weight, increasing charge density stretches the polymer chains through increasing
electrostatic repulsion between charged units, thereby increasing the viscosity of the polymer
solution.
Figure 2: Stages in the bridging mechanism: (i) Dispersion; (ii) Adsorption; (iii) Compression
or settling down (see inset); (iv) Collision

 Phipps Bird Six-Place Stirrer
 Aluminum sulfate solution
 Pipettes/syringes
 Volumetric cylinders
 0.15, 0.5, 1 and 2-liter beakers
 Turbidimeter
 Burettes
 Salt
Determine the turbidity and pH of the raw water sample.

Add 1 liter of raw water into each of the six beakers of the laboratory stirrer.
0.25%, 0.5%, 1.0%, 1.5%, 2.5% and 4.0% of 1% Al2 (SO4) 3 solution were added to 6 water
samples respectively. At approx. the same moment, add Alum solution into each beaker and
stir at approx. 400 rpm for 1 minute. (Describe your observations in lab exercise book)
Dose of the coagulant (ml) can be calculated by the formula:
Where D – dose of coagulant, ml; V – volume of water solution, ml; C1 – required concentration
of coagulant, % mass; C2 – initial concentration of coagulant solution, % mass.
Decrease the speed to approx. 30 rpm and allow the sample to mix for a period of 10 minutes.
Observe any changes including the flocs formation speed, appearance, size, compactness in
the suspended matter in the sample.
At the end of the mixing period, turn off the stirrer and let the flocs settle for about 20 minutes.
With a syringe, carefully remove the supernatant (clear liquid) from each beaker in the 150mL
beaker. And determine the turbidity and pH of each of the samples.
Report content
Add all results in a Table format. (Describe your observations in lab exercise book)
Plot turbidity and pH as a function of coagulant dosage (x-axis = dosage; y-axis = turbidity or
pH).
Determine the optimum dosage of coagulant based on the plot.
Coagulant Coagulant concentration

Raw water temperature PH value of raw water
Turbidity of raw water（NTU） Volume (L)
Experimental equipment
parameters
Beaker # Raw 1 2 3 4 5 6
mL
Dosage
mg/L
Turbidity（NTU）
pH value of all samples
Time of the beginning of formation
flocs, min
Time of the beginning of
precipitation flocs, min

Discussion issues
1. What is the best coagulant dosage and why?
2. Does the pH value have a significant change during the coagulation?
3. Why did the pH change with dosage? (What made them change?)
References
1. Jiang, Jia-Qian (2015-05-01). "The role of coagulation in water treatment". Current

Opinion in Chemical Engineering. Nanotechnology • Separation engineering. 8: 36–
44. doi:10.1016/j.coche.2015.01.008.
2. Chekli, L.; Eripret, C.; Park, S. H.; Tabatabai, S. A. A.; Vronska, O.; Tamburic, B.; Kim, J.
H.; Shon, H. K. (2017-03-24). "Coagulation performance and floc characteristics of
polytitanium tetrachloride (PTC) compared with titanium tetrachloride (TiCl4) and ferric
chloride (FeCl3) in algal turbid water". Separation and Purification Technology. 175: 99–
106. doi:10.1016/j.seppur.2016.11.019.
3. Ramavandi, Bahman (2014-08-01). "Treatment of water turbidity and bacteria by using a
coagulant extracted from Plantago ovata". Water Resources and Industry. 6: 36–
50. doi:10.1016/j.wri.2014.07.001.
4. a b Ayekoe, Chia Yvette Prisca; Robert, Didier; Lanciné, Droh Gone (2017-03-
01). "Combination of coagulation-flocculation and heterogeneous photocatalysis for
improving the removal of humic substances in real treated water from Agbô River (Ivory-
Coast)". Catalysis Today. Heterogeneous Photocatalyis from fundamentals to possible
applications. 281, Part 1: 2–13. doi:10.1016/j.cattod.2016.09.024.
5. A. Koohestanian; M. Hosseini & Z. Abbasian (2008). "The Separation Method for
Removing of Colloidal Particles from Raw Water" (PDF). American-Eurasian J. Agric. &
Environ. Sci. 4 (2): 266–273. ISSN 1818-6769. See p. 267.

3.2 Filtration
Lab purpose
Understand the procedures of rapid filtration and backwashing. Understand the relationship of
the bed expansion and flow rates.
Theory
The water coming from the sedimentation tank are not pure and may contain some very fine
suspended particles and bacteria. To remove these remaining impurities, the water is filtered
through the beds of fine granular material, such as sand, anthracite coal etc. The process of
passing the water through the beds of such granular materials is known as Filtration.
Rapid sand filters use relatively coarse sand and other granular media to remove particles and
impurities that have been trapped in a floc through the use of flocculation chemicals - typically
alum. The unfiltered water flows through the filter medium under gravity or under pumped
pressure.
This filtering process is determined by two basic physical principles. First, relatively large
suspended particles get stuck between the sand grains as they pass the filter medium
(mechanical straining). Second, smaller particles adhere to the surface of the sand grains
caused by the effect of the van der Waals forces (physical adsorption). A chemical filter-aid
(i.e. coagulant or flocculant) might be added to promote additional adhesion.
During these processes more and more particles accumulate in the filter medium, increasingly
causing filters clogging and decreased performance. When the filter effluent turbidity is greater
than a treatment guideline, initial filtering performance can be re-achieved through
backwashing by reversing the flow and causing the treated water to flow upwards through the
bed. The bed is expanded and the accumulated debris and particles are separated in the
surface water. Often, air is injected additionally to support the cleaning process. As soon as
most particles are washed out and the backward flowing water is clear, the filter is put back to
the operation.
 Filter device
 Coagulants
 0.02-liter volumetric cylinders
 0.2-liter beakers Turbidmeter
 Thermometer

The filter bed was washed with wash rate of 12 - 15 L / (sꞏm2) for 1 minute in order to remove
the filter layer bubbles.
Measure the diameter of the filter and height of the bed. Measure the turbidity and temperature
of raw water. The filtration starts with a flow rate of 8 m/h.
The turbidity of effluent is measured at 1, 3, 5, 10, 20, 30 min.
Increase the flow rate to 16 m/h, and then measure turbidity water turbidity at 1, 3, 5, 10, 20,
30 min.
Backwash with different backwashing intensity (6, 9, 12, 14, 16 L/(sꞏm2). The height of the
expanded bed was measured.
Report content
Table 1: Filter with different flow.
Diameter of Filter Height of the filter
(mm) bed (cm)
Water temperature
Coagulant
(OC)
Dosage of coagulant
（mg/L）
Turbidity of
Filtration rate(m/h) Flow(L/h) Filtration time (min)
effluent (NTU)
0
1
3
8 5
10
20
30
0
1
3
16 5
10
20
30

Table 2: Filter expansion with different backwashing intensity and flow.
Height of the filter Backwashing water
bed (cm) temperature (OC)
Backwashing Backwashing flow Height of expanded Filter expansion rate
intensity (L/h) bed (cm) (％)
(L/（sꞏm2）)
6
9
12
14
16
Report all results in a Tables 1 and 2.

Plot the effluent turbidity as filtration time (x-axis = filtration time; y-axis = effluent turbidity).
Plot the height of expanded bed as backwashing intensity (x-axis = backwashing intensity; y-
axis = the height of expanded bed).
Discussion issues
1. What is the impact of the air bubble on the filter and backwashing performance?
2. When the turbidity of the raw water is constant, what measures can be taken to reduce
the effluent turbidity of the initial filtrated water?
3. Why should backwashing intensity not be too high?

3.3 Rapid Sand Filtration and Ultrafiltration
Experiment 1: Direct filtration of a model suspension in a rapid sand filter.
Theory
Rapid sand filters are used in water treatment for the removal of particles from large amounts
of water. The filter effectiveness depends on a number of factors that can be divided
schematically in transport processes and adhesion to the filter grains. Several mechanisms
are responsible for the transport of the solid particles to the surface of the filter grains. Their
individual contributions depend primarily on the size of the particles which shall be retained.
Experimental observations and model calculations have shown that particles on the order of 1
micrometer are most difficult to separate. For raw waters which contain a significant proportion
of this fraction, it is advisable to combine the filtration with preceding coagulation/flocculation.
The process is called direct filtration when the coagulant is added by flash mixing and
flocculation takes place in an upstream basin or a long pipe. Under certain conditions, it may
also be sufficient to dose the coagulant just before the filter inlet. Flocculation then occurs in
the supernatant of the filter and in the pores of the filter bed, and the process is called
flocculation filtration.
Successful removal of particles further requires that attractive surface forces become effective
upon contact between the solid particles or flakes and the filter grain in order to prevent
detachment of the particle by hydrodynamic forces. For an improved adhesion small amounts
of organic polymers are often added to the water to be filtered. With increasing filter load the
separation efficiency decreases. At the same time the flow resistance in the filter bed
increases, and thus the head loss increases at constant flow. Once the solids concentration in
the effluent exceeds the allowable value or the maximum head loss is reached, the filter bed
must be cleaned by backwashing with air and/or water. The solids are then separated from the
backwash water by sedimentation.
Rapid sand filters are usually operated downstream at a constant rate. They are often designed
with two or three layers with different grain diameters in order to obtain more homogeneous
distribution of the filter load and the head loss profile. In this case, coarse, light filters grains
are forming the top layer and fine, heavy grains are arranged in the sublayer. The use of
materials of different density is necessary to prevent the layers from mixing during
backwashing.
For optimizing filter operation two aspects have to be considered: Firstly, the largest possible
amount of water shall be treated during each cycle while the cleaning requirements must be
met. Secondly, the backwash water demand should be kept low. For final optimization, tests
at the technical scale plant are necessary. However, pilot filter experiments can be used for a
comparison of removal efficiencies and head loss characteristics of different filter materials.
Tasks
A pilot filter is loaded with a kaolin suspension. The particle removal efficiency shall be
examined without and with dosing of a coagulant. Furthermore, the development of the
head loss profile inside the filter bed is to be observed as a function of time.

Experimental approach
The raw water tank of the pilot plant (Figure. 1) contains a suspension of 100 mg/L kaolin in
tap water that is stirred continuously for homogenization. The rest of the system is filled with
tap water. In the beginning of the experiment pump P1 is turned on and the flow rate is adjusted
to 20 L/h at the rotameter R1. The needle valve at the effluent of the filter is adjusted in a way
that the supernatant in the filter remains constant. Then the head loss profile for the unloaded
filter bed is registered.
After 25 min of the test period, the dosing pump P2 is turned on, and a solution of aluminum
sulfate (concentration 1 g/L Al) is dosed at a flow rate of 1 mL/min.
The following parameters are measured during the experiment:
Turbidity in the filter effluent: continuously, with readings at every 5 minutes.

Turbidity in the feed: after 10, 30, 60 and 90 minutes (taking samples from the
recycle flow to the raw water tank).
Head loss profile: every 30 minutes until the end of the experiment.
Total head loss: every 10 minutes until the end of the experiment.
By controlling the rotameter R1 a constant flow rate has to be ensured. The increasing filter
resistance must be compensated for by opening the needle valve in the filter effluent.
At the end of the filter run, backwashing is demonstrated. With the help of the rotameter R2
the backwash water flow rate is determined when the filter bed starts to become fluidized.
iser pipes
iser pipes
e valve
way valve
ent
water
Figure 1: Sketch of the pilot filter

Discussion issues
1. What is the filter velocity and the backwash velocity, both related to the entire cross-
sectional area of the filter?
2. a) The effluent turbidities shall be listed in a table and plotted as a function of time.
b) What are the initial filter parameters obtained for the two experimental phases
(without and with coagulant addition) when turbidity is used as a surrogate parameter
for the concentration of kaolin?
c) Which average filter loading kaolin (in mL/L) is obtained at end of the test?
3. a) The head loss profiles shall be plotted as a so-called Michau-diagram.

b) Which differences can be seen here between the two filter layers?
4 a) The total head loss shall be plotted as a function of time.

b) What is the value of the constant b in the simplified head loss equation?
Density of kaolin = 2.65 g/cm3

Experiment 2: Cross-flow ultrafiltration of a model suspension.
Theory
Pressure driven membrane processes, including microfiltration, ultrafiltration, nanofiltration

and reverse osmosis, are used not only for treating small amounts of water, but also for
throughputs of >100,000 m3/d. As a process with a cut-off in the colloidal range, ultrafiltration
is used for pretreatment prior to a reverse osmosis stage, for particle removal from spring
waters, and for the treatment of reservoir water. In wastewater treatment, it has become an
established method for separating dispersed colloidal solids and oil droplets from suspensions
or emulsions.
By concentrating the retained material on the feed side of the membrane, deposits can quickly
form that will increase the head loss and change the separating properties of the membrane.
Here the principle of cross-flow filtration in which the concentrate flows at high speed over the
membrane, can limit these scaling and fouling effects. However, in order to come to significant
yields, such systems are usually operated intermittently, that is, the concentrate is recirculated
until the desired final concentration is reached. Tubular membrane bundles or rotating disks
are primarily suited for this purpose.
Important design parameters are the flux, the operating pressure, and the cross-flow velocity.
It is advisable to carry out tests with the water to be treated and a pilot membrane unit in order
to verify these parameters. In addition, periodic membrane cleaning by flushing or back-
flushing, possibly also by mechanical means, and chemical cleaning after longer time intervals
must be optimized.
Ultrafiltration systems in water treatment are, however, preferably operated in the dead-end
mode because of energetic reasons. Here, the retained materials must be removed from the
membrane surface during short purge and backwash intervals.
Task
A model suspension of polystyrene latex beads is to be concentrated by cross-flow

ultrafiltration. The flux of a capillary tube membrane shall be determined as a function of the
trans-membrane pressure.
A stock suspension of polystyrene latex beads (mean particle size about 0.1 microns) at a
concentration of 1 g/L is given. From that, suspensions of 5 mg/L and 10 mg/L in a 0.002 molar
solution of NaHCO3 shall be prepared and filled in the storage tank of the test apparatus (Figure
1). At the beginning of the test, the initial turbidity of the suspension is to be determined.

Apparatus I:
The gear pump P1 is turned on and adjusted to a concentrate stream of 95 L/h with the valve
at the rotameter R1 being open. At a constant setting of the pump the pressure on the
concentrate side is then increased gradually by closing the rotameter valve in accordance with
the following schedule:
t / min 0-5 5-10 10-15 15-20 20-25

p / bar 0.5 0.75 1.0 1.25 1.5
For each pressure stage, the amount of filtrate is measured using graduated cylinders and a
stopwatch. At end of the test, the turbidity of the collected filtrate is measured.
Figure 1: Sketch of Apparatus I for the investigation of cross-flow ultrafiltration.
Apparatus II:
Valve 3 is opened and valve 2 is initially kept closed. After switching on the pump, valve 2 is
opened slowly while the filtrate is fed back into the storage tank. Subsequently, the setting of
the excess pressure p2 at the concentrate outflow takes place at valve 3 according to the
following scheme:
Operating 1 2 3 4 5
point
p2 / bar 0.5 0.75 1.0 1.25 1.5
At each operating point, the pressure values p1 and p3 are read, and the amount of filtrate is
determined by means of a graduated cylinder and a stopwatch. At the end of the test, the
turbidity of the collected filtrate is measured.

Additional task:
100 mL samples are prepared from the colloidal stock suspension with the following
concentrations: 5 mg/L; 10 mg/L; 20 mg/L; 30 mg/L and 50 mg/L. Then the respective turbidity
values are measured.
Discussion issues and Evaluation
1a. The experimental data are to be listed in a table.

b. From the measured turbidities of the different colloidal suspensions a turbidity-vs-
concentration calibration curve shall be derived.
From Apparatus I:
2a. The flux shall be plotted as a function of the excess pressure in the inlet.
b. What is the value of the maximum flux?
3. What is the approximate rejection of the membrane for the colloidal material?
4a. What is the value of the minimum cross-flow velocity?
b. Which maximum ratio of filtrate flow: concentrate flow has been reached?
From Apparatus II:

5a. From the pressure values p1, p2 and p3, the trans-membrane pressure (TMP) shall be
calculated by the following formula:
TMP = [(p1 + p2) / 2] - p3
The flux shall then be plotted as a function of TMP.
b. What is the value of the maximum flux?

6. What is the approximate rejection of the membrane for the colloidal material?
Additional literature
 DVGW code of practice W 213-3 (2005), W 213-5 (2013) and W213-6 (2013)
Filtration processes for particle removal
- Rapid sand filters
- Membrane filtration
- Control by turbidity and particle analysis
 Cheryan, M.: Ultrafiltration and Microfiltration Technomic Publ. Co., Lancaster (PA)
1998
 Melin, T., Rautenbach, R.: Membranverfahren – Grundlagen der Modul- und Anlagen-
auslegung (Membrane Processes – Fundamentals of Module and Plant Design), 3. ed.,
Springer, Berlin 1997

3.4 Adsorption on Activated Carbon
Theory
Numerous dissolved organic substances, in particular hydrophobic species, can be efficiently
removed from water by activated carbon adsorption. When describing the relevant processes,
one must distinguish between the equilibrium and the kinetics of adsorption.
Adsorption equilibrium refers to the stable state which is established between the
concentration of a substance in water and the surface loading of activated carbon after a
sufficiently long contact time. The relationship between concentration and loading is called
adsorption isotherm. It is dependent on the properties of the individual substances and on the
type of activated carbon used. Isotherms cannot be predicted theoretically but must be
determined experimentally. In multi-component systems, it also comes to competing effects
between different substances. Earlier it was necessary to determine experimental data for the
respective mixtures in order to describe multi-solute adsorption. Today, models are available
that allow to predict multi-component equilibria, based on the adsorption parameters of the
individual components. Most important for the adsorption capacity of an activated carbon is its
large internal surface area. Therefore, the adsorption process involves the transport of
molecules from the bulk solution flowing through the packed bed of activated carbon granules
to the adsorption sites inside the particles.
The first transport step is the diffusion through the hydrodynamic boundary layer around the
activated carbon granules. This external mass transfer, also called film diffusion, can be
determined experimentally quite simply and accurately. When knowing both the diffusion
characteristics of the substance, the hydrodynamic conditions and the outer surface of the
activated carbon granules, the film diffusion coefficient can also be calculated from empirical
mass transfer correlations.
Several mechanisms may contribute to the mass transfer inside the carbon granules. Firstly,
the molecules diffuse in the pore water, on the other hand they are believed to be transported
in the adsorbed state when local loading differences exist on the activated carbon surface.
Parameters describing the internal mass transfer cannot be predicted but must be determined
from experimental data.
Experiment 1: Determination of a single-solute isotherm.
Task
For the substance 4-hydroxybenzoic acid (HBA) the equilibrium loadings on activated carbon
Norit ROW 0.8 S shall be determined at various residual concentrations. From the data, the
parameters of the Freundlich isotherm equation are to be derived.
From a stock solution of 1 g/L of HBA, test solutions with different initial concentrations
according to Table 1 are prepared by dilution with distilled water. The pH value is adjusted with

HCl to pH < 3 in order to prevent pH effects on adsorption. Aliquots of 200 mL of each solution
are then distributed to 5 Erlenmeyer flasks.
500 mg of finely ground activated carbon, to be stored in a desiccator, are weighed and
dispersed in 0.5 L of water with the aid of a high-speed stirrer. The suspension volumes given
in Tab. 1 are then withdrawn with a pipette and added to the corresponding solute samples.
This type of metering is more accurate than weighing and adding dry activated carbon for
amounts of activated carbon <20 mg. The flasks are sealed and continuously mixed on a
shaker for at least 24 hours.
(In order to save time, the isotherms are prepared for this task one day in advance.)
Table 1: Experimental conditions for the determination of the HBA isotherm
Isotherm HBA initial concentration Dosage

(mg/L) (mL of activated carbon suspension)
Concentration of the suspension: 1 g/L
1 10 0 2 4 8 12
2 20 0 3 6 10 15
3 30 0 4 8 12 18
4 40 0 5 10 15 20
After the mixing period, the activated carbon particles are allowed to settle for 1-2 hours. Then
20 mL of samples are taken from each flask with a syringe and filtered through 0.45 μm
membrane filters. The HBA concentrations are determined photometrically at a wavelength of
255 nm. For isotherm 2, 3 and 4 it is necessary to dilute the filtrates by 1:2 (isotherm 2), 1:3
(isotherm 3) and 1:4 (isotherm 4), respectively.
Evaluation
1. The surface loading values for each isotherm point shall be calculated using the mass
balance equation and listed in a table together with the residual HBA concentrations.
2. The data for all four isotherms shall be depicted in a log-log graph.
3. The parameters KF and n of the Freundlich isotherm equation are to be determined from
all data points by regression analysis using the Excel software. Outliers have to be
identified and omitted here.
4. The Freundlich isotherm shall also be included in the diagram. Which measurement points
show the largest deviation, and why?

Experiment 2: Determination of the internal mass transfer coefficient.
Task
For HBA as a model substance, diffusion inside the granules of activated carbon Norit ROW
0.8 S shall be investigated. From the experimental data, the effective internal diffusion
coefficient is to be determined.
In a beaker, 5 L of a HBA solution with a concentration of 10 mg/L are prepared and acidified
to pH <3. Then two initial samples are taken. A representative granular activated carbon
sample of about 250 mg is weighed and dispersed in distilled water in a beaker. The water is
briefly heated to the boiling point in order to remove the air from the pores of activated carbon
granules and cooled again. The activated carbon particles are then transferred to the screen
basket of a turbine stirrer (see Figure 1).
At the beginning of the test, the stirrer is immersed into the HBA solution and set at a stirring
speed of 200 min-1. Then samples of 10 mL each are taken according to the following schedule:
Sample-no. 1 2 3 4 5 6 7 8
t (h) 0.25 0.5 1 2 4 6 23 25
After the test is finished, the HBA concentrations are measured photometrically in each of the
samples.
Figure 1: Apparatus for determining concentration-vs-time curves.

Evaluation
1. The measured concentrations shall be listed in a table, and the ratios of residual to initial
concentration shall be plotted as a function of the square root of time.
2. The correlation of Gnielinski is to be applied to calculate the external mass transfer

coefficient ßL for a superficial velocity of 100 m/h that is assumed to be an approximate
value for the hydrodynamic conditions in the screen basket.
The following parameters shall be used:

Bed porosity ԐF = 0.42
Average particle diameter dP = 1.18 mm
Kinematic viscosity (water, 20° C) ν = 1 * 10-6 m2/s
Diffusion coefficient (HBA, 20° C) DL = 0.77 * 10-9 m2/s
3. The effective internal diffusion coefficient DS shall be determined by fitting the calculated
concentration-vs-time curve of the film-surface diffusion model to the measured data. (A
computer program for that purpose is available in the Laboratory for Water Technology.)

Experiment 3: Adsorption of breakthrough curves of a single-solute in a fixed-bed
granular activated carbon column
The following simulation is to be carried out in a computer pool:
1. For the adsorption of HBA in a fixed-bed of Norit ROW 0.8 S, the equilibrium parameters
KF and n, and the kinetic parameters ßL and DS, respectively, shall be used to predict the
breakthrough at different filter velocities. The operating conditions are as follows:
Feed concentration c0 = 10 mg/L

Filter cross sectional area AF = 1 m2
Filter bed length L = 2 m
Bulk density of the carbon ρF = 374 kg/m3
Filter velocity vF= 5; 10 and 15 m/h
The mass transfer coefficient ßL = f (Re, Sc) is to be calculated for each filter velocity
using the correlation of Gnielinski.
2. What is the actual residence time of the water in the filter bed?
3. When do the effluent concentrations reach 10% of the feed value, and how many bed
volumes have then passed the filter?
4. Which percentage of the maximum available adsorption capacity has approximately be
used in each case?
Additional literature
 DVGW code of practice W 239 (2011)

Removing organic substances in drinking water treatment by adsorption on activated
carbon.
 DVGW code of practice W 651 (M) (2013)
Feed systems for powdered activated carbon in drinking water treatment.
 Sontheimer, H., Crittenden, J.C., Summers, S.R.
Activated Carbon for Water Treatment. DVGW Research Center, Karlsruhe 1988

3.5 Membrane methods for cleaning water solutions: ultrafiltration,
nanofiltration, reverse osmosis
Lab purpose
Studying the process of separation of water solutions by the reverse osmosis method and
effect of pressure on selectivity and permeate flux.
The objects of the study are model water solutions of various salts that undergo desalination
and of concentrating by the reverse osmosis method.
Theory
Membrane processes are widely used both in various technological applications and in the
field of environmental protection for the purification, separation and concentration of a variety
of water solutions. Also, membrane processes are used to create combined technological
schemes for wastewater treatment and for the organization of closed water circulation
systems.
Ultrafiltration, nanofiltration, reverse osmosis are the main membrane methods of separation
of liquid systems. These methods are used to remove particles whose size is less than 1 μm.
The operating principle of a typical membrane module represents in Figure 1. The stream of
feed water divided into two constituents: the first is a stream of purified water (permeate) and
the second is a stream saturated with impurities (concentrate or retentate).
Figure 1: Schematic representation of a typical membrane module.
The main advantages of membrane methods:

• High stability of cleaning with changing parameters of the feed solution;
• Significant reduction in production areas;
• Reduction of reagent costs;
• Enough low power consumption;
• Modularity of the equipment, allowing increasing productivity easily;
• Full automation of processes;
• Possibility of economical reception of drinking water from the sea.

Ultrafiltration
Ultrafiltration is the process of membrane separation of liquid mixtures under pressure. This
process based on the difference in molecular weights or the molecular dimensions of the
components of the mixture that is separated. Separation occurs under the action of pressure
difference on both sides of the membrane. The ultrafiltration units assemble from tubular
ceramic elements, spiral elements, and hollow fibers. The pore size of the UF membranes is
0.1 to 0.01 μm.
Ultrafiltration used to separate systems in which the molecular weight of the dissolved
components is much greater than the molecular weight of the solvent. Since the osmotic
pressure of high molecular compounds is less than the working pressure of the liquid,
ultrafiltration carried out at low pressures of 0.1 - 0.5 MPa.
Nanofiltration
Nanofiltration is the process of water admixtures separation by the membrane having a higher
dense and lower-permeable selective layer than in microfiltration membranes. Accordingly,
nanofiltration membranes in comparison with microfiltration membranes have higher selectivity
and higher operating pressure for a given productivity. Monovalent ions (cations and anions)
retained by nanofiltration membrane insignificantly, while their selectivity to multivalent ions
and large ions is high. The working pressure in nanofiltration processes is usually in the range
from 0.5 to 1.0 MPa.
Reverse osmosis
The reverse osmosis method consists of filtering solutions under high pressure through
semipermeable membranes that pass the solvent and completely or partially retain molecules
or ions of dissolved substances. If the solution and the solvent separated by a semipermeable
membrane, a spontaneous transition of the solvent into the solution takes place. This
phenomenon called osmosis (Figure 2a). The pressure at which equilibrium sets in (Figure 2b)
called osmotic pressure, When high pressure is applied at the side of the solution that is greater
than the osmotic pressure, the transfer of the solvent will be carried out in the reverse direction
(reverse osmosis), and the dissolved substance is retained partially or completely (Fig. 2c).
Figure 2 - The scheme of occurrence of reverse osmosis: a - direct osmosis, P <π; b - osmotic
equilibrium, P = π; c - reverse osmosis, P> π; 1 - pure water; 2-membrane; 3 -water solution
The pressure π in the solution, which causes the passage of the solvent through the
membrane, is called the osmotic pressure. The value of the osmotic pressure for solutions is
determined by the Van't Hoff equation:
(3.4.1)
i - the isotonic coefficient; T - the absolute temperature of the solution, K; R - the universal gas
constant (8,310 J/mol ∙ K); C - the concentration of dissolved salt (g/m3); M - the molecular
weight of the salt (g/mol).
The isotonic coefficient is calculated by the formula:

(3.4.2)
α - the dissociation degree of the dissolved salt; n - the number of ions that are formed upon
the dissociation of the dissolved salt.
The higher the concentration of the solution to be purified, the higher the osmotic pressure
drop and the greater the hydrodynamic pressure required to realize the water purification.
Osmotic pressure of solutions can reach tens of megapascals (MPa). The operating pressure
for reverse osmosis should be significantly greater because the productivity determined by the
driving force of the process which is the difference between working pressure and osmotic
pressure. The operating pressure in reverse osmosis processes is usually in the range of 1.0
to 10.0 MPa. For example, at an osmotic pressure of 2.45 MPa for seawater, which contains
3.6% of salts, the operating pressure in desalination plants should be maintained at 6.85-7.85
MPa.
The methods of ultrafiltration, nanofiltration and reverse osmosis have much in common, in
particular, in the field of apparatus design, but there are enough differences (Table 1.).
Table 1: Comparison of ultrafiltration, nanofiltration and reverse osmosis.

Process Ultrafiltration Nanofiltration Reverse osmosis
Pore size, μm 0,1 -0,001 0,01 -0,001 0,001 -0,0001
Operating
0,1-0,5 0,5-1,0 1,0-10,0
pressure, МPа
Macromolecules, Low-, high-
High-molecular organic
proteins, molecular organic
Separation substances, multivalent
polysaccharides, substances, ions,
ions
viruses molecules
Energy Low Moderately low (0,2 -0,9 Moderate

consumption (0,1 -0,2 kWh / m3) kWh / m3) (0,9 -3,7 kWh / m3)

The main parameters that characterize the process of membrane separation of liquid solutions
are selectivity (φ) and permeate flux (G). Selectivity - the ability of the membrane to have
different permeability for different components of the mixture to be separated.
The selectivity of the separation process using semipermeable membranes is determined by
the formula:
С  С С
  fs p
 100  (1 
p
)  100
С С
fs fs
, %, (3.4.3)
where: Cfs and Cp - the concentration of the feed solution and permeate, respectively.
Permeate flux of the membrane is the amount of matter passing through the unit surface of the
membrane per unit time. Permeate flux of the membrane calculated by the formula:
V
G ,
F   m3/m2s, (3.4.4)
where: V - the volume of permeate, m3; F - working area of the membrane, m2;  - the time, s.
The permeate flux and degree of cleaning affected by several parameters, namely: applied
pressure; temperature; yield of permeate; concentration of salts in the source water.
Membrane plant - a set of devices and technical equipment that provides the process of
membrane separation. The membrane installation includes two main elements:
1. Device for creating fluid pressure (pump);
2. A membrane apparatus that provides the necessary membrane surface.
These elements are basic in the installation scheme for the membrane separation process
(Figure 3).
Figure 3: The scheme of installation for the membrane separation process [1].

Membrane apparatus is a device for performing a process of membrane separation of the feed
solution into a filtrate (permeate) and a concentrate (retanate), which includes structural
elements for introducing the feed water and for removing permeate and concentrate.
Membrane apparatus should have a high packing density - the surface of semipermeable
membranes per unit volume of the apparatus, be easy to assemble and operate. When
processing solutions and colloidal systems, it is necessary to create conditions for their uniform
distribution to reduce the effect of concentration polarization. According to the method of
packing membranes, apparatuses divided into four main types:
 with flat sheet membrane elements;
 with tubular membrane elements;
 with membrane elements of the spiral type;
 with membranes in the form of hollow fibers.
All these devices usually assembled from separate, so-called, membrane elements, the design
of which characterizes this apparatus.
 Installation of reverse osmosis "Ecosoft 2500 MO-F3A" with a module of storage tanks.
 Glasses for sampling.
 Laboratory conductometer MP 521, ULAB (Сonductometer MP 521, ULAB)
 Reagents for preparation of model solutions.
Figure 4: The scheme of the experimental setup [2]: 1 - tank for the feed solution and
accumulation of permeate; 2 - tank for accumulation of concentrate; 3 - installation of reverse
osmosis "Ecosoft 2500 ME-F3A"; 4 - input of the feed solution for treatment; 5 – permeate
output; 6 - concentrate output; H1, H2 - centrifugal pumps; В1-В6 - ball valves; P1, P2 - float
level gauges.

 Prepare a model solution by dissolving the salt in water (the type of salt and its
concentration in the model solution is set by the teacher). The volume of the solution
must be at least 80 dm3.
 Install the reverse osmosis "EcoSoft 2500 MO-F3A" in accordance with the instructions.
Set the initial working pressure level Prab = 0.4 MPa on the manometer.
 After three minutes of operation at a given operating pressure, take samples of
permeate and concentrate for analysis.
 Using a laboratory conductometer, in accordance with the instructions attached to this
instrument, determine the salt content of the feed solution (Sfs), permeate (Cp) and
concentrate (Cc) in ppm (mg / dm3) and write the data in Table 2.
 According to the indications of rotameters, determine the permeate (Ip) and concentrate
(Ic) consumption in dm3 / min. Write the data in Table 2. Calculate the flow rate of the
feed solution using the formula:
Ifs = Ip + Ic,
Write the result to the table 1.
 Repeat the operations on items 3-6, setting the working pressure level Pworking = 0.5;
0.6; 0.7 and 0.8 MPa and write the results in Table 2.
 Calculate the membrane selectivity by the formula (3), the permeate flux (G) according
to the formula (4) and record the data in Table 3.
Report content
Table 2: The results of an experimental investigation of salt solution.
Рworking, Ip , Ifs,
Сp, mg/dm3 Ic, dm3/min Сc , mg/dm3 Сfs, mg/dm3
(МPа) dm3/min dm3/min
0.4
0.5
0.6
0.7
0.8

Table 3: The results of calculation of membrane parameters.
Рworking,
0.4 0.6 0.7 0.8
(МPа) 0.5
φ,%
G,
m3/m2∙s
Discussion issues
Using the data in Table 2:

1. Plot the curve of function of permeate flow and salt concentration in permeate upon working
pressure Ip = f (Pworking) and Cp = f (Pworking).
2. Plot the curve of function of concentrate consumption and concentration of salt in the
concentrate on the working pressure Ic = f (Pworking) and Cc = f (Pworking).
3. Plot the graph of the flow rate of the feed solution on the operating pressure Ifs = f
(Pworking).
Using the data in Table 3:
1. Plot the graph dependences of selectivity and permeate flux on the working pressure φ =
f (Pworking) and G = f (Pworking).
2. Describe the shape of the obtained curves and formulate the conclusions.
3. Explain how results obtained results are consistent with theory.
4. To explain how it is possible to reduce the amount of waste that is generated during the
purification of salt solutions by the reverse osmosis method.
References
1. Physico-chemical methods of water purification. Management of water resources. Edited
by І.М. Astrelin and H.Ratanavira. Project “Water Harmony”, Private limited company
“Drukarnya Volf”., 2015. - 578 p.
2. Methodical instructions to the laboratory practice course on the theoretical basis of
chemistry and water technology / Contributors: VS Gevod, N.G. Borisova - Dnipropetrovsk:
DNVZ UDKhTU, 2016. - 68 p.

4 Operation
4.1 Microscopy of activated sludge
Microscopy
Biodegradable substances are the main compound of the organic load in domestic wastewater.
These substances are working as
In municipal wastewater, degradable organic substances make up a large part of the burden.
These substances have a nutrient function for heterotrophic microorganisms and are degraded
by microorganisms in water bodies into which wastewater is discharged. Existing species grow
and new species can be established according to the nutrient supply. Since this biodegradation
of organic substances is associated with a consumption of respiratory oxygen, this "self -
purification of the water bodies" is limited.
Because of the increase in sewerage since the industrial revolution (from around 1850),
especially in densely populated areas, technical wastewater treatment became necessary. For
example, sewage treatment plants have been in operation since around 1900, and biological
sewage treatment plants have been increasing since about 1950, exploiting the degradation
activity of microorganisms.
Normally the wastewater in a wastewater treatment plant flows first through a screen for the
removal of coarse material, then through a grit trap in which mineral particles settle and then
into a primary clarifier in which solids are deposited. Substances with a higher density sink to
the bottom, substances that are lighter than water accumulates on the surface.
Screen Grit Primary

Pumping Chamber Clarifier
Primary
Sludge
Sludge
Treatment
Figure 1: Flow scheme of a mechanical wastewater treatment.
After this mechanical treatment, the wastewater still contains dissolved organic substances
and on a low-level organic particles as well, which are degraded in the following biological
stage.
Here are mostly activated sludge basins (or trickling filters) in use. In activated sludge basins,
colony-forming bacteria and organic and inorganic wastewater substances form activated
sludge flocs that are barely visible to the naked eye.

Figure 2: Electron microscopic figure of an activated sludge floc (8 mm equals to 1 µm;).
Only at high magnifications, individual bacteria forming a colony are visible in the flocs. The
bacteria excrete a gelatinous matrix (EPS, extracellular polymeric substances) in which they
remain embedded, even after cell division; fine filaments, which are visible only in the electron
microscope, link the individual cells. (The actual activated sludge floc then also includes
inclusions and mounting, mostly unicellular, organisms.)
Extracellular polymeric
Bacteria cell substances
Filament
Figure 3: Section across a colony of bacteria.

Bacteria required for wastewater treatment are found in the wastewater itself or they establish
themselves in wastewater and in wastewater treatment plants due to abiotic factors that are
suitable for them; only their number is insufficient. Thus, biological purification processes were
able to prevail only when it was possible to increase the biomass in activated sludge by a
sludge recycling at the beginning of the 20th century. This kind of wastewater treatment is the
most common way of purifying wastewater.

Aeration tank Secondary clarifier
Inflow
Effluent
Air (O2)
Surplus sludge
Return sludge
Figure 3: Biological wastewater treatment with the return of sedimentation sludge into the
aeration tank
The shape of the activated sludge flocs and the kind of species, as well as the individual density
of the growing community of organisms (biocenosis), are dependent on many abiotic factors
such as temperature, oxygen concentration, pH, nutrient composition and concentration, the
presence of possible pollutants and hydraulics. Biotic factors such as feeding behaviour,
predator - prey relationships or parasitism also influence the morphology of the flocs.
Thus, from the microscopic picture and its potential changes, valuable conclusions can be
drawn on wastewater composition and operating conditions. By regularly examining the
activated sludge of a treatment plant (for example 2 to 3 times a week) and recording the
observed details, changes can be detected that go beyond normal and constant fluctuations.
Early reactions before manifesting disturbances are possible.
For a reliable assessment of the state of a wastewater treatment plant, the microscopic image
should never be used alone, moreover the combination of physical measurements and
chemical and biological determinations are useful.

Practice of microscoping
The microscope
Figure 4: Microscope and its units.
By using a phase contrast device, delicate structures such as e.g. flagellums become more
visible; they appear black against a gray background. For the determination of many activated
sludge organisms, a phase contrast is mandatory. To do this, you must insert a suitable loose
slide into the slot provided for each objective (10x or 40x) in the condenser. When changing
the lens, do not forget to change the slider!
The preparations
Aqueous preparations are examined directly and alive. For this purpose, a not too large drop
of the sludge is placed on an object carrier (for example with a glass rod or pipette) and
carefully covered with a cover glass. The cover glass is not allowed not "float" (deglaze excess
liquid with paper). It is not allowed to push the cover glass over the sample; The flocs are
otherwise transformed into "sausages" and can no longer be judged.

The investigation
The activated sludge sample is examined using the microscopy form (next pages). As a rule,
the preparation and consideration of several preparations are necessary to complete the form
for a sludge sample. There are several reasons for this: examining multiple grab samples from
the sampling container allows a better overview of the content: organisms may burst or change
due to lack of oxygen (under the coverslip!). By this a correct determination becomes
impossible. The worst case the preparations can dry out.
The evaluation of the microscopic image is in accordance with the microscopy form.
First, a species list (tax list) is created qualitatively on the microscopy form. Here, the order of
the organisms should correspond to the system. Then the levels of frequency are estimated.
For frequency levels, the attributes "rare" for a single find and "plentiful" for such a high density
of the organism that there is hardly any room for other species. are assigned. Remaining levels
are distributed between these attributes. In case of doubt, the trophic position of the species
should be considered. To classify e.g. predatory species, which are at the top of the food
pyramid and thus exert a relatively high pressure on the entire ecosystem, tend to be slightly
higher. Bacterial or detritus eaters, which are the basis of the food pyramid, are classified rather
lower.
Generally, the findings are compiled on the microscopy form and show a consistent picture of
the operating state of the wastewater treatment plant. In actual WWTP investigations, physical,
chemical, hydraulic and other biological parameters are included in the overall interpretation.

--------------------Microscopic Investigation--------------------
WWTP: Date:
Place of sampling:
1. Morphology of the sludge flocs
Size small (<150μm) medium (150 - 500μm) large (>500μm)
Shape rounded irregular elongated
Structure compact irregular fluffy
Edge sharp (smooth) mid-sharp diffuse (rugged)
2. Bacteria (estimated frequency)
rare multiple frequent very frequent plentiful
free bacteria
filamentous bacteria
3. Mono and multi-cellular organism
rare multiple frequent very frequent plentiful

Assessment of the flocs
Four different parameters describe the morphology of activated sludge flocs: size, shape,
structure and edge.
First, the size is estimated: small flocs have a diameter below 150 µm, medium flocs between
150 and 500 µm and large flocs have a diameter of about 500 µm. As an aid for the estimation,
an exact length measurement serves us at the beginning, which we can perform in a
microscope with the eyepiece micrometer integrated there.
The size of the flocs has an effect on their settling behavior in the secondary clarifier. Very
small flocs are too light to sediment and leave the sewage treatment plant with the treated
wastewater. Such "pin-point" flocs may e.g. caused by heavy metal poisoning or by excessive
shear forces. Very large flocs have too little metabolically active surface relatively. Flocs of
medium size are desirable, possibly weighted by attached animal organisms.
The shape of activated sludge flocs is often irregular. Rounded shapes indicate high sludge
age, elongated shapes may be a hint for industrial wastewater. Round flocs sediment better
than elongated ones.
With regard to their structure, flocs of a sludge can have compact as well as loose areas; their
percentage can be stated in%. Often floc edges are loosely structured and the fluff centers are
compact. These areas usually contain older bacterial material; they appear brown or dark
brown in the microscope and are often rounded. Loose, visually bright and delicately
translucent areas are formed by young, newly grown bacterial colonies or foothills of these
colonies; they are metabolically active.
The assessment of the sharpness of the floc edge is essential, as it allows direct conclusions
about the sludge age via the growth activity of colony-forming bacteria.
Rounded, compact, dark flocs usually have a smooth, sharp edge. They consist of older, less
metabolically active organisms (high sludge age). Fissured, fuzzy floc margins consist of
young, rapidly growing bacterial colonies, which can be recognized in the phase contrast at a
400-fold magnification in a regular punctuation: these are the individual bacterial cells. Such
fluffy structured, blurred floc edges indicate a low sludge age and thus a high metabolic activity.
Figure 5: Bacteria floc with a high sludge age.

Figure 5 shows a medium-sized activated sludge floc that has a rounded shape and a compact
structure. This sludge has a good sedimentation behavior. The edge of the floc is sharp; you
could easily draw the outline of the floc with a pen in one go. This sharp edge indicates a high
sludge age. This is confirmed by the monocellular and multicellular organisms that occur on
and beside the floc.
Figure 6: Bacteria floc with a low sludge age.
The activated sludge flocs in Figure 6 are also medium in size but have an elongated shape
and an irregular structure. Compact areas, especially towards the edge of the flocs, turn into
fluffy areas. The edge itself is diffuse. This fuzziness or outgrowth of the flocs towards the edge
is caused by the growth of young bacterial colonies, whose individual cells are so clearly visible
only in sludges with a low sludge age. In this example, load sizes and hydraulic parameters
were used to calculate a sludge age of only 2 days. The sedimentation behavior is not as good
as with the first sludge (SV = 180 ml / l).
Bacteria
The individual densities of all organisms are recorded in a semi-quantitative method. Their
frequency is estimated on a five-point scale, and the terms "rare, multiple, frequent, very
frequent, or mass" are given.
The qualitative detection of the organisms takes place on different levels. Monocellular and
multicellular animal organisms, as well as the few occurring plant organisms, are determined
on the basis of morphological characteristics - if possible up to the species - with specific
literature. In bacteria, this approach is not possible; they are morphologically too similar. In
order to determine bacterial genera or species very costly floral analyses must be carried out.
The result that could be achieved would not justify the high cost of routine activated sludge
testing. For normal controls of wastewater treatment plants, it is sufficient to assign the bacteria
occurring to some functional groups that can be identified morphologically by light microscopy
(Figure 7).

Mono or cell structure Description Examination for
wastewater treatment
Single free bacteria Mostly mobile, predominantly cocci or sticks, Causing turbid effluent,
many species, e.g. from the generic group negative
Bacillus, Pseudomonas; (1 - 3 µm)
Spirochaetes Very mobile, fine spirally wound filaments -
often buckled
Colonies of bacteria Recognized by the regular puncturing, often Floc forming, positive
at floc edges, numerous species, e.g.
Zoogloea ramigera
Filamenteous bacteria Filamentous cell aggregates, individual cells Dominant behavior

often not recognizable, some species worsens sedimentation
branched. Typical representatives: behavior, negative
Spaerotilus natans, type 021 N, Nocardia sp.
Spirilla Very large (e.g., 10 µm long), rigid body, indicator for lack of
spirally wound, fast-floating, microaerophilic, oxygen, negative
e.g. Spirillum volutans (PHB granules as
reserve substance), Thiospira sp. (Sulfur
bacterium)
Sulphur bacteria to SO42-, in H2S oversupply elemental S is indicator for lack of

incorporated into cells, highly refractive, e.g. oxygen and in addition
Beggiatoa alba, Thiothrix nivea, Thiospira sp. indicator for aus H2S
negative
Figure 7: Functional groups of bacteria in activated sludge systems
In the form "Microscopic investigation" single bacteria and filamentous bacteria are listed
separately and must be estimated separately in their frequency because of their importance
for the treatment plant operation.
Free bacteria are understood as round, rod-shaped, curved or spiral (spirochaetes) single
cells. They appear black in the phase contrast setting of the microscope against a light
background. They are detached from any flock dressings and burden the wastewater treatment
process. They can be mobile or immobile.
Bacterial colonies are often fast-growing bacteria that blur the edge of a flake. They are then
recognized by a regular puncture, the individual fresh, young cells. They often belong to the
generic group "Zoogloea sp." and are also referred as "tree bacteria". In the activated sludge
form, they are detected by the characterization of the edge of the floc.
The frequency determination of filamentous bacteria is of major importance in the case of
the sludge bulking problem. At least one preparation must be completely screened at 100x
magnification. The respective frequency level is compared with internationally uniformly used
photos; This is to ensure a comparability of all findings worldwide (EIKELBOOM & VAN
BUIJSEN, 1987). There is one photograph each for the filament categories 0 and 1 (rare and
multiple) used in the filamentous bacteria (Figure 8). For the categories 2, 3 and 4 (frequently,
very frequently and in large numbers), there are two photos each (Figure 9 to Figure 11), with
thinner or thicker filamentous bacterial species.
Activated sludges with a high proportion of filamentous species among the bacteria have a
poor sedimentation behavior. The sludge may under certain circumstances not be adequately
thickened and does not reach the dry matter contents necessary for a satisfactory cleaning. In
the case of hydraulic turbulences, e.g. due to stormwater, a large part of the biomass can be
lost with the effluent of the sewage treatment plant.
Figure 8: Activated sludge of the filament category 0 (rare) (left) and category 1 (multiple)
(right)
Figure 9: Activated sludge of the filament category 2 (frequent) - left side: sludge with
multiple thick filaments right side: sludge with a few thick filaments

Figure 10: Activated sludge of the filament category 3 (very frequent) - left side: sludge with
multiple thin filaments right side: sludge with multiple thin filaments
Figure 11: Activated sludge of the filament category 4 (plentiful) - left side: sludge with plenty
of thin filaments right side: sludge with plenty thick filaments
A significant group of bacteria in activated sludge are spirillas; they are in the shape of a short,
rigid spiral (unlike spirochaetes). They are important indicators of a lack of oxygen. Some of
them belong to the group of sulphur bacteria, which can also be filamentous. Sulphur bacteria
are ever indicators for hydrogen sulfide (H2S).

Organism Size Food Generation Movement Indicator for
time
green algae 3-100 m c-autotroph h prn. flagellum light
Funghis 10-50 m c-heterotroph h no low pH

Plants
wide
Flagellates 5-70 m bacteria or h flagella high load

particular and (imbalance)
10-20 m dissolved
organic matter
Amoebae 10-200 m bacteria or h Pseudopodia high load

unicellular organism
particular and (industry)

30-100 m dissolved
organic matter
Ciliates 15-2000 m bacteria, algae, h/d clia/cirri depending on

detritus, species different
30-200 m flagellates, water qualities
Ccliates
Suctories 50 m - Flagellaten, d no high sludge age,

1 mm Ciliaten low toxicity
Rotifers 50 m filter feeder, d with rotifer high sludge age

- bacteria, organ and foot,
3 mm detritus, small fixed on
Animals
preys surfaces
Gastrotrichs 0.4 organic d creeping, high sludge age

- particles, small winding,
1,5 mm preys floating
Nematodes 0,2 - 5 mm detritus, small d winding (turgor) high sludge age

multicellular organism
preys, algae
Annelids 1 - 20 mm detritus d creeping high sludge age

bacteria (bristles)
Tardigrades 50 m - plants, d running high sludge age

1.2 mm nematodes, (3 pairs of leg)
rotifers
Miltes 1 - 4 mm predatory d/weeks creeping, high sludge age

4 pairs of legs
Crustacea 1 - 5 mm Phytoplankton d floating low load
Insects, larva: 4 mm larva: biofilm weeks Larva: creeping biofilm in trickling

Psychoda filters
Imago: 3mm Imago: flying
Figure 12: Organisms in activated sludge and biofilms in trickling filters
Vegetable Organism
An overview of organisms occurring in activated sludge systems and biofilms in trickling filters
is given in Figure 12.
Plants are relatively rare in activated sludge systems. Green algae, which use light for
photosynthesis, are not specifically used for wastewater treatment in our latitudes.
Nevertheless, biofilms grow on the upper areas of tanks edges, where green algae have a high

proportion. These can be detached and found randomly in the microscopic picture. In biofilms
of open trickling filters, green algae can grow in quantities and make up a large part of the
biofilm.
Funghi hyphae are more frequent than algae in activated sludge systems. They contribute to
the degradation of organic compounds with their heterotrophic metabolism. Preferentially they
grow at low pH-values and - similar to filamentous bacteria - can cause a bad sedimentation
of the activated sludge. The difference to filamentous bacteria is there 10 times higher width.
Animal unicellular organism

Flagellates
Usually, Flagellates occur in a normal activated sludge with several species. On the one hand,
they are food competitors for bacteria by uptaking dissolved organic substances, on the other
hand, they eat organic particles, for example bacteria cells. They themselves are often eaten
by ciliates. Flagellates are frequent in highly loaded sludge. Most species swim very fast with
the help of one or more flagella, which can sometimes be seen in the microscope, even in
phase contrast, only after prolonged observation.
Four frequent types are known:
1. Fitted with floating and dragging flagellum
Unicellular organism
Dragging flagellum
Activated sludge floc
Floating flagellum
Pleuromonas jaculans
2. Fitted with floating flagellum stretched forward
Floating flagellum
Peranema sp.
To the generic group Peranema belong very large (40 - 70 m) flagellates, which swim slowly.
They are metabolites, that means, they can change their shape.

3. Fitted with collar and flagellum
cell
flagellum
collar
stall
Codosiga sp.
Flagellates with collar and flagellum my occur solitarily (like demonstrated in the sketch) or as
a colony. In comparison to the two species described before they are more rare.
4. fitted with multiple symmetrical arranged flagella
Trepomonas agilis
The generic group Trepomonas is easily recognizable by its staggering swimming motion. The
species are indicators of a lack of oxygen.
Amoebae
Amoebae occur in the soil, in the water and as parasites. In balanced activated sludge
systems, they are almost always found in low numbers of individuals. They are often found in
heavily loaded sludge systems or in sludge systems that are not in equilibrium. They move
very slowly, if at all, and are often overlooked by inexperienced eyes.
Two morphological groups of amoebae are easy to distinguish:
1. Naked amoebae, which evert fake feet out of their body plasma and are using this for
movement of taking up their food.
pseudopodium
inclusion
Cell nucleus

Chaos sp.
Ectoplasm (jellylike)
Endoplasm (non jellylike) with inclusion
Vahlkampfia sp.
Filopodium
Astramoeba sp.
2. Shell amoebae, which excrete a shel produces from body's own organic substance. In
this shell foreign objects (grains of sand, diatom shells) can be incorporated or
reinforced with silica plates. An opening, from which the fake feet can reach out, is
located in the shell.
Top view Cross-section
Pseudopodium-often unvisible
Arcella sp.
Mineral particles
Difflugia sp.

Ciliates
Ciliates are very sophisticated and highly differentiated protozoa. Many types have mouth and
anus, they have a precursor of a nervous system and muscle fibers, their eyelashes are similar
to them in human bodies. Ciliates have a simple excretory system for dietary fibers.
Furthermore, they have the possibility of the exchange of genetic material between partners,
which serves as a genetic refresher and they possess a mechanical and chemical defense
systems, which is used to ward off enemies. Approximately 7,500 species are known so far.
Of these, about 20 to 30 species occur in activated sludge systems. Many species are good
indicator organisms for various abiotic factors; thus, ciliates are an essential group of
organisms for characterizing the operating conditions in bioreactors or for the determination of
the flow water quality.
Three systematic groups can be identified:
1. Holotriche ciliates, which are eyelashed entirely:
Cell mouth
Prorodon sp.
Strong Cilies at mouth(predactor)
nucleus
Contractile vacuole
Trachelophyllum pusillum
Angular mouth
Glaucoma sp.
Cross section Top view
Mouth weir
Chilodonella sp.

2. Peritriche ciliates, sessile "vorticellas" whose eyelates is reduced to wreaths around
the mouth.
Epistylis sp.
3. Spirotriche ciliates, which eyelatches transmuted to legs - similar to motion organels.
These legs are used for running over activated sludge flocs
Front or back view Top view
ribs
mouth
Cirri
Aspidisca costata
Oral fissure
Cirri
Euplotes sp.

Suktorians
Suktorians are welcome organisms in activated sludge. They are indicator organisms for a
high sludge age and all species are predatorily and are on top of the food pyramid. In addition,
they are very sensitive to toxic impacts.
Tentacle
stalk
Tocophrya sp.
Multicellular organism
All multicellular organism are indicators of a high sludge age.
Rotifers feed by filtration; they are frequently found in harmonic activated sludge. They eat
individual free bacteria and thus cause desired clear processes from the sewage treatment
plant.
Gastrotrichs are closely related to the rotifers, but their occurrence is rare.
Nematodes are also known from the soil or as parasites. In activated sludge they occur
occasionally only.
Annelids can become abundant in over-aged activated sludge and then eat enormous
amounts of activated sludge due to their size, so that their mass and thus their age decrease
rapidly. A nitrification can thus come to a standstill and the degradation can be reduced.
Tardigrades are organisms of uncertain systematic status; they probably regressed from the
group of insects.
Mites can occur in over-aged activated sludge.
Crustaceans and insect larvae can occur in secondary sedimentation basins and in biofilms
of trickling filter.
Additional microscopic examinations
In addition to a microscopic examination of activated sludge, further microscopic examinations can
provide additional information on the entire treatment plant operation or can be used
specifically for disturbances in the search for causes.
Figure 13 shows useful sampling points.

Sewer Primary Treament Activated Sludge Polishing Waterbody
Pondgsteich
Sewer Primary Sludge Surplus sludge Sediment or Biofilm,

biofilm Secondary Sludge free Water Sediment or
free water
Digestor Alternative for activated sludge:

Trickling Filter
Digested Sludge Effluent Trickling Filter

Biofilm
Figure 13: Potential sampling points for samples used for microscopical investigations.

Figure 14: Unhabited sinking and floating matter in water (Liebmann, 1962)
1 wool fiber
2 rat hair
3 human hair
4 hemp fiber
5 cotton fiber
6 and 7 paper fiber
8 epidermis rest of a grass leaf
9 spiral fiber of plant tissue
10 pine pollen, fresh
11 pine pollen, dead
12 stems or root remnants
13 softwood fibers
14 softwood residue
15 starch grain from the potato
16 potato cells
17 leaf underside with stomata
18 butterfly scales
19 chitin residue
20 Body ring of an insect
21 Down stream of a feather
22 sponge needle
23 worm bristles
24 meat muscle fiber (undigested)
25 soot particles
26 hard coal
27 brown coal
28 iron oxide hydrate
29 sulfuric iron
30 grain of sand
31 pieces of glass
32 oil drops
33 air bubble

References
1. Bayrisches Landesamt für Wasserwirtschaft (1999): Das mikroskopische Bild bei der
biologischen Abwasserreinigung, München (in German)
2. Berger, Foissner, Kohlmann (1997): Bestimmung und Ökologie der Mikrosaprobien nach
DIN 38410; Gustav Fischer Verlag, Stuttgart (in German)
3. Eikelboom, D.H., Van Buijsen, H.J.J. (1992): Handbuch für die mikroskopische
Schlammuntersuchung; 3. Aufl. Hirthammer, München (in German)
4. Eikelboom (2000): Process control of activated sludge plants by microscopic investigation;
IWA publishing;
https://www.iwapublishing.com/sites/default/files/ebooks/9781900222297.pdf

4.2 Optimizing recovery of RO under different salinity
Lab purpose
The goal of this laboratory work is to study a practical approach for optimizing the operation of
RO systems, maximizing permeate flow, salt rejection and recovery while minimizing operating
and maintenance costs, that means minimizing operating pressure and membrane fouling.
Theory
Initially designed for seawater desalination, reverse osmosis (RO) is widely applied in water
treatment today: from small-scale systems under kitchen sinks in households to large plants
producing water for cities, meeting new challenges to conserve water, control environmental
pollution and reclaim both water and valued resources.
Due to recent advances in membrane science, modern RO membranes offer high rejection
rates at reasonably high permeability together with good fouling resistance, reduced energy
consumption and operational costs. Reverse osmosis systems are operated today as single-
standing units or in combination with other processes like ion exchange, electro-deionization
etc.
This laboratory guideline is designed as the test protocol that provides scale-up data such as
permeate flux and permeate quality as a function of feed pressure and system recovery. The
test method consists of determining the desalinating ability and permeate flow rate of the
reverse osmosis unit. It is useful for simulation of water treatment efficiency and suited for both
general environmental engineering courses as well as special courses on mass transfer,
separation processes or unit operations.
Typically, reverse osmosis covers the wide range of applications by raw water salinity – from
50 mg/L and up to 50 000 mg/L. Basically, RO is a pressure-driven membrane separation
process, the driving force resulting from the difference of the electro-chemical potential on both
sides of the membrane. Molecular separation size for RO is below 200 Da, therefore it retains
salts to a high extent.
Figure 1: Membrane processes spectrum [1]
In comparison with conventional porous membranes (Microfiltration and Ultrafiltration), RO

membranes are dense. Separation by pore membranes (MF, UF) is based on a sieving effect,
while differences in solubility and diffusivity are responsible for the selectivity of solution-
diffusion membranes (NF, RO) [2]. Asymmetric polymer or composite membranes are used
today providing rejection of dissolved salts typically at 95% to greater than 99%, depending on
polymer type, feed composition, temperature and unit design.
.
Figure 2: Mass transport through membranes: (a) pore-flow and (b) solution-diffusion [3]
The simplified solution-diffusion model describes performance of RO membrane in terms of

permeate flow (under constant feed pressure) and salt rejection or salt passage:
(1)
(2)
where: A is the membrane permeability coefficient

B is the membrane rejection coefficient
pi – average concentrate/permeate side osmotic pressure
S – wetted surface area
TCF – temperature correction factor
FF – fouling factor
P – pressure
The membrane selectivity or salt rejection is dependent on several factors and increases with
the decrease of:
o degree of dissociation: less dissociated compounds are better rejected
o ionic charge: polyvalent ions are better rejected than monovalent ions
o molecular weight
o non-polarity: less polar substances are rejected better
o degree of hydration: higher hydrated ion are better rejected
RO membranes are typically operated in the cross-flow mode in the pressure range 5-70 bar
and up to 120 bar in special cases. In the cross-flow mode the feed water is pumped “parallel”
(tangentially) to the membrane surface, and the permeate is withdrawn diagonally to it. This is
the principal difference from the dead-end operation similar to coffee filter.
The main advantage of the cross-flow is that the formation of cake layer on the membrane
surface is diminished because there is a continuous turbulent flow over the membrane. This
can be done with membrane modules of spiral-wound, tube, plate, cushion or disc-tube type.

Figure 3: Cross-flow and dead-end filtration [4]
The most widely applied RO module type in water treatment is spiral-wound. It consists of
several membrane bags that are wound helically with one spacer each around the permeate
collecting tube and spacers inside the bags, allowing permeate to flow between the
membranes. The bags are glued at three sides and attached to the perforated permeate
collection pipe at the open side. The cylindrical module resulting from the whirl is supplied to
the front with the feed, which flows through the module in an axial direction. While the feed
flows through the space outside of the membrane bags resulting from the feed spacer, the
withdrawn permeate flows in the membrane bags helically to the permeate collection pipe [5].
Figure 4: Spiral-wound membrane element [6]
Several membrane elements can be connected in series or in parallel and arranged in a

membrane stage this way, together with necessary pumps, valves etc.
Figure 5: From membrane element to membrane stage [5]

The key terms used in reverse osmosis are:
 Permeate – purified product water produced by RO unit
 Concentrate – flow of concentrated salts from the RO unit
 Membrane flux – the rate of permeate transported per unit of membrane area in liters
per square meter per hour (L/m2h)
 Recovery – the percentage of feed converted to permeate, typically maximized while
preventing precipitation within membrane system
 Salt rejection – the percentage of solute concentration removed from the feed by the
membrane
 Salt passage – opposite to rejection
Flux and rejection are the key performance parameters of RO and under specific conditions
are intrinsic properties of the membrane, influenced by pressure, temperature, recovery and
feed water salinity.
From a practical point of view, the task of optimization of RO system operation is to maximize
permeate flow, salt rejection and recovery, while minimizing operating and maintenance
costs, that means minimizing operating pressure and membrane fouling.
Typically, RO unit performance is characterized by two parameters: permeate flow and quality
referenced to a given feed water composition, operating pressure and recovery. The goal of
RO system adjustment is to minimize operating pressure, while maximizing permeate quality
and recovery at required permeate flow. The feed pressure needed to produce the required
permeate flow for a given membrane depends on the designed permeate flux that has to be
limited to minimize fouling. A system designed with high permeate flux rates is likely to
experience higher fouling rates and more frequent chemical cleaning.
In typical single-module system, the feed water from the high-pressure pump flows to the feed
inlet, the permeate stream leaves the system at no more than 0.3 bar over atmospheric
pressure and the concentrate leaves the module at essentially the feed pressure, but with
account of 0.3-2 bar pressure drop from feed inlet. The concentrate flowrate is controlled by
the concentrate valve and should not exceed the designed value. To achieve this, while
increasing the system recovery to more than 50%, concentrate recycling is organized. A part
of the concentrate goes to drain, while the other part is recycled and added to the suction side.
A high fraction of the concentrate being recycled helps reducing element recovery, while
keeping the system recovery reasonably high.
Figure 6: Typical single-module system [7]
Equipment, materials and methods

Recommended RO units are Ecosoft MO6000 or MO10000 (provided to the partners through
the Water Harmony projects), but any other similar unit is suitable.
Portable or laboratory TDS-meter or conductivity meter
Portable or laboratory pH-meter

The RO unit diagram and the principle of its operation is illustrated here:
Figure 7: RO unit diagram (unit of 10 m3/d capacity is equipped with two membranes) [8]
Raw water is fed through the polypropylene sediment filter (5 μm) to prevent particulate
clogging of the membrane. High pressure pump feeds the pre-treated water into the membrane
module, where spiral wound RO membrane element is installed. Recycle flow control throttles
concentrate flow maintaining pressure inside the membrane module. Permeate exits the
membrane module through the permeate outlet. Part of the concentrate is bled to drain; the
rest is fed back to suction end of the high pressure pump via recycle line. Flow rates in recycle
line and drain line can be manually adjusted with respective flow controls. Treated water is
collected in a water tank. Float switch mounted in the tank halts the machine when the
maximum level of permeate is reached. When the float switch is high, the controller first runs
forward flush rinse then brings the machine to a halt. During forward flush, the membrane
module is rinsed with raw water for 60 seconds.
Table 1: Standard operational parameters of the RO unit [8]
Unit capacity, m3/d 6 10

Recycle flow rate, LPM 13-15 8.2-11.2
Drain flow rate, LPM 1.2-1.7 2.2-3.0
Permeate flow rate, LPM 3.5-4.5 6.5-9.0
Pressure in the membrane module, MPa 0.1-1.0 0.8-1.0

(a) Unit capacity 6 m3/d
b) Unit capacity 10 m3/d
Figure 8: Permeate flow rate – salinity curves for the RO unit [8]
Model water
The major water types being treated by RO/NF can be roughly characterized from the total
dissolved solids (TDS) content that can be modeled with NaCl (technical grade).
Water type Low saline Medium saline High saline
Tap water Ground water Brackish water Brackish water
TDS, mg/L 500 1500 5000 10000

General principle
The optimum operating pressure is determined by adjusting the feed pressure until the desired
permeate quality and permeate flux rate are obtained. Sufficient feed flow should be
maintained to ensure a low recovery rate (< 5%) as the membrane flux rate is increased.
Permeate and concentrate streams are recycled back to the feed tank during the first test. The
feed pressure at which the optimum permeate flux and permeate quality is obtained is the feed
pressure used for the second test, determining the recovery rate.
Before Starting the RO Unit

1. Prepare model water solutions of different concentration, using NaCl and measuring final
solution TDS with portable TDS-meter. Recommended concentration: 0.5, 1.5, 5, 10 g/L.
2. Calculate flow configuration:

Recovery, % 30 50 75 80
Permeate flow rate, L/h Should be selected for the specific RO unit
Drain flow rate, L/h
3. Make preliminary checks to make sure all fittings are tight, all components are operational,
and the feed solution is at the proper concentration. Before energizing the high-pressure pump,
the low-pressure switch must be off for start-up to complete the circuit past the low-pressure
cutout. Energize the high-pressure pump momentarily to check proper rotation.
RO Unit Operation
1. Ensure recycle and drain flow controls are fully open before starting the RO unit
2. Run the permeate tube to drain for the duration of the first run of the RO system
3. Switch on main circuit breaker to start the machine. After the controller starts up and RO
unit starts operating, tighten drain flow control until drain rotameter reading meets
experimental specifications.
4. Start turning down recycle flow control. This will raise pressure in the membrane module
shown on pressure gauge. Stop when pressure in the membrane module meets
experimental specifications.
5. After setting proper operating pressure, readjust drain flow rate (if it deviates in the process)
to ensure that system operates with proper recovery according to the experimental
conditions.
6. Make sure that the permeate flow rate and drain flow rate conform to your recovery
calculation.
7. Let the RO unit run for 15 minutes discarding permeate and concentrate to drain. Watch
pressure and flow rate readings to make sure these do not exceed requirements.
8. After 15 minutes, switch to experimental solution.
9. Record permeate flow rate for 3-5 different pressures for pure water.
Cautions
1. Take care not to exceed 1.6 Mpa in membrane module at any time. If membrane pressure
rises above the upper limit in specification, open recycle flow control to bring it down.
2. Turn flow control knobs smoothly when regulating recycle and drain flow. Do not make rapid
turns or apply disproportionate force as this can damage the RO unit.
Shutdown
Shut down by adjusting the by-pass valve or throttling valve to reduce the pressure, depressing
the stop buttons on the high-pressure pump motor and the booster pump motor, and shutting
off the feed supply valve (shutoff valve). When high concentrations (>5000 mg/L) are used, it
is best to flush the reverse osmosis device with the feed solution to remove the high salt
concentration in the device. This can be done by opening the concentrate flow control valve
for approximately 10 min with at least 345 kPa (50 psi) feed pressure. Allow the pressure to
reach zero before disconnecting the reverse osmosis device or carrying out maintenance on
the piping system. Take care to ensure that the membranes are kept wet at all times and are
properly sanitized or winterized, or both (based on supplier’s recommendations) for long-term
storage (more than 5 days).
Data collection and processing
The following data should be recorded during experimental operation:

 Date and time of RO unit operation
 Pressure drop per filter cartridge
 Feed temperature
 Feed, permeate and concentrate pressure
 TDS of the feed, permeate and concentrate streams
 pH of the feed, permeate and concentrate streams
 Observations
The performance of an RO unit is influenced by the feed composition, feed pressure,
temperature and recovery. For example, a feed temperature drop of 4°C will cause a permeate
flow decrease of about 10%, which is normal.
In order to distinguish between such normal phenomena and performance changes due to
fouling or problems, the measured permeate flow and salt passage have to be normalized.
Normalization is a comparison of the actual performance to a given reference performance
while the influences of operating parameters are taken into account. The reference
performance may be the performance specified by the membrane element producer in the
Technical Data Sheet. Normalization has to be done following ASTM D 4516 standard
(Appendix A).
Process modeling and data cross check

Membrane producers usually provide calculation software for modeling separation processes
for their products. Therefore, it is possible to calculate RO operation parameters and
crosscheck experimental data. For Filmtec membranes used in Ecosoft RO units, it is possible
to do calculations in WAVE software that is available for free use https://www.dow.com/en-
us/water-and-process-solutions/resources/design-software
It is necessary to input raw water TDS, membrane module type and RO unit operation
parameters to the WAVE software and generate detailed report with hydraulic and
concentrations results. The report will contain flow, TDS and pressure values for feed,
concentrate and permeate flows.

Report content
Results report shall include graphs of dependencies of permeate flux and salt rejection vs.
pressure, recovery and feed concentration. The results obtained in the laboratory should be
compared with calculation results from the modeling software (WAVE in case of Filmtec
membranes)
Interpretation of results should be given with reference to the solution-diffusion model. For
example, for temperature that is not considered as varying factor in this guideline, the results
would have gotten the following interpretation:
o If the temperature increases and all other parameters are kept constant, the permeate
flux and the salt passage are increasing due to (i) decrease of water viscosity and (ii)
increase of polymer permeability.
o The graphs would have had the following look:
Questions
1. What are the objectives of the RO application testing?
2. Which quantities are to be set and measured in this work?
3. Where can you read the values of pressure and flow?
4. From where will you sample the solutions?
5. What is the purpose of bypass and safety valve? Where are they located?
6. Describe the procedure before starting and turning off the pump.
7. What pressure must not be exceeded and why?
Literature (recommended for reading)

Baker, Richard W., Membrane Technology and Applications, 2nd Edition, John Wiley & Sons,
Ltd. (2004), Chapters 4 & 5.
Membrane Technology for Wastewater Treatment, introduction and overview chapters, open
access book https://www.fiw.rwth-aachen.de/neo/index.php?id=386
Geankoplis, Christie J., Transport Processes and Separation Process Principles, 4th Edition,
Prentice-Hall (2003), Chapters 6 & 13
References
1. Water Planet http://www.waterplanet.com/technology/
2. Rautenbach et al. Perspektiven der Membrantechnik bei der Abwasserbehandlung; ISBN
3-921955-24-6, Aachen
3. SOUZA, V. C. and QUADRI, M. G. N.. Organic-inorganic hybrid membranes in separation
processes: a 10-year review. Braz. J. Chem. Eng. [online]. 2013, vol.30, n.4
4. Spectrum portal, Repligen Corporation http://spectrumlabs.com/filtration/Edge.html
5. Pinnekamp, J., Frledrich, H.: Membrane Technology for Wastewater Treatment, RWTH
Aachen.
6. NALCO. Reverse Osmosis.
https://www.slideshare.net/NelsonIzaguirre1/reverse-osmosis-module
7. Dow Filmtec Technical Manual
8. Ecosoft MO6000/MO10000 User Manual
Instructor’s notes
With increasing effective feed pressure, the permeate TDS will decrease while the permeate
flux will increase.
Pressure:
Recovery:
Recovery is the ratio of permeate flow to feed flow. In the case of increasing recovery, the
permeate flux will decrease and stop if the salt concentration reaches a value where the
osmotic pressure of the concentrate is as high as the applied feed pressure. The salt rejection
will drop with increasing recovery.
Feed salt concentration:

4.3 Impact of the main parameters on the effectiveness of coagulation and the
determination of the optimal dose of coagulant
Lab purpose
Study of regularities of the formation of the contact medium in the process of coagulation
clearing the object of research and determining the optimal dose of coagulant based on waste
alumina production.
Theory
Salts of aluminum and ferrum are most often used for coagulation purification of natural and
sewage as coagulants.
The technology of water purification by coagulants consists of the following basic operations:
pre-clarification, water purification, preparation and mixing of coagulant, discoloration and
clarification.
In the case of the polydisperse composition of suspended matter, especially in the presence
of coarse-dispersed coarse particles (sand, lobes of ore and non-metallic minerals), the
wastewater is pre-clarified in horizontal tangential and aeration sand-traps with a circular or
straight-line motion of water. Smaller mineral or organic particles are also separated by settling
or filtration on slow filters filled with sand and gravel. Prefiltering on microfilters can be done in
front of slow sand filters, before water treatment with coagulants or before rapid sand filters.
As sedimentation structures, sedimentation ponds, horizontal tanks and their combination, as
well as vertical, horizontal, radial, tubular, lamellar, etc. are used. The purified water is
suspended if the alkaline reserve is insufficient for satisfactory hydrolysis of the coagulants.
Hydrogen and sodium carbonate. In the course the pH values are maintained within (6.5-
7.5). It also contributes to reducing the residual content of aluminum and ferrum in purified
water and reducing its corrosive properties. The clarification and discoloration of muddy waters
with increased hardness by coagulants is expedient to carry out at high pH, and of colored soft
waters - at reduced pH. Especially important is the order of the introduction of reagents. When
the flotation reagents are added into the colored water before the addition of coagulants, the
process of coagulation and cleaning quality has been deteriorated. In water, there is an
increased content of stained substances. It is better to discolor the water in the case by adding
of flotation reagents after of coagulants. Since some of the colored matter manages to absorb
at the time of the formation of hydroxides. Organic substances contained in water in the form
of humates of sodium, at low pH values are hydrolyzed to form negatively charged particles.
The latter interact with positively charged polynuclear hydroxcomplexes (micelles). Therefore,
in the case of treatment with coagulants of highly colored waters, they are subjected to flotation
after coagulant administration.
One of the most important technological parameters of the process of water purification by
coagulation is the dose of coagulant, its optimum value depends on the properties of the
disperse system: temperature, the number of suspended and colloid dispersed substances,
color, ionic composition of the dispersion medium, pH and other physico-chemical parameters.
In the case of an insufficient dose of coagulant, the desired effect of cleansing is not achieved,
and in the case of excess - in addition to over-consumption of a high-value reagent, in some

cases the effectiveness of coagulation may deteriorate. With a decrease in the temperature of
treated water, the dose of coagulant increases significantly, especially in the case of muddy
waters. With decreasing water turbidity, the temperature is lower. From the considerable
content of suspended matter, they, coated with a "casing" of colloidal particles of aluminum
hydroxide, coagulate, preventing the formation of long chain bridges from spherical particles.
As a result, a smaller dose of coagulant is required. For high-infused waters with an increase
in their alkalinity, the dose of coagulant increases, for turbid - decreases.
To coagulate quickly and throughout the volume of purified water, it is necessary to mix the
reagents vigorously over a short period of time (1-2 minutes in the case of wet and no more
than 3 minutes - dry dosage of reagents) in hydraulic or mechanical mixers. The mixing of the
coagulant with water should occur so that a large number of small aggregates originally formed
on the surface, on which chemoresubbed charged polynuclear hydroxocomplexes of aluminum
having high activity relative to the impurities to be purified. In a one-stage technological
scheme, the coagulant is added in the immediate proximity of the filters.
It is desirable to introduce the reagent into a relatively small amount of purified water, and then
quickly mix it with the rest (separate coagulation). An increase in the initial concentration of
coagulant contributes to the intensification of the coagulation process due to an increase in the
partial concentration of coagulant in the treated volume of water (concentrated coagulation).
Sometimes we recommend the ratio of volumes of treated and untreated water to 1: 1.5. In the
case of concentrated coagulation, the cost of ferrous sulfate is reduced by 20-30%, as well as
the turbidity and color of water are reduced.
Effective process is fractionated (fractional or partial) coagulation of water, in which the
coagulant is added to the water to be purified in two or more portions or successively
introducing different coagulants. In this case, polydisperse aggregates of the coagulant are
formed, as well as the period of formation of positively charged polynuclear hydroxocomplexes,
resulting in coagulation intensified. The recommended optimal time interval between the
introduction of individual parts of the coagulant is 90 - 120 seconds. In the case of discoloration
of water, the first dose of coagulant should be half the total.
Periodic coagulation is based on a combination of methods of concentrated and fractionated.
The feeding periods of increased doses of coagulant alternate with periods of complete
cessation of coagulation. As a result of such treatment of low-purity water for a two-stage
scheme, the cost of coagulant is reduced by 30 - 40%, the degree of discoloration of water
rises. Deeper removal of coloring impurities is due to lower pH values during the feeding of
increased doses of coagulant.
Coagulation inactivation is also achieved by recirculation of coagulant (coagulated curvature).
The essence of the method is to supply part of the spent siege to the zone of dosage of fresh
portions of the coagulant. This helps to accelerate the process and form denser flakes. The
application of this method is effective for the intensification of coagulation of low-wasting
waters, while significantly (up to 30%) the costs of coagulant are reduced.
The process of formation of flakes successfully occurs with a slow and even mixing of the
disperse system, which helps to agglomerate the small flakes into larger ones that easily
deposit. It is especially necessary to mix at low temperatures of treated water (below 5 ° C).
During the mixing, the growth of the particles is accelerated as a result of their collision, the
interconnection increases and solid flakes are formed. Mixing positively influences the
formation of flakes in the event that the particles have reached a certain size as a result of the
Brownian motion (spherical aggregates larger than 0.02 microns and larger).

The mixing of water should not be too intense to prevent the destruction of flakes. In order to
ensure optimal mixing conditions in front of the septic tanks, flush cameras are often arranged
in which the vertical or horizontal movement of water is provided by means of partitions or
whirlpool devices.
The formed coagulant flakes, together with the adsorbed impurities, are separated from the
water to be purified during clarification by means of defending, filtering, centrifuging or floating.
In the practice of water preparation, the suspended matter is first separated by settling, and
then the drains are filtered. Horizontal, vertical or radial tanks are usually used.

Equipment
Flasks are nominal volume of 100 cm3 - 6 pcs, volume 50 cm3 - 6 pcs; the flask measures a
nominal volume of 1000 cm3; the flask measures a nominal volume of 250 cm3 - 2 pcs; spatula;
Pipettes with a nominal volume of 1, 2, 10 cm3; sampler; photoelectrocolorimeter type KFK -
2; cuvettes; scales analytical, litmus paper.
Reagents
A mixture of ammonium molybdate, sulphatic acid, stybium chloride and tartaric acid. Up to
300 cm3 of palladium acetate is stirred with 144 cm3 of concentrated sulfuric acid, ppm. It is
then cooled to 20 oC and, when stirred, add a solution of 10 g of sulfamic acid NH2SO8H pda.
and 100 cm3 of distillate solution, 12.5 g of ammonium molybdate (NH4)2Mo7O34 ꞏ 4H2O in 200
cm3 of distillate solution, 0.235 g of sitbium chloride of SbCl2 pda a. and 0.6 g of tartaric acid
odd.a. in 100 cm3 of distillate. The icons are kept in a glass of orange glass.
Ascorbic acid. 10% solution. Dissolve 10 g of ascorbic acid in bistystilate and bring the volume
of solution to 100 cm3 in a measuring dish. Store the solution in the cold, stable for about 30-
40 days.
Potassium phosphate monosubstituted - standard solution.
Basic solution: Dissolve 0,7165 g of КН2РО4 pda, dried for 2 hours at a temperature of 105 оС,
in a distillate. Add 2 cm3 of chloroform and bring the volume of the solution with distillate to 1
dm3 in a volumetric flask. 1 cm3 of this solution contains 0.389 mg of phosphate.
Working solution 1: Make 10.0 cm3 of the basic solution to 250 cm3 of distillate. Use freshly
prepared solution. 1 cm3 of this solution contains 0.0156 mg of phosphate.
Working solution 2: Make 50,0 cm3 working solution 1 to 250 cm3 distillate. Use freshly
prepared solution. 1 cm3 of this solution contains 0.00312 mg of phosphate.
Plotting of the calibration graph

In a volumetric flask with a capacity of 50 cm3 are placed 0; 1; 2.5; 5; 10; 25 cc of working
solution 2. Add 2 cc of a solution of the mixture, 0.5 cc of 10% solution of ascorbic acid to each
flask. The mixture is brought to the mark with a distillate and stirred. The first bulb (without the
content of the working solution 2) - "idle experiment". After 20-25 minutes photometrically in a
cell 10 mm in wavelength 670 nm and based on the data we build a calibration graph "Optical
density - concentration of phosphates". An example of the calibration graph is shown in
Figure1.

Calibration
1
0,8
Optical density, А
0,6
0,4
0,2
0
0 0,02 0,04 0,06 0,08 0,1
СРО4, mg
Figure 1: Calibration graph

Determination of the efficiency of removal of phosphates by coagulation method,

selection of the optimal dose of coagulant.
Experiment 1: Coagulation at pH 9 (NaOH substrate reagent).
For study use working solution 1. In 4 cylinders of coagulation pour 100 cm3 of the test solution.
Then add a dose of coagulant (a solution of sulfate or iron chloride) to the pipette so that its
content in water is 25, 50, 75 and 100 mg / dm3. Using a solution of sodium hydroxide, the pH
of the solution is adjusted to 9-10. After mixing the three-fold turning of the closed cylinder
caps, the time of entering the coagulant in water is noted. After 30 minutes, water from each
cylinder is filtered through a "blue ribbon" filter in conical flasks. To analyze the coagulation
efficiency, the filtrate is diluted similarly to the preparation of the solution for preparation of the
calibration: 5 cm3 from each cylinder in a 100 cm3 flask and brought to the label with a distillate.
From the obtained solutions, an aliquot of 5 cm3 is drawn into a 50 cm3 flask, 2 cm3 of a mixture
solution, 0.5 cm 3 of ascorbic acid is added and distilled to the label. After 20-25 minutes,
photometrically, and on the basis of the obtained value, the residual concentration of
phosphate in the solution is found on the graph.
WARNING! Time to settle solutions before photometry should be the same for all
experiments.
Experiment 2: Coagulation with the addition of CaO as a subletting reagent.
For study use working solution 1. In 4 cylinders of coagulation pour 100 cm3 of the test solution.
Then add a dose of coagulant (a solution of sulfate or iron chloride) to the pipette so that its
content in water is 25, 50, 75 and 100 mg / dm3. On technical scales weigh 0.3-0.5 g CaO.
After the introduction of the coagulant and CaO, the solution is mixed with a three-fold reversal
of closed cylinder cams. Mark the time when the coagulant is injected into the water. After 30
minutes, water from each cylinder is filtered through a "blue ribbon" filter in conical flasks. To
analyze the coagulation efficiency, the filtrate is diluted similarly to the preparation of the
solution for preparation of the calibration: 5 cm3 from each cylinder in a 100 cm3 flask and
brought to the label with a distillate. From the obtained solutions, an aliquot of 5 cm3 is drawn
into a 50 cm3 flask, 2 cm3 of a mixture solution, 0.5 cm3 of ascorbic acid is added and distilled
to the label. After 20-25 minutes, photometrically, and on the basis of the obtained value, the
residual concentration of phosphate is found on the directory.
Report content
Removal degree of phosphates, %:
Cv  Ck
X   100 %
Cv ,
where Сv – initial concentration, g/dm3;
Ck – final concentration, g/dm3.
Make conclusions about the effectiveness of the dose of coagulant and the conditions for
coagulation.
Discussion issues
1. Provide the conditions (mode) of the formation of the contact environment in the
suppressed state.
2. How does the characteristic of treated water affect the efficiency of the process?
3. Under what conditions depends on choosing the type of coagulant?
4. Give the types of luminaries and the principle of their operation.
5. Describe the loading of filters used in water treatment technology
6. Types of filters and the principle of their operation.
References
1. Comprehensive Surface Water Treatment Rules Quick Reference Guide: Systems Using
Conventional or Direct Filtration, USEPA, Office of Water (4606), EPA 816-F-04-003
August 2004
2. Eastman, John; “Colloid Stability,” Colloid science; principles, methods and applications.
Terance Cosgrove Ed., Blackwell Publishing, 2005
3. Edney, Daniel, Introduction to the Theory of the Streaming Current Meter, Application Note,
Accurate Measurements NZ Ltd.
4. Eikebrok, Bjørnar, “Water Treatment By Enhanced Coagulation - Operational status and
optimization issues,” SINTEF
5. Emelko, M, et. al., “A review of Cryptosporidium removal by granular media filtration,”
Journal of the American Waterworks Association, Vol. 97, No. 12, Dec 2005, p 101
Engelhardt, Terry, Calculation of CT Values for Compliance with Drinking Water
Regulations, Hach Company, 2008.
6. Enhanced Coagulation and Enhanced Precipitative Softening Guidance Manual, United
States Environmental Protection Agency, Office of Water (4607), EPA 815-R-99-012,
May, 1999.
4.4 The dynamic exchange capacity of cation exchanger
Lab purpose
The purpose of this lab work is to determine the dynamic exchange capacity of cation
exchanger, until the penetration of Ca2+ ions into the filtrate and to regenerate cation in co-
current mode by the solution of sodium chloride and to wash up it by distilled water.
Theory
The essence of ion exchange is the solid (liquid) substance (resin) absorbs anions or cations
from electrolyte solution, changing it on equivalent quantity of other ions with the same charge
sign. There is the classification on anion exchangers and ampholytes. The exchanged ions are
classified on cation and anion exchangers.
There are artificial high molecular organic ion exchangers. Poly electrolytes are almost
undissolved in water and other solvents. The possibility of ion exchanging is caused by
availability of active ion groups with moveable ions (counter ions) in ion exchangers. They can
be exchanged on ions of environment. If this active groups have acidity character, ion
exchangers (cation exchangers) are capable to change moveable hydrogen or sodium ions on
other cations from electrolyte solution. If the functional groups have alkalinity properties, ion
exchangers change moveable hydroxide ions on other anions.
The certain polymer determines the space structure of ion exchanger. Cells consist of the
matrix. These are high molecular, ion exchange materials and moveable ions, which are almost
insoluble in water and other solvents. The last one’s caused the charge of certain signal. The
ion exchange process can be described by reversible equilibrium heterogeneous chemical
reactions of double exchange:
H[Kat] + Na+ = Na[Kat] + H+;
2Na[Kat] + Ca2+ = Ca[Kat]2 + 2Na+;
[An]OH + Cl- = [An]Cl + OH-;
2[An]OH + SO42- = [An]2SO4 + 2OH-; where [An] – anion exchanger; [Kat] – cation exchanger.
The most important characteristic of ion exchange sorbents is full exchange capacity. This is
the theoretic number of ionic groups, containing in the single mass or volume of the ion
exchanger.
The capacity is quantity of sorbed ion by the specific mass or volume of the ion exchanger in
equilibrium conditions (mmole/g, mmole/L). It can be determined in static and in dynamic
conditions. The static exchange capacity is determined by contacting the mass of ion exchange
material with certain volume of the investigated solution, until the equilibrium. The dynamic
exchange capacity is the quantity of ions, absorbed by the same mass of ion exchange at
continuous flowing of the electrolyte solution through the layer of ion exchange resin, until the
penetration of ions. The full dynamic exchange capacity is determined by flowing of solution
through the column of knowing quantity of the ion exchanger, until the composition of filtrate
and inlet solution become the same. The ion exchange reactions are reversible. That’s why, if
the concentration of ions H+, Na+, OH- is increased extremely, the equilibrium moves to the
left. This property is used at the regeneration of ion exchangers.

Equipment: volumetric flasks (100 cm3); volumetric cylinders (250 cm3) – 2 ones; pipettes
(100 cm3); glass columns; bulbs for the titration (volume 250 cm3).
Reactants: cation exchanger; working solutions of CaCl2 (8 % mass) and NaCl (8 % mass),
ammonia buffer solution; the indicator – chrome dark blue; 0.01 mole/L solution of Trilon B;
argentum nitrate solution.
The installation of the dynamic exchange capacity of cation exchanger determination (fig 1.)
consists of ion exchanger column-filter 1, presented by glass tube (diameter 15…25 mm); the
bottom part of it includes glass diaphragm 2 for supporting of ion exchange resin layer 3. The
cation exchanger quantity in column is 30 g. The glass taps and Mor’s clamp on resin tube is
designed for the regulation of liquid rate in the bottom part of column. From above the column
is closed hermetically by resin tube with hole 5. For entering the investigated solution of CaCl2,
NaCl solution to the column for regeneration of ion exchanger and water for washing up, the
glasses 6 – 8 are set up. The reactants are entered for score of hydrostatic pressure from it.,
The volumetric cylinder 9 is used, for the determination of solution quantity, moving through
column.
Experiment 1. Ion exchange filtration of calcium chloride solution

The filtration rate of solution through the cation exchange filter is regulated at the beginning.
Distilled water is passed through the filter from top to bottom. The regulation of flow rate is
carried by regulated device 4. The time is determined by passing 20…25 cm3 of water,
accumulated in volumetric cylinder 9. When liquid flow achieves necessary rate, the tap is
closed. The linear rate of filtration should be approx. 5 m/hour.
Figure 1: The installation for the determination of dynamic exchange capacity of cation
exchanger: 1 – column-filter; 2 – glass diaphragm; 3 – ion exchange resin; 4 – clamp or tap; 5
– stopper with hole; 6 – 8 – glasses with solutions and water; 9 – volumetric cylinder

When the speed of liquid pouring has been determined, the solution of calcium chloride with
molar concentration of equivalents 3.5 mmole/L is filtrated. The column with ion exchanger,
instead of glass with distilled water 8, the glass 6 with CaCl2 is connected. Then the tap is
opened and filtrate is picked up by portions (100…110 cm3). The concentration of calcium
cations is determined in it by complexonometric method. The filtration is stopped when the
concentration of calcium ions in filtrate achieves above 0,05 mmole/L. After this the summary
volume of filtrate has been passed through the ion exchange layer can be determined.
Figure 2: The photo of installation for the determination of dynamic exchange capacity of cation
exchanger
Experiment 2. The regeneration and washing up of the cation exchanger

The glass 7 with solution of sodium chloride (mass fraction – 8 %) is connected to the column.
It is passed through the layer of ion exchanger. The volume of NaCl solution should include
the summary volume of CaCl2 solution, passed through the column with the same rate. Then
excess of salt solution is washed up by connecting glass 8 with distilled water to the column.
It is washed up, while water for washing would not contain chloride ions (sample with argentum
nitrate). It’s necessary to use not less than 1000 cm3 of distilled water.
Report content
The determination of content of calcium ions in filtrate is based on the reversible reaction
Ca2+ + H2Y2- = CaY2- + 2H+,
where H2Y2- - anion of Trilon B.

The reaction charged in alkaline buffer mixture (at pH 10), neutralized hydrogen ions, for
moving the equilibrium to the formation of complex CaY2-.
100 cm3 of the filtrate is picked up (105...110 cm3). Then 5 cm3 of ammonia buffer solution and
2...3 drops of indicator are added into conical bulb with capacity 250 cm3 by pipette or
volumetric flask. The liquid is mixed and titrated by Trilon B solution from crimson to violet blue
color.
The content of calcium ions is calculated by the formula:
where is the concentration of calcium ions in filtrate, mmole/L; V1 – volume of Trilon B

solution spent for the titration, cm3, is the concentration of Trilon B solution, mole/L; V
–volume of water sample, cm3.
The experimental data is recorded to the table.
Table 1: The experimental data

The volume of The concentration of
Volume of sample,
Number of sample electrolyte passed calcium ions in
cm3 3
through the filter, cm filtrate, mmole/L
It’s necessary to build filtration curve in coordinates “electrolyte volume passed through the
filter – calcium ions concentration in filtrate” and to determine the filtrate volume until the
penetration of calcium ions by it. The penetration concentration of calcium ions equals 0,05
mole/L.
The dynamic exchange capacity is calculated by the formula
where DEC is the dynamic exchange capacity of cation exchanger, mmole/g; Vpen is the volume
of filtrate until the penetration of calcium ions, cm3; is the concentration of calcium ions
in filtrate, mmole/L; m is mass of cation exchanger, g.

Example - Determination of dynamic exchange capacity of
cation exchange (by graphic method)
The concentration of calcium ions in filtrate,
5
mmole/L
0
0 500 1000 1500 2000 2500
Volume of filtrate, cm3
Discussion issues
1. Explain the essence of ion exchange. Write down equations of reactions.
2. Describe the structure and properties of artificial ion exchangers (moveable ions, exchange
capacity, swelling, acidity-alkalinity properties, osmotic stability etc.).
3. How the dynamic exchange capacity of ion exchangers can be determined?
4. What the essence of the regeneration of ion exchangers?
5. Call the spheres of using ion exchange in water treatment processes. What are its
preferences and drawbacks comparing with other methods?
6. Present the principal scheme of softening and desalination of water by ion exchange
method. What requirements are to water quality in different branches of industry?
References
1. Water Treatment: Principles and Design/ John C. Crittenden, R. Rhodes Trussell, David
W.Hand. – Printed in the United States of America. – 2005. – 1948p.
2. Zagorodni A. Ion Exchange Materials: Properties and Applications/ Zagorodni A. - Elsevier
Science, 2006. – 496p.

4.5 Evaluation of the carbonic-acid equilibrium
Lab purpose
At performing of this lab work, you will study the alkalinity and buffering capacity of several
types of water: surface water, groundwater (mineral water) and sea water. You will observe
shifting of carbonate content vs. the source of water sampling. You will get experience to
calculate the kind of alkalinity in different waters. You will determine the buffering capacity in
selected water samples by titration method.
Theory
Of all acid-base systems, the most important is the system encompassed CO2, , and
Interconversions of these carbonate containing compounds cover a huge scale of
nature, from the operation of the global carbon cycle till the pH balance of blood cellular fluids,
the hardness of raw water and its alkalinity. The water used everywhere and in particular to
cool equipment. The reason for frequent violations and sometimes accidents with the work of
cooling equipment is the unsatisfactory quality of the cooling water, namely its instability.
Instability can be of two types: aggressive and which lead the formation of precipitation in the
pipelines. Therefore, it is very important to control the content of carbonic acid and its
derivatives in water.
The carbonate system embraces virtually all of the environmental compartments: the
atmosphere, hydrosphere and lithosphere. The complementary processes of photosynthesis
and respiration drive a global carbon cycle. In this cycle, carbon passes slowly between the
atmosphere and the lithosphere, and more rapidly between the atmosphere and the
hydrosphere as shown on Figure 1.
Figure1: Schematic representation of carbonate cycle in Nature.

At present, the volume-percent of CO2 in atmospheric air is about 0.04%, leading to a partial
pressure of ≈ 4*10−4 atm. In a crowded and poorly ventilated room, PCO2 can be as high as 100
ꞏ10−4 atm. About 54ꞏ1014 moles (2.4ꞏ1011tones) per year of CO2 is removed from the
atmosphere by photosynthesis, divided about equally between land and sea. All of this, except

0.05% comes back to biosphere by respiration (mostly by microorganisms). The remainder
comes into sedimentary part of the geochemical cycle where it can remain for thousands to
millions of years.
Since the advent of large-scale industrialization around 1860, the amount of CO2 in the
atmosphere is increasing continuously. Most of this is due to fossil-fuel combustion; Large-
scale destruction of tropical forests, which has accelerated greatly in recent decades do
exacerbate this effect. At present about 30-50% of the CO2 released into the atmosphere by
combustion remains there; the remainder enters the hydrosphere and into the soil of
lithosphere as shown on Figure 1.
The oceans have a large absorptive capacity for CO2 by virtue of its transformation into
bicarbonate and carbonate in a slightly alkaline aqueous medium, and they contain about 60
times as much inorganic carbon as it is in the atmosphere. In addition to atmospheric CO2,
there is a carbon input to oceans from streams. This input is in the form of CO3 2-, which derives
from the weathering of rocks and terrestrial carbonate sediments, and gives rise to the acid-
base reaction:
H2CO3 + CO32- 2HCO3¯ (1)
In such a way, the Earth ocean, is the site of a gigantic acid-base titration in which atmospheric
acids (mainly CO2 but also SO2, HCl and other acids) interacts with carbonates of mineral rocks
and particles of wind blown dust. Carbon dioxide is slightly soluble in water. Its solubility in the
range of 0oC - 20oC decreases from 0.077 mol/liter to 0.039 mol/liter.
In the presence of gaseous CO2, dissolved CO2 exchanges with CO2 gas according to
equations:
CO2(g) + H2O CO2(aq) + H2O (2)
and
CO2(aq) + H2O H2CO3 (3)
where (g) and (aq) refer to the gaseous and aqueous phases, respectively.
Dissolved carbon dioxide consists mostly of the hydrated oxide CO2(aq), i.e.(CO2ꞏH2O). It
coexists with a small portion of carbonic acid. The relationship is following:
[CO2(aq)] ≈ 650 [H2CO3] (4)
Concentration of CO2(aq) far exceeds that of dissolved H2CO3 as shown by equation 4. The
total concentration of [CO2aq] and [H2CO3] in water considered as concentration of dissolved
CO2, so:
[CO2aq] + [H2CO3] = a. (5)
In this mixture, H2CO3 dissociates according to equations:
H2CO3 H+ + HCO3 - (6)
and
H+ + (7)
where the equilibrium conditions are quantified by the dissociation constants:

K1 = (8)
and
K2 = (9)
The values of dissociation constants K1 and K2 at 250C in the fresh surface water are equal to
4.498ꞏ10-7 and 4.79ꞏ10-11 consequently.
Here we have to emphasize, that although the hydrogen ion, H+, commonly exist in hydrated
state to form H3O+ it adopted to write the hydrated hydrogen ion as H+ since the hydrated
structure does not enter the chemical models. The [H+] concentration generally given as a pH
value, defined as the negative logarithm of [H+], i.e.:
pH = -log [H+] (10)
In such a way the total concentration of dissolved inorganic carbon (CT) in the any water
sample defined by the sum:
CT = [CO2aq] + [H2CO3] + [ ]+[ ]=a+b+c (11)
Where:
[CO2aq] + [H2CO3] = a;
[ ]= b,
[ ] = c.
Concentration of each depicted carbon-containing substances in different waters and

relationships among these substances are functions of the partial pressure of CO2 in the
atmosphere, the concentration of different minerals dissolved in water, and of water
temperature.
So, the studying of carbon composition of water, whether it concerns freshwater, sewage water
or seawater, is a complicated task. To be able to get success one need to know the properties
of the following compounds:
● gaseous CO2 (CO2 g) with its partial pressure in the atmosphere;
● dissolved CO2 (denoted by CO2aq.);
● dissolved carbonic acid, H2CO3 with concentration “a” = [H2CO3] + [CO2aq.];
● dissolved bicarbonate, with concentration “b” = [ ];
● dissolved carbonate, with concentration “c” = [ ];
● total dissolved inorganic carbon, with concentration CT = a + b + c;
● concentration of other solutes which affect the degree of dissolution and ionization of
carbonate containing compounds in water;
Quantitative analysis of carbonate system in different waters is based on calculation “effective”
concentration of [CO2aq], [H2CO3], [ ], [ ] and their influence on each other as
function of pH, temperature (oC) and concentration of all inorganic solutes present in water.

Performing this work, the specialists operate with activity (a) of dissolved substances instead
of their formal concentration. Activity is much smaller of concentration in the presence of
essential amounts of salts dissolved in water. The basic analytic equations are following:
(12)
Here the activity coefficient γ ˂1 (γ = 1 when solutes concentrations are very low.)
When seawater or brackish water is studied, more practical is to describe the relation between
actual and theoretical concentration by the apparent solubility constant like shown here:
(13)
Respectively, the thermodynamic and the apparent dissociation constants of the first
dissociation step of carbonic acid expressed as follow:
(14)
and
(15)
And the constants of the second step of dissociation are expressed as:
(16)
and
(17)
Having these expressions, the relative concentrations of [H2CO3], [HCO3-] and [CO32-] one can
found by next manner:
(18)
(19)
(20)

So that:
(21)
And the fractional concentration of [H2CO3], [HCO3-] and [CO32- ] can be given in terms of the
total carbon content like shown below:
(22)
(23)
(24)
The magnitudes of K0, K1 K2 of carbonic acid in freshwater, and K0' K1', K2' in mineralized water
are differed essentially.
The difference in values of K0 , K1, K2 and K0', K1', K2' leads to distribution of H2CO3 and
of its dissociation products in fresh water and in seawater like shown on fig.2.
Figure 2: Distribution of the carbonic acid fractions as percentages of the total carbon content,
CT for temperatures of 50C and 25 0C and for salinities of 0% and 3,5 % vs. pH.
On this figure arrows with indexes of “1” and “2” depicts of maximal concentration when
salinity of water is zero and 3,5% consequently. One can see also the pH values when the
concentration of H2CO3 and becomes gone as well as that pH values when the
concentration of these substances reaches of maximum. So, Figure 2. show in details the
difference in distribution of carbonic acid fractions vs. pH, salinity of water and its temperature.

The sum of the concentration of carbonic acid and its fractions in water named the
alkalinity of water.
The alkalinity of water represents its ability to neutralize inputs of acids. For natural water the
alkalinity is most important in the context of inputs of acids caused by human activities; e.g.,
acid rain, acid mine drainage, excessive fertilization with ammonium etc.
Alkalinity is an important consideration in waters used for drinking and industrial applications,
as well as in the cases of raw water and wastewater treatment. Most microorganisms can
function best at neutral pH values. So, when the pH of water shifts outside of optimal range
(e.g., through denitrification, nitrification, anaerobic fermentation reactions) the run of water
treatment processes can be affected disastrously. The oxidation of Fe2+ that results when
anaerobic ground waters are exposed to air can cause a marked acidification of water that are
poorly buffered with alkalinity; As the result arisen acidity enhance the pipes corrosion.
The simplest method to study alkalinity of water is its acid titration up to certain endpoints
taking into account that alkalinity (AT) is a quantity of , following from
the preservation of electroneutrality in the water samples where the metal-ion concentrations
(Na+, Ca++, Mg++, etc.) and pH are constant, i.e.:
AT = [ ] + 2[ ]+[ ] -[ ] (25)
In (25) participation of other weak acids may also be included in the interest of high precision,
such as humus acids in freshwater, or borate, [ in seawater.
In many cases [H+] and [ ] are negligibly small compared of concentration of carbonate
species. So, in these cases the sum of [ ] and 2[ ], determined by an acid titration is
referred to as the total alkalinity (AT) defined as:
AC = [ ] + 2[ ] = b + 2c (26)
If water contains and carbonate, or it is in contact with calcite, the dissociation
equilibrium of calcite affects the carbon chemistry also:
CaCO3 + (27)
where the ions concentrations of [ ] and [ ] are limited by the solubility of the product.
The principle of alkalinity evaluation describes the following thought experiment, which
illustrated by Figure 3.

Figure 3: The principle of carbonate containing compounds distribution in water.
First of all, let us assume that X moles of solid bicarbonate was added to pure water. Some of
dissolved part of this salt would react with protons of water to form H2CO3, and some would
dissociate to form like shown below:
H2CO3 + O (28)
H H2 O (29)
These reactions are written according to the usual convention for acids, as dissociation
reactions, but in the particular system under consideration, reaction (29) would proceed in
reverse.
The alkalinity is usually expressed as mg/L of CaCO3.
In the case of the equivalent of calcium carbonate, alkalinity expressed as Eq/L.
The equilibrium {H+} of bicarbonate solution obtained, would lie midway between the meaning
of equilibrium constants for reactions 28 (pKa1 = 10-6.35) and 29 (pKa2 = 10-10.33). So, equilibrium
pH would be 8.3 at this condition (low ionic strength - low solute concentration in water). Being
identified in Fig. 3 by blue arrow this pH would correspond to maximal concentration of .
This pH also identified as the point satisfying the balance in the reactions:
(30)
(31)
If the same X moles of Na2CO3 (instead of NaHCO3) were added to water, some of the
carbonate ions would react with protons to form bicarbonate (Reaction 29 in reverse), and this
consumption of protons would raise the pH value. The resultant pH would lie at the point where
Equation (32) is valid:
CO3Total = (32)

This pH identified in Figure 3 as pH (approx. pH 10.5 and higher) and satisfies the proton
condition when H2O and make up the balance:
2[H2CO3] + [ ] + [H+] = [OH-]. (33)
Similarly, pH CO2 defined as the pH of a solution in which X moles of CO2 dissolved. This pH
is approximately 4.7. (Note that the exact pH values depend on the concentration X of CO2 in
water.).
In this explanation, all categories of carbonate alkalinity defined in terms of three pH values
(pH , pH , pH CO2).
In such a way:
Total alkalinity defined as the amount of acid required reduce the pH of studied water
to pH of dissolved CO2; i.e. the amount of acid required to convert of all bicarbonate and
carbonate ions into H2CO3.
Carbonate alkalinity is the amount of strong acid required to lower the pH of a sample
to pH At this point the carbonate is converted to bicarbonate, and at lower pH it would
be converted to H2CO3.
Caustic alkalinity is the amount of strong acid required to lower the pH of a sample to
.
pH Such alkalinity must result from bases stronger than .
Similarly, the categories of acidity may defined as:
Mineral acidity is the amount of strong base required to raise the pH of a sample to
pH CO2 (stronger acids than H2CO3 must be present);
Carbon dioxide acidity is the amount of strong base required to raise the pH of a
sample to pH
Total acidity is the amount of strong base required to raise the pH of a sample to
pH
To evaluate constituent parts of alkalinity stipulated by , , is recommended to
use the Table 1.
Table 1: Alkalinity Relationships.
Volume of titrant required to Alkalinity due to:
reach pH=8.3 (P) and pH=4.5
(T) endpoints OH−
P=0 0 0 T
P˂1/2T 0 2P T-2P
P=1/2T 0 2P 0
P˃1/2T 2P-T 2(T-P) 0
P=T T 0 0
In this table: P- phenolphthalein alkalinity (pH = 8,3); T- total alkalinity (pH = 4,5).

 If P = 0 (volume of titrant to reach phenolphthalein endpoint equal to zero) the alkalinity
due to hydroxyl and carbonate ions is equal to 0. In this case the alkalinity due to
bicarbonate ion is equal to the total alkalinity (T).
 If P ˂1/2 of T, the alkalinity due to hydroxyl ions is 0. In this case the alkalinity due to
carbonate ions is 2P. And alkalinity due to bicarbonate ions is equal to the total alkalinity
minus 2 times phenolphthalein alkalinity, i.e. T-2P.
 If P = 1/2T than the alkalinity due to hydroxyl ions is 0. The alkalinity due to carbonate ions
is 2P. And alkalinity due to bicarbonate ions is equal to 0.
 If P ˃ 1/2T than the alkalinity due to hydroxyl and carbonate ions is 2P. The alkalinity due
to bicarbonate ions is equal to 0.
 If P = T than the alkalinity due to hydroxyl ions is equal to total alkalinity (T). In this case
the alkalinity due to bicarbonate ions and carbonate ions is 0.

Equipment
 Calibrated pH meter
 Stirring plate;
 Micro magnetic stir bar;
 Clean polyethylene bottles
 Burette with Burette stand and porcelain tittle;
 Volumetric pipets with elongated tips;
 Pipette bulb, or Pipette pump;
 Syringe equipped with a filter holder and a fine-pore filter;
 Conical flask (Erlenmeyer flask);
 250 ml measuring syringe;
 Standard flask;
 Wash bottle;
 Beakers.
Chemicals required
 Standard sulfuric acid 0.02M;
 Phenolphthalein;
 Sodium thiosulfate 0,01M;
 Activated carbon;
 Methyl orange (or Bromocresol green);
 Distilled water.

 You should get the samples of water from original sources. If this is not possible, the
artificially prepared solutions can used.
 Appropriate samples should include river or lake water, groundwater (if possible from
a well, and if not, a sample of bottled mineral water—preferably from a natural source),
seawater (if not available, it can be prepared synthetically or obtained from a
commercial source).
 The samples must be collected in clean polyethylene bottles, and analyzed as soon as
possible after sampling. Otherwise, the values may vary substantially.
 Estimated time required to complete the experiment depends on the number of
samples analyzed (approx. 5–20 min per sample per analysis). All experiments may
carried out in one or two sessions.
 Experimental part consists of measures the pH, alkalinity and buffering capacity of
water samples and compares the values among them as well as with those of tap and
distilled or deionized water. The alkalinity will be measured by titration with dilute
sulfuric acid up to two specific pH values: 8.3; and 4,5 and by obtaining of titration
curves using pH meter. This will make possible to calculate the different contributions
of species responsible for the alkalinity and evaluate the buffering capacity of water
samples. The acid–base indicator phenolphthalein is used to indicate the pH = 8.3
endpoint and to obtain the P-alkalinity. Results can best interpreted in the light of
algorithm described in Table 1.To identify the amount of acid required to reach the 4,5
endpoint, methyl orange or bromocresol green indicators should be used.
 The total amount of acid required to reach endpoint 4.5 indicates total alkalinity. For a
more accurate titration, the pH searching must followed with a pH meter. Buffering
capacity of water samples evaluated as quantity of acid required for shifting pH of one
liter of studied water to one unit of pH.
 You will compare the amount of titrant added and the pH of each sample, determining
the alkalinity and buffering capacity.
Safety Measures
The titrants should not come in contact with the skin or eyes because they may be corrosive.
You must consider all the safety measures normally taken when handling this kind of reactants.
In case of spillage of an acid solution or of skin contact, wipe clean with a clean cloth and wash
thoroughly and abundantly with water (sprinkle the table or surface with sodium bicarbonate).
All of the residues generated in this experiment can be disposed of down the drain once they
have been neutralized.
Sampling and titration
1. Collect samples (or prepare them synthetically) of: (1) river or lake water, (2) ocean water,
(3) mineral water (or if available, use groundwater), (4) tap water, and (5) deionized water.
If automatic titration equipment is available, use them, otherwise use a calibrated pH meter
throughout the entire experiment and wait until stable readings obtained.
2. Measure the pH and temperature of each sample. If the pH is above 8.3, determine the P-
alkalinity. If it is not, only the T-alkalinity (i.e., methyl orange or bromocresol green alkalinity at
pH = 4.3).
3. To prevent masking of the endpoint when using colored indicators, make sure the sample
is colorless and free of turbidity. To have this condition, filter the sample prior to titration and if
the color were a problem, add a small amount of activated carbon (prior to filtration). Filtration
can be done by means of a syringe equipped with a filter holder and a fine-pore filter. To

eliminate any free chlorine that might interfere with the titration, add a drop or two of 0.01M
sodium thiosulfate.
4. Place burette in a stand, rinse it with a small amount of the acid titrant, and fill it to the
desired mark.
5. With a volumetric pipet, take a 10-mL portion of the water sample and place it in a 25- or
50-mL beaker. Place a micro magnetic stir bar inside the beaker and put the beaker on a
stirring plate. Immerse the bulb of the pH electrode in the liquid sample, without touching the
stir bar. Measure the pH.
6. Add 2 to 3 drops of the phenolphthalein indicator (e.g., with a Beral pipet). Observe if any
color appears.
7. If upon adding the P indicator there is no color, then the P-alkalinity is zero. In this case,
immediately add 1 to 2 drops of the methyl orange (or of the bromocresol green) indicator and
start titrating to the endpoint.
8. If a pink color appears upon adding the phenolphthalein indicator, titrate drop wise (in 0.1 or
0.2 mL increments), stir gently, and record the resulting pH and volume of titrant added. Note
the pH reading at the point when the color of the indicator disappears (it must be close to 8.3).
Then, immediately add 2 to 3 drops of the methyl orange (or the bromocresol green) indicator,
and continue the titration until the exact 4.5 endpoint is reached (the solution turns Salmon in
the case of methyl orange or yellowish in color, in the case of the bromocresol green indicator).
9. Repeat the same technique for each water sample.
To get titration curves you will carуfully add drops of standard acid to various samples and
record the resultant pH values. The data that you obtain will look similar to that shown in Figure
4 There are regions where the pH changes rapidly with each acid addition, and regions where
the response is much less.
Figure 4: Titration curve. Endpoints of the titration are shown in the plot (point z for total
alkalinity, point y for carbonate alkalinity, point x for caustic alkalinity).

You can determine from this titration curve the approximate endpoints (x, y, z) for the
various alkalinity species (see discussion above).
LABORATORY REPORT
Alkalinity and Buffering Capacity of Water

Name __________________Section______________ Date__________________
Instructor_______________ Partner____________________________________
Experimental Tittle:__________________________________________________
Objectives_________________________________________________________
Flow sheet of procedure______________________________________________
Waste containment procedure__________________________________________
PART “A” DETERMINATION OF ALKALINITY

Origin of samples:
№1________________________________________________________________
№2________________________________________________________________
№3________________________________________________________________
№4________________________________________________________________
№5________________________________________________________________
Visible characteristics of the sample (color, odor, suspended solids present, etc.)
№1________________________________________________________________
№2________________________________________________________________
№3________________________________________________________________
№4________________________________________________________________
№5________________________________________________________________
Precautions observed during the sampling. Sampling procedure for each sample:
№1________________________________________________________________
№2________________________________________________________________
№3________________________________________________________________
№4________________________________________________________________
№5________________________________________________________________

EXPERIMENTAL DATA
1) Initial pH of the samples:

№1________________________________________________________________
№2________________________________________________________________
№3________________________________________________________________
№4 Tap water_______________________________________________________
№5 Deionized water____________________________________________________
Analysis of obtained results

1. From the experimental data obtained with each sample tested, report the alkalinity values
and the concentration of each alkalinity species as mg/L of CaCO3.
To calculate the alkalinity, apply the following formula:
𝑽 𝐚𝐜𝐢𝐝,𝐦𝐋 𝐌,𝐚𝐜𝐢𝐝 𝐬𝐨𝐥𝐮𝐭𝐢𝐨𝐧 𝟓𝟎 𝐠/𝐚𝐪𝐮𝐢𝐯𝐚𝐥𝐞𝐧𝐭 𝐂𝐚𝐂𝐎𝟑 𝟏𝟎𝟎𝟎 𝐦𝐠/𝐠
Alkalinity (mg CaCO3/L) =
𝑽 𝐨𝐟 𝐬𝐚𝐦𝐩𝐥𝐞,𝐦𝐋
Fill the table 2 of Report.

Table 2: Alkalinity structure of studied samples.
Sample P-Alkalinity as M- Alkalinity as Total Alkalinity as
CaCO3 CaCO3 CaCO3
№1
№2
№3
№4
№5
To determine the approximate concentration of hydroxide, carbonate and bicarbonate ions in

each sample use the algorithm described in Table 1 of theoretical part.
The dominant species at pH = 4.5 are assumed to be bicarbonate and carbonate, and when
OH− ions are present, no bicarbonate ions can be present. It is also assumed that [H+] is not
relevant in alkaline pH values, and that one half of the carbonate ions present become
neutralized at the pH = 8.3 endpoint. Fill the table below based on the results of titrations
performed by you.

Alkalinity Relationships.
Volume of titrant required to Alkalinity due to:
reach pH=8.3 (P) and pH=4.5
(T) endpoints OH−
P=0 0 0 T
P˂1/2T 0 2P T-2P
P=1/2T 0 2P 0
P˃1/2T 2P-T 2(T-P) 0
P=T T 0 0
Carry out titration with using of pH meter

Plot the titration curves of different water samples and designate the points of pKa and titration
endpoints
The titration data should be assembled in separate table like shown below and after that plotted
as pH (y axis) vs. acid added (x axis) curves for different studied samples of water.
Table 3: Example titration data of 50 mL natural water containing carbonate species. The titrant
is 0,01 M H2SO4.

Table 4: Titration data of 50 mL(100mL) sample of____________ (natural water , seawater,
tap water, mineral water) containing carbonate species. The titrant is 0,02 M H2SO4.
Acid added(mL) pH Acid added (mL) pH Acid added(mL) pH
Prepare the plots of pH (y axis) vs. acid added (x axis) for different studied samples of water
(natural water , seawater, tap water, mineral water) containing carbonate species like shown
in Figure 5.
Figure 5: Example of plotting titration curve and designation of inflection points on it.
The inflection points on the titration curve correspond to the pKa for the carbonate system
(equivalence points are 6.35 and 10.33 consequently) and the equivalence points (pH
pH and pH CO2).

 Compare the titration curves obtained on different water samples.
 Explain why the resulting curves have a different shape.
 Calculate the buffering capacity of different water samples using titration plots.
 Collect the results obtained in the table 5 like shown below
Table 5: Buffering capacity of different water samples at chosen pH values.

Sample pH range Buffering capacity( M of acid
added per unit of pH in 1L of
water sample)
№1
№2
№3
№4
№5
Report content
1. In the Introduction you should explain global importance of carbonate system and why
water`s alkalinity is of interest t why the alkalinity of water is of interest to specialists from
different fields of knowledge.
2. In the Results section, insert the tables and figures described under headings: Origin of
samples, Visible characteristics of the samples (color, odor, suspended solids present, etc.),
Precautions observed during the sampling, Initial pH values of samples, Alkalinity
measurement data obtained by titration up to chosen endpoints (without using of pH meter),
Titration curves and results of there analysis
Explain the differences of obtained results related to different samples of water. Why your
measured values for alkalinity components are different in different samples? What effect does
atmospheric CO2 have on the alkalinity and the carbonate species distribution in water
samples? Present tables listing the alkalinity structure for all the samples that you measured.
Discuss and compare these results and discuss whether any of these samples would be
susceptible to acidification from acid rain. Evaluate whether there is a relationship between
alkalinity and pH. Discuss the results of buffering capacity measurement.
Discussion issues:
1. Why the distilled water or DI water considered to have no alkalinity and buffering capacity?
2. Why the sample of seawater exhibit high alkalinity and buffering capacity?
3. What is the reason of surface water buffering capacity?

Test yourself with answers to the following questions:
1. The alkalinity of water is an indication of:

a) Base neutralizing capacity;
b) Acid neutralizing capacity;
c) Quantity of base present;
d) Quality of base present.
2. Alkalinity is present due to all except:
a) Bromates;
b) Phosphates;
c) Silicates;
d) Chlorides.
3,Alkalinuity is not caused by:
a) Carbonate ions;
b) Bicarbonate ions;
c) Hydroxyl ions;
d) Chloride ions.
4.The phenolphtalein alkalinity is present then the pH of that water will be more than:
a) 8.3;
b) 9.3;
c) 7.3;
d) 6.3.
5.Alkalinity of natural water is manly due to presence of:
a) Bicarbonates;
b) Bromates;
c) Phosphates;
d) Silicates.
6.The bicarbonate equivalence point of rain water normally occur at pH:
a) 2.5;
b) 3.5;
c) 4.5;
d) 5.5.
7.What is ppm?
a) Parts per meter square’
b) Parts per meter’
c) Parts per million’
d) Parts per millimeter.
8.The normality of acid used in the titration is:
a) 0.2N;
b) 0.02N;
c) 0.002N;
d) 2.0N.
9.A standard solution is a:
a) Solution of accurately known strength;
b) Solution of accurately known pH;
c) Colored solution;
d) Colorless solution.

References
1. APHA Standard methods for the examination of water and wastewater 20th edition. Method
2320.
2. Methods for chemical analysis of water and wastes EPA 600/4-79-020, USEPA, method
310/1.
3. Dickson, A.G., C.L. Sabine and J.R. Christian (2007). Guide to best practices for ocean
CO2 measurements (A.G. Dickson, C.L. Sabine and J.R. Christian, 191 pp.
4. Keeling, C.D. and T.P. Whorf (2006). Atmospheric CO2 records from sites in the air
sampling network. In Trends: A Compendium of Data on Global Change. CDIAC, Oak
Ridge National Laboratory, U.S. Department of Energy, Oak Ridge,Tenn., U.S.A
5. Millero, F.J., F. Huang, T. Graham and D. Pierrot (2007). The dissociation of carbonic acid
in NaCl solutions as a function of concentration and temperature. Geochim. Cosmo chim.
Acta 71:46-55.
6. Andersen. C. B. “Understanding Carbonate Equilibria byMeasuring Alkalinity in
Experimental and Natural Systems,” Geochem. Educ. 2002, 50, 389–403.
7. Dunnivant, F. M. Experiment 21 (Determination of Alkalinity of Natural Waters) in:
Environmental Laboratory Exercises for Instrumental Analysis and Environmental
Chemistry; Wiley-Inter science: New York, 2004.

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WATER HARMONY

Lab course guide www.waterh.eu Page 1/128

Lab course guide www.waterh.eu Page 2/128

1 Good Lab Practices .......................................................................................................................................... 1

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1.1 Quantitative analysis. Spectrophotometric determination of Fe ions in

4Fe+2 + O2 + 2H2O = 4Fe+3 + 4OH-

4Fe+3 + 3H2O = Fe(OH)3 + 3H+

Lab course guide www.waterh.eu Page 1/128

Quantitative analysis of Fe in water samples can be carried out by atomic absorption

2NH2OH + 4Fe3+ = N2O + 4Fe2+ + H2O + 4H+.

In this reaction, n is equal to 3.

Lab course guide www.waterh.eu Page 2/128

A spectrophotometer (photo-colorimeter) is an instrument designed to accurately measure the

Light source → Monochromatic unit → Cell → Detector → Meter

Figure 1: Calibration graph

Equipment and reagents

 Spectrophotometer UV-Vis or photo-colorimeter for use at 510 nm, providing a

Tasks performance order

1. Preparation of reference solutions

Lab course guide www.waterh.eu Page 5/128

Concentration Volume of Volume of Volume of Volume of Volume of the

2. The samples preparation

3. Determination of total iron:

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5. Determination of ferrous iron

The Use of Spectrometer

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Solution % Transmittance Absorbance (A=-logT)

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1. What is the purpose of using the hydroxylamine hydrochloride in procedures of preparation

3. Why the "blank" sample is necessary at performing of colorimetric measurements?

4. How to determine Fe+3 concentration in water samples using phenanthroline method?

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Equipment and reagents

Tasks performance order

Experiment 1. Determination of alkalinity of the water samples.

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Experiment 2. The determination of general acidity of water.

MOA (CaCO3, mg/L)=

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Total permanent water hardness is calculated with the following formula:

So total permanent water hardness expressed as equivalent of CaCO3 can be calculated

Experiment 4. The determination of pH of water.

In chemistry, pH (/piːˈeɪtʃ/) is a logarithmic scale used to specify the acidity or basicity of

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Figure 1: Digital pH meter

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Conductivity (or specific conductance) of an electrolyte solution is a measure of its ability

Figure 2: Digital Conductivity meter

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Experiment 6. The determination of turbidity of water.

Experiment 7. The determination of SUVA.

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Calculation of acidity of water